Yashwant V. Pathak, Hemant K. S. Yadav - Nasal Drug Delivery - Formulations, Developments, Challenges, and Solutions-Springer (2023)

Yashwant
V. Pathak
Hemant K. S. Yadav Editors
Nasal Drug
Delivery
Formulations, Developments,
Challenges, and Solutions
Nasal Drug Delivery
Yashwant V. Pathak • Hemant K. S. Yadav
Editors
Nasal Drug Delivery

Formulations, Developments, Challenges,
and Solutions
Editors
Yashwant V. Pathak Hemant K. S. Yadav
Taneja College of Pharmacy School of Pharmacy
University of South Florida Suresh Gyan Vihar University
Tampa, FL, USA Jaipur, Rajasthan, India
ISBN 978-3-031-23111-7 ISBN 978-3-031-23112-4 (eBook)

https://doi.org/10.1007/978-3-031-23112-4
© The Editor(s) (if applicable) and The Author(s), under exclusive license to Springer Nature
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To the loving memories of my parents and
Dr Keshav Baliram Hedgewar, who gave
proper direction to my life, to my beloved
wife Seema who gave positive meaning and
my son Sarvadaman who gave a golden
lining to my life.
I would like to dedicate this book to the
loving memories of Ma Chamanlaljee,
Ma Lakshmanraojee Bhide and Ma
Madhujee Limaye who mentored me
selflessly and helped me to become a good
and socially useful human being.
Yashwant V. Pathak
I would like to dedicate this book to my
Parents Shri. B S Yadav and Kamla Yadav for
making me what I am today, to my strength
and companion my wife Sharmishtha, my
daughter Khyati who made both of our life
complete and my teachers who always kept
me motivated.
Hemant K. S. Yadav
Preface
In recent years, interest in using nasal passage as drug absorption site has received
increased attention from formulation scientists. The nasal passage, even though
offering a small surface area of the body as compared to other absorption passages
such as the gastrointestinal tract or skin, still shows significant possibility of drug
absorption at a quicker rate. Another application is the possibility of delivering
drugs to the brain using this passage.
Nasal delivery has come up as a promising approach to deliver diverse therapeu-
tic agents from small drug molecules to biomacromolecules like peptides and pro-
teins and genes to treat various disorders of the central nervous system including
depression, epilepsy, migraine, schizophrenia, Parkinson’s disease, Alzheimer’s
disease, and brain tumor.
Different approaches have been studied in order to facilitate the delivery of dif-
ferent drugs into the brain; among all these approaches, intranasal administration
has gained special interest. Nose-to-brain delivery provides a direct pathway of drug
delivery to the brain without the need to permeate the BBB, potentially avoiding
adverse effects that could occur when the drug is systemically absorbed.
The book comprehensively covers the anatomy and physiology and pharmacol-
ogy of the nasal passage and its use as a drug absorption site. It discusses various
drug delivery systems and polymeric applications for nasal drug delivery. It pro-
vides in-depth information on nasal drug delivery and predicts a potential market in
the global scenario. It describes challenges and approaches to overcome drug
absorption via the nasal route. The volume also covers various formulations like
nanosuspensions, niosomes, and vaccines, which effectively deliver drugs via the
nasal route.
This book, Nasal Drug Delivery: Formulation and Development, Challenges
and Solutions, contains 19 chapters written by academicians and researchers from
the USA, India, UAE, Italy, Turkey, Korea, Belgium, Ghana, Brazil, South Africa,
West Indies, and Serbia.
We are extremely indebted to all the authors who took lot of efforts to complete
the chapters on time and ensure that all the aspects related to nasal drug delivery
vii
viii Preface
formulation and challenges are covered comprehensively in their respective

chapters.
We are extremely thankful to Ms. Carolyn from Springer for encouraging and
helping us to edit this book, as well as other Springer employees who supported this
book, and the printing press employees who helped to get this book in the print
book form.
We would like to state the support of our family members during the editing of
the book.
We would certainly appreciate our readers’ feedback, which will help us in future
projects.
Tampa, FL, USA Yashwant V. Pathak

Jaipur, Rajasthan, India Hemant K. S. Yadav
Contents
1 An Overview of the Anatomy and Physiology of Nasal

Passage from Drug Delivery Point of View�� 1
Hemant K. S. Yadav, Allyson Lim-Dy, and Yashwant V. Pathak
2 Pharmacological and Clinical Problems with Special
Focus on Nasal Drug Delivery�� 15
Misha Mathur and Yashwant V. Pathak
3 Drug Absorption via the Nasal Route: Opportunities
and Challenges �� 25
Seth Kwabena Amponsah and Ismaila Adams
4
Factors Affecting the Design of Nasal Drug Delivery System�� 43
Jéssica Bassi da Silva, Maria Vitoria Gouveia Botan,
and Marcos Luciano Bruschi
5 Challenges in Targeting Nasal Passage and Nose-to-Brain
Delivery via Nanoemulsions�� 59
Shiv Bahadur and Kamla Pathak
6
Potential Targeting Sites to the Brain Through Nasal Passage�� 83
Mershen Govender, Sunaina Indermun, Pradeep Kumar,
and Yahya E. Choonara
7
Biomedical Applications of Nanocarriers in Nasal Delivery�� 101
Namdev Dhas, Soji Neyyar, Atul Garkal, Ritu Kudarha,
Jahanvi Patel, Srinivas Mutalik, and Tejal Mehta
8
Delivery of Vaccines via the Nasal Route �� 127
Seth Kwabena Amponsah and Emmanuel Boadi Amoafo
9 Overview on Nanocarriers for Nasal Delivery�� 141
An
Sunita Dahiya and Rajiv Dahiya
ix
x Contents
10 Nose-to-Brain
Delivery of Peptides and Proteins�� 169
Meltem Ezgi Durgun, Gamze Çamlık, İsmail Tuncer Değim,
and Yıldız Özsoy
11 Novel
Mucoadhesive Polymers for Nasal Drug Delivery�� 189
Ljiljana Djekic
12 Novel
Approaches in Nasal In Situ Gel Drug Delivery �� 235
Cinzia Pagano, Luana Perioli, and Maurizio Ricci
13 Nasal
Delivery of High Molecular Weight Drugs: Recent
Trends and Clinical Evidence �� 253
Emine Kahraman, Sevgi Güngör, and Yıldız Özsoy
14 Niosomes-Based
Drug Delivery in Targeting the Brain
Tumors Via Nasal Delivery�� 279
Mahmoud Gharbavi, Sepideh Parvanian, Milad Parvinzad Leilan,
Shabnam Tavangar, Maedeh Parchianlou, and Ali Sharafi
15 Nanosuspension
– A Novel Drug Delivery System via
Nose-to-Brain Drug Delivery�� 325
Hemant K. S. Yadav and Raghad Zain Alabdin
16 Nasal
Delivery of Micro and Nano Encapsulated Drugs�� 339
Muhammad Sarfraz, Sara Mousa, Ranim Al Saoud,
and Raimar Löbenberg
17 Different
Strategies for Nose-to-Brain Delivery
of Small Molecules �� 361
Smita P. Borkar and Abhay Raizaday
18 Is
There a Global Market and Opportunities for Nasal
Drug Delivery? Recent Trends in Global Nasal Delivery Market �� 381
Abdullah Abdelkawi, Jean Pierre Perez Martinez,
and Yashwant V. Pathak
19 Nasal
Drug Delivery System: Regulatory Perspective�� 393
Sudhir Sawarkar and Julie Suman
Index�� 417
Contributors
Abdullah Abdelkawi Taneja College of Pharmacy, University of South Florida,

Tampa, FL, USA
Ismaila Adams, PhD Department of Medical Pharmacology, University of Ghana
Medical School, Accra, Ghana
Raghad Zain Alabdin, PhD Department of Business Development, Gulf
Pharmaceutical Industries (Julphar), Ras Al Khaimah, UAE
Department of Pharmaceutics, College of Pharmacy, RAK Medical and Health
Sciences University, Ras Al Khaimah, UAE
Ranim Al Saoud, PhD College of Pharmacy, Al Ain University, Al Ain Campus,
Al Ain, United Arab Emirates
Emmanuel Boadi Amoafo, B Pharm Department of Pharmaceutical Sciences,
North Dakota State University, Fargo, ND, USA
Seth Kwabena Amponsah, PhD Department of Medical Pharmacology,
University of Ghana Medical School, Accra, Ghana
Shiv Bahadur, PhD Institute of Pharmaceutical Research, GLA University,
Mathura, Uttar Pradesh, India
Smita P. Borkar College of Pharmacy, JSS Academy of Technical Education,
Noida, Uttar Pradesh, India
Arvind Gavali College of Pharmacy, Jaitapur, Satara, Maharashtra, India
Maria Vitória Gouveia Botan, PhD Department of Pharmacy, State University of
Maringa, Maringa, PR, Brazil
Marcos Luciano Bruschi, PhD Department of Pharmacy, State University of
Maringa, Maringa, PR, Brazil
Gamze Çamlik, PhD Department of Pharmaceutical Technology, Faculty of
Pharmacy, Biruni University, Istanbul, Turkey
xi
xii Contributors
Yahya E. Choonara, PhD Wits Advanced Drug Delivery Platform Research Unit,

Department of Pharmacy and Pharmacology, School of Therapeutic Sciences,
Faculty of Health Sciences, University of the Witwatersrand, Johannesburg,
Gauteng, South Africa
Rajiv Dahiya, PhD School of Pharmacy, Faculty of Medical Sciences, The
University of the West Indies, St. Augustine, Trinidad and Tobago
Sunita Dahiya, PhD Department of Pharmaceutical Sciences, School of Pharmacy,
University of Puerto Rico - Medical Sciences Campus, San Juan, PR, USA
Jéssica Bassi da Silva, PhD Department of Pharmacy, State University of Maringa,
Maringa, PR, Brazil
İsmail Tuncer Değim, PhD Department of Pharmaceutical Technology, Faculty
of Pharmacy, Biruni University, Istanbul, Turkey
Namdev Dhas, PhD Department of Pharmaceutics, Institute of Pharmacy, Nirma
University, Ahmedabad, Gujarat, India
Department of Pharmaceutics, Manipal College of Pharmaceutical Sciences,
MAHE, Manipal, India
Ljiljana Djekic, PhD University of Belgrade-Faculty of Pharmacy, Department of
Pharmaceutical Technology and Cosmetology, Belgrade, Serbia
Meltem Ezgi Durgun, PhD Department of Pharmaceutical Technology, Faculty
of Pharmacy, Istanbul University, Istanbul, Turkey
Atul Garkal, Ph Department of Pharmaceutics, Institute of Pharmacy, Nirma
Mahmoud Gharbavi, PhD Nanotechnology Research Center, Ahvaz Jundishapur
University of Medical Sciences, Ahvaz, Iran
Mershen Govender, PhD Wits Advanced Drug Delivery Platform Research Unit,
Faculty of Health Sciences, University of the Witwatersrand, Parktown,
Johannesburg, Gauteng, South Africa
Sevgi Güngör, PhD Department of Pharmaceutical Technology, Faculty of
Pharmacy, Istanbul University, Istanbul, Türkiye
Sunaina Indermun, PhD Wits Advanced Drug Delivery Platform Research Unit,
Emine Kahraman, PhD Department of Pharmaceutical Technology, Faculty of
Pharmacy, Istanbul University, Istanbul, Türkiye
Ritu Kudarha Department of Pharmaceutics, Manipal College of Pharmaceutical
Sciences, MAHE, Manipal, India
Contributors xiii
Pradeep Kumar, PhD Wits Advanced Drug Delivery Platform Research Unit,
Milad Parvinzad Leilan, PhD Department of Medical Biotechnology, School of
Medicine, Zanjan University of Medical Sciences, Zanjan, Iran
Allyson Lim-Dy Taneja College of Pharmacy, University of South Florida,
Tampa, FL, USA
Raimar Löbenberg, PhD Faculty of Pharmacy and Pharmaceutical Sciences,
Katz Centre for Pharmacy & Health Research, University of Alberta, Edmonton,
AB, Canada
Jean Pierre Perez Martinez Taneja College of Pharmacy, University of South
Florida, Tampa, FL, USA
Misha Mathur University of South Florida, Taneja College of Pharmacy,
Tampa, FL, USA
Tejal Mehta, PhD Department of Pharmaceutics, Institute of Pharmacy, Nirma
Sara Mousa, PhD College of Pharmacy, Al Ain University, Al Ain Campus, Al
Ain, United Arab Emirates
Srinivas Mutalik Department of Pharmaceutics, Manipal College of
Pharmaceutical Sciences, MAHE, Manipal, India
Soji Neyyar, PhD Department of Pharmaceutics, Manipal College of
Pharmaceutical Sciences, MAHE, Manipal, India
Yıldız Özsoy, PhD Department of Pharmaceutical Technology, Faculty of
Pharmacy, Istanbul University, Istanbul, Turkey
Cinzia Pagano, PhD Department of Pharmaceutical Sciences, University of
Perugia, Perugia, Italy
Maedeh Parchianlou, PhD Zanjan Pharmaceutical Biotechnology Research
Center, Zanjan University of Medical Sciences, Zanjan, Iran
Sepideh Parvanian, PhD Faculty of Science and Engineering, Åbo Akademi
University & Turku Bioscience Center, Turku, Finland
Jahanvi Patel, PhD Department of Pharmaceutics, Institute of Pharmacy, Nirma
Kamla Pathak, PhD Faculty of Pharmacy, Uttar Pradesh University of Medical
Sciences, Saifai, Etawah, Uttar Pradesh, India
Yashwant V. Pathak, PhD Taneja College of Pharmacy, University of South
Florida, Tampa, FL, USA
Faculty of Pharmacy, Airlangga University, Surabaya, Indonesia
xiv Contributors
Luana Perioli, PhD Department of Pharmaceutical Sciences, University of

Abhay Raizaday College of Pharmacy, JSS Academy of Technical Education,
Noida, Uttar Pradesh, India
Maurizio Ricci, PhD Department of Pharmaceutical Sciences, University of
Muhammad Sarfraz, Ph.D. College of Pharmacy, Al Ain University, Al Ain
Campus, Al Ain, United Arab Emirates
Sudhir Sawarkar, PhD QRServes Global LLC, Sharjah, United Arab Emirates
Ali Sharafi, PhD Zanjan Pharmaceutical Biotechnology Research Center, Zanjan
University of Medical Sciences, Zanjan, Iran
Julie Suman Next Breath, Halethorpe, MD, USA
Shabnam Tavangar, PhD. Department of Medical Biotechnology, School of
Medicine, Zanjan University of Medical Sciences, Zanjan, Iran
Hemant K. S. Yadav, PhD School of Pharmacy, Suresh Gyan Vihar University,
Jagatpura, Jaipur, Rajasthan, India
Chapter 1
An Overview of the Anatomy
and Physiology of Nasal Passage
from Drug Delivery Point of View
Hemant K. S. Yadav, Allyson Lim-Dy, and Yashwant V. Pathak
Abstract This chapter covers the anatomy and physiology of the nasal passage
with special focus on utilizing the nasal passage for delivering drugs for disease
treatment. The total length of the nasal cavity is 120–140 mm and the total surface
area is about 160 cm2. Also, the total volume is nearly 15 ml. The flow of air and
particles in the cavity is controlled by structures within it. The sense of smell is
controlled by the nasal cavity’s olfactory area. Nasal blood flow can be affected by
a variety of causes. The vasomotor response of the nose is influenced by a variety of
stimuli, both locally and generally. Changes in ambient temperature and humidity,
topical application of vasoactive medications, external compression of major veins
in the neck, trauma, and inflammation are all examples of local factors. This chapter
discusses further details about how this nasal passage can be sued for drug delivery
and as an absorption surface for the drugs whether small molecules or macro
molecules.
Keywords Nasal · Drug Delivery · Intranasal · Targeting and physicochemical
1 Introduction
1.1 Nasal Drug Delivery Systems [1]
There are many attractive reasons to introduce a drug via the nasal route of admin-
istration as a convenient accessible route which achieves localized and systemic
actions rapidly and manageably. Some of these reasons, that made many pharma-
ceutical companies and researchers investigate further about this route of adminis-
tration, are: the rapid onset of drug action potential, hepatic and gastrointestinal
H. K. S. Yadav (*)
School of Pharmacy, Suresh Gyan Vihar University, Jagatpura, Jaipur, Rajasthan, India
A. Lim-Dy · Y. V. Pathak
Taneja College of Pharmacy, University of South Florida, Tampa, FL, USA
© The Author(s), under exclusive license to Springer Nature Switzerland AG 2023 1

Y. V. Pathak, H. K. S. Yadav (eds.), Nasal Drug Delivery,
https://doi.org/10.1007/978-3-031-23112-4_1
2 H. K. S. Yadav et al.
metabolism avoidance, noninvasiveness and ease of access, no sterilization require-

ments compared to parenterals leading to lower costs, non-irritative, and can be
used for prolonged periods. The most attractive part in this route of administration
is the olfactory region, which is considered as a direct physiological link to the cen-
tral nervous system. Thus, the nasal route of administration has a high level of
potential for administering the drugs to effectively treat many diseases such as
Alzheimer’s disease, epilepsy, brain tumors, and many others. To appreciate this
potential and digging deeper into a clear picture of the idea of locally, systemically,
and nose-to-brain drug administration, it is very important to review the physiology
and anatomy of the nasal route, nasal route applications, challenges, and the factors
affecting the formulations and drug absorption via the nasal route.
1.1.1 Nasal Anatomy and Physiology
The total length of the nasal cavity is 120–140 mm and the total surface area is about
160 cm2. Also, the total volume is nearly 15 ml. By the nasal septum, the nasal cav-
ity is divided into two and extends posteriorly to the nasopharynx. The nasal vesti-
bule is the first part of the nasal cavity which opens to the face through the nostril
(Fig. 1.1) and it is the narrowest part with an area of 30 mm2 and it contains vibris-
sae (hairs) that filter the inhaled particles that are larger than 10 μm. The atrium is
an intermediate region between the vestibule and the respiratory region. The respi-
ratory region, the nasal conchae, or turbinates are convoluted projections from the
nasal septum dividing it into three sections: the superior, middle, and inferior nasal
turbinates. These folds provide the nasal cavity with a very high surface area com-
pared to its small volume. The epithelial cells in the nasal vestibule are pseudostrati-
fied columnar epithelium with ciliated, non-ciliated, basal, and mucus-secreting
goblet cells.
Hundreds of motile cilia which are located on each of the ciliated cells and the
serous and seromucous glands are providing the mucus support and nasal secre-
tions. Olfactory region is located at the top of the turbinates and its area is about
12.5 cm2 composed of nonciliated pseudostratified columnar epithelium traversed
by 6–10 million olfactory neurons that are passing from the nasal cavity through the
cribriform plate to the olfactory bulb of the brain. Figure 1.1 is a schematic diagram
of a sagittal section of human nasal cavity.
2 Physiology of the Nose
The flow of air and particles in the cavity is controlled by structures within it. The
sense of smell is controlled by the nasal cavity’s olfactory area. The following fac-
tors should be considered when describing the functions of the nose: airway, olfac-
tion, effects on speech, air conditioning, reflex functions, and other common factors.
1 An Overview of the Anatomy and Physiology of Nasal Passage from Drug Delivery… 3
Fig. 1.1 Schematic diagram of a sagittal section of human nasal cavity showing lateral wall of the
nasal cavity (a), and cross-section through the middle of the nasal cavity (b), the respiratory epi-
thelium (c), and the olfactory epithelium [1]
2.1 Airway
One of the most essential components of airflow resistance is the nose. It has been
estimated that it provides between 30% and 50% of total resistance to inspiration.
Because the anterior nares’ cross-sectional diameter is much smaller than the poste-
rior choana’s, inspiratory air currents differ significantly from expiratory air cur-
rents. The former is directed upward into the nasopharynx, crossing the superior
surface of the inferior turbinate and the main surfaces of the middle turbinate. The
horizontal position of the front nares, the smooth anterior ends and surfaces of the
turbinates, and the form of the septum all contribute to this upward movement. The
expiratory airway differs in that it starts at a bigger posterior choana close to the
nasopharynx and travels through the nose to a relative constriction of the anterior
nares. This forms a huge central eddy or recurrent stream that flows into the inferior
meatus before joining the main nasopharyngeal stream. The airflow pattern used to
induce olfaction is slightly different. Sniffing starts the process. The air is redirected
superiorly into the olfactory cleft, where it stimulates the neuroepithelial
membrane. The olfactory area of the human nose is restricted to the roof of the nasal
cavity, the superior section of each superior turbinate in the lateral wall of the nose,
and the upper one-third of each nasal septum.
2.2 Olfaction
The olfactory area of the human nose is restricted to the roof of the nasal cavity, the
superior section of each superior turbinate in the lateral wall of the nose, and the
upper one-third of each nasal septum. The membrane has the histologic appearance
of a thick, pseudostratified, columnar nonciliated epithelium. Supporting cells,
basal cells, and olfactory receptor cells are the three types of cells found there. The
receptor cells are found in the vicinity of the supporting cells. Their population is
believed to be between 10 and 20 million people. They are oval in shape and operate
as both peripheral sensory receptors and neuronal cell bodies with processes. Each
olfactory cell’s central elongated end functions as a continuous strand or thick axon
that is encased by the basal cells. As an unmyelinated nerve axon, the filaments
breach the foundation membrane and become immediately continuous. The fibers
of the olfactory nerve are formed by these axons, which eventually terminate in the
glomeruli of the olfactory bulbs.
2.3 Effects on Speech
Vocal resonance is provided by the nose. It is self-evident that nasal blockage can
alter a person’s voice for any reason. The medical word for nasal resonance caused
by a nasal obstruction is rhinolalial clausa. When the vibrating air column moves
from the larynx to the pharynx, mouth, and nose, rhinolalial aperta occurs. This is
most typically seen in those who have a cleft palate. The vibrating air column of air
flowing from the larynx through the nose produces only the nasal consonants M,
N, and NC.
2.4 Air Conditioning
One of the most well-known functions of the nose is to humidify and heat the air
that is inhaled. The volume and rate of airflow, as well as the specific vascular nasal
mucosa, are critical in keeping the temperature of the inspired air within a narrow
range from the portal of entrance to the alveoli. Because the inspiratory air comes
into contact with a moist and warm nasal mucous membrane, the process of condi-
tioning this air changes very quickly. This quick change is due to the high tempera-
ture and water vapor pressure gradients. Expiration is, in some ways, the inverse of
inspiration. Some of the air that has been cooled by the preceding inspired air is
ejected from the dead space in the tracheobronchial tree and passes across the
mucosa of the pharynx and nose. This membrane absorbs the heat and moisture
from the air. In a 24-hour period, a healthy adult living in a temperate climate would
lose between 300 and 400 mL of water and between 250 and 350 kcal in expired air.
These values will rise as a result of increased physical activity and living in dry,
chilly settings. The humidification of the air happens at the same time as the heating
of the inspired air. Moisture is mostly obtained through the physical process of fluid
transudation through the mucosal epithelium. A lower volume is provided by epi-
thelial gland secretions and goblet cells in the nasal membrane. The actual volume
of fluid required to achieve this high level of saturation will be influenced by the
ambient air’s temperature and relative humidity. The daily volume of secretion and
transudate from the nose is estimated to be 1000 cc. Three-quarters of this is thought
to be used to saturate the inspired air, with the remaining going to the ciliary mecha-
nisms that clean and purify the air [2].
2.5 Reflex Functions
The nose is the source of a plethora of reflex functions. They are divided into two
groups: those begun by the sense of smell and those initiated by trigeminal nerve
ending terminals. Reflexes connected to digestive movements are a common exam-
ple of olfactory group functions. The olfactory centers stimulate the salivary, gas-
tric, and pancreatic glands reflexively. Changes in the lower respiratory tract,
particularly the larynx, trachea, and tracheobronchial tree, variations in the heart
rate, changes in pulmonary ventilation, and reflex sneezing are all examples of fifth
nerve reflexes.
2.6 Common Factors
Nasal blood flow can be affected by a variety of causes. The vasomotor response of
the nose is influenced by a variety of stimuli, both locally and generally. Changes in
ambient temperature and humidity, topical application of vasoactive medications,
external compression of major veins in the neck, trauma, and inflammation are all
examples of local factors. The natural mucous membrane of the nose reacts to keep
the nasopharynx and tracheobronchial airway in a homeostatic state. Congestion of
the nasal mucous membrane and restricted airflow are caused by para-
sympathomimetic medications, which also increase the volume of nasal secretions.
Cocaine causes considerable contraction of these erectile regions through constric-
tion of the capillaries and contraction of the cavernous tissue in the nose. On the
other hand, sympathomimetic medications that have been administered locally to
the mucosa will cause shrinking. Depending on the medicine, this impact can last
anywhere from 1 to 3 hours. Adrenaline and ephedrine are common medicines that
cause this reaction. Nasal blood flow is affected by a variety of elements, which
have been recognized for a long time. In their monograph on the nose, Holmes et al.
explain the emotional factors that are associated with nasal symptoms. Fear elicited
a “sympathetic” reaction from the nasal mucous membrane, but frustration, shame,
and anxiety engorged the mucous membrane and elicited a “parasympathetic”
response. They demonstrated these effects by comparing emotional tone to biopsies
of the inferior turbinate mucosa taken from people who had been through various
stages of emotional conflict [3].
3 Nasal Airflow
The human nose’s physiological activities, such as filtering and conditioning inhaled
air, respiratory feedback, and the perception of smell and discomfort, all require
adequate airflow. The anatomical structures of the nose, as well as the respiratory
situation, determine nasal airflow patterns. Human nasal anatomy varies substan-
tially from person to person, which is unsurprising. Furthermore, nasal illnesses
such as inflammation, allergies, sinusitis, and polyps can all obstruct nasal airflow
[4]. On the other hand, due to its position and the anterior nasal valve, air flows
superiorly into the nares. The airstream then passes into the nasopharynx, turning
roughly 90° posteriorly. After passing via the pharynx and larynx, the airstream
rotates inferiorly 90° and passes through the trachea, eventually reaching the lungs.
The anterior nasal valve is the narrowest part of the upper airway, positioned
1.5–2 cm posterior to the anterior nares. The upper airway’s narrowing allows for
close contact between the airstream and the mucosal surfaces. Evaporation of fluid
from the mucosal blanket causes humidification. The humidity level in the air is
between 75% and 80%. Contact between air and the abundant blood supply of the
nasal membranes, particularly the inferior turbinate mucosa, warms inspired air
to 36 °C.
Adults condition about 14,000 liters of air per day, which necessitates over 680 g
of water, or around 20% of our daily water intake. Sniffing is also an important
aspect of nasal airflow because it forces air into the superior nasal vault, where it can
better contact the olfactory mucosa [5].
4 Abnormal Nasal Physiology
Inhaled irritants and environmental allergens are the leading causes of nasal mem-
brane inflammation, for example, various chemicals, perfumes, cigarette smoke,
and other noxious odorants.
Nonallergic rhinitis, also known as vasomotor rhinitis, is caused by autonomic
nervous system dysfunction or alterations in blood flow caused by iatrogenic or
drug-related factors. Increases in blood flow or parasympathetic tone, as well as

decreases in sympathetic tone, cause nasal congestion and discharge. Nasal conges-
tion and discharge are reduced when blood flow is reduced, the parasympathetic
system is suppressed, and the sympathetic system is stimulated. Nasal systems may
be affected by supplemental female hormones or hormonal changes induced by
pregnancy or menstruation. Nasal physiology may be affected by medicines used to
treat hypertension or heart malfunction [6].
Nasal physiology is also influenced by anatomical abnormalities, which can impact
congestion, drainage, and olfaction in different ways. Airflow into the nasal cavity can
be affected by septal deviation and increased turbinates, changing it from a laminar to
a turbulent pattern (see the images below). Turbulent airflow irritates nasal mem-
branes even more, resulting in increased nasal discharge and congestion [6].
The most frequent cause of transient loss of smell is turbinate hypertrophy, which
can occur as a result of an upper respiratory infection (URI) or an allergic reaction.
The sense of smell is vital for quality of life, taste, and detecting smoke and other
potentially life-threatening odorants.
The primary sources of nitric oxide in the upper airways are nasal cavities and
paranasal sinuses (NO). Although the specific function of NO in nasal physiology is
unknown, it is assumed to be involved in host defense, ciliary motility, and a better
ventilation-perfusion ratio in the lungs through auto-inhalation. Low NO concentra-
tions have been found in illnesses including primary ciliary dyskinesia, cystic fibro-
sis, and acute and chronic maxillary sinusitis, whereas high NO concentrations have
been found in upper airway infection, allergic rhinitis, and nasal polyposis [6].
5 Tests of Nasal Physiology
Tests of nasal physiology include studies of airflow, ciliary function, and olfaction.
Nasal Endoscopy
Fiber optic cables and a light source are used in nasal endoscopy to see the nasal and
sinus cavities, which would otherwise be hard to see. Visualization is the best
approach for assessing a wide range of medical issues affecting the nose and sinuses
since it is less intrusive. A nasal endoscope is used to perform a variety of typical
surgical operations in the nose cavity [7].
Rhinomanometry
During exclusive nasal breathing, rhinomanometry aims to measure nasal airflow
and total nasal area. A nasal catheter is inserted into the nasopharynx to acquire dif-
ferential pressure readings. Nasal resistance testing evaluates all resistive compo-
nents of the nasal airway, from the anterior nares to the nasopharynx, and is sensitive
to even minor changes in airway diameter. This approach has been proven to be
effective in recording changes in nasal patency as a result of pharmaceutic or surgi-
cal treatments. It’s a somewhat invasive procedure that takes a long time to execute
and needs patient’s help [7].
Acoustic Rhinometry
Acoustic rhinometry is a relatively recent method of determining the cross-sectional
area of the nose and the volume of the nasal cavity by analyzing incident and
reflected sound during a brief pause in nasal breathing. This method has also been
verified and may be used to document changes in nasal patency as a result of phar-
maceutic or surgical treatments. It is a minimally invasive procedure that takes only
a few minutes to complete and requires little patient participation [7].
In a variety of clinical settings, rhinomanometry and acoustic rhinometry can be
used to assess nasal patency. Either test can be used to assess nasal airflow in gen-
eral and to compare premorbid circumstances to changes that may occur following
medicinal or surgical treatment. Furthermore, these tests can be used to compare
nasal passageways in order to plan medical or surgical procedures [7].
Clement et al. conducted a critical study and found that among the three tests—
active anterior rhinomanometry (AAR), four-phase rhinomanometry (4PR), and
acoustic rhinometry—AAR is the best way of objectively evaluating nasal patency.
While acoustic rhinometry, which evaluates different characteristics than AAR, has
limits and cannot be used in substitute of AAR, the researchers found that it may be
used as a supplementary test. Furthermore, Clement and colleagues claimed that
while 4PR may be capable of providing extra data, the open technical and mathe-
matical issues associated with it have yet to be fully resolved [7].
CT/MRI
Congenital, inflammatory, benign, and malignant diseases in the sinonasal area are
best evaluated using CT. CT and MRI offer the benefit of revealing anatomic fea-
tures that would otherwise be hidden, as well as displaying exquisite anatomic
detail [7].
Saccharin Test
The saccharin test is used to determine the mucociliary clearance time in the nose.
A drop of saccharin is put behind the inferior turbinate and brushed back into the
nasopharynx. As the cilia sweep the saccharin posteriorly, the patient should detect
a pleasant flavor. Poor ciliary function is indicated by delayed taste perception [7].
UPSIT
There are several olfactory tests available, but the most popular is the University of
Pennsylvania Smell Identification Test (UPSIT). The test consists of 40 questions,
each of which releases a distinct odor when scratched. The patient is asked to iden-
tify the odor that has been emitted by the examiner. This test can help with the
diagnosis of a variety of diseases, including Alzheimer’s, Parkinson’s, Huntington’s,
and others [7].
Nitric Oxide
Exhaled nitric oxide measurements may be useful as non-invasive objective tech-
niques for the assessment and management of normal nasal physiology and nasal
and sinus diseases in the future, according to a recent study.
Computational fluid dynamic studies of nasal airflow and physiology have

increased knowledge of the complicated nasal architecture and the consequences of
illness and surgery on physiology, according to research by Leong et al. and Liu
et al. [7].
6 Factors Affecting Intranasal Delivery
Intranasal delivery includes numerous benefits like non-invasiveness, convenient

self-administration, and quick absorption. Nevertheless, diverse factors influence
the restrictions and potency of IN drug delivery. Some include volume, permeabil-
ity, absorption, and mucus.
Volume and Permeability
The volume that can flow through a human’s nasal cavity is limited. Only 25–200 μL
of drugs is permitted [8]. This small amount hinders drugs from being efficiently
transported. Permeability is also crucial for the drug to enter the blood or brain.
However, the limited volume prevents drugs from having ample permeability in the
nasal cavity.
Mucus and Absorption
Mucus or nasal secretion performs a significant role in absorption. When mucus is
swallowed, it may carry the drug’s molecules to the gastrointestinal tract [9]. This
would prevent the drug from reaching the CNS. Hence, the drug must pervade the
mucus to be properly transported. Mucus promotes absorption if the drug remains
longer in the nasal cavity and is not inadequately transported.
7 Nasal Passage Targeting the CNS
The intranasal route functions as a direct passageway from the nasal cavity to the
brain. Drug(s) travel this route via olfactory or trigeminal nerves during IN delivery.
Molecules of a drug proceed along these nerves to their corresponding origins in the
brain: the cerebrum and pons. Subsequently, the drug disseminates in the brain
through the intranasal route’s two pathways, intracellular and extracellular. The
intracellular pathway carries drugs to the olfactory bulb with olfactory neurons and
various cells. The olfactory bulb directly connects and transfers the drug to the cen-
tral nervous system (CNS). The extracellular pathway transports drugs from the
nasal epithelium to the lamina propria until it externally travels along the axon. The
drug eventually reaches the CNS and fluid movement helps it to incrementally dis-
perse. Altogether, both pathways allow efficient transportation for nose-to-brain
drug delivery [9, 10].
8 Barriers to Drug Transport from Nose to Brain
Three important barriers that influence intranasal drug delivery and formulations
are the nasal epithelium, blood-brain barrier (BBB), and blood-cerebrospinal fluid
barrier (B-CSF-B). The latter two barriers can potentially be bypassed by IN drug
delivery.
Nasal Epithelium
The nasal epithelium is a crucial barrier to comprehend for nose-to-brain drug deliv-
ery. Its natural leaky behavior allows for swift absorption. However, the epitheli-
um’s compact junctions can restrict a drug’s impact due to the limited permeation of
the membrane [9]. Permeation enhancers can be utilized to overcome this barrier
and promote drug delivery to the CNS.
Blood-Brain Barrier (BBB)
The blood-brain barrier is a sensitive barrier that contains the basal lamina and
endothelial cells. Its limited access protects the central nervous system from xeno-
biotics or neurotoxic substances. This restriction defends the brain yet limits the
direct delivery of drugs to the CNS. Thus, many CNS drugs have been ineffective
due to the BBB. Drug transport from the nose to brain can bypass the barrier via the
intranasal route and molecular diffusion [9, 11].
Blood-Cerebrospinal Fluid Barrier (B-CSF-B)
Another determining barrier in CNS drugs is the blood-cerebrospinal fluid barrier
(B-CSF-B). It consists of cerebrospinal fluid (CSF) that encompasses the brain.
B-CSF-B protects the brain from detrimental substances by regulating the move-
ment of CSF. Like BBB, numerous drugs cannot pervade, and drug delivery via the
nasal cavity can avoid the B-CSF-B barrier.
8.1 Mucociliary Clearance
Mucociliary clearance is a defense mechanism in the lungs that can affect IN drug
delivery. Gravity and absence of cilia in the olfactory mucosa cause mucociliary
clearance to arise [12]. It also varies with environmental conditions and certain
illnesses. Mucociliary clearance rate determines the absorption of drugs, with a
lower rate being more optimal. Mucoadhesive agents can be employed to lower
the rate. Adding these agents to standard nanoemulsions can enhance the absorp-
tion and distribution of a drug in the brain. Mucoadhesive substances like chito-
san allow for better retention time and affect the mucus in the nasal cavity.
Furthermore, the viscosity of mucus and clearance rate are indirectly propor-
tional. Increasing the viscosity of mucus or drug formulation can decrease clear-
ance rate. Drug formulations with high viscosity such as gels are effective at
assisting drug absorption. It provides longer retention time with the nasal mucosa,
which can help increase absorption.
8.2 Physico-Chemical Properties of the Drugs
Proper drug formulations are necessary for efficacy and preventing irritation. There
are diverse forms of IN drugs—liquid, gels, nasal sprays—but they must maintain
standard properties. Several components include pH level, temperature, solubility,
and particle size.
pH Level and Temperature
pH levels and temperature can alter the mucus layer when the drug travels intrana-
sally [13]. The pH level must range from 5.0 to 6.5 to prevent irritation in the nasal
mucosa [10]. Temperature is another property that maintains drug stability and
potency. Mucociliary clearance rate can be influenced if the nasal cavity’s tempera-
ture is not within the range of 5–50 °C [13].
Solubility
Solubility can also affect drug absorption. Drugs travel quickly in the nasal cavity;
thus, there is insufficient time for a drug to be dissolved and effectively absorbed
[9]. This occurs because nose-to-brain drug delivery bypasses several phenomena:
dilution and first pass effect.
Pires and Santos suggest a formulation strategy that can improve low soluble and
less potent drugs. They express that these drugs require formulations and systems
that strengthen bioavailability and brain-targeted delivery. Using nanotechnology
like nanosystems is part of their strategy. Nanosystems can carry the drugs to certain
cells and tissues that may promote solubility through enhancing agents [10].
Particle Size
Particle size is an important property since the volume the intranasal route can carry
is limited. Particle sizes of a nanoemulsion can determine the drug’s retention time
and permeation in the nasal cavity and brain. A study by Ahmad et al. illustrated that
nanoemulsion (NE) particles approximately the size of 100 nm or smaller remained
longer in the nasal cavity. These 100 nm particles could travel through the intracel-
lular pathway, but only a particular amount. 900 nm NE particles were also tested;
they were able to move along the intranasal route yet were hindered by mucociliary
clearance and diffusion [14]. Particle sizes affect the drug potency but must be
minuscule to prevent irritation and poor absorption.
9 The Sensitivity of the Nasal Mucosa as a Limiting Factor
The nasal mucosa’s rapid clearance and tight junctions limit absorption and perme-
ability. Therefore, the sensitivity of the nasal mucosa acts as a guide for creating
drug formulations. Ideal formulations must have permeation and mucoadhesive
enhancing agents, higher viscosity, and certain pH levels. Nevertheless, fundamen-
tal characteristics of a drug can hinder exemplary formulations. For example, lipo-
philic drugs offer better bioavailability than certain hydrophilic drugs (peptides)
[13]. The latter exhibits low permeability along the nasal mucosa. If drugs cannot be
efficiently absorbed, they can easily be eliminated by mucociliary clearance.
Enzymatic degradation in the nasal mucosa is another concerning factor with pep-
tides. This degradation reduces the original dosage’s effect on the brain.
Implementing enzyme inhibitors can assist the peptides. It will not alter the pep-
tides’ bioavailability but can partially defend them from proteolytic enzymes [13].
The sensitivity of the nasal mucosa establishes various limitations, but alternatives
and enhancers are available.
10 Impact of Delivery Instructions, Patient Compliance,

and Body Position
Instructions and delivery systems can heavily affect patient compliance and drug
efficacy. Many common delivery systems like nasal sprays and drops require certain
steps and body positions. In an observational study by Rollema et al., they inter-
viewed 64 patients to determine how many accurately followed the patient informa-
tion leaflet (PIL) for intranasal corticosteroid sprays (INCS). Six percent of patients
followed all, while less than half had proper positions. These observations show that
instructions were either disregarded, unclear, or improperly instructed. Another
study by Trabut et al. illustrates their findings regarding body positions during self-
administration. Body positions are crucial because they can target specific areas of
the nasal cavity. They proposed that several positions—Lying Head Back and
Lateral Head Low—provide better alternatives to traditional ones like Head Down
and Forward. Head Down and Forward is unfavored because it causes higher dis-
comfort and poorly targets the middle meatus or middle turbinate. Results from
both studies exhibit the importance of instructions and body positions. Patients
should be properly advised since efficacy can be influenced by these factors [15, 16].
11 Conclusion
Intranasal drug deliveries are convenient, effective, and constantly improving with
new formulations and research. The easy accessibility through the intranasal route
implements IN drug deliveries as an appealing alternative to invasive administra-
tions. Administrating via this route allows drugs to bypass common, intricate barri-
ers and phenomenon: blood-brain barrier (BBB), blood-cerebrospinal fluid barrier
(B-CSF-B), and first-pass effect. Although there are many advantages, numerous
limitations hinder substantial efficacy. Major limitations include nose anatomy and
physiology, nasal inflammation, permeability, absorption, and mucociliary clear-
ance. New drug delivery formulations and systems should be constructed around
these diverse factors. Nasal tests and assessments should also be conducted to
ensure a patient receives optimal treatment. Non-invasive administrations display

higher patient compliance; however, patients should be fully informed of the recom-
mended instructions and positions for self-administration. Clinicians must be
responsible for providing this information and any applicable alternatives. Overall,
nose-to-brain drug delivery is a promising approach that can help alleviate various
illnesses.
References
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4. Zhao K, Jiang J. What is normal nasal airflow? A computational study of 22 healthy adults. Int
Forum Allergy Rhinol. 2014;4(6):435–46.
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0300-0729)
6. Archer SM. Nasal Physiology: Overview, Anatomy of the Nose, Nasal Airflow (medscape.
com), 2021.
7. Freeman SC, Karp DA, Kahwaji CI. Physiology, Nasal. 2022 May 8. In: StatPearls [Internet].
Treasure Island (FL): StatPearls Publishing; 2022 Jan–. PMID: 30252342.
8. Gao H. Progress and perspectives on targeting nanoparticles for brain drug delivery. Acta
Pharm Sin. 2016;6(4):268–86.
9. Erdő F, Bors LA, Farkas D, Bajza Á, Gizurarson S. Evaluation of intranasal delivery route of
drug Administration for Brain Targeting. Brain Res Bull. 2018;143:155–70.
10. Pires PC, Santos AO. Nanosystems in nose-to-brain drug delivery: a review of non-clinical
brain targeting studies. J Control Release. 2018;270:89–100.
11. Crowe TP, Greenlee MHW, Kanthasamy AG, Hsu WH. Mechanism of intranasal drug delivery
directly to the brain. Life Sci. 1973;(195):44–52.
12. Jain H, Prabhakar B, Shende P. Modulation of olfactory area for effective transportation of
actives in CNS disorders. J Drug Deliv Sci Technol. 2022;68:103091.
13. Dholakia J, Prabhakar B, Shende P. Strategies for the delivery of antidiabetic drugs via intra-
nasal route. Int J Pharm. 2021;608:121068.
14. Ahmad E, Feng Y, Qi J, Fan W, Ma Y, He H, Xia F, et al. Evidence of nose-to-brain delivery of
nanoemulsions: cargoes but not vehicles. Nanoscale. 2017;9(3):1174–83.
15. Trabut S, Friedrich H, Caversaccio M, Negoias S. Challenges in topical therapy of chronic
rhinosinusitis: the case of nasal drops application – a systematic review. Auris Nasus Larynx.
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corticosteroid sprays. J Asthma Allergy. 2019;12:91–4.
Chapter 2
Pharmacological and Clinical Problems
with Special Focus on Nasal Drug Delivery
Misha Mathur and Yashwant V. Pathak
Abstract The production of medications has seen many advancements. More dis-
eases can now be managed or even treated and a variety of symptoms can be elimi-
nated. While these options exist, many come with a long list of side effects with a
range of severity. Furthermore, many diseases still do not have treatment options,
such as neurodegenerative diseases like Alzheimer’s.
A method that has shown to be successful in addressing these concerns is nasal
drug delivery. Due to the manner in which it is introduced to the body, it decreases
the chance of many metabolic side effects such as nausea and vomiting. Furthermore,
the proximity of the nose to the brain would allow certain drugs to cross the blood-
brain barrier and enter the brain to address neurodegeneration. The nasal cavity was
also very beneficial in testing for COVID-19. It provided a convenient and non-
invasive way to test for COVID-19. However, there are certain constraints on drugs
that can be delivered nasally, depending on their properties. Yet, there have been
parallel developments on absorption enhancers, components mixed with the medi-
cation that can increase the absorption. Overall, introducing medication through the
nose has demonstrated a variety of benefits such as a decreased chance of side
effects, lower cost, and higher bioavailability.
Keywords Nasal passageway · Drug delivery · Blood-brain barrier · COVID-19 ·

Absorption enhancer · Bioavailability · Nasal to brain delivery
1 What Is the Nasal Passageway?
The nasal cavity, the nose, has two openings called external nares, commonly known
as nostrils. The internal nares are the open area between the nasal cavity and phar-
ynx. The nasal passage specifically is a subset of sinuses, which are hollow spaces
around the nose, cheek, and forehead. These three regions have four sinus cavities.
M. Mathur · Y. V. Pathak (*)

University of South Florida, Taneja College of Pharmacy, Tampa, FL, USA
e-mail: ypathak1@usf.edu

https://doi.org/10.1007/978-3-031-23112-4_2
16 M. Mathur and Y. V. Pathak
Firstly, the frontal sinuses are located at the base of the forehead, closely above the
eyebrows. The sphenoid sinuses are on the back of the head, close to the optic nerve.
The largest sinus is the maxillary sinuses which are on both sides of the nose under
the eyes. Ethmoid sinuses have three layers topped on each other. This tends to be
the location of the noticeable effects of a sinus infection. Understanding the sinuses
is a major part of understanding the nasal passageways since anything breathed in
will travel through all the sinuses. Furthermore, blockage of any part of the sinuses
will result in obstructed airflow [19]. The cells that line the nasal cavity possess tiny
hair, called cilia. The goblet cells also produce mucus in the nose. The mucus traps
certain pathogens and the cilia are responsible for clearing them from the nose [6].
2 What Is the Blood-Brain Barrier?
The information that humans and animals perceive and the actions they do are under
the control of the nervous system. One part of the nervous system, the central ner-
vous system, consists of the brain and the spinal cord and is known as the processing
center [2]. Since this is such a vital part of the body, the movement of materials such
as ions, molecules, and cells is tightly regulated by the cerebral endothelial cells
[20]. This is referred to as the blood-brain barrier. The endothelial cells in the cen-
tral nervous system form tight junctions to not allow large molecules in, such as
drugs and other solutes which have been quantified by the blood-brain barrier’s high
trans-endothelial electrical resistance [20]. The cerebrospinal fluid is hydrophobic.
Since hydrophobic and hydrophilic substances cannot interact, the blood-brain bar-
rier places an extra emphasis on blocking hydrophilic compounds from entering the
brain tissue [3]. Trials demonstrated successful delivery of these particles through
the blood-brain barrier when the particles were dissolved in an aqueous phase [21].
While the exact mechanism is not fully understood, it has been determined that if
medication is delivered to the olfactory region of the nares, the medication can
travel to the brain.
3 Properties of Nasal Passages
In terms of drug delivery, the nasal passageway can be categorized into the respira-
tory and the olfactory area. The respiratory area is located low in the nostrils, while
the olfactory area is located higher up in the nares. The epithelium in both catego-
ries is able to absorb molecules. While the exact mechanism is not fully understood,
it has been determined that if medication is delivered to the olfactory region of the
nares, the medication can travel to the brain by the olfactory neurons. From what is
understood, this is done by the trigeminal nerves [21]. By observing the movement
of stained dye that is administered nasally, it has been shown that the dye travels
through the middle concha, the maxillary sinus, and the choana, prior to reaching
2 Pharmacological and Clinical Problems with Special Focus on Nasal Drug Delivery 17
the trigeminal nerve. Overall, the dye was seen to reach the brain within 10 minutes
[8]. The steps of mucosal absorption are drug release, penetration, permeation, and
absorption [15].
4 Current Relevance with COVID-19
The connection between the nasal passageway with the rest of the body has been
extremely emphasized recently by SARS-CoV-2, the virus responsible for
COVID-19. In this virus, the primary point of entry was the nasal cavity. From
there, the virus had the ability to spread to the lungs which potentially developed
into more serious and even fatal conditions, such as pneumonia. The theory that the
nasal cavity is the point of origin can be explained due to ACE2, a cell surface
receptor utilized by the virus to get into cells, being present in larger quantities in
the nasal lining than in the lower airway cells. Research on the mechanism of this
virus has also highlighted the importance of TMPRSS2, a transmembrane protease
that allows the virus to reach the cells in the airway. Overall, these findings have
demonstrated that the upper airway is more susceptible to outbreaks [7].
The fact that the nasal passageway housed the markers that were needed to detect
COVID-19 was extremely beneficial. In nasal cavity swabbing, the swab goes to the
anterior nasal cavity which was important because this technique was quick, pain-
less, and could be done with basic medical training [10]. At the start of the pan-
demic, some people reported injury to the base of their skull. However, it has been
demonstrated that the individual swabbing will be able to successfully swab if they
go in the anterior nasal cavity at an angle of less than 30° while the patient’s head is
tilted back [12]. It was important to understand that the rapid antigen test which was
done after a swab was submitted was a successful strategy in testing individuals in
resource-poor countries [10]. Overall, the nasal passageway allowed for an easily
accessible and non-invasive route for COVID-19 detection (Fig. 2.1).
5 Benefits of Utilizing the Nasal Passageway
There are multiple options by which medications can be delivered to individuals,

such as intravenous, intramuscular, and many more. The oral passage is one of the
most common ways. However, there are certain circumstances in which the utiliza-
tion of the nasal passageway would be a more effective form of delivery than inges-
tion. The nasal cavity allows direct access to the bloodstream via the vascular
network for substances that can cross mucous membranes. This method allows for
rapid onset because the medication is directly absorbed into the systemic circula-
tion. Some medications can reach the brain from the nasal mucosa in less than
10 minutes. On the other hand, a medication taken orally must first go through the
liver and get metabolized to a certain degree prior to entering the bloodstream. The
Fig. 2.1 Ideal route of nasal swab for COVID-19 testing [12]
metabolism of the medication would not allow the proper dosage to enter the blood-
stream so physicians may have to prescribe higher dosages to account for the metab-
olism of the drug. Additionally, the process of metabolism takes many hours which
means that the medication would not reach the bloodstream for a couple of hours.
In contrast, since delivery via the nasal cavity is easily absorbed and does not have
to follow the longer absorption process that oral medications must, a smaller dosage
would suffice because the ingested amount would not have to be increased to take
metabolism into account. In turn, a smaller dose will result in less cost of the medi-
cation and a decreased chance of unintentional consequences to the body because a
higher dosage is correlated with a higher probability of developing side effects.
The nasal passageway is also effective at delivering medication across the blood-
brain barrier due to the proximity of the two regions, which results in a faster impact
of the medication administered [17]. Devices like nebulizers and mucosal atomiza-
tion devices (MAD), also known as nasal sprays, are extremely efficient at quickly
getting the medication absorbed into the bloodstream because these devices expel a
soluble mist in the mucosa [23]. These delivery methods are straightforward and
non-invasive, making them more convenient than other delivery methods. This con-
venient nature of the sprays leads to higher compliance in long-term therapy [18].
Many diseases that medicine does not have cures for are neural degeneration dis-
eases. This is largely due to the difficulty of delivering medication to the brain since
the delivery is blocked by the blood-brain barrier. Given that certain medications,
such as macromolecules and low molecular weight drugs are delivered via the olfac-
tory and trigeminal nerve pathways through the nasal passageway, they can easily
enter the blood [14]. Furthermore, research shows that the proximity of the nose and
the brain is pertinent in explaining how this passageway can bypass the blood-brain
barrier and deliver medication to cerebrospinal fluid [11]. Utilization of these pas-
sageways has prospective implications for finding treatments to neural diseases,
such as Alzheimer’s disease. Researchers have been able to reverse neurodegenera-
tion in a mouse model with Alzheimer’s disease [5] (Fig. 2.2).
Overall, medication delivered via the nasal passageway is easy to deliver, non-
invasive, and has decreased side effects. For example, there is a lower chance of
vomiting if the medication was delivered orally due to a lower dosage. Delivery via
the nasal passageway also demonstrates rapid onset. For example, nanoparticles
delivered via the nasal passageway had reached the nasal bulb in under 5 minutes [21].
Fig. 2.2 Flow of medication via nasal spray delivery [9]

It also allows for higher bioavailability and cost-effectiveness because fewer prod-
ucts being used correlate with lower prices.
6 Possible Barriers to the Utilization

of the Nasal Passageway
While there are many benefits of utilizing the nasal passageway, it is not the most
ideal delivery mechanism for all scenarios. Instead, the reason for delivery and the
medication being delivered highly influences the preferred route of administration.
In instances where it is pertinent to get the medication in the bloodstream right
away, intravenous delivery is more effective since it would have a 100% bioavail-
ability, which is a measure of the amount of medication that reaches the blood-
stream divided by the amount administered, which is also understood as the
proportion of the administered dose that enters the bloodstream and is available to
the site of action [23]. Furthermore, the nasal lining has a thick layer of mucus,
which is often composed of many enzymes. This may degrade the medication which
would result in an ineffective cure [21]. It is also imperative to ensure that the cilia,
and thus the nasal cavity, is not irritated by the drug.
The nasal passageway also doesn’t allow a large amount of medicine to be
administered when compared to taking something by mouth. Magnetic resonance
imaging (MRI) has determined that the average adult nostril opening is
357.83 ± 108.09 mm2 and the average nasal volume of an adult is
16,449.81 ± 4288.42 mm3 [21]. This small area and volume tend to only allow the
entry of nanoparticles. Even particles of 900 nm were not successful in reaching the
brain. On the other hand, a dosage too small like 100 nm results in a very small
amount reaching the brain [21].
However, in cases where the medication has a large molecular size, has poor
membrane permeability, or is likely to be degraded by aminopeptides, absorption
enhancers and enzyme inhibitors may be utilized. These work by increasing the
solubility of the drug, decreasing the surface tension of the mucus, and decreasing
enzymatic activity so that the medication is not broken down. The first possible bar-
rier is during absorption because medication would have to go through many layers,
such as the mucus layer, epithelial layer, interstitium, and capillary endothelium [4].
The nasal tract has a thinner and more permeable mucus layer than the respiratory
tract which allows the medication to be more readily absorbed. Hydrophobic mol-
ecules can cross the epithelial cell membrane via a concentration gradient but
hydrophilic molecules would require a transport system to cross the bilayer. Once
the medication has traveled across this physical barrier, there are other obstacles
that must be overcome as well. Surfactants provide support to the body by acting as
absorption enhancers. Specific examples include fatty acids, non-ionic surfactants,
and bio-surfactants, which assist the medication in crossing the epithelial layer dur-
ing nasal absorption [4].
Another obstacle that a medication must overcome is enzyme degradation. The

nasal passageway has enzymes such as proteases and nucleases that may degrade
the medication before it can be absorbed. Protease inhibitors have been effective in
animal trials, especially when it was delivered alongside absorption enhancers, such
as sodium glycocholate. A study was conducted on rats to demonstrate this relation-
ship in terms of insulin absorption through the lungs. Protease inhibitors enhanced
the absorption between 24.0% and 66.7% for a 90-minute timeframe [4]. Overall,
these mechanisms allow a higher bioavailability of the drug in the patient, which
will allow it to be more successful [15].
7 Properties of an Effective Nasal Delivery Drug
While the nasal passageway is a beneficial route of drug delivery, it is not always
feasible or necessary due to the characteristics of the drug. One important character-
istic to consider is size. When comparing the nasal passageway and the nose to other
common delivery pathways such as oral intake, there is a stark difference in the
amount of liquid that each can hold. Thus it is only feasible to use intranasal deliv-
ery when the particles are less than 10 nm, ideally 25–200 μL [23]. This is similar
to the characteristic that the quantity of the administered drug should be of low
molecular weight. To take this idea a step further, it is important to be cognizant that
if the delivered amount is smaller, the smaller amount should be more potent to
compensate for a lack of quantity. More specifically, medications that are capable of
being potent in nanomolar quantities are ideal for intranasal delivery. Some exam-
ples of therapeutics that have been successfully delivered intranasally are neutro-
phils, neuropeptides, cytokines, and polynucleotides [5] (Fig. 2.3).
The nasal passageway has been determined to be an effective manner in deliver-
ing mRNA. One application of this idea is seen in tumor vaccinations to cure cancer.
Since mRNA rapidly degrades, this delivery system is only effective if the mRNA
is encapsulated in nanoparticles. This idea has already been successful in mice. The
survival rates were 14.5–23 days for mice that were intranasal immunized while it
was only 7–13 for mice that had mRNA delivered in a naked form. Furthermore, 2
out of 10 ended the 40-day study while still being tumor-free [16].
When analyzing which medications are fit to be delivered intranasally, it is also
important to consider the physical properties. One property would be the solubility
of the medication. The nasal mucosa and the blood-brain barrier are cell membranes
with a lipid bilayer. Thus, for a medication to pass the membrane, it must be able to
dissolve. In other words, the medication must be lipophilic [23]. It is also notewor-
thy to consider the pH. When looking at the pH of the nose, there are different
measurements necessary for the anterior and posterior nasal cavity. However, the
pH of these regions varies depending on the individual. Thus, a study was done in
which the original pH of the participant’s nose was measured, and then the pH is
measured again after nasally administering different pHs of liquids. The average pH
of the anterior nose is 6.4, while the average pH of the posterior of the nasal cavity
Fig. 2.3 Characteristics of a successful nasal delivery drug [15]
is 6.27. The nasal anterior pH is much more sensitive than the posterior nasal cavity
pH [22]. For example, when a spray that has a pH of 7.2 is used, the average anterior
pH of 6.4 increased to 7.06 while the pH in the posterior of the nose did not change.
Ideally, nasal sprays are made to mirror the slightly acidic pH of the nose. Some
nasal sprays are manufactured without maintaining this ideology which leads to
some sprays having a high pH. However, a high pH can result in fungal infections,
stinging, and a decrease in the effectiveness of steroids [13]. Overall, the chemical
properties of the medication contribute to whether or not the medication will be well
tolerated, adequately absorbed, or effective [23].
8 Conclusion
Over the past decades, technology has greatly improved and has allowed for the
synthesis and production of a variety of medications. However, the increase in the
number of medications has consequently seen an increase in side effects and there
are still many diseases that have no cure. The nasal passageway has the ability to
overcome both of these obstacles in healthcare. Medications delivered via the nasal
route have lower dosages which decrease the chances of many side effects.
Furthermore, the proximity of the nose to the brain allows certain medications to
cross the blood-brain barrier. This opens a door to further research in neurological
diseases for which there is no way to deliver medication to the brain.
9 Future Trends
As described by COVID-19, a virus that originated in the nasal cavity can spread to
the lungs. Thus, studies in the future can extrapolate and assume that medication
administered through the nose could also make it to the lungs and be beneficial [1].
While the general idea of utilizing nasal passageways to administer some types
of medication seems extremely effective, it is more challenging to fully gauge if any
of the medication delivered this way would result in detrimental side effects. Various
trials have been done in mice but due to the varying physiology between humans
and various animals is very profound so it is unclear to assume a direct correlation
of results [15]. However, once this is better understood, another possibility is work-
ing on ensuring that medication delivered via the nasal passageway remains at the
nasal mucosal surface so that it can eventually penetrate the blood-brain barrier.
More research and discovery on this may shed some light on cures for neurological
disorders [21].
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mb.ca/old/extranet/eipt/files/EIPT-055.pdf
Chapter 3
Drug Absorption via the Nasal Route:
Opportunities and Challenges
Seth Kwabena Amponsah and Ismaila Adams
Abstract Drug administration via the nasal route appears to be another reliable
way of getting drugs into systemic circulation. The nasal route has easy access,
large surface area, is well vascularized, and circumvents first-pass metabolism.
Currently, there is a lot of attention on nasal delivery of drugs. This route has been
found to aid rapid absorption of drugs into systemic circulation. The mucosa found
in the nasal cavity has been shown to aid absorption of bioadhesive drug delivery
devices. Microspheres, liposomes, and gels expand readily when they come into
contact with the mucosa in the nasal cavity. Furthermore, to enhance absorption of
drugs, a number of methods have been used to extend residence time in the nasal
cavity. However, prospects of using the nasal route as a means of getting drugs into
systemic circulation face a number of challenges. Some of which include barriers in
the mucosa and toxicity which may be associated with the excipients used. Drug
absorption enhancers are currently being explored to improve intranasal drug deliv-
ery. Drug absorption via the nasal route (opportunities and challenges) is discussed
in this chapter.
Keywords Absorption · Challenges · Enzyme inhibitors · Nasal route · Surfactants
1 Introduction
Over the years, nasal administration of drugs has had several systemic applications.
Some of the applications include management of pain, allergy, infections, osteopo-
rosis, and sexual dysfunction [1, 2]. The nasal route also serves as a topical site for
administration of drugs in the management of nasal congestion associated with
allergic rhinitis [3]. Decongestants and antihistamines are examples of drugs admin-
istered topically in the treatment of rhinitis [4, 5].
S. K. Amponsah (*) · I. Adams

Department of Medical Pharmacology, University of Ghana Medical School, Accra, Ghana
e-mail: skamponsah@ug.edu.gh

https://doi.org/10.1007/978-3-031-23112-4_3
26 S. K. Amponsah and I. Adams
A number of studies have explored the nasal route in the administration of drugs
for cardiovascular indications [6]. In individuals with angina pectoris, administra-
tion of propranolol via the nasal route has been found to improve tolerance during
exercise [7]. Patients with perioperative hypertension and hypertensive crises have
been treated with intranasal nifedipine [7]. Nitroglycerin given intranasally has
been proven to reduce hypertensive episodes after endotracheal intubation [8].
Intranasal administration of drugs for cardiovascular diseases, according to the
aforementioned studies and others, might be used in clinical settings when fast and
intermittent therapeutic effects are needed, and could possibly replace parenteral
drug administration [9, 10].
In 1991, it was discovered that it was possible to deliver drugs to the central
nervous system (CNS) via the nasal route [11]. Subsequently, there has been a surge
in nasal drug delivery systems, particularly nose-to-brain delivery [12]. Therapeutic
proteins currently exist for some CNS diseases [8]. These agents have the potential
to improve efficacy while minimizing adverse effects [8]. Additionally, intranasal
drug delivery systems have been employed in the management of diabetes-mediated
cerebral degeneration and Alzheimer’s disease [9, 10]. Glioblastoma is currently
being managed with anticancer drugs administered via the nasal route [13, 14].
Several other drugs could be administered through the nasal route. Indeed,
administration of peptide hormones (calcitonin, desmopressin, insulin, and gluca-
gon) is also possible via the nasal route [15, 16]. Inhaled insulin is currently avail-
able on the market [17]. Analgesics and some rescue drugs (e.g., naloxone) depend
on rapid absorption via the nasal mucosa [18]. In addition, intranasal triptans have
been used for migraines, fentanyl for pain, and ondansetron for nausea [19–21].
Vaccines may benefit from intranasal administration as well [22]. Intranasal vac-
cination can provide wider protection because of lymphoid tissues found in the
nasal cavity [23]. Intranasal vaccination can bring about mucosal and systemic
immunity [24]. Furthermore, the nasal route provides cross-protection against dif-
ferent types of viruses, such as influenza, which might help with the creation of
“universal vaccinations” [25]. The nasal route has also been linked with possible
vaccination for hepatitis B [26].
The anatomical position and physiological features (surface area, innervation,
and blood supply) of the nose make it an external conduit to the lungs, and an appro-
priate route for the topical and systemic administration of drugs [27–30]. When
systemic distribution is required, intranasal administration provides excellent bio-
availability. The nasal route has some advantages over the oral route. These include
rapid onset of drug action, fast attainment of therapeutic drug levels, and the fact
that small drug doses can achieve required therapeutic effect [31]. Despite its advan-
tages, administration of drugs via the nasal route could be plagued by mucociliary
clearance, metabolic obstacles (peptides/proteins), and inadequate drug deposition
to the targeted site [32]. As a result of the aforementioned, there has been minimal
progress with new drug candidates for nasal route administration [15, 33].
3 Drug Absorption via the Nasal Route: Opportunities and Challenges 27
2 Challenges Associated with Absorption of Nasal Drug

Delivery Systems
There are a number of challenges that could potentially affect the use of the nasal
route for systemic drug delivery. Some of these challenges include barriers against
mucosal drug absorption and toxicity associated with intranasal administrations.
2.1 Barriers Against Mucosal Drug Absorption
Absorption of the active pharmaceutical ingredient (API) of a drug would require

that it moves across a number of barriers before reaching circulation. These barriers
include mucus layer (surfactant), epithelial layer, basement membrane, and capil-
lary endothelium [34], as shown in Fig. 3.1.
Administered drugs usually get deposited on the first barrier: mucus layer [35].
There could be ciliary action that can remove the drug from the absorption site [36].
Thus, knowledge of the thickness of the mucus layer and clearance is important in
the development of drug delivery systems [37–39]. The nasal tract has a thin mucus
layer. This layer is highly permeable compared to other mucosal surfaces [40, 41].
Drug clearance from the nasal cavity, which is determined by nasal mucociliary
clearance, is required for drug absorption via the nasal route.
The second barrier is the epithelial layer. This layer is made up of pseudostrati-
fied columnar cells that are linked together by tight junctions [42]. A number of
drugs are absorbed primarily through transcellular diffusion, which occurs when
they pass through the epithelial cell membrane. A concentration gradient allows
small hydrophobic molecules to partition across biological membranes. To cross the
lipid bilayer, hydrophilic molecules usually require a selective transport system.
Paracellular mechanism can aid the absorption of large and polar drugs, and the
Fig. 3.1 Barriers against mucosal drug absorption via the nasal route
tight junctions usually act as barriers [43]. Absorption across the capillary endothe-
lium is necessary for APIs that have to reach systemic circulation. Strategies to
overcome these aforementioned barriers to improve absorption are thus relevant.
2.2 Toxicity Associated with Intranasal Applications
When developing a pharmacological formulation for intranasal administration,

safety is paramount. Usually, large molecules (peptides and proteins) would require
absorption enhancers. Other excipients serve as mucoadhesives which can extend
contact time with the nasal mucosa. Excipients can greatly reduce the safety of the
final therapeutic product [44–46]. Therefore, the toxicological implications of a
drug formulation must also be examined in vivo.
Benzalkonium chloride, which is used in cosmetics and various nasal formula-
tions, is a good illustration of the toxicological importance of preservatives [47].
The safety of benzalkonium chloride is debatable. Some research report that benzal-
konium chloride has no harmful impact in vivo [47]. The use of benzalkonium chlo-
ride in medical products for nasal use is between 0.02 and 0.33 mg/mL; occasionally
cilia toxicity in vitro and in vivo has been reported [47–49].
For CNS conditions such as epilepsy, psychosis, and glioma, the use of
nanotechnology-based drug delivery systems has to be done with caution [50, 51].
Oxymetazoline is a nasally administered sympathomimetic that is used as an anes-
thetic and in the treatment of epistaxis. It is noteworthy, however, that the unfavor-
able side effects of intranasal oxymetazoline are independent of the route of
administration [52].
3 Drug Absorption Enhancers as Opportunity for Improving

Nasal Drug Delivery
There are a number of permeation enhancers that are known to improve nasal and
pulmonary drug administration. Some of these include surfactants, tight junction
modulators, protease inhibitors, cyclodextrins, and cationic polymers [36, 53, 54].
3.1 Surfactants
Surfactants are amphiphilic molecules with lipophilic and hydrophilic residues

[55]. Surfactants can improve absorption by disrupting the cell membrane through
membrane protein leaching, opening tight junctions, and can prevent degradation of
drugs by enzymes [56]. Surfactants that can be used as absorption enhancers include
phospholipids, bile salts and their derivatives, fatty acids, non-ionic surfactants, and
alkyl glycosides [57–59].
3.1.1 Phospholipids
Natural pulmonary surfactant could be a mixture of about 90% phospholipids and

10% proteins [60]. The primary function of this surfactant is to reduce surface ten-
sion [61]. Phospholipids have also been shown to improve absorption of drugs [60].
Dipalmitoylphosphatidylcholine (DPPC), for example, is a major component of
lung surfactant [62]. DPPC has been used to improve absorption of parathyroid
hormone from the lungs [63]. Phosphatidylcholines, sphingomyelin, phosphati-
dylinositol, phosphatidylglycerol, and neutral lipid have also been used as absorp-
tion enhancers [35, 64]. In a diabetic rat model, phospholipid hexadecanol tyloxapol
(PHT) was tested as an absorption enhancer for recombinant human insulin in the
lungs [65]. To further investigate its absorption potential, PHT was tested in vitro on
Calu-3 ALI (air-liquid culture) cells [66, 67]. The recombinant human insulin was
found to permeate the cell layer more efficiently in vitro [66, 67]. It can be postu-
lated that PHT interacted with tight junctions and enhanced absorption via the para-
cellular route [35].
3.1.2 Bile Salts and Their Derivatives
Bile salts and their derivatives have the ability to enhance drug absorption.
Glycodeoxycholate, salts of cholate, taurodeoxycholate, and taurocholate have been
investigated as absorption enhancers for nasal and pulmonary drug administration
[57, 68–70]. One of the most commonly used bile salts that has the ability to enhance
bioavailability, particularly insulin, is sodium taurocholate [71, 72]. Despite the fact
that bile salts and derivatives have shown promise as absorption enhancers, their
safety is a major concern. The impact of inhaled bile salts as absorption enhancers
on surfactant function has been studied in vitro and in vivo. Bile salts were found to
impair surfactant function in vitro and caused lung irritation in vivo [70].
3.1.3 Fatty Acids
Polyunsaturated fatty acids (PUFA) have been studied as nasal and pulmonary drug
absorption enhancers [73, 74]. PUFA has tight junction modulatory action and
improves drug penetration across epithelial cell barriers [75, 76]. Although the spe-
cific mechanism is uncertain, research has shown that PUFA may affect membrane
permeability by increasing membrane fluidity or act via Ca2+-dependent tight junc-
tion processes [77–79]. Absorption of fluorescein isothiocyanate was improved
when arachidonic acid was added [80]. Often, medium-chain fatty acids like capric
and lauric acid are used as absorption enhancers [81, 82].
3.1.4 Non-ionic Surfactants
Non-ionic surfactants are relatively non-toxic in nature [83]. Alkylglycosides (AGs)

are non-ionic surfactants that include groups like glucose, sucrose, and maltose that
are linked to alkyl chains of varying lengths [84]. The two most widely used AGs
are N-lauryl-b-d-maltopyranoside and tetradecyl maltoside [85, 86]. At extremely
low doses, these two agents have demonstrated significant nasal absorption aug-
mentation characteristics [54]. Also, AGs could be used to improve insulin, calcito-
nin, and glucagon absorption via the nasal route [87]. The exact mechanism behind
these in vivo absorption-boosting actions of tetradecyl maltoside is unknown. AGs
may have direct effect on epithelial cells, most likely through the paracellular route
[88]. Despite their ability to improve absorption, AGs are toxic to airway epithelial
cells, most likely due to a membrane-damaging action [89].
Another non-ionic surfactant, poloxamer 188, has been extensively studied for
its role in intranasal drug delivery [90]. Addition of poloxamer 188 to nano-cubic
vehicles for intranasal administration has been shown to affect their flexibility [91].
An insulin formulation that contained sucrose cocoate (0.5%) was found to increase
plasma insulin level [92].
The non-ionic surfactants cremophor EL and laurate SE have been evaluated as
intranasal absorption enhancers in rats [56]. Cremophor EL was found to open
Caco-2 cell tight junctions when used as an excipient in taxol infusions, ritonavir
oral gelatin capsules, and oral solutions [93], hence, could be a good intranasal drug
absorption enhancer.
3.1.5 Biosurfactants
Biosurfactants are surface-active chemicals produced by living organisms like bac-

teria, fungus, and yeast [94]. Biosurfactants are non-toxic, ecologically friendly,
and biodegradable in most cases [95]. Previously, biosurfactants were studied as
medication absorption enhancers. Rhamnolipids are a kind of biosurfactant that
have been studied extensively [96]. Rhamnolipids have been shown to affect epithe-
lial permeability across Calu-3 and Caco-2 cells [97].
3.1.6 Animal-Derived Surfactants
There have been a number of studies that have used animal-derived surfactants to
aid drug absorption. Gentamicin and polymyxin E were delivered into the lung of a
newborn rabbit using poractant alfa, an animal-derived surfactant [98]. The bacteri-
cidal activities of gentamicin and polymycin E were enhanced in vivo [98]. At rest-
ing transpulmonary pressures, poractant alfa decreased surface tension at the
air-liquid interface on alveolar surfaces during breathing and stabilized alveoli
against collapse [99].
3.2 Enzyme Inhibitors
A significant number of enzymes are found in airway surfaces (fluids and mucus),
which may destroy active constituents of drugs before absorption [100].
Aminopeptidases and proteases account for the majority of enzymes found in the
nasal pathway and lungs [101, 102]. Drugs that are peptides, proteins, and nucleic
acids are particularly susceptible to this metabolism. Thus, methods to protect drugs
against breakdown by these enzymes within nasal cavities may be required.
Bacitracin, leupeptin, soybean trypsin inhibitor, bestatin, and phosphoramidon are
some of the protease inhibitors that have been explored as absorption enhancers in
nasal or pulmonary medication delivery systems [103, 104]. Rats were given insulin
with a protease inhibitor by intratracheal injection during a trial [105]. The hypogly-
cemic and hypocalcemic response of insulin from the lungs was examined. Within
90 minutes of administration of insulin, the plasma concentration of glucose reached
a minimum baseline (24.0–66.7%) in the presence of the protease inhibitors [105].
Bacitracin (20 mM) has also been shown to be effective in increasing insulin pulmo-
nary absorption [35].
Neutrophil-specific serine proteases, neutrophil elastase (NE), are released into
the lung lumen during infection or inflammation [106]. It is hypothesized that
excessive NE builds up in the pulmonary fluid of individuals with persistent lung
infection, and this has the tendency to reduce inhaled drug absorption [107]. A
rapid-acting NE inhibitor, EPI-HNE-4, has been used in patients with cystic fibro-
sis [108].
3.3 Cationic Polymers
Cationic polymers usually have cationic substances integrated into their side chains
[109]. Examples of cationic polymers include cationic gelatins, cationic pullulans,
polyethylenamine, chitosan, and poly-L-arginine [110]. Negative-charged insulin in
neutral fluids can interact with cationic polymers and improve insulin absorption
[111]. A good contact can assist insulin to get to the cell surface; conversely, a bad
interaction can prevent insulin from getting to the cell surface [112].
Polyethylenamine, a cationic polymer, has shown promise as a drug carrier in the
nasal cavity [113]. Polyethylenamine is known to improve insulin absorption in the
lungs of rats [114]. Another cationic polymer, spermined dextran (SD), has been
investigated as an absorption enhancer for drugs intended for pulmonary adminis-
tration [115]. SD was found to boost insulin absorption and FD-4 permeability
through Calu-3 cells [115]. The mechanism of action of SD is unknown, however,
it is thought that the molecule causes the opening of tight junctions, allowing water-
soluble drugs to pass between cells [116].
Cationic polyelectrolytes such as chitosan can also enhance absorption [117].
Chitosan and its derivatives have been employed in the development of
mucoadhesive polymers because of their biocompatibility, biodegradability, and

low toxicity [118]. They are ideal as pharmaceutical excipients due to the aforemen-
tioned properties. Chitosan has mucoadhesive characteristics because it interacts
electrostatically with mucin chains that are negatively charged [119]. This mucoad-
hesion increases API absorption by prolonging the period of residence of the drug.
On the other hand, chitosan derivatives at physiological pH are poorly soluble in
water and this restricts their use [120]. Due to the fact that chitosan has good muco-
adhesive capabilities, various derivatives have been created to address solubility
problems [121]. Chitosan oligomers, for example, have high water solubility than
normal chitosan and have been studied for their ability to improve absorption via the
nasal route or lungs [122]. A 0.5% (w/v) chitosan hexamer seemed to be more effi-
cient than other chitosan oligomers at the same concentration in increasing inter-
feron pulmonary absorption when compared to control. O-palmitoyl chitosan,
which is made from chitosan and palmitoyl chloride, has been shown to have better
mucoadhesive and absorption-boosting characteristics [123, 124].
In rats, sperminated pullulans have been found to improve insulin pulmonary
absorption [125]. When a 0.1% (w/v) solution of sperminated pullulans was admin-
istered with insulin concurrently in vivo, insulin absorption improved [125].
Pulmonary delivery of salmon calcitonin in rats was investigated using both posi-
tively and negatively charged gelatin microspheres [126]. The pharmacological
impact of salmon calcitonin was considerably greater in the positively charged gela-
tin microspheres than the negatively charged ones [126]. Subsequently, salmon cal-
citonin was administered in positively charged gelatin microspheres, and this lead
to a greater pharmacological effect.
3.4 Polyamines
Polyamines have also been studied for their ability to improve drug absorption
[127]. In mammalian cells, the polyamines spermine and spermidine are abundant
[128]. Reports suggest that polyamines, notably spermine and spermidine, may sig-
nificantly enhance absorption of insulin and other water-soluble agents without
causing tissue membrane injury [129, 130]. The opening of epithelial tight junctions
is thought to be responsible for the absorption-enhancing action of spermine [36].
In rats, sperminated dextrans enhanced insulin pulmonary absorption and FD-4
penetration across Calu-3 cell monolayers in vitro [131].
3.5 Tight Junction Modulators
Tight connections decrease gaps between neighboring epithelial cells [132]. Drug
absorption via the paracellular route can be improved by modulating tight junctions.
Large macromolecules cannot be transported by paracellular transport, which is
limited to substances with molecular radii of less than 11 nm [133]. Nonetheless,
low-molecular-weight hydrophilic drugs have limited bioavailability [134]. Some
peptides, including thyrotropin-releasing hormone, desmopressin, and octreotide,
have been found to be absorbed in between tight junctions [135, 136]. Tight junc-
tion modulators that target tight junction proteins like claudin are 400 times more
effective than other treatments in opening these tight junctions [137].
4 Conclusion
Intranasal drug formulations may have local and systemic indications. Suitability of
the nasal route for local and systemic drug delivery is based on its rich blood supply,
large surface area, and avoidance of pre-systemic drug metabolism. Drugs could be
administered via the nasal route for the management of rhinitis and pain, among
others. Over the last few years, there has been an interest in intranasal drugs for
Table 3.1 Absorption enhancers and their potential mechanisms of action

Absorption
enhancers Example(s) Possible mechanisms References
Phospholipids Phosphatidylglycerol, May interact with tight [53, 138,
phosphatidylinositol, junctions and enhance 139]
phosphatidylethanolamine absorption via the
paracellular route
Bile salts and Deoxycholate, glycocholate, Modulation of tight [100, 138]
derivatives glycodeoxycholate junctions
Fatty acids Capric acid Contraction of actin [78, 79]
microfilaments and
dilatation of tight junctions
Non-ionic Poloxamer 188, Direct effect on the [84, 140]
surfactants tetradecylmaltoside epithelial layer, most likely
through the paracellular
route
Biosurfactants Rhamnolipids Enhance epithelial [97]
permeability across Caco-2
and Calu-3 monolayers
Animal-derived Poractant alfa Decrease surface tension at [99]
surfactants the air-liquid interface
Enzyme Nafamostat mesilate, aprotinin, Prevent enzymatic [103, 105]
inhibitors bacitracin breakdown within nasal
cavity
Cationic Poly-L-arginine, Cationic Alteration of tight junctions [120, 136,
polymers pullulans, chitosan, 141]
polyethylenamine
Polyamines Spermine, sperminated dextran Open tight junctions [115, 128]
Tight junction Claudin Open tight junctions [75]
modulators
diabetes, cardiovascular, and CNS diseases. Despite its merits, administration of

drugs via the nasal route could be affected by barriers against mucosal drug absorp-
tion, and inadequate drug deposition to the targeted site. To improve absorption via
the nasal route, surfactants, protease inhibitors, cationic polymers, cyclodextrins,
and tight junction modulators (as summarized in Table 3.1) can be used.
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Chapter 4
Factors Affecting the Design of Nasal Drug
Delivery System
Jéssica Bassi da Silva, Maria Vitoria Gouveia Botan,

and Marcos Luciano Bruschi
Abstract Over the recent years, many strategies have improved the delivery of
bioactive agents to the brain, improving the treatment of several pathologies.
Nonetheless, the design of new formulations is highly dependent on the capacity of
the drugs to permeate the blood-brain barrier (BBB) and reach a significant effect
on the neurological disorders. Therefore, different approaches have been studied in
order to facilitate the delivery of different drugs into the brain; for instance, intrana-
sal administration has gained special interest. The nose-to-brain delivery provides a
direct pathway of drug delivery to the brain without the need to permeate the BBB,
potentially avoiding adverse effects that could occur when the drug is systemically
absorbed. Although it may overcome BBB, there are alternative barriers that have to
be circumvented for nose-to-brain route and different strategies have been mas-
sively studied. This chapter provides a comprehensive overview of factors related to
the physiology of nasal cavity, the drug, and the formulation that can affect the
development of formulations for nose-to-brain drug delivery.
Keywords Design of nasal drug delivery · Blood-brain barrier · Nasal cavity ·

Drug formulation · Nasal absorption · Nose-to-brain drug delivery
1 Introduction
The drug administration by nasal route has been used for more than three decades
aiming at the therapy of systemic and local diseases, such as perennial and allergic
rhinitis, inflammations, pains, and also microbial infections [1, 2]. Nowadays, the
J. B. da Silva · M. V. G. Botan · M. L. Bruschi (*)

Department of Pharmacy, State University of Maringa, Maringa, PR, Brazil
e-mail: mlbruschi@uem.br

https://doi.org/10.1007/978-3-031-23112-4_4
44 J. B. da Silva et al.
use of this pathway for the delivery of challenging bioactive compounds is of great
interest among the scientific community and in the pharmaceutical industry [3, 4].
During the last two decades, this administration route has been used for systemi-
cally acting bioactive agents that are difficult to deliver via routes other than paren-
teral and for nose-to-brain drug delivery. Moreover, the nasal administration of
vaccines and other preparations for prevention against infections constitute an
important and efficient strategy [5], which is being highlighted with the COVID-19
pandemic, for example.
The nasal administration constitutes an important strategy for the rapid onset of
drug action. Many characteristics of nasal region synergically enhance the perme-
ation of bioactive agents administered by this route, such as the relatively large
surface area (high number of microvilli), the presence of a porous endothelial mem-
brane, and a highly vascularized epithelium [4]. Conventional relatively lipophilic
low-molecular–weight bioactive agents can be absorbed with improved degree of
efficiency across the different regions of nasal cavity, resulting in suitable availabil-
ity at the action site. The nasal route enables to achieve similar blood level profiles
for many bioactive agents compared to the intravenous administration [1].
In this context, the nasal drug delivery can enable many possibilities [2, 3]:
• Local delivery for the treatment of nasal allergy, congestion, and/or infection.
• Systemic delivery for crisis treatments when a rapid onset is necessary.
• Systemic delivery for long-term therapy (daily administration).
• Systemic delivery of peptides and proteins (when difficult to administrate).
• Vaccine delivery using antigens (whole cells, split cells, surface antigens, and
others) and DNA.
• Central nervous system access for reaching local receptors and/or to circumvent
the blood-brain barrier.
The function and physiology of the nasal cavity, the metabolism (e.g., the pres-
ence of cytochromes P-450, especially in the epithelium of the olfactory region),
and the patient’s pathological state must be deeply understood during the develop-
ment of nasal formulations. Physicochemical characteristics of drug (e.g., charge,
molecular weight, and lipophilicity), anatomical and physiological variables (e.g.,
membrane transport, deposition, enzymatic degradation, and mucociliary clear-
ance), and also formulations characteristics (e.g., pH, osmolarity, viscosity, concen-
tration, volume and type of dosage form) constitute important factors that impact
the design of nasal drug delivery system.
Conventional relatively lipophilic low-molecular-weight bioactive molecules
have been shown to be efficiently absorbed across the nasal cavity. However, large
hydrophilic molecules (e.g., peptides and proteins) often display less efficient
absorption by this route of administration. Therefore, compounds of hydrophilic
nature, large molecular size, enzymatic degradation, and fast nasal mucociliary
clearance (movement away from the absorption site in the nasal cavity) are chal-
lenges for the development of nasal drug delivery formulations [1].
Therefore, many physicochemical and technological strategies have been used
for overcoming these drawbacks. The modification of the permeability of nasal
4 Factors Affecting the Design of Nasal Drug Delivery System 45
membrane using absorption enhancers (e.g., bile salts, surfactants, cyclodextrins,

phospholipids, and fatty acids), the use of micro and nanostructured systems, and
mucoadhesive and/or environmentally responsive systems are some examples [1].
This chapter provides a comprehensive overview of the factors that can affect the
development of formulations for nasal drug delivery. The factors related to the phys-
iology of nasal cavity, the drug, and the formulation are discussed. In addition, the
design, optimization, and challenges are also considered.
2 Factors Related to Nasal Anatomy and Physiology
The development of new medicines for nasal application with the aim of delivery
drugs is a challenge, given that the nasal cavity and the body have mechanisms able
to hinder the absorption of biologically active agents administered at this place.
Among the factors related to physiology that may impair the absorption there are
mucociliary clearance, blood flow, enzymatic degradation, and the physical condi-
tion of the nasal mucosa [4, 6].
The ciliary mucosa present in the nasal cavity is responsible for mucociliary
clearance. This process works for the protection and maintenance of the physiologi-
cal environment, constituting the main defense mechanism against foreign bodies
(i.e., bacteria, dust, and other types of allergens) that can settle in the nasal cavity;
however, they can also remove the formulation administered in this location. When
a preparation is administered through the nasal route, the mucociliary clearance
eliminates around 50% of it in 15–30 min, fostering a shorter contact time of the
drug with the mucosa, which considerably reduces its absorption [7, 8]. The nasal
blood flow also plays a great role in drug absorption. The nasal mucosa is highly
vascularized and the blood flow will depend on vasoconstriction and vasodilation of
the blood vessels. Better blood circulation makes it easier for drugs to be absorbed
and distributed through the system [9, 10].
Although intranasal administration has the advantage that the drugs do not
undergo the first-pass effect, considering the molecules are absorbed by trigeminal
pathways present in the nasal cavity or by the systemic route, directly reaching the
bloodstream, some drugs can still suffer due to the presence of metabolizing
enzymes present in the nasal mucosa. For instance, proteins and peptides can suffer
alteration by proteases and aminopeptidases. Meanwhile, the cytochrome P450 is
responsible for the metabolization of some drugs, such as progesterone, cocaine,
and decongestants [6, 11, 12].
Finally, changes in the physical conditions of the nasal cavity can also change
drug absorption. Colds, allergies, and nasal infections can impair absorption because
in these cases the mucociliary clearance is increased, which reduces the time the
formulation remains in the mucosa. Inflammation or irritation of the mucous mem-
brane causes swelling and also changes the conditions of drug absorption. Still
related to nasal secretions, the circadian rhythm influences them. Some studies have
revealed that the secretion and clearance rates of nasal cavity are reduced at night,
thus altering the permeating of drugs. In addition, some diseases can change the pH
of the mucosa and the viscosity of the mucus as well [4, 10–12].
3 Factors Related to the Biologically Active Agent
The physicochemical characteristics related to the biologically active agent also

influence the development of a formulation. The three main characteristics involved
in the passage of bioactive agents through the nasal mucosa are molecular weight,
lipophilicity, and degree of ionization. Regarding molecular weight, molecules up
to 400 Da are still freely and easily transported; however, larger molecules are more
difficult to absorb. Generally, drugs with molecular weight above 1000 Da exhibit
low ability to penetrate the physiological barrier. A high molecular weight limits the
paracellular passage of a drug through tight junctions. In these cases, the best alter-
native is to use special drug delivery systems, such as permeation enhancers, for
example, cyclodextrins and surfactants, or nanosized systems, in order to improve
the bioavailability of the drug [7, 8].
The partition coefficient of the biologically active agent is another factor that
impacts drug absorption. Due to the lipophilic nature of the biological membranes,
the more a molecule is lipophilic, the more easily it will pass through physiological
barriers. However, a massive lipophilicity makes the drug difficult to dissolve in the
aqueous environment of the nasal cavity. Thus, the drug must have an ideal lipo-
philic and hydrophilic balance to be absorbed correctly. Lipophilic molecules freely
cross the mucosa, while hydrophilic ones need to use the paracellular route for this.
An example of a lipophilic drug that has good nasal absorption and about 80% bio-
availability is fentanyl, in addition to its rapid absorption, very similar to its intrave-
nous administration. Other studies have shown that other hydrophobic drugs such as
naloxone, buprenorphine, and testosterone are highly absorbed nasally in animals’
models [5, 6, 8, 13].
The pKa is also an important parameter that must be considered in the biologi-
cally active agent. In the case of weak electrolytes, the absorption is dependent on
the degree of ionization and it is greater for non-ionized species. However, for polar
drugs, the partition coefficient is the major factor that influences the permeability
through the nasal mucosa, and there is a quantitative relationship between this
parameter and the nasal absorption constant. In this context, several studies have
shown that the drug concentration on cerebrospinal fluid (CSF) rises with the
increase of the lipophilicity or of the partition coefficient of the drugs [4, 6].
The polymorphism of the biologically active agent is also a factor that should be
also considered, as it affects the rate of dissolution and solubility of drugs, which
affect their absorption. This is because polymorphs have different solubilization
profiles in body fluids. Thus, drugs that exhibit many polymorphisms will have dif-
ferent absorption profiles due to varying rates of solubilization and absorption
across biological membranes. It is suggested to study this characteristic of the active
ingredient so that there are no surprises in the development of a formulation intended

for nasal application [11, 13].
The drug’s solubility is also critical and, since nasal secretions are aqueous in
nature, it is important that the drug has sufficient aqueous solubility for adequate
dissolution. Furthermore, it not only limits the drug absorption itself, but it can also
limit the ability to develop a product if the drug is not sufficiently soluble in the
desirable vehicles. Considering that formulations intended for nasal administration
are, in most cases, solutions and that it is not possible to administer high dose vol-
umes, drugs with low solubility end up becoming an obstacle in the treatment. It is
also important to consider the water activity of the nasal surface during the absorp-
tion of the bioactive agent, since this can result in the water-bioactive agent interac-
tion that conduces to an improved solubility [6, 9, 11].
4 Factors Related to the Formulation
The World Health Organization (WHO) has reported central nervous systems disor-
ders, such as depression, migraine, Alzheimer’s, and Parkinson’s disease as the
most debilitating in humans [14, 15]. Over the recent years, many strategies have
improved the delivery of therapeutic agents to the brain [16]. However, the design
of new formulations is highly dependent on the ability of the drugs to adequately
permeate the BBB to reach a significant concentration on the brain; therefore, intra-
nasal administration has gained special interest in this field [17].
As the main barriers avoid the entrance of external substances into the brain,
there are the BBB and the blood-cerebrospinal fluid barrier (BCSF). When a bio-
logically active compound is administered through oral or intravenous route, it
firstly needs to cross the BBB to reach the brain [7]. The blood capillaries on the
BBB are covered with endothelial cells tightly connected among themselves by
tight junctions, which constitute a strong barrier to the passage of plenty of com-
pounds and make many studied drugs to fail on treating brain disorders in preclini-
cal studies [7]. The nose-to-brain delivery enables a direct pathway of drug delivery
to the brain without the need to cross the BBB. Therefore, adverse events that usu-
ally occur when the bioactive agent is systemically absorbed can be avoided.
Despite to overcome BBB, there are alternative barriers that have to be circum-
vented for nose-to-brain drug delivery systems. The critical factor in this field is the
ability of the drug to achieve the upper and posterior nasal regions, such as the
olfactory region [15, 18]. Furthermore, a tiny amount of a drug administered via
intranasal route can reach the brain (ca. 0.1 to 1%) [19, 20], which is often related
to the small volume that can be applied in the nasal cavity (ca. 25 to 200 μl) [7].
Thus, strategies and technologies to improve nose-to-brain drug delivery have been
massively studied. Considering there are two pathways for this delivery, [1] indirect
pathway: from respiratory mucosa evolving blood capillaries, systemic circulation,
and BBB passage; and [2] the direct pathway: from trigeminal nerve and olfactory
epithelium, there are different strategies aiding to enhance both (Fig. 4.1). Promising
Fig. 4.1 Potential pathways for the nose-to-brain drug delivery (BBB blood-brain barrier)
properties for nose-to-brain formulations are often related to their ability to get into
contact with the mucosal surfaces, can pass through the mucus barrier, and to
improve the extracellular/paracellular diffusion or intracellular absorption. For
example, the strategy of using protein inhibitors, surfactants, or even nanosized
vehicles may improve the nose-to-brain delivery [15, 18]. Herein, we will discuss
the most used strategies and their applications in this field.
4.1 Strategies and Technologies
4.1.1 Permeation Enhancers
Permeation enhancers are compounds used to facilitate the permeation of drugs

across biological membranes. Although lipophilic drugs are able to easily permeate
nasal mucosa, some hydrophilic substances (i.e., peptides) frequently find it diffi-
cult to permeate. Therefore, permeation enhancers may contribute to the permeation
of drugs that cannot effectively permeate biological surfaces by themselves [21].
Among the constituents of this class, the surfactant agents are frequently used
due to their ability to disrupt the nasal barrier, which involve irritative effects to the
mucosa in some cases [7]. Non-ionic surfactants such as Cremophor EL, poloxamer
188, and laurate sucrose esters have been used in nose-to-brain preparations [22,
23]. Horvát and collaborators successfully facilitate the permeation of a hydrophilic
molecule of high molecular weight (dextran – 4.4 kDa) by using sodium hyaluro-
nate and Cremophor RH40. Combining mucoadhesive polymer and a surfactant
agent, the system prolonged the contact time of the preparation with the mucosa,
and enhanced the drug permeation [22].
Moreover, cyclodextrins, lipids, and different polymers have also been reported
enhancing the permeation of different drugs across nasal mucosa [7, 19]. For
instance, chitosan presents the ability of opening tight junctions in the nasal epithe-
lium. Rassu and collaborators mixed chitosan with β-cyclodextrin for the
development of nose-to-brain microparticles objecting the delivery of deferoxamine

mesylate [15, 24]. Besides the chitosan effect, β-cyclodextrin has demonstrated the
ability to disrupt the lipid bilayer by creating inclusion complexes with the lipids in
the mucosa membrane, increasing fluidity and enhancing permeation [19, 24].
Strategies improving the penetration are particularly interesting when the drug is
absorbed via direct pathway. Besides the use of permeation enhancers, which favor
extracellular/paracellular diffusion, cell-penetrating peptides may be added to the
formulations in order to improve intracellular absorption. These peptides, also
known as protein transduction domains, have been reported as small sequences of
amino acids able to cross the biological membranes and help drug internalization
[25]. In a nose-to-brain drug delivery system, Kamei and collaborators successfully
improved the delivery of insulin administering it with penetratin, mainly through
the olfactory bulb way. While L-penetratin leads to high insulin levels in the plasma,
D-penetratin demonstrated to increase insulin delivery to the brain, with low sys-
temic absorption [26]. Moreover, the delivery of a siRNA and dextran in rats were
also demonstrated via nose-to-brain by polymeric micelles coated with a cell-
penetrating peptide known as TAT. This cell-penetrating peptide is derived from
HIV-Tat and modified amphiphilic block copolymers of poly (ethylene glycol) and
poly (ε-caprolactone), which have demonstrated high ability to form stable com-
plexes with pDNA and siRNA [27].
Eutectic mixtures are known for their ability to enhance drug transportation
across biological membranes [28]. For instance, Li and collaborators developed a
mixture of borneol/menthol to enhance the permeation of cobrotoxin via olfactory
epithelium. Interestingly, they demonstrated cobrotoxin could not permeate through
this pathway out of the eutectic mixture [29]. Moreover, aiding migraine treatment,
Khan and co-workers described the intranasal administration of zolmitriptan using
a eutectic system, which achieves high levels of brain delivery in comparison to
intranasal instillation of the drug powder and intravenous injection of the drug solu-
tion [28].
4.1.2 Protein Inhibitors (Enzyme and Glycoprotein)
Several enzymes may be found in the nasal cavity, being able to promote drug deg-
radation. For instance, CYP450 isoforms, transferases, and carboxylesterases are
reported to constitute the nasal environment and their inhibition has improved the
stability of the compounds topically applied. Therefore, the use of these agents
increases the amount of drugs available to be effectively delivered to the brain [7].
The inhibition of proteases, by α-aminoboronic acid derivatives, has avoided the
degradation of peptides, albeit its real impact on nose-to-brain delivery is not com-
pletely known [30]. Dhamankar and Donavon have shown the enhancement of the
permeation of melatonin via nasal administration, when it is formulated with flu-
voxamine, a CYP450 inhibitor [31].
There are P-glycoproteins in the BBB, nasal mucosa, and olfactory epithelium
that can act as membrane transporters. When a drug is the substrate of this
transporter, it performs the efflux of the drug from the brain, losing nose-to-brain
delivery property. In order to overcome this, Shingaki and collaborators used cyclo-
sporine A and the inhibitor or the P-glycoproteins, improving the permeability of
verapamil, a known substrate of this protein [32]. Graff and co-workers also dem-
onstrated the P-glycoproteins effect on the transportation of diazepam, verapamil,
and antipyrine drugs to the brain [33]. As P-glycoproteins inhibitors, the properties
of pantoprazole and elacridar have been reported, Hada et al. demonstrated this
mixture enhancing imatinib mesylate delivery to the brain [34].
4.1.3 Nanostructured Systems
Nanostructured pharmaceutical platforms have been extensively utilized in the

nose-to-brain delivery of drugs. Many studies have proposed nanoparticles covered
with some additives, trying to overcome the limitations of this route of administra-
tion. Lectin has been used for coating nanoparticles for increasing the affinity with
mucin and improving the residence time of the system in the nasal cavity [35].
Bioactive agents displaying polar characteristics and high molecular weight usually
display a low nasal permeability, and the use of micro/nanoencapsulation has been
shown to successfully increase the drug permeation with this type of profile [24].
Even considering their small diameter, for nose-to-brain delivery, the relatively
large size (ca > 100 nm) of the nanoparticle preparations is often a disadvantage,
since it could exceed the diameter of the filia olfactoria, fostering low transportation
through the olfactory pathway [36]. If the nanoparticles are fully transported to the
brain via olfactory epithelium or if the bioactive agents released from the nanostruc-
tures can diffuse into the brain is not totally comprehended [15]. However, many
investigations have used nanosized materials to improve nose-to-brain delivery. For
instance, Gao and colleagues used wheat germ agglutinin on the surface of
poly(ethylene glycol)-poly (lactic acid) nanoparticles to enhance the delivery of a
vasoactive intestinal peptide and fluorescent probe to the brain [35]. They reported
the nanoparticles reached the brain’s levels 5- to seven-fold higher than the admin-
istration of the raw drug in solution [35].
Recently applied in the nose-to-brain field, the lipid particulates and liposome
systems are also nanostructured preparations. For instance, Singh and co-workers
demonstrated higher bioavailability of rizatriptan via intranasal than via intravenous
route by using solid lipid nanocarriers [37]. Moreover, increased haloperidol brain/
blood levels have been observed when it was intranasally administered using lipid
nanocarriers [38]. Liposomes ranging from 40 to 10,000 nm have been explored for
nose-to-brain delivery, the majority of them prepared with phosphatidylcholine and
cholesterol [39].
Besides solid, colloidal nanoparticles have also been recently studied to drive the
drugs on this route. Polymeric micelles are colloidal structures composed of surfac-
tant copolymers. With a diameter lower than 100 nm, they are able to cross BBB
[40] and to carry drugs of different polarities, self-assembling in a hydrophobic core
and a hydrophilic shell [41]. Pokharkar and collaborators explored the poloxamer
407 micellar-based system for neurotherapeutics nose-to-brain delivery. When

entrapped in micelles, the bioactive agent lurasidone could reach the brain tissue
crossing through the BBB via trigeminal and olfactory and nerves [42].
4.1.4 Mucoadhesive and Mucus-Penetrating Formulations
The cilia in the nasal and respiratory regions act as a barrier to the entrance of exter-
nal particles. They are motile and perform the movement of the mucus from the
nose to the oropharynx, with a clearance of 10–15 min [43]. When a drug enters into
the nasal cavity, they are trapped in the cilia and mucus secretions, tending to be
eliminated after a few minutes [44]. Therefore, some approaches have been
described to overcome this inconvenience, trying to increase the retention time of
the preparation in the nasal mucosa. For instance, there are dosage forms containing
inhibitors of the mucociliary clearance or composed of mucoadhesive or mucus-
penetrating polymers. Although some classes of drugs (i.e., α-adrenergic agonists)
have been explored regarding the reduction of the cilia beat frequency, in an attempt
to delay the mucociliary clearance [45], mucoadhesive polymers are the most used
in this way.
When Horvát and colleagues reported dextran nose-to-brain delivery by a muco-
adhesive and permeation enhancer formulation, they did not demonstrate by which
pathway the delivery occurred. Although alone the excipients could not increase the
delivery of dextran to the brain, together they massively enhanced the drug avail-
ability in the olfactory bulb and frontal cortex. The authors linked the observation to
the mucoadhesive property of the sodium hyaluronate [22]. In another study, the
mucoadhesive property of chitosan, covering microparticles, was responsible for
increasing the residence time of a drug in the mucosal surface, increasing also its
permeation [24].
Although most of the mucoadhesive polymers are not selective for the olfactory
epithelium, and could be dispersed in other mucosal surfaces (e.g., olfactory
mucosa, respiratory mucosa) [35], the use of mucoadhesive materials has been
related to the inhibition of the mucociliary clearance. Since they increase the viscos-
ity and adhesive profile of the formulations, they improve adhesiveness to the mucus
and delay mucociliary clearance [15, 46, 47]. Moreover, mucoadhesive polymers
can consolidate the adhesion of pharmaceutical formulations to the mucosa with
chemical and physical interactions. Therefore, hydrogen-bonding, electrostatic and
ion-dipole interactions between excipients and mucosa play great role on the
increased period of time of the formulation on the surface of the mucosal membrane
together with polymer chain interpenetration into the mucus gel [48]. In pursuit of
this, the use of chitosan, poly (acrylic acid) derivatives, cellulose derivatives, and
other mucoadhesive polymers facilitate nose-to-brain delivery when incorporated to
new drug delivery systems [49].
Performing mucociliary inhibition, the use of thermoresponsive polymers, such
as poloxamers, has also been described in the literature. They constitute liquid prep-
aration during storage and application and become gel once in contact with nasal
mucosa [50], increasing the local viscosity and, consequently, inhibiting mucocili-
ary movements.
Mucoadhesive dosage forms are typically designed using cationic and anionic
polymers capable of interacting with the mucosal surfaces [49]. Meanwhile, mucus-
penetrating dosage forms are able to effectively penetrate the mucus barrier and
subsequently be accumulated in the epithelial surface [51]. Therefore, PEGylated
dosage forms, that contain poly (ethylene glycol) (PEG) on the surface, have been
described to optimize transmucosal drug delivery, besides they increase systemic
circulation time of nanoparticles. They are often prepared through the use of block-
copolymers containing PEG as one of the blocks [52], or through the use of agents
as poloxamers [53] or functionalized phospholipids that generates PEGylated lipo-
somes [54]. For instance, Porfiryeva and collaborators studied mucoadhesive
(Eudragit®) and mucus-penetrating PEG nanoparticles for nose-to-brain haloperi-
dol delivery and found that the non-mucoadhesive PEGylated nanoparticles demon-
strated more pronounced in vivo effects than the mucoadhesive carriers [49].
4.1.5 Other Strategies
Vasoconstrictor drugs have been used as an additional tool to limit the systemic
absorption of drugs applied in the nasal mucosa, enhancing their retention at the site
of application. Dhuria and colleagues, for example, added 1% phenylephrine hydro-
chloride to systems containing two neuropeptides (hypocretin-1 or L-Tyr-D-Arg).
By this, the drug concentration into the plasma was reduced by 65% and 56%,
respectively, after 30 min of intranasal administration in comparison to the admin-
istration of the drug alone [55]. However, when ephedrine was co-administered with
an angiotensin agonist, no reduction of systemic absorption was observed [16].
Besides chemical additives, physical methods also have been demonstrated to
improve nose-to-brain drug delivery controlling the localization of the drug. The
technology based on applying of a magnetic force to drive magnetic particles to a
target site of the body (magnetophoresis) can be used for the development of nasal
drug delivery systems [56]. Xi and collaborators, in nose-to-brain field, have
increased the efficiency of the drugs crossing the olfactory region and being
straightly transported to the brain by using ferromagnetic microspheres [20].
Another physical strategy was used by Chen and colleagues who used focused ultra-
sound sonication in rats to study its effect on the transport of brain-derived neuro-
trophic factor to the brain. The same method was previously used to facilitate the
BBB permeation for drugs administered via intravenous pathway [57]. The com-
parison between intranasal drug administration with and without ultrasound appli-
cation demonstrated improvement of the localization of the drug in a specific area
of the brain [57].
5 Design and Optimization
The controlled drug delivery field has its origin in the 1960s to 1980s, and a high
number of systems, carriers, and devices were developed to be administered by dif-
ferent routes and for different purposes [58]. The concept and technology of nasal
drug delivery systems improved during the last three decades. Different strategies
have been applied for the development of improved nasal medicines [59].
Biologically active agents of natural and synthetic origin have been investigated
for nasal delivery in each more specialized formulation with the aim to control the
delivery, reduce the adverse effects, increase the safety and patient compliance, and
improve the therapeutic. A considerable knowhow and information have been
obtained from scientific and technological studies. The development of new nasal
delivery carriers and innovations has been used for system design and efficient clini-
cal translation to nasal products [2].
Therefore, aiming for the efficient design of nasal delivery systems, innovative
strategy approaches for specific bioactive agents are being investigated. They are
composed of new nasal enhanced delivery technologies, design of carriers for avoid-
ing the drug enzymatic degradation, modulation of system’s physicochemical prop-
erties, and also systems for nose-to-brain drug delivery [2, 19].
The investigation of potential advantages and limitations of nasal route is a fun-
damental step during the design of a system for nasal drug delivery. Among the
main variables, the ones described below should be highlighted [4]:
• The degradation of drug in the gastrointestinal tract (by acidic or enzymatic deg-
radation) and/or the hepatic first pass can be avoided.
• It is possible to acquire rapid absorption and onset of drug effect.
• Higher bioavailability can be reached and using lower doses of drug.
• It is a route of easy access and non-invasive, which can improve the patient
adherence to the treatment.
• It is possible the direct transport into systemic circulation and central ner-
vous system.
• Offers lower risk of overdose.
In this context, we can also conclude that the design of nasal systems does not
have any complex formulation requirement. However, some limitations can turn it
into an endeavor. The volume of formulation that can be delivered by this route is
limited to 25–200 μl, and molecules of high molecular weight cannot be delivered
through the nasal route (mass cut off ~1 kDa). Irritation of mucosa and pathological
conditions can affect the nasal region and also the drug delivery. The intrinsic
defense mechanisms, such as mucociliary clearance, ciliary beating, and the enzy-
matic barrier, can affect the residence time of the system and the drug
permeability.
Therefore, for a biologically active agent to be successfully administered through
the nasal cavity, these challenges should be overcome. A better understanding of
permeation pathways is necessary for the formation scientist. An effective
correlation must be established between the physicochemical properties of the bio-

active and formulation with that of permeation rate. It is also necessary to perform
extensive research for alternatives at the molecular level for the increase of the drug
permeation through the nasal mucosa without compromising normal function [4].
This process will lead to the optimal design of formulation for nasal drug delivery
and can cut down the experimental efforts involved as well.
Therefore, for achieving safe and efficient intranasal drug medicines, some strat-
egies to overcome nasal delivery barriers should be considered. In the design of a
nasal formulation, three main cooperative points should be taken into consider-
ation [2]:
• The bioactive agent (chemical structure, chirality, molecular size, potency, lipo-
philicity, solubility, and ionization).
• The physicochemical characteristics of the carrier (pH, components, enhancers,
viscosity, charge and solubility) and dosage form (nano or microparticulate sys-
tems, solution, powder, emulsion, gel, etc.)
• The administration device (single or multi-dose, simple or sophisticated).
During the development of nasal products, it is fundamental to think about the
quality of the process. The ability to identify and control the variables of the phar-
maceutical formulation can ensure this objective and the utilization of the pharma-
ceutical Quality by Design (QbD) is fundamental. QbD is a systematic approach for
development. It begins with predefined aims and emphasizes product and process
understanding and process control, based on sound science and quality risk man-
agement [60]. QbD constitutes a culture shift from knowledge exchange to knowl-
edge integration, and it can improve the safe assurance, effective drug supply to the
consumer, and also ensures significant improvement of manufacturing quality per-
formance [58].
The International Conference on Harmonisation of Technical Requirements for
Registration of Pharmaceuticals for Human Use (ICH) has the pharmaceutical
development guideline Q8 (Q8r2 is the updated version) and it can enable the appli-
cation of scientific approaches and quality risk management for development of
nasal products and its manufacturing process [61].
Over the design of a nasal drug delivery system, it is necessary to achieve the aim
of developing a quality formulation and its manufacturing process to consistently
deliver the intended performance of the formulation. Thus, the use of tools that can
provide information from pharmaceutical development studies constitutes a basis
for quality risk management [58].
In this context, the use of experimental design constitutes an important strategy
for formulation of nasal products. The more important statistical activity is the plan-
ning of the experiments where the data is obtained for analysis. The suitable way to
plan an experiment leads to significant and trusted numbers from which it is possi-
ble to get conclusions [62].
The essence of good experimental design is to plan an experiment in such a way
that it can provide exactly the kind of information that the formulation researcher is
looking for. Therefore, the factors affecting the development must be considered in
full and the more advantageous techniques should be used. In this context, the first
step is to accomplish a trial and discard non-significant variables, so as not to waste
time and resources with them in the laboratory.
The use of Design of Experiment (DoE) is a good strategy to achieve this goal.
DoE enables the determination of the influence of one or more variables on another
variable of interest. For example, the composition and type of polymeric material
can directly influence the mucoadhesive and rheological properties of a semi-solid
nasal delivery system [2, 62]. Thus, during the planning of any nasal formulation
development, the first thing that must be done is to decide what factors and responses
are of interest. With the answers indicating which were the statistically significant
and the most influential variables, it will be possible to achieve an optimized nasal
drug delivery formulation.
6 Concluding Remarks
Considering the many advantages, benefits, and interest in nasal drug delivery, it
was observed an increased interest for the development of formulations and a high
number of novel nasal products have reached the market. Most of these products
comprise formulations designed for crisis treatments, sleep disorders, acute pain,
panic attacks, nausea, heart attacks, and Parkinson’s disease. Furthermore, medi-
cines for the treatment of long-term illnesses (i.e., diabetes, growth deficiency,
osteoporosis, fertility treatment, and endometriosis) for nasal administration are
also available in the market. The research and development of nasal delivery sys-
tems have gained higher evidence with the strategy of nose-to-brain drug delivery
and important progress has been observed. It enables the bioactive agents to achieve
a rapid and efficient concentration in the central nervous system, since this route is
able to circumvent the blood-brain barrier. Besides the treatment of several neuro-
logical diseases, many formulations have also been applied to prevent and the treat
infectious diseases. Therefore, considering the growth interest in this area and the
development of novel nose-to-brain formulations, factors that affect their design
must be considered in order to overcome physiological/drug conditions that could
impair the availability of the carried drug into the brain.
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Chapter 5
Challenges in Targeting Nasal Passage
and Nose-to-Brain Delivery via
Nanoemulsions
Shiv Bahadur and Kamla Pathak
Abstract Nasal drug delivery has been considered an important approach for brain
targeting since the last decade. The intranasal route has gained attention as a poten-
tial route of drug administration for the treatment of CNS disorders. The intranasal
route can be more effective than other conventional routes for the treatment of CNS
disorders. Presently, several strategies with novel approaches have been used to get
nose-to-brain delivery of drugs. The management of neurological disorders is still
challenging despite the enormous development of various strategies for drug deliv-
ery. The main limiting factor for drug therapeutics is the blood-brain barrier (BBB).
Nanoemulsion has shown a promising formulation tactic among other nanocarrier-
based drug delivery systems which can deliver the higher drug to the brain through
the intranasal route compared to the conventional drug delivery systems.
Nanoemulsions consist of emulsions and stabilized with surfactants and co-
surfactants having small droplet size and large surface area. Hence, nanoemulsion
can be an alternative drug delivery to oral to avoid some constraints such as enzy-
matic degradation, low solubility of drugs and low bioavailability. There are several
nanoemulsion-based formulations that have been explored for brain targeting and
results indicate the significant enhancement of bioavailability of various drugs for
CNS diseases. Therefore, the present review highlights the several aspects of nano-
emulsion as a potential carrier for brain targeting through intranasal
administration.
Keywords Nanoemulsion · Nose-to-brain delivery · CNS disorder · Alzheimer’s

disease · Parkinson’s disease
S. Bahadur
Institute of Pharmaceutical Research, GLA University, Mathura, Uttar Pradesh, India
K. Pathak (*)
Faculty of Pharmacy, Uttar Pradesh University of Medical Sciences, Saifai,
Etawah, Uttar Pradesh, India

https://doi.org/10.1007/978-3-031-23112-4_5
60 S. Bahadur and K. Pathak
1 Introduction
The brain is the most complex organ in the body which is protected by the skull and
separated from the blood circulatory system [26]. There are two major types of cells
in the brain such as neurons and glia. Neuron cells have the most important function
in the brain. Hence any disruption or imbalance in neuronal cell function may cause
neurological diseases in the brain [1]. The death of the neuron due to any region is
most commonly known as neurodegenerative disease [55]. The functions of neuro-
nal cells get disturbed due to neurodegenerative disease. Various symptoms are
associated with these diseases such as losses in memory and thinking ability, diffi-
culty in body movement and intelligence of individuals [90]. The neurodegenerative
disease could be from different areas of the brain such as the cerebellum, brainstem
and hippocampus. The most commonly known neurodegenerative disorders are
Alzheimer’s and Parkinson’s diseases [81].
The nanotechnology-based drug deliveries have shown various potential advan-
tages over other delivery systems [80]. Higher concentration of drugs may be
reached to the predetermined site of action through targeted drug delivery. Unwanted
adverse effects can be also minimized by targeted drug delivery system. BBB has
been considered as a potential barrier for drug delivery to the brain. Therefore, sev-
eral approaches have been applied for the drug targeting to the brain such as physi-
ological approach (e.g., pseudonutrients, chimeric peptides, ligand binding
proteins), pharmacological approach (e.g., chemical drug delivery, liposomes,
nanoconjugates, nanoparticles) and invasive approach (e.g., intracerebral implants,
BBB disruption) [46, 77]. These strategies play a very significant role not only in
the treatment of CNS diseases but also in the diagnosis of various diseases.
Nanotechnology can be seen as a potential problem solver for the therapy and diag-
nosis of several CNS diseases like epilepsy, AD, psychosis, migraine and PD [3,
53]. Nose-to-brain delivery system has been most widely explored since the last
decades. Drugs can be directly delivered to the brain through the nasal cavity.
Hence, nasal administration can be a choice of route for drug delivery to the brain
in the treatment of several CNS disorders. Implications of novel drug delivery sys-
tems have added several advantages for the nose to brain targeting [53, 77].
Nanoemulsions (NEs) are nanosized which are most commonly used for the tar-
geting of drugs to get the maximum concentration of drug at the desired site of
action with minimum adverse effects [49]. Several research reports showed the vari-
ous benefits of NEs which are most efficient drug delivery systems for nose-to-
brain [13].
The globule size of NEs ranges from 100 to 200 nm [83]. NEs are thermody-
namically stable and no gravitational separation occurs due to kinetic stability. NEs
have small globule size due to reduced attractive force between the small-sized
droplets [65]. The drug transport pathways from nose-to-brain delivery have been
depicted in Fig. 5.1.
The NEs are stable for the physical and chemical variations such as pH and tem-
perature because less amount of surfactants are required for the formulation
5 Challenges in Targeting Nasal Passage and Nose-to-Brain Delivery via Nanoemulsions 61
Fig. 5.1 Drug transport pathways from nose-to-brain delivery
development. Hence, NEs are most appropriate due to having several applications.
Nanoemulsification is known to protect and increase the bioavailability of bioactive
compounds and has been observed in several studies.
Further, NEs have significant encapsulation efficiency for both lipophilic and
hydrophilic compounds [15]. The present chapter highlights on NEs as a significant
drug delivery system for treatment of neurodegenerative diseases through intranasal
administration. Further several research reports have been discussed along with the
related challenges and future prospects.
2 Composition of Nanoemulsion for Nasal Administration
NEs are composed of two immiscible liquids in which one is lipophilic and second
is aqueous with an emulsifier. In the aqueous phase, the lipophilic component is
distributed. The core-shell structure is present in both the o/w and w/o nanoemul-
sions. In an o/w nanoemulsion system, the amphiphilic shell is made up of surface-
active molecules, whereas the lipophilic core is made up of non-polar molecules.
Monoacylglycerols, diacylglycerols, triacylglycerols, and free fatty acids make up
the oleaginous phase of a nanoemulsion [13]. The oily phase may also be consti-
tuted of non-polar essential oils, lipid substitutes, mineral oils, waxes, oil-soluble
vitamins, weighting agents, and various other lipophilic components. The physical
properties of the oil phase components, such as density, refractive index, viscosity,
interfacial tension, and phase behaviour, influence the formation, stability, and func-
tional qualities of nanoemulsions [4]. However, because of their low cost, availabil-
ity, and functionality, long-chain triacylglycerols are favoured for NE formation. A
polar solvent and a cosolvent compose the aqueous phase of a NE. The polarity,
rheology, phase behaviour, interfacial tension, and ionic strength of a nanoemulsion
are all determined by this factor. Water is the most common polar solvent, while
carbohydrates, protein, alcohol, and polyols are utilized as cosolvents. Ostwald rip-
ening (increase in mean droplet size over time), flocculation, coalescence, and grav-
itational separation can cause the aqueous and oil phases to separate. A stabilizer
agent can be added to nanoemulsion to prevent this. The stabilizers can form a
monolayer, multilayer, or solid particulate nanoemulsion depending on how they
are distributed on the particle. Emulsifiers, weighting agents, ripening retarders, and
texture modifiers are just a few of the stabilizers that are used [15].
2.1 Surfactants
Surfactants are one of the most significant components of NE, as they help to reduce
surface tension, and prevent globule coalescence, and phase separation. Surfactants
should have the ability to dissolve maximum amounts of drug(s). Further, they help
in stabilization of NE formulations and small globule size. As a result, surfactants
can impact medication penetration through the nasal mucosa, either by changing the
fluidity or by disrupting the epithelial layers’ tight junction. Several studies report
that the globule size decreases on increasing the surfactant concentration [29]. The
lower the globules size, the higher will be permeation and ultimately higher drugs
will be delivered to the brain. The type and concentration of surfactant have signifi-
cant effects on the drug permeation on the nasal mucosa. The changes in the struc-
tural integrity of nasal mucosa by surfactants are critically questionable for the
toxicity issues. Hence, as a result, surfactant concentrations are kept as low as fea-
sible in order to maintain a balance between medication penetration and harmful
effects [13, 43].
2.2 Co-surfactant
Surfactants employed in nanoemulsion formulations are often single-chain surfac-

tants that may not reduce desirable interfacial tension [89]. Hence, co-surfactants
are substances that aid the surfactant in lowering surface tension [12]. By entering
into the hard layer of surfactants, co-surfactants provide flexibility to the interfacial
layer, breaking interfacial layers and imparting fluidity, which aids in the emulsifi-
cation process as well as formulation stability. For the development of stable NE,
combinations of surfactant and co-surfactant have a very significant role [2]. The
ternary phase diagram is a widely used method for determining the working range
and optimum concentrations of oil, surfactant, and co-surfactant. When the concen-
tration of co-surfactant is increased, the globule size decreases, and the drug con-
centration rises. These are some most commonly used co-surfactants such as
polyethylene glycol (PEG), Transcutol-P® and ethanol in NE formulation for intra-
nasal drug delivery permeation [13, 89].
2.3 Oil
The solubility of novel chemical compounds is a crucial issue, as it impacts the

pharmacokinetic and pharmacodynamic aspects of medications. Hence drugs are
usually solubilized in the lipid phase of the NE and the solubility of drugs increases
with increase in the lipophilicity of oils [73]. The solubilizing capacity of oils
decreases in the order of vegetable oils > medium-chain triglycerides > medium-
chain mono and diglycerides [86]. The solubilization capacity is determined not
only by the oils, but also by a delicate balance with the oil’s emulsification zone as
determined by phase diagram studies. Generally, the globule size of NE increases
with increase in oil concentration in the formulation [17]. Higher globule size of
NEs reduces drug permeation from nasal mucosa. Thus, optimal concentration of
oils has to be selected to get sufficient solubility of drugs. Some oils have permeation-
enhancing properties and they increase the drug permeation through the nasal
mucosa. In the study, it was found the polar lipids are present in butter oil which
have the key role in the permeation of quetiapine fumarate through nasal mucosa by
transcellular and paracellular pathways [36].
3 Factors that Influence Nanoemulsion Transport from Nose

to Brain
NE offers better permeation through nasal mucosa compared to the other conven-
tional drug delivery [16]. Surfactants and co-surfactants have a permeation-
enhancing effect aside from that; there are some other characteristics of nanoemulsion
which have significant role in the transport of drugs to the brain. NEs may be a
viable option for nose-to-brain delivery because they meet all of the desirable NE
characteristics [13].
3.1 Globule Size
The globule size of nanoemulsion is one of the most important features for drug
permeation from the nasal mucosa. The most common pathway for nose-to-brain
drug transport is through either trigeminal or olfactory pathways. The typical diam-
eter of an olfactory axon in different species is about 200 nm; however, it varies
from 100 to 700 nm in humans [13, 67]. As a result, the size range of new formula-
tions should be less than 200 nm to allow drug absorption across transcellular chan-
nels. Ahmad et al. [8] reported that quercetin loaded mucoadhesive NE having
100 nm typical globule size was found to be higher rate and extent of drug absorp-
tion than the typical globule size of 700 nm reaching the olfactory bulb via the olfac-
tory and trigeminal nerve. However, NEs having globule size more than 200 nm
have also shown the efficient transport of drugs into the brain by intranasal admin-
istration. Further, the retention time on the nasal mucosa is also affected by droplet
size, which is an important factor for drug delivery to the brain. The less retention
time resutls the lowers the drug absorption. The larger droplet size can be cleared
out from nasal mucosa more easily by mucosal clearance. NEs having average glob-
ule size >200 nm were cleared from the nasal mucosa after 4 h of intranasal admin-
istration, whereas formulations having globule size 80 and 200 nm showed more
retention time 16 and 12 h respectively on nasal mucosa. As a result, the globule
size of NE plays an important role in medication targeting the brain via intranasal
delivery [7, 8, 13].
3.2 Zeta Potential
The zeta potential of NE is linked with the stability of formulations. A zeta potential
value of 30 mV indicates stability [93]. Several investigations have found that zeta
potential is also essential in medication retention on nasal mucosa. The positively
charged globules get firmly attached to the nasal mucosal layer which contains neg-
atively charged mucin [84]. However, the majority of the reported NEs for nose-to-
brain transport have a negative zeta potential. The mechanism of NE adherence on
nasal mucosa in relation to electrical charge has not been considered enormously.
Therefore, the effects of zeta potential on drug permeation across nasal mucosa
need to be considered for the development of formulations for nose-to-brain drug
delivery [13].
4 Methods of Preparation of Nanoemulsion
The presence of multiple nanoscale droplets in the NEs increases the surface area.
Therefore, a significant quantity of energy is required to generate additional surface
area. Thus, NE creation is not self-sustaining and necessitates the application of
energy [7, 8]. The amount of energy required to produce nanoemulsions (ΔG) is
determined by the following equation:
G A T S (5.1)

where ΔA denotes a rise in the interfacial area, γ denotes surface tension, and TΔS
denotes dispersion entropy.
NEs can be made using either high or low energy processes (Fig. 5.1). The size
of the globule is determined by the constituents, operating conditions, and tech-
nique of manufacture. Mechanical devices are used in high-energy methods to dis-
turb the oil phase, allowing it to interact with the water phase and create smaller oil
droplets. The mechanical device’s enormous tension interrupts the oil phase. The
Fig. 5.2 Different methods of preparation of nanoemulsion
NEs are created using low-energy methods by changing the temperature or compo-
sition of the oil-water system, with the energy input coming from the chemical
potential of the ingredients. The low-energy methods involve minimal energy gen-
eration, and hence are appropriate methods for preparing NEs of heat-labile actives
[20]. The methods of preparation of NE have been depicted in Fig. 5.2.
The traditional nanoemulsification formulation entails the breakdown of bigger
droplets or the inversion of solvents. New emulsification technologies are being
developed at an increasing rate in order to broaden the range of material formula-
tions and operating conditions while simultaneously lowering manufacturing costs.
To manufacture nanoscale emulsions, a bottom-up strategy based on condensation
has been developed. The technique is simple, quick, scalable, and energy-efficient,
and it has the potential to be used in processed meals [49]. The vapour condensation
method was used to create Pickering nanoemulsions. Pickering eliminates the issues
associated with surfactant desorption and Ostwald ripening [54].
5 Intranasal Delivery of Nanoemulsion for CNS Disorders
The several NE formulations have been developed for the nose-to-brain delivery.
The NE formulations for intranasal administration of drugs are usually O/W emul-
sions. Several preclinical studies showed that CNS administration via the nasal
mucosa outperforms intravenous administration. NE can be created in a variety of
ways, including the use of oil, surfactants, co-surfactants, and water [13]. The major
components of the NEs have significant role in drug penetration through
nasal mucosa.
Intranasal NE has been recognized as a potential drug delivery system for direct
nose-to-brain delivery. NEs with enhanced retention time on nasal mucosa are able
to drug targeting to brain bypassing BBB. Safety and toxicity of NE are one of the
major issues that should be considered for long-term uses. Several intranasal NEs
have been researched for the therapy of CNS illnesses such as migraine, Alzheimer’s
disease, and Parkinson’s disease. Thus, several fundamental studies need to be con-
sidered in the creation of NE for administration from the nose to the brain. The
several characteristics of nanoemulsion make them more appropriate for nose-to-
brain transfer. Mucoadhesive agents reduce the mucociliary clearance in the nasal
mucosa. Table 5.1 shows several examples of intranasal NEs and their potential
effects for neuronal diseases. Nasal medication administration may be an option to
oral therapies for brain targeting. NEs have several characteristics for nose-to-brain
delivery for CND diseases. Several studies suggest that intranasal route NEs led to
better results than intravenous administrations. However, clinical trials of NE for-
mulations are still needed to show that they are appropriate for clinical use. Hence,
there is utmost requirement of clinical study of NEs for intranasal drug delivery for
CNS diseases [6, 13].
NEs in the field of nanomedicine, formulations are becoming increasingly sig-
nificant. Their properties (high-surface nanodroplets) make them ideal for nose-to-
brain administration. To impede nasal clearance, mucoadhesive polymers might be
added to their formula. Because it is mucoadhesive and has penetration-enhancing
characteristics on nasal mucosa, the introduction of chitosan as an extra excipient
serves a dual purpose. Nasal administration of NEs is a promising technique for
nose-to-brain medication delivery and CNS targeting for neurodisease treatment.
Clinical trials of these formulations, however, are still needed to demonstrate their
efficacy in clinical practice. To improve the performance of NEs, a lot of work needs
to be done. The use of other excipients may be considered in the future [15].
The primary obstruction in drug delivery to the brain is the BBB that does not
allow the drug to attain therapeutic levels in the brain leading to a significantly low
CNS bioavailability. Consequently, several strategies are being approached for the
local delivery of active therapeutics to the brain including many invasive methods
that are risky and induce neurotoxic effects. While many of them cannot be consid-
ered appropriate for chronic treatments, there is a pressing need for methods that
can bypass BBB. Of the several methods, one significant approach is nose-to-brain
delivery [9].
For many systemic medications and vaccinations, nasal delivery has gained pop-
ularity as an alternative to injections and oral administration. The nasal mucosa,
which is highly vascularized and immunogenic, may provide advantages in terms of
speed of action, bioavailability, and patient compliance. Migraine, smoking cessa-
tion, acute pain alleviation, nocturnal enuresis, osteoporosis, and vitamin B12 defi-
ciency are among the conditions for which the method is now used. Cancer therapy,
epilepsy, psychosis, rheumatoid arthritis, neurological disease, and insulin-
dependent diabetes are some of the other therapeutic areas where nasal administra-
tion has potential. Because of the high total blood flow, porous endothelium
membrane, and vast surface area, intranasal delivery has been shown to carry
Table 5.1 Recent report of nanoemulsion-based therapeutics for nose-to-brain delivery through
in-vivo and in-vitro
Drug Therapy for Study model(s) Relevant therapeutic outcomes Ref.
Donepezil Alzheimer’s In vitro drug The permeation of donepezil [39]
disease diffusion study was found to be significant
Ex vivo drug through intranasal NE. The
permeation study polymers can be used as an
Tolerability study effective strategy to improve
through in vitro and the bioadhesion and drug
in vivo models penetration through nasal
mucosa, which enhances the
bioavailability of donepezil
Rivastigmine Alzheimer’s In vitro drug release Rivastigmine-loaded NE [48]
disease study showed significantly higher
Ex vivo diffusion drug concentration in the brain
study than the solution. The
In-vivo optimized formulation was
pharmacokinetic and devoid of nasal ciliotoxicity
biodistribution study
in the rat
Nasal ciliotoxicity
studies in goat nasal
mucosa
Resveratrol Parkinson’s In vitro drug release Diffusion controlled release of [52]
disease study resveratrol was till 6 h with flux
Ex vivo diffusion of 2.86 mg/cm2 h through sheep
study nasal mucosa. The drug level in
In vivo drug the brain from intranasal
biodistribution study resveratrol mucoadhesive NE
in Wistar rat’s brain was higher than the resveratrol
solution. Bioavailability was
seven times higher through this
approach
Selegiline Parkinson’s In vitro drug release Selegiline NE showed 3.7-fold [37,
disease study more penetration than the drug 38]
Ex vivo diffusion solution. Haloperidol-induced
study Parkinson’s disease in animals
Behavioural with selegiline intranasal NE
activities of showed significant
Parkinson’s disease improvement in behavioural
in Wistar rats activities in comparison to
conventional drug delivery
Letrozole Epilepsy In vitro and ex vivo Intranasal administration of NE [78]
drug release study. showed the prolonged drug
The behavioural release profile as compared to
seizure, biochemical suspension. High concentration
and of drug was found in the brain
histopathological
study were
performed
(continued)
Table 5.1 (continued)
Amiloride Antiepileptic In vitro drug release Bioavailability and brain- [69]
study targeting efficiency with
Ex vivo diffusion efficacy of developed amiloride
study NE was enhanced through nasal
In vivo administration
pharmacodynamic
and pharmacokinetic
study in Wistar rats
Zolmitriptan Migraine In vitro Zolmitriptan mucoadhesive NE [5]
mucoadhesion study showed higher permeability
Ex vivo drug coefficients than the solution
permeation studies through the nasal mucosa. In
In vivo vivo study of zolmitriptan
pharmacokinetic and mucoadhesive NE showed
biodistribution higher AUC0–8 and shorter Tmax
studies in the brain in comparison to
intravenous and nasal solution
Rizatriptan Migraine In vitro drug Ex vivo drug diffusion defined [44]
diffusion study controlled release with 86% in
Nasal irritation 4 h. Brain targeting through
study on sheep nasal intranasal NE
mucosa. In vivo (AUC = 302.52 μg min/g) was
brain targeting more than intranasal gel
potential (AUC = 115 μg min/g) and
intravenous route
(AUC = 109.63 μg min/g)
Cyclosporine-A Neuroprotective In vitro drug The brain/blood ratios of [80]
diffusion study cyclosporine-A by intranasal
In vivo brain uptake and intravenous were found to
study be 4.49 and 0.01, respectively.
Cyclosporine-A NE can be
used for direct nose-to-brain
delivery bypassing the BBB.
Kaempferol Neuroprotective Ex vivo diffusion The drug concentration through [47]
and anti-tumour study intranasal NE was found to be
In vivo drug 4–5 fold higher than solution.
biodistribution study Ex vivo permeation and in vivo
in Wistar rats biodistribution studies showed
higher drug concentration in the
brain with chitosan NE through
intranasal administration
compared to NE and
kaempferol solution
(continued)
Ziprasidone Antipsychotic Ex vivo diffusion Higher drug diffusion of [14]
hydrochloride study ziprasidone NE than solution
In vivo was found. Pharmacodynamic
pharmacodynamic study revealed the superiority
study in Wistar rats of mucoadhesive NE than NE
Nasal ciliotoxicity in locomotor activity and paw
studies in goat nasal test. Formulation was devoid of
mucosa acute nasal ciliotoxicity
Quetiapine Antipsychotic In vitro dissolution Higher drug transport efficiency [45]
study (DTE%) via intranasal NE
In vivo drug
distribution study in
Wistar rats
Reproduced from Bahadur et al. [15]
Abbreviations: GS Globule size; PDI Polydispersity Index; ZP Zeta potential; DC diffusion
coefficient
medications noninvasively from the nose to the brain in minutes. Intranasal medica-
tion administration can deliver a wide range of therapeutic substances (small mol-
ecules and macromolecules) to the CNS. When delivered nasally, several CNS-active
drugs are more efficacious and provide therapeutic effects in lower dosages. It does
not necessitate any therapeutic agent change, nor does it necessitate the medicine
being paired with any carrier [6, 13].
5.1 Nanoemulsion in the Treatment of Alzheimer’s Disease
Alzheimer’s disease (AD) has become the most common and progressive form of
dementia; 60–80% reports of dementia are due to Alzheimer’s disease. As the age-
ing population is increasing the occurrence of this disease has also been rising for
decades which ultimately results in the financial and emotional burden for society
and family [91]. While ageing is one of the important correlations with the AD, in
the last 30 years researchers are focusing on the role of genetics and other agents.
They discovered many factors beyond keeping the age as the factor. Intranasal drug
delivery has been accounted as one of the promising routes for drug delivery system
for targeting brain disorders. The nasal mucosa is directly connected to the brain
parenchyma and CSF. Delivery through the brain via the intranasal route trans-
ported the drug through various mechanisms such as olfactory transport and regi-
mental transport [33]. NEs have several significant applications in bioavailability
and solubility enhancement, and could be delivered by various routes such as oral,
parenteral, nasal and ocular [11]. NEs of two polyphenols namely resveratrol and
curcumin have demonstrated enhanced drug concentration in the brain through
intranasal administration [52, 85]. NEs have garnered great attention in dosage
design due to favourable features such as optical clarity, increased surface area, ease
of preparation, etc. There are two major problems in the conventional drug delivery
such as low bioavailability and non-compliance. To overcome these disadvantages
NEs have been used as a carrier for CNS drug delivery in AD, stroke, neurodegen-
eration, etc. [35]. Alzheimer’s disease (AD) is a neurological condition that causes
psychological and behavioural problems. Several drugs like acetylcholinesterase
inhibitors usually fail due to poor solubility and inability to cross BBB which ulti-
mately leads to lower bioavailability.
Novel drug delivery systems (NDDS) include design, production and character-
ization in the nanoscale delivery system. These NDDS include nanoemulsion, solid
lipid nanoparticles, nanospheres, etc. which can be a potential method for delivering
drugs to the brain via various routes especially intranasal route [89].
Ahmad et al. [7, 8] studied NE of coumarin-6 for intranasal administration. The
particle size of about 100 nm results in longer retention of time and lower mucocili-
ary clearance than the other conventional delivery. NE with more than 900 nm can-
not be transported through the olfactory region. With the help of confocal microscopy
the translocation of 100 nm in the nasal cavity was assured, that was followed by
trigeminal nerve with depleted intensity [7, 8, 34].
5.2 Nanoemulsion in the Treatment of Parkinson’s Disease
Parkinson’s disease (PD) has surpassed Alzheimer’s disease (AD) as the second
most common chronic and progressive neurological disease. PD has a high impact
both socially and economically on the suffering population [41]. Dopamine synthe-
sis decreases as dopamine neurons in the ventral tegmental region and substantia
nigra of the brain degenerate. There are a variety of clinical symptoms linked with
Parkinson’s disease patients, including motor, non-motor, and mixed motor symp-
toms. Among them non-motor is the most significant are rigidity, tremor and brady-
kinesia [88]. Several researches have reported nanoformulations such as NEs for
improvement of drug delivery for PD [71]. NEs have been very well recognized for
extended drug delivery from nose to brain by olfactory pathway. Mustafa et al. in
2012 developed ropinirole loaded and studied different parameters. The developed
formulations were evaluated for several physiochemical parameters including par-
ticle size and viscosity. Further, ex-vivo, in vitro, and in-vivo experiments were also
conducted, and the concentration of ropinirole was found to be greater in the brain
than conventional formulation [68]. Kumar et al. [37] developed selegiline-loaded
NEs by using principles of QbD. The behavioural investigation in rat model with
intranasal injection of selegiline NE revealed a considerable improvement in selegi-
line concentration in the brain [37, 38].
5.3 Nanoemulsion in the Treatment of Migraine
Migraine is a headache illness defined by moderate to severe intensity of pain

attacks, which results in many autonomic dysfunction disabilities such as nausea,
vomiting, gastric stasis, effort, small bowel, photophobia, and so on. Because of the
many symptoms associated with migraine disorder, such as vomiting and nausea,
oral medication delivery is not appropriate. For migraine treatment, parenteral and
nasal medication delivery are the best options. The innovative medication delivery
is in the form of a spray of sumatriptan (OptiNoseTM), which is being tested in a
phase II clinical trial for migraine therapy. The delivery has a quick commencement
of action since it is deposited in the olfactory region and subsequently travels
through the nose to the brain. A measured dose actuated pump is used to measure
the dose. This device is inhaled from the nasal cavity by activating the nozzle, which
releases the medicine into the olfactory area and then to the action site [82].
Migraine can be treated with a variety of natural and synthetic medications.
Natural drugs such as ergot derivatives, alkaloids, and synthetic pharmaceuticals
such as analgesics and anti-inflammatory drugs are examples of natural and syn-
thetic drugs. These drugs have been demonstrated to be effective in the treatment of
migraines. Abdou et al. [5] has studied the zolmitriptan-loaded intranasal nano-
emulsions. In vivo pharmacokinetic and biodistribution experiments revealed that
the solution had higher permeability coefficients than the nasal mucosa.
5.4 Nanoemulsion in the Treatment of Psychosis
Our study group provided one of the earliest examples in the literature of the use of
NEs for intranasal administration, in which NEs were used to convey risperidone.
This medication is available in oral formulations (tablets and oral solutions),
although it has a low bioavailability due to first-pass hepatic metabolism. Risperidone
NEs were synthesized with Capmul MCM as the oily phase and Tween 80 as the
surfactant. In addition, mucoadhesive NE was created by combining NEs with chi-
tosan [57]. In vivo investigations in Swiss albino rats using technetium (99mTc)
labelled formulations revealed faster and bigger drug transport into the CNS follow-
ing intranasal delivery of the mucoadhesive NEs compared to plain NE delivered
intranasally, intravenously, and as a solution [50]. Analogous results were obtained
for NEs loaded with olanzapine. The positive results were attributed to the increased
nasal retention duration caused by the presence of chitosan [50, 51, 56, 57]. Another
study created buffered mucoadhesive NEs filled with ziprasidone hydrochloride.
Mucoadhesive NE had a 1.79 times higher diffusion coefficient than ordinary
NE. The pharmacodynamic trials, which were free of acute nasal toxicity, produced
remarkable results in the locomotor activity test and the paw test. Thus, a safe and
effective NE for intranasal delivery of ziprasidone was created, despite the fact that
pharmacokinetic and biodistribution studies would have provided proof of evidence
in this situation. Intranasal NE of the drug is justifiable as it is commercially
available as oral capsule formulation (Geodon® and Zeldox®) that exhibits low
bioavailability due extensive first-pass metabolism of drug [14].
Quetiapine fumarate is an atypical antipsychotic medication that is taken orally
as tablets. Due to weak water solubility and a significant first-pass effect, quetiapine
oral treatment has a low bioavailability (5–15%) [45]. Hence, different formulations
are preferable. Following nasal delivery, O/w NEs were created to target the medi-
cation directly into the brain. In an in vivo tissue distribution study in Wistar rats,
post-intranasal delivery of quetiapine fumarate-loaded NE resulted in a lower T
max than intravenous administration. Because of its higher drug transport effi-
ciency, NE appears to be a potential method for brain-targeted delivery of quetiap-
ine fumarate [75].
5.5 Nanoemulsion in the Treatment of Epilepsy
Presently most of the antiepileptic drugs prescribed are orally in different dosage
forms such as tablet, solution, suspension, capsule and controlled release tablet and
capsule. For the rapid action, there is only one choice for the parenteral routes.
While several routes have been explored such as buccal, rectal and sublingual but
they have various limitations for the fast action. One report suggests that one-third
of epileptic patients have resistant to the presently marketed formulations of antiepi-
leptic drugs [22]. This resistance may be due to not achieving desired drug concen-
tration in the brain. There may be several regions for not reaching drug to the brain
such as hepatic drug metabolism, high plasma protein binding, efflux transporters in
GIT and BBB, drug-drug interaction, etc. These factors may be responsible for the
restricting effective transport of antiepileptic drugs and lower concentration of ther-
apeutic agents in the brain [72, 92]. The patients having drug resistance are treated
by intracerebral or intracerebro-ventricular delivery system for the management of
epilepsy [31]. Various studies have been performed with modern technologies for
the treatment of epilepsy such as nanoformulations through transdermal and intra-
nasal routes [27, 32].
Some research reports showed that intranasal NEs can be an alternative drug
delivery system for antiepileptic drugs. The nanoemulsion of phenytoin was pre-
pared and evaluated for biopharmaceutical parameters such as nasal toxicity and
drug release profile. The developed formulation showed that NEs were stable hav-
ing globule size of less than 20 nm. Release study showed 100% drug was released
within 48 h. The nasal toxicity study was performed with optimized formulation
and found to have safe. The formulations can be sterilized by filtration method
through 0.22 μm [54]. Further, preclinical studies are required to validate safety and
efficacy of nose-to-brain delivery. Jain et al. have developed amiloride-loaded NEs
for nose-to-brain delivery. The globule size of NEs was found to be in the range of
9.41 ± 1.23 to 10.71 ± 1.09 nm. The toxicity study of mucoadhesive NE was per-
formed in sheep nasal mucosa and found to be safe [70]. Hence, intranasal NEs can
be an alternative drug delivery system for the management of epilepsy.
6 Recent Patents on Nose-to-Brain Delivery

for CNS Disorders
Nanocarrier-based drug delivery system has been most widely explored for the
treatment of several neurological diseases. Various patents have been reported on
nanocarriers such as nanoparticles, nanoemulsion, solid lipid nanoparticles, lipo-
somes, and multiple dosage forms (Table 5.2) [74]. Improvement in therapeutic
interventions for the treatment of individuals with CNS disease should be empha-
sized by clinicians and pharmaceutical industries. Therefore, significant clinical
data should be collected and investigated for the development of products [75, 79].
7 Current Challenges and Future Prospective

for Intranasal Nanoemulsion
Intranasal NE may be an effective strategy for the drug targeting to the brain. Nasal
mucociliary clearance has been considered as one of the major limiting factors but
it can be resolved by developing mucoadhesive formulations of NEs. There are
some other challenges which can create obstacles for the successful product devel-
opment. Generally, in labs nanoemulsions are prepared by addition of excipients
along with sonication and these methods cannot be used for the large-scale produc-
tion of nanoemulsions. But fortunately, some new techniques have been explored
for the preparation of nanoemulsion formulation at industrial scale like high-
amplitude ultrasound, high-pressure homogenization, high shear mixing, etc.
Marketed NE formulations (Limethasone®, Vitalipid®, Diprivan® and Ropion®)
are evident of these methods but still not explored for nose-to-brain targeting [76].
Apart from issues of preparations of formulations, nasal irritation and chances of
nasal mucosa damage which can be appeared by repeated administration of NEs
have been considered as major limitation for nasal drug delivery. Several studies
showed that surfactants have efficient permeation properties but they also can dam-
age the structural integrity of nasal mucosa [62]. The repetitive administration of
formulation on nasal mucosa may cause irritation and bleed [87]. Several researches
have been reported on the NEs for the nose-to-brain delivery and they show signifi-
cant results. Hence NEs can be the most suitable drug delivery systems for the treat-
ment of CNS disease through intranasal administration [40].
In various researches, toxicity study has been performed either as short-term
duration (2–4 h) or single dose study in nasal mucosa for animals. Although, no
toxicity was found in several short-term studies by intranasal NEs [58]. While for
treatment of CNS diseases like as epilepsy, PD, AD, therapy is required for long
time [23]. Hence, long-term toxicity studies are required in animals on intranasal
NE. Surfactant concentration should be selected minimum which can be another
approach for the reduction of toxicity. There are so many surfactant and co-surfac-
tants which are not approved by USFDA but most commonly being used in the
Table 5.2 Patents for various nanotechnology-based dosage forms for treatment of

Alzheimer’s disease
Active
ingredient/ Publication
Patent no. compositionCentre point/main outcome year/granted Ref.
US 2013 Metallic ions
Metal NPs for diagnosing AD using 2013 [19]
8349293B2 magnetic resonance imaging (MRI)
US 2014 Cerium oxide Polymeric NPs of cerium oxide with 2014 [18]
8877207B2 antibody specific for amyloid β
embedded for better targeting in AD
US 2009 Nutritional Food supplement, nutritional mixture for 2009 [64]
0252796 A1 supplement improving the status of AD disease
manufactured using microfluidizers
US 2011 Multiple Nanoemulsion can improve the 2011 [21]
0045050A1 therapeutic bioavailability for vast therapeutic
agent segment
EP 2550020 Metal ions and Reverse micelle system for improved 2015 [63]
B1 lipids targeting using metal ions and various
lipids
US 2015 Model drug Liposomes with specific lipids that can 2015 [60]
0017235A1 reduce amyloid b plaques in AD
WO 2009 Model drug Liposomes with specific lipids content 2009 [61]
150686A1 that high binding capacity to reduce
amyloid b plaques in AD
WO 2014 Model drug Peptide conjugated liposomes for 2014 [10]
076709A1 targeting in AD
CA 02203513 Selegiline Selegiline liposome for better targeting by 2001 [66]
parenteral route and improved permeation
in case of transdermal delivery for
treatment of AD
US 2015 Curcuminoid The formulation makes the curcuminoid 2015 [24]
9192644B2 SLN stable at basic pH and improves the
concentration in AD patient brain
US 2006 Cholinesterase Delivery of cholinesterase inhibitors via 2006 [30]
0018839A1 inhibitors various dosage forms through nasal and
ophthalmic route to improve the targeting
in AD and ocular disorders
US 2014 Pharmaceutical CNS agent complexed with transport 2014 [94]
0100282A1 CNS agent moiety for effective transport through
intranasal route in neurological disease
US 2015 NSAIDS Intranasal NSAIDS for improved 2015 [42]
0086616A1 neuro-protection in case of AD using
various nano-dosage forms
EP 2332570 Glatiramer Proteosomes and nanoemulsion for GA 2011 [25]
A1 acetate (GA) through nasal route and parenteral route
to improve neuroinflammation in AD
267/ Glatiramer Proteosomes and nanoemulsion for GA 2007 [59]
KOLNP/2007 acetate (GA) through nasal route and parenteral route
to improve neuroinflammation in AD
Reproduced from Pathak et al. [75]
development of NE formulations [13]. Thus, for the development of intranasal NE,

we need to select approved surfactant and co-surfactants along with considering
their concentration limits. These approaches can be very helpful in the commercial-
ization of intranasal NE for nose-to-brain delivery [37, 38]. Several products have
been approved by FDA for various pathologies through intranasal administration
which has been represented in Table 5.3 [28].
Table 5.3 FDA-approved intranasal products for various pathologies [28]

Dosage Approval
Proprietary name Active ingredient form Applicant holder Year
DDAVP Desmopressin acetate Solution Ferring 1982
Pharmaceuticals
Inc
Tyzine Tetrahydrozoline Solution Fougera 1982
hydrochloride Pharmaceuticals
Inc
Beconase Beclomethasone dipropionate Metered GlaxoSmithKline 1987
monohydrate spray
Synarel Nafarelin acetate Metered Pfizer Inc 1990
spray
Nicotrol Nicotine Metered Pfizer Inc 1996
spray
Astelin Azelastine hydrochloride Metered Mylan Specialty 1996
spray LP
Imitrex Sumatriptan Metered GlaxoSmithKline 1997
spray
Nasonex Mometasonefuroate Metered Merck Sharp and 1997
spray Dohme Corp
Migranal Dihydroergotamine mesylate Metered Bausch Health US 1997
spray LLC
Cromolyn sodium Cromolyn sodium Metered Bausch and Lomb 2001
spray Pharmaceuticals
Inc
Butorphanoltartrate Butorphanol tartrate Metered Mylan 2001
Inc
Flunisolide Flunisolide Metered Bausch and Lomb 2002
Inc
Ipratropiumbromide Ipratropium bromide Metered Bausch and Lomb 2003
spray Inc
Nascobal Cyanocobalamin Metered Endo 2005
Inc
Omnaris Ciclesonide Metered CovisPharmaBv 2006
spray
(continued)
Dosage Approval
Proprietary name Active ingredient form Applicant holder Year
Patanase Olopatadine hydrochloride Metered Novartis 2008
Corp
Calcitoninsalmon Calcitonin salmon Metered ApotexInc 2008
spray
Sprix Ketorolac tromethamine Metered Zyla Life Sciences 2010
spray US Inc
Lazanda Fentanyl citrate Metered BtcpPharma LLC 2011
spray
Qnasl Beclomethasonedipropionate Metered Teva Branded 2012
monohydrate aerosol Pharmaceutical
Products R and D
Inc
Zomig Zolmitriptan Metered AstraZeneca 2013
spray Pharmaceuticals LP
Nasacort allergy24 Triamcinolone acetonide Metered Sanofi Aventis US 2013
hour spray LLC
Natesto Testosterone Metered Acerus 2014
gel Pharmaceuticals
Corp
Flonase allergy Fluticasone propionate Metered GlaxoSmithKline 2014
Relief spray
Rhinocort allergy Budesonide Metered AstraZeneca 2015
Narcan Naloxone hydrochloride Metered Adapt Pharma 2015
spray Operations Ltd
Onzetraxsail Sumatriptan succinate Powder Currax 2016
Kovanaze Oxymetazoline hydrochloride; Metered St Renatus LLC 2016
tetracaine hydrochloride spray
Xhance Fluticasone propionate Metered Optinose US Inc 2017
spray
Tosymra Sumatriptan Spray Upsher Smith 2019
Laboratories LLC
Spravato Esketamine hydrochloride Spray Janssen 2019
Pharmaceuticals
Inc
Nayzilam Midazolam Spray UCB Inc 2019
Baqsimi Glucagon Powder Eli Lilly and Co 2019
Valtoco Diazepam Metered NeurelisInc 2020
spray
Gimoti Metoclopramide Metered Evoke PharmaInc 2020
hydrochloride spray
8 Conclusion and Future Prospective
The present chapter covers the various aspects of intranasal drug delivery of nano-
emulsion in the management of several neurological disorders. Due to the limited
entry of therapeutic agents into the brain, drug delivery to the brain has become a
challenging task. NEs are composed of two immiscible liquids with surfactantSur-
factants and co-surfactants having low globule size with thermodynamic stability.
NEs have unique feature to increase the solubility and stability of drugs along with
their good interaction with the endothelial membrane of the brain. NEs have been
found to be a favourable delivery system for the brain targeting of drugs through
intranasal administration. While clinical data with robust studies are required for the
validation of in-vivo data.
Several in-vivo studies indicate that NEs may be an encouraging approach for
drug delivery for brain targeting by intranasal administration for the treatment of
CNS diseases. Hence, intranasal drug delivery could be an alternative route of drug
administration for the management of neuronal diseases. Several research reports
showed that better drug concentration in the brain was found through intranasal
NEs. The intranasal Nanoemulsions can be used as alternative drug delivery system
for the management of neuronal diseases. Hence, further extensive researches are
required for the confirmation of safety, efficacy and toxicity of NEs formulations
through clinical studies for commercialization. NEs could be most suitable for the
treatment of neurological disease through intranasal administration. They have abil-
ity to cross the BBB and desired drug concentration may be achieved in the brain.
Various studies showed significant indications that nose-to-brain products are likely
to emerge commercially included for several neurodegenerative diseases.
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Chapter 6
Potential Targeting Sites to the Brain
Through Nasal Passage
Mershen Govender, Sunaina Indermun, Pradeep Kumar,

and Yahya E. Choonara
Abstract Delivery to the central nervous system (CNS) has posed a major challenge
in both therapeutics and diagnostics. Although nose-to-brain delivery has gained
momentum as a result of the advantages it provides, it is not without its shortcomings.
Anatomical positions, delivery formulation constituents and cellular targeting have
been determined to be important considerations in optimal nasal drug delivery. This
chapter seeks to highlight the numerous anatomical challenges that face nose-brain
drug delivery platforms and will discuss the pathways that have been effectively
utilized for targeted CNS delivery through the nasal cavity.
Keywords Intranasal · Nose-to-Brain Delivery · Central Nervous System

Targeting · Blood Brain Barrier · Nasal Pathways
1 Introduction
Disorders such as epilepsy, multiple sclerosis, cerebrovascular diseases, Parkinson’s

disease, Alzheimer’s disease and brain tumours have been noted to affect the periph-
eral and central nervous system (CNS) in multiple ways [1]. Although many treat-
ments and therapies exist for CNS disorders, some are complex and sometimes
ineffective in eliciting an appropriate pharmacological response. Effective interven-
tions such as oral, topical, and intravenous treatments, deep brain stimulation, sur-
geries and rehabilitation therapies each come with their own disadvantages and
limitations. Furthermore, invasive strategies such as catheter infusions;
M. Govender · S. Indermun · P. Kumar · Y. E. Choonara (*)

Wits Advanced Drug Delivery Platform Research Unit, Department of Pharmacy and
Pharmacology, School of Therapeutic Sciences, Faculty of Health Sciences, University of the
Witwatersrand, Johannesburg, Gauteng, South Africa
e-mail: Yahya.Choonara@wits.ac.za

https://doi.org/10.1007/978-3-031-23112-4_6
84 M. Govender et al.
mini-pump-assisted intracranial delivery; electromagnetic force-field techniques;

precise ultrasound methods; and intracerebroventricular or intraparenchymal injec-
tions are acute treatment options which pose many risks and have the potential to
induce neurotoxic effects at the site of delivery [2].
Even though there are numerous advances in the drug delivery and neuroscience
fields, drug delivery to the CNS is still eluded by the microvascular blood-brain bar-
rier (BBB), thus rendering a cure elusive. The BBB, which is critical in the mainte-
nance of CNS homeostasis, is formed by the tight junctions that join specialized
CNS endothelial cells [3–5]. These cells are positioned at the blood-nervous tissues
interface as well as between the blood and the cerebrospinal fluid, where the blood-
cerebrospinal fluid barrier (BCSFB) is formed [6–8] (Fig. 6.1).
BCSFB cells are composed of choroid plexus epithelium cells, as well as the
arachnoid epithelium, which line the cerebral ventricles and the brain vasculature in
the subarachnoid space, respectively [8]. In comparison to the BCSFB, the BBB has
a larger surface area as well as a faster blood flow rate, and as a result is considered
to be the primary obstacle to brain permeability [7].
A continuous, non-fenestrated basal lamina is conferred by the endothelial cells
in addition to interacting with several perivascular elements such as pericytes, astro-
cytes, and perivascular macrophages which contribute to the barrier [6]. The tight
junctions that are present in the paracellular space between adjacent cells are imper-
ative in determining the BBB’s properties [5].
Hydrophilic molecules and drugs have limited permeability due to the junctions
consisting of several specific transmembrane proteins such as junction adhesion
molecules, claudin and occludin [9]. Low pinocytic activity, an enzymatic barrier as
well as several drug efflux mechanisms, including transmembrane proteins
expressed by the specialized CNS endothelial cells and other multidrug resistance
proteins, further support the physical barrier between the CNS and the blood and is
responsible for the removal of exogenous substances such as micro-organisms and
toxic compounds from the brain circulation [10]. These specialized CNS endothe-
lial cells also function to prevent fluctuations in the bloodstream, thereby ensuring
that synaptic transmission is maintained [6].
Either circumventing the BBB or optimizing systemic drug delivery has therefore
been the primary focus of CNS drug delivery. Permeation of approximately 100%
of macromolecules and 98% of low molecular weight drugs have been estimated to
be hindered by the BBB even though low molecular weight drugs not exceeding
400 Da, along with high lipophilicity, are typically considered as favourable
permeation factors [11]. Molecular physicochemical properties such as the
molecular mass, lipid solubility and charge further govern transportation across the
BBB. Since the cell membrane is unionized and consists of anionic phospholipids,
which confer a negative charge, basic molecules are favoured over acidic molecules.
The preferred drug molecule should possess an octanol: water partition coefficient
(log p) value near 2 (1.5–2.7) and have a molecular weight of less than 400–500 Da.
Notably, for each polar functional group or pair of hydrogen bonds that the drug
molecule possesses, the permeability across the BBB is reduced by one log unit
6 Potential Targeting Sites to the Brain Through Nasal Passage 85
Fig. 6.1 The three main barriers in the central nervous system (CNS), namely the meningeal or
arachnoid barrier, the choroid plexus barrier and the blood-brain barrier (BBB). The arachnoid and
choroid plexus barriers separate the blood from the cerebrospinal fluid (CSF), and the BBB sepa-
rates the blood from the interstitial fluid (ISF). At each site, the barrier is mainly formed by tight
junctions that seal off the paracellular space. The blood-brain barrier possesses an intricate archi-
tecture of basement membrane, mural and glial cells that work in synergy to maintain the barrier’s
integrity and regulate its permeability in response to neuronal needs. (Reproduced from Razzak
et al. [8] © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution (CC BY)
license (http://creativecommons.org/licenses/by/4.0/))
with the cumulative number of hydrogen bonds for appreciable permeability being
less than 8–10 [12].
Consequently, nose-to-brain delivery has been explored as a feasible drug
delivery route to the brain utilizing the nasal passages and tissue. This delivery
pathway allows for simplified administration, a fast onset of action, patient
compliance and a reduced systemic exposure of the delivered agent [4, 13, 14].
2 Drug Delivery Pathways
Understanding the morphological and structural properties of the potential targeting

sites in the nasal cavity is pertinent in the engineering of nose-to-brain delivery
systems. The nasal cavity, as the starting of the respiratory system, is vital in olfac-
tion processes, dust and particulate filtration, as well as the regulation of the tem-
perature and humidity of inhaled air [4].
2.1 The Nasal Cavity
The nasal cavity is longitudinally bisected by the nasal septum and extends from the
nostrils to the nasopharynx [13, 15] (Fig. 6.2). It has a length of 12–14 cm [13, 16,
17] and is 5 cm in height [16] with a reported surface area of between 150 and
200 cm2 [13, 18–20] and a total volume of 13–25 ml [13, 17, 18, 21].
The highly vascularized nasal mucosa lines three turbinates, i.e., the superior,
middle and inferior nasal turbinates, which extend laterally from the wall of each
nasal compartment, functioning to warm, filter and humidify inhaled air [13, 19].
The turbinates furthermore have an approximate surface area of about 160 cm2 [15].
Each half of the nasal cavity consists of three regions: the vestibule (∼0.6 cm2)
[22], which is located immediately at the nostril, extending from the nostrils to the
inferior turbinate [15, 23]; the olfactory region (2–12.5 cm2) [13, 19, 22]; and the
respiratory region. The vestibule region contains stratified squamous epithelium,
and its mucosa contains hairs, sweat and sebaceous glands [15]. Non-ciliated tran-
sitional epithelium separates the squamous cells from the respiratory epithelium and
the respiratory epithelium from the olfactory epithelium, respectively [15]. The
relatively small surface area of the vestibule region, as well as the non-ciliated cell
surfaces, makes this a poor drug absorption site [24] with the respiratory and olfac-
tory mucosa an intended drug absorption site [15]. The nasopharynx-associated
lymphoid tissue (NALT), containing immune cells, is interconnected to the tonsils
and the local lymph nodes [15].
During inhalation, air enters the nostrils, flows into the flexible nasal valve via
the nasal vestibule and into the main chamber [16, 19, 25]. Despite the nasal cavity’s
high vascularization and large surface area, its anatomic configuration results in
only 15–20% of inhaled air reaching the olfactory region [25, 26].
Fig. 6.2 Anatomy of the human nasal cavity. Squamous mucosa (green) is located at the frontal
parts of the nasal vestibules. The three turbinates (inferior, middle and superior) humidify and
warm the inhaled air. The area covered predominantly with respiratory mucosa is labelled in blue.
The olfactory mucosa (yellow) is located next to the cribriform plate at the skull base down to the
superior turbinate. Nasally transmitted substances can cross the cribriform plate via different path-
ways to enter the brain. Nasopharynx-associated lymphatic tissue (NALT) is located in close prox-
imity to the tonsils at the nasopharynx. (Reproduced from Gänger and Schindowski [15] © 2018
by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distrib-
uted under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://
creativecommons.org/licenses/by/4.0/))
2.2 The Respiratory Region and Epithelium
The respiratory region accounts for the largest area of the nasal cavity, occupying a
total surface area of approximately 80–90% or ~130 cm2 due to its ciliated pseu-
dostratified columnar epithelium lining [13, 21, 22, 24]. Its high degree of vascular-
ization, along with a large microvillus- covered surface area, renders the respiratory
region a major site for systemic drug absorption [21, 25, 27]. Furthermore, blood
supply is received from the maxillary artery via an arterial branch and is innervated
by the trigeminal nerves, a potential targeting pathway for nose-to-brain deliv-
ery [27].
Four distinct cell types comprise the respiratory epithelium, viz. the ciliated and
non-ciliated columnar cells, the goblet cells and the basal cells [4]. These cells func-
tion in facilitating intracellular water and ion exchange, mucus secretion and clear-
ance and mucosal humidity regulation, in addition to the coordinated cilial sweeping
motion with an approximate frequency of 1000 S/min [20, 22, 25].
A double-layered viscoelastic mucus gel covers the respiratory epithelium,

comprising of a network of mucins, water, salts, proteins and some lipids. Its
viscoelastic and adhesive properties confer protective abilities against inhaled
irritants and particulates [28, 29]. The sweeping motion of the epithelial cilia tips
together with continuous mucus secretion allowing for the entrapped inhaled
irritants, particulates and microbes to be transported along the nasal passage and
pharynx, at an approximate rate of 1–30 mm/min [19], until enzyme- and acid-
mediated lysis in the stomach occurs. This pathway refers to the process of
mucociliary clearance [20, 21, 25]. The respiratory mucus layer is additionally
estimated to be renewed every 1 to 20 min [17].
2.3 The Olfactory Region and Epithelium
The olfactory region is situated under the cribriform plate, which is the horizontal
bone that separates the brain and the nasal cavity, occupying 2 ~ 12.5 cm2 or approx-
imately 1.25–10% of the nasal cavity [13, 19, 22, 24, 25]. This highly perforated
structure allows for nerve endings to enter the nasal cavity and is thus recognized
extensively as a viable nose-to-brain drug delivery route for various CNS disease
treatments [22, 30].
Ciliated chemosensory pseudostratified columnar epithelium furthermore lines
the olfactory region of the nasal cavity (Fig. 6.3). The olfactory mucosa, which is
surrounded by the respiratory epithelium, is located on the surface of the superior
turbinate. The olfactory mucosa is also located bilaterally on the nasal septum [4].
The ophthalmic artery branches provide the blood supply to this region [27] and the
olfactory epithelial cilia is longer (over 50 μm) than that of the respiratory epithe-
lium and is non-motile [22].
Mucus clearance occurs as a result of continuous mucus secretion by the
Bowman’s glands, gravitational and mechanical forces, and the solvent drag effect,
with the mucus turnover being over several days [19, 20]. The olfactory mucosa is
also highly innervated, containing the autonomic nerve fibres, the olfactory axon
bundles and the maxillary branch of the trigeminal nerve.
The olfactory epithelium has been noted to consist of several distinct cell types
including the sustentacular cells which is the most abundant olfactory epithelium
cell type. These are microvilli-possessing columnar cells that function to provide
both mechanical and metabolic support to the olfactory epithelia, whilst also func-
tioning to regulate the ionic environment of the overlying mucus [4, 13, 22]. These
cells also function to catabolize inhaled xenobiotics as they exhibit high enzymatic
activity [19].
The small, conically shaped basal cells are also situated in the basement
membrane and provide mechanical support to the other cells [18] and due to their
ability to differentiate into other cell types, specifically olfactory sensory neurons
(OSNs), function to continuously replace dead cells [13].
Fig. 6.3 Structure and composition of the olfactory mucosa. In the posterior part of the nasal
cavity, the olfactory mucosa, together with the olfactory epithelium (OE) and the lamina propria
(LP), represents the first contact zone of environmental cues towards the human body. Within the
OE, the mature olfactory sensory neurons (OSNm) are projecting their axons towards the olfactory
bulb (OB), where they form glomeruli with the dendrites of mitral cells. The axons of OSNms are
enclosed by olfactory ensheathing cells (OECs) and olfactory nerve fibroblasts (ONFs). The axons
together with the OEC and ONF form the olfactory nerve bundles (ONBs) in the lamina propria.
The OE further consists of sustentacular (SUS) and mucus-producing Bowman’s glands (BGs). In
the middle part of the OE are the immature ORN (ORNi). The OE is surrounded by a layer of
immature basal cells, the globose (GBC) and horizontal basal cells (HBCs). The lamina propria
(LP) is separated from the OE by a basal lamina (BL). Further, the LP also contains blood vessels
(BVs) and lymphatic vessels (LVs). (Reproduced from Keller et al. [32] © The Author(s) 2021.
This article is an open access article distributed under the terms and conditions of the Creative
Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/))
The terminal branches of the trigeminal nerve also consist of brush cells. The
surface of these cells displays a tuft of blunt, squat microvilli (approximately
120–140 per cell) which contain filaments that extend into the cytoplasm of the cell
[31]. In the respiratory system, the brush cells are characterized by the absence of
the distinctive terminal situated immediately beneath the microvillous border, dif-
ferentiating them from those present in the gastrointestinal system [31]. The brush
cells and their associated microvilli function to provide sensory mucosal stimula-
tion [22].
2.3.1 Olfactory Sensory Neurons
The olfactory sensory neurons (OSNs), which are interspersed between the
sustentacular cells [18], are non-myelinated neurons that are enclosed by specialized
ensheathing cells. The OSNs are responsible for the process of olfaction, as they are
exposed to the inhaled air [15]. Dendrites from the OSNs also extend into non-
motile cilia and terminate into externally exposed mucus [13]. Unmyelinated axons
bundle into profuse axon bundles that are enwrapped by the olfactory ensheathing
cells and nerve fibroblasts, entering the cribriform plate of the ethmoid bone through
its foramina to further penetrate further into the brain [4].
Due to their inherent function, OSNs have a lifespan of 1 month. OSN systematic
apoptosis protects the brain from any infections [15]. Basal stem cells further ensure
tissue maintenance-related cell death or recovery of the olfactory mucosa after
injury [15]. Two types of different basal stem cells are located in the olfactory
mucosa that differentiate into OSNs [33]. When the OSN dies, gaps in the epithelial
layer are formed until an OSN can regrow into that space, resulting in the delayed
formation of tight junction during this time [34]. Beneath the epithelium layer, the
Bowman’s glands (for production and secretion of mucus), axons, lymphatic ves-
sels, blood vessels, and connective tissue are contained [35].
3 Drug Delivery Pathways/Brain Targeting Sites via

the Intranasal Route
Successful intranasal administration of small lipophilic compounds occurs when

drugs bypass mucociliary clearance in the vestibular region and migrate to posterior
regions of the nasal cavity, where they are absorbed through contact with the respi-
ratory epithelium. Subsequent absorption into the systemic circulation via the blood
or lymphatic system then occurs. However, both increased systemic exposure as
well as hepatic metabolism is still necessary for BBB delivery through this transcel-
lular delivery pathway [4, 24]. Direct drug transport to the brain occurs via the
olfactory or trigeminal pathways with indirect drug transport occurring via the sys-
temic pathway [18, 22, 24, 36] (Fig. 6.4).
3.1 The Olfactory Pathway
The olfactory epithelium lends itself to three possible drug transport pathways: the
transcellular pathway; the paracellular pathway; and the olfactory nerve pathway
(Fig. 6.5). Passive diffusion or endocytosis governs the transcellular pathway, where
the drugs move across the respiratory epithelium and the sustentacular cells in the
olfactory epithelium to the OSNs and peripheral trigeminal neurons.
The intracellular and transcellular transport pathways are responsible for drug
delivery to the various regions of the brain. The intracellular route transports the
drug from the olfactory nerve to the olfactory lobe and from the trigeminal nerves
to the brain. This pathway is primarily responsible for the lipophilic drug transport
and is also referred to as the intraneuronal route of drug transport. This drug
(a) Mucocilliary clearance

First-pass metabolism
Respiratory epithelium Systemic circulation BBB
Nasal Cavity Trigeminnal nerve Brain
Olfactory epithelium
Olfactory nerves
CSF
(b)
Olfactory
bulb
Cribriform Olfactory axon

Plate
Olfactory
Lamina
Propria neuron
Basal cells
Olfactory
epithelium
Cilia
Mucus layer
i) Extracellular ii) Intracellular iii) Transcellular

pathway pathway pathway Intranasal
Administration of
drugs or drug
formulations
Fig. 6.4 (a) The potential drug transport routes leading to brain uptake following intranasal
administration (b) Schematic representation of various possible mechanisms involved in direct
nose-to-brain drug transport from the olfactory region. (Reproduced from Hong et al. [24] © 2019
by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distrib-
uted under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://
creativecommons.org/licenses/by/4.0/))
transport route from the nasal cavity to the brain is relatively slow, reaching the
CNS after 24 hours [37, 38].
Hydrophilic drugs are transported between the sustentacular cells via the
paracellular pathway, where appreciable bioavailability is achieved with drugs
having a molecular weight of 1000 Da or more. The olfactory nerve pathway
involves the uptake of the drug via the neuronal cells with transport to the olfactory
bulb being facilitated through intracellular axonal transport [17, 40].
The olfactory neurons also transport drugs into the olfactory bulb via the
intracellular axonal channel [41]. The diameter of the olfactory axon is approximated
to be 0.1–0.7 μm [4], providing a targeted size range for nanoparticles or molecules
Fig. 6.5 Schematic overview of several molecular pathways after intranasal drug administration.
CSF cerebrospinal fluid. BBB: blood-brain barrier. (Reproduced from Tashima [39] © 2020 by the
author. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under
the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creative-
commons.org/licenses/by/4.0/))
to be appropriately delivered. Both extracellular and intracellular mechanisms

govern drug transport through the olfactory pathway, with the paracellular route
transporting hydrophilic drugs and passive diffusion transporting lipophilic
drugs [2].
Using the olfactory pathway, wheat germ agglutinin-horseradish peroxidase was
shown to be concentrated (140 nM) in the olfactory nerve and glomerular layers of
the olfactory bulb [42]. Additionally, for the treatment of Alzheimer’s disease,
recombinant human nerve growth factor was delivered to the brain achieving con-
centrations of 3400 pM and 660–2200 pM in the olfactory bulb and in other brain
regions, respectively [43]. Nanogold-labelled insulin was also noted to reach the
anterior regions of the olfactory bulb 30 min after administration [44].
3.2 The Trigeminal Pathway
The trigeminal nerve functions to provide chemo- and thermo-sensory stimuli to the
nasal, oral and ocular mucosa [45]. Since the dorsal nasal mucosa is innervated by
the trigeminal nerve, the trigeminal nerve pathway is a potential delivery pathway
(although not widely used) for drug delivery to the brain, particularly the frontal
brain and olfactory bulb [36].
Labelled fluorescein isothiocyanate (FITC)-insulin was intranasally administered
in female Sprague-Dawley rats for its distribution and delivery pathways into the
brain via the trigeminal nerves [46] (Fig. 6.6). Upon excision of the nerves from the
base of the skull, at the point in which the V1V2 branches exit the anterior lacerated
Fig. 6.6 FITC-insulin was intranasally administered and imaged in the trigeminal nerve 30 min
later (b, e, h). Axons in the trigeminal nerve were labelled with the pan-neuronal marker Neuro-
Chrom (a, d, g). The merged images (c, f, i) show FITC-insulin in the perineural spaces of the
trigeminal nerve. These data suggest FITC insulin reaches the CNS along perineural spaces of the
trigeminal nerve. Scale bar = 10 μm. (Reproduced from Lochhead et al. [46] © The Author(s)
2019. This article is licensed under a Creative Commons Attribution 4.0 International License.
This article is an open access article distributed under the terms and conditions of the Creative
Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/)))
foramen, FITC-insulin localization was observed in the endoneurium, with stronger

signals detected in the perineurium and epineurium of the trigeminal nerve.
In a study by Wang and co-workers [47], a self-assembling two-component
supramolecular hydrogel was evaluated as a vehicle for L-DOPA delivery. The sys-
tem was evaluated as a potential treatment of Parkinson’s disease using a hemipar-
kinsonian rat model. The gel was shown to display 4.1 times more L-dopa uptake in
the brain. Additionally, more L-dopa was present in the blood (2.1 times) at 10 min
after intravenous administration of an equivalent dose. Radioactivity after 10 min in
various regions of the brain was evaluated following nasal administration of the
hydrogel (Fig. 6.7). 3H-labelled L-DOPA was readily distributed throughout the
brain with the greatest detection in the trigeminal nerve, which accounted for more
than 30% of the brain’s total radioactivity.
Fig. 6.7 Brain distribution of [3H]l-DOPA hydrogel at 10 min post intranasal administration. (a)
Dorsal view of the mouse brain and dissection guidance of different brain segments, the olfactory
bulbs (OB), the cerebrum (CB 1&2), the brain stem (BS), cerebellum (CE), spinal cord (SP), and
trigeminal nerves (TN). (b) % of [3H]l-DOPA uptake in different brain segments. At the experi-
mental endpoints, whole body perfusion with 0.9% saline was performed and studied tissues were
dissected and proceeded for liquid scintillation counting. Results are expressed as % uptake nor-
malized to total [3H]l-DOPA detected in these collected tissues. Data are expressed as mean ± SD,
n = 3. (Reproduced from Wang et al. [47] © 2021 The Authors. Advanced Science published by
Wiley-VCH GmbH. This article is an open access article distributed under the terms and condi-
tions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/
by/4.0/))
3.3 The Lymphatic Pathway
Olfactory nerves from the lamina propria of the olfactory region, terminating in the
olfactory bulb, give rise to extracellular pathways such as the perivascular, perineu-
ral and lymphatic pathways [2, 4, 13].
In a study by Furubayashi and co-workers [48], intranasal administration of
methotrexate to the cervical lymph nodes (CLNs) via the nasal mucosa was studied
in Wistar rats. The study concluded that the delivery of methotrexate was attributed
more to the direct nasal–CLN pathway as opposed to the direct blood–CLN path-
way with a direct transport percentage of 74.3%. Although these pathways are not
new, research on them are few and far between [13, 49].
3.4 The Systemic Pathway
Drugs that are lipophilic with low molecular weights are most suited for delivery via
the systemic pathway. This pathway avoids hepatic first-pass metabolism as drugs
absorbed from the vascular epithelium membrane of the nasal mucosa and the lym-
phatic system are transported directly into the systemic circulation [2, 20, 26].
Table 6.1 Overview of drug delivery pathways related to the nasal cavity
Examples with supporting
Drug delivery route related to different nasal mucosa clinical data
Local Predominantly squamous and respiratory Decongestants, local
administration mucosa anaesthetics,
glucocorticoid [50, 51]
Systemic delivery Predominantly respiratory mucosa Calcitonin, sumatriptan,
desmopressin [52–54]
Intranasal NALT and immune cells in all mucosal Seasonal flu vaccine [55, 56]
vaccination types
CNS delivery Olfactory mucosa: olfactory neuronal Oxytocin, insulin [57, 58]
(N2B) bundles;
Respiratory mucosa/olfactory mucosa:
trigeminal nerve endings
Reproduced from [15] © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is
an open access article distributed under the terms and conditions of the Creative Commons
Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/)
Table 6.1 summarizes the many nose-to-brain pathways, delivery routes and their
targeting compartment.
4 Factors Affecting Nasal Absorption
Although nasal delivery offers many advantages, its biggest limitation is the
restriction of dose size/volume, allowing only 50–250 μL per nostril at a time,
making drugs requiring high dose obsolete from nasal delivery. Molecules exhibiting
weights >1000 Daltons further display poor absorption through the nasal mucosa.
Alterations in nasal secretions such as tonicity and pH (generally 4.5 ~ 6.5) as a
result of flu, cold, allergies and other pathological conditions pose a challenge to
nasal drug delivery and may exaggerate drug effects [24, 35, 59].
Furthermore, inner nasal surface enzymes such as exonuclease and endonuclease
as well as aldehyde hydrogenase, epoxide hydrolase, glutathione S-transferase and
carboxylesterase contribute to protein and peptide degradation and drug metabolism
[35, 60–62]. In addition to the afore-discussed physicochemical properties affecting
BBB transversion, poor nasal mucosa penetration capacity and tonicity-related
rapid mucociliary clearance result in poor nose-to-brain drug transport [59]. Dosage
form factors of drug concentration, osmolality, surfactant type and viscosity also
have an effect on nasal absorption and toxicity [63]. Preservatives such as benzalko-
nium chloride have further been noted to sensitize the nasal mucosa [64], while
head position during nasal administration has been determined to affect the extent
to which drug deposition occurs as well as localization within the nasal cavity
[24, 65].
5 Conclusion
Despite the mentioned restrictions and limitations, research into nose-to-brain

delivery has resulted in promising platforms with the potential to significantly
improve the delivery of drug molecules to the brain. This has been achieved through
the targeting of specific nasal pathways, dependent on the properties of the drug
molecule and the delivery system, with effective drug delivery occurring in various
parts of the brain. With research into this field of study, new and more effective
systems can be developed which will seek to further enhance the treatment of CNS
conditions.
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Chapter 7
Biomedical Applications of Nanocarriers
in Nasal Delivery
Namdev Dhas, Soji Neyyar, Atul Garkal, Ritu Kudarha, Jahanvi Patel,

Srinivas Mutalik, and Tejal Mehta
Abstract Non-invasive drug delivery is an emerging way to target a wide range of

therapeutics. One of them, i.e., the nasal drug delivery has shown promising results
in delivering small and large molecules, genes, peptides, and proteins. This drug
delivery system targets the drug to the brain by direct nose-to-brain and/or indirect
nose-to-blood-to-brain routes. Nanocarriers play a vital role in the nasal delivery
system due to their small size which provides ease in targeting. Numerous strategies
have been explored by researchers for the transportation of drugs from nose to target
and found excellent results in therapy. It was observed that polymeric and lipidic
nanocarriers have numerous applications in targeted drug delivery, gene delivery,
and vaccine delivery. Moreover, they are also used for diagnostics as well as ther-
anostics purposes. This chapter briefly summarizes the different types of nanocarri-
ers used for nasal delivery with their characterization techniques. Further, the
biomedical applications of nanocarriers via the nasal route are discussed in detail.
Keywords Nasal drug delivery · Biomedical application · Nanocarriers · Nose-to-

Brain · Ploymers
N. Dhas
Department of Pharmaceutics, Institute of Pharmacy, Nirma University,
Ahmedabad, Gujarat, India
Department of Pharmaceutics, Manipal College of Pharmaceutical Sciences, MAHE,
Manipal, India
S. Neyyar · R. Kudarha · S. Mutalik
Department of Pharmaceutics, Manipal College of Pharmaceutical Sciences, MAHE,
Manipal, India
A. Garkal · J. Patel · T. Mehta (*)
Department of Pharmaceutics, Institute of Pharmacy, Nirma University,
Ahmedabad, Gujarat, India
e-mail: tejal.shah@nirmauni.ac.in
https://doi.org/10.1007/978-3-031-23112-4_7
102 N. Dhas et al.
1 Introduction
Clinical utilization of nanomaterial is turning out to be progressively significant in

the treatment of several diseases and diagnostics along with theranostic applica-
tions. In medicine, nanocarriers improve the pharmacokinetics, bioavailability, and
efficiency of many medicines or contrast agents by assuring greater hydrophilicity,
less interaction with cellular proteins and plasma, and better accumulation in target
tissues. The ability of nanoparticles to localize (or be targeted) in a precise manner
to the site of action and reduce or eliminate harmful side effects is the most promis-
ing application of nanocarriers in medicine [1–4]. The Food and Drug Administration
(FDA) and the European Medicines Agency (EMA) have already approved over 50
nano carrier-based formulations for intravenous, intramuscular, topical, oral, intra-
bronchial, subcutaneous, administration as a result of nanomaterials research.
Liposomes and polymeric nanoparticles (NPs) are the most clinically authorized
nanomedicines; however, the number of nanomaterials approved by the FDA for
medical uses is still small [4–8]. Apealea®/Paclical®, which contains micellar
paclitaxel PTX (PTX formed with the surfactant XR17) and is used to treat Fallopian
tube cancer, primary peritoneal cancer, and epithelial ovarian cancer, is an example
of a polymeric nanomedicine [9]. The safety and therapeutic efficacy of novel nano-
materials were recently confirmed in multiple clinical investigations, the majority of
which focused on anti-cancer medication nanocarriers [10]. Another PTX nanofor-
mulation, NK105, a “core-shell-type” polymeric micellar NPs formulation used in
patients with metastatic or recurrent breast cancer, has already completed phase 3 of
clinical trials. Additionally, numerous distinct nano-based systems have been cre-
ated in connection with the severe acute respiratory syndrome coronavirus 2 (SARS-
CoV-2) pandemic that could be effective in the treatment of individuals with the
coronavirus disease-19 (COVID-19) [11]. Clinical trials for a full-length recombi-
nant SARS-CoV-2 glycoprotein NPs vaccine adjuvanted with Novavax’s saponin-
based Matrix M (NVX-CoV2373, NCT04368988) or Remdesivir’s inhaled NPs
formulation have recently begun (developed by NeuroActiva, NCT04480333, data
from clinicaltrials.gov). There are consistent huge requests on new, progressed,
multifunctional nanomaterials to be utilized in the eventual fate of medication. In
addition, to this, these fabricated nanocarriers need to be delivered via a proper
route through which an efficient pharmacological action can be obtained. For the
same, route of administration also plays a significant role ineffective treatment.
Each administration routes have their advantages and disadvantages. Nasal delivery
has garnered more attention among conventional and non-conventional delivery
routes. The nasal route exhibits numerous merits, for instance, non-invasiveness,
first-pass metabolism can be avoided, the nasal route provides large surface area as
a site of absorption, lesser systemic exposure, rapid and fast absorption, avoids tox-
icity to healthy cells to a greater extent, more specifically when the drug needs to be
transported to the brain, the nasal route bypasses the blood-brain barrier (BBB)
[12]. However, along with its merits, it also possesses several disadvantages, rapid
7 Biomedical Applications of Nanocarriers in Nasal Delivery 103
Frontal sinus
B
Upper nasal turbine
Middle nasal turbinate
Nasal cavity Nasal septum Sphenoid sinus
Lower nasal turbinate

Vomeronasal
organ Nasal choanae
Oral cavity
Tongue
Nasal cavity Histology
Fig. 7.1 The anatomy of the human nasal cavity. (a) Histological section of the human nasal cav-
ity. (b) Schematic view of the internal structure of the human nasal cavity. (Open access article
distributed under the terms of the Creative Commons CC-BY license, which permits unrestricted
use, distribution, and reproduction in any medium, provided the original work is properly
cited) [13]
mucociliary clearance results in a short residence time of nanocarrier at the site of

absorption ultimately leading to lesser bioavailability, the nasal cavity is small
hence it limits dose. Figure 7.1 illustrated the anatomy of the human nasal cav-
ity. [13].
Although the exact mechanisms by which therapeutic moieties go from the
nose to the brain are unknown, the olfactory pathway plays an important role. The
pathway involves the olfactory bulb, lamina propria, and olfactory epithelium.
Figure 7.2 illustrates the intranasal drug transport through the olfactory route to
the CNS by intracellular and extracellular pathways [14, 15]. The second signifi-
cant pathway, the trigeminal nerve pathway, enters the brainstem through the pons
by innervating the nasal cavity, whereas it enters the forebrain through the cribri-
form plate, allowing drugs/therapeutic moieties to reach the caudal and rostral
portions of the brain. This connection of the nasal route and brain opened several
avenues for research and attracted several researchers to narrow down their
research orientation. Figure 7.3 illustrates the trigeminal nerve pathway.
Additionally, the nasal route can be significant for the delivery of nanocarrier-
based systems in which several drug molecules such as peptides, nucleotides, and
proteins can be entrapped through which enzymatic degradation can be avoided in
the nasal cavity [16]. Nanocarrier-based systems which are the most promising
technology among the advanced technologies and controlled release systems are
discussed in the present chapter.
104 N. Dhas et al.
Fig. 7.2 Intranasal drug transport through the olfactory route to the CNS by intracellular and
extracellular pathways. The drug is taken up by OSNs, which project to the olfactory bulb. The
extracellular route is between the SCs, where the drug passes through the tight junctions (TJs),
paracellular cleft, the lamina propria, perineural space, and ultimately to the subarachnoid space
where it is transported to distal targets around the CNS. Abbreviations: SC Support cells, OSN
olfactory sensory neurons, OEC olfactory unsheathing cells, ONF olfactory nerve fibroblasts.
(Ref: Modified from [14, 15])
Fig. 7.3 The trigeminal nerve pathway, The initial processes at intracellular and extracellular
mechanisms of intranasal drug transport to the CNS. Intracellular shows pinocytosis/endocytosis
(1), trafficking of the endosome to Golgi apparatus (2), sorting with the Golgi stacks (3), and axo-
nal transport toward olfactory bulb (4). The extracellular pathway shows the movement of the drug
to the paracellular space and translocations through an absent tight junction (TJ) (6), and finally
translocation to the lamina propria through the paracellular cleft (7). Abbreviations: olfactory sen-
sory neuron (OSN), supporting cells (SC), endosome (EN), Golgi apparatus (GA), exosome (E),
and tight junction (TJ). (Ref: Modified from [14, 15])
2 Types of Nanocarriers
2.1 Polymeric Nanocarriers
The polymeric nanocarriers are usually derived from natural as well as synthetic
origin, with a size range from 10 to 1000 nm [17]. The polymeric nanocarriers
exhibit several advantages like outstanding intracellular uptake and prolonged and
controlled delivery of therapeutic moiety. In a study, uptake of PLA, chitosan, and
PLGA nanoparticles (NPs) by the olfactory ensheathing cells were evaluated. The
PLGA NPs showed better uptake than PLA and chitosan [18].
While treating neurological diseases like schizophrenia, the drug should be pres-
ent in the brain cells for an extended duration of time to get a better therapeutic
response. Some of the biological obstacles like first-pass metabolism, low bioavail-
ability, and frequent dosing are making conventional delivery systems of less usage.
Shah and his co-workers formulated quetiapine (QT)-loaded chitosan-based poly-
meric NPs and microemulsions for nasal delivery. The polymeric nanoparticles
(PNP) were manufactured by ionic gelation and chitosan-based mucoadhesive
microemulsion (MME) through the water titration technique. The loading efficiency
was higher (95%) in MME compared to PNP (65%). The reason for poor drug load-
ing in PNP could be due to pH-dependent and poor aqueous solubility of quetiapine.
MME showed 1.3-folds of higher diffusion compared to PNP as it is hydrophilic.
Permeation-enhancing property of chitosan in MME made them possess superior
diffusion characteristics than NPs. The pharmacokinetic study in Sprague-Dawley
rats showed better concentration in the brain and about 1.9-fold higher bioavailabil-
ity in MME over PNPs, attributed due to bypassing of the blood-brain barrier and
transporting via an olfactory route. MME (i.n.) showed 1.2–2.0-folds of blood/brain
ratio than that of QTNP. The biodistribution of the formulation was evaluated using
gamma scintigraphy, which is a non-invasive imaging technique. The formulations
were labeled with technetium-99 m via a direct labeling approach. Thin-layer chro-
matography was used to confirm radiolabeling efficiency of the formulation. The
formulations were then administered via i.v. and i.n. route and compared their radio-
activity. MME formulation administered via i.n. route showed the highest radioac-
tivity. Pharmacokinetic study along with gamma scintigraphy study demonstrated
that brain delivery of quetiapine was notably better in MME compared to PNP. The
findings of in vivo biodistribution and imaging study confirmed the outstanding
potential of MME as a candidate for the delivery of quetiapine via the non-invasive
intranasal route [19].
Mucus-penetrating NPs can reach the brain via intranasal administration. Junior
et al. developed PEGylated polycaprolactone (PCL) NPs loaded with bexarotene to
overcome reduced epithelial permeation, enzymatic degradation, and rapid muco-
ciliary clearance. All the formulations exhibited spherical structure and an average
diameter of 100 nm, and PDI less than 2. Particle diameter and morphology were
unchanged on PEGylation with different concentrations of PCL-PEG such as 1, 3,
5, and 10% w/w. 5% and 10% of PCL-PEG exhibited sufficient homogeneity and
106 N. Dhas et al.
stability in artificial nasal mucus. 98.8% of mucus-penetrating activity was observed

in 5% concentration and 99.5% in 10%. Fluorescence microscopy confirmed that
the presence of PEG on the surface of NPs has no role in changing the uptake by
RMPI 2650 cells. Translocation of the system into the brain was confirmed by fluo-
rescence tomography for 5% PCL-PEG. The concentration of drug-loaded 5%
PCL-PEG in the brain was threefold higher than drug dispersion and threefold than
non-PEGylated NPs. The increased distribution and retention of formulation in the
brain followed by intranasal administration is expected due to enhanced mobility
and stability of the 5% PCL-PEG. All these data indicate that 5% of PCL-PEG is
effectively promoting permeation of NPs through the mucus, bypassing the clear-
ance and finally increasing the concentration of drug in the brain cells [20].
In another study, Masjedi and his co-workers developed chitosan NPs loaded
with sumatriptan succinate for intranasal delivery. The drug-loaded nanoparticles
were prepared by the ionic gelation technique. The drug loading efficiency was rela-
tively high due to the establishment of a hydrogen bond between drug molecules
and chitosan. In vitro release study suggested more than 50% drug release in 24 h.
The in vivo parameters such as drug targeting efficiency (DTE%) and drug transport
percentage (DTP %) showed 493.39% and 79.79%, which indicates direct and fast
delivery of sumatriptan to the brain via nasal cavity [21]. Sclachet et al. evaluated
the biodistribution of polymeric nanoparticles of poly (methyl methacrylate), chito-
san, and poly (vinyl alcohol) for brain delivery via intranasal (i.n.) and intravenous
(i.v.) administration. The drug concentration in the off-target region was found to be
less in i.n. compared to i.v. The results suggest that both delivery routes exhibited
differential accumulation in the brain region and could be utilized for various medi-
cal conditions [22]. Joachim et al. explored gelatin NPs (GNPs) as a carrier for the
intranasal delivery of osteopontin (OPN) for the therapy of ischemic stroke. GNPs
can potentiate the neuroprotective activity of OPN on intranasal administration. The
increased efficacy of OPN was attributed due to the presence of GNP which protects
OPN from degradation by protease, sustained drug release, and higher concentra-
tion of drug in brain regions [23]. Muntimadugu et al. designed a poly (lactide-co-
glycolide) (PLGA) nanocarrier for the brain delivery of tarenflurbil (TFB), which
has failed in the clinical trial (phase 3) due to its poor brain penetration. The formu-
lation showed particle size less than 200 nm, which assured the transcellular trans-
port of the drug through the olfactory axons. In vitro drug release study confirmed
the prolonged residence time at the targeting site. DTE and DTP results indicated
that the drug was capable of direct transportation to the brain after the administra-
tion of polymeric nanoparticles via the olfactory pathway [24].
2.2 Lipidic Nanocarriers
Lipid-based nanocarriers are one of the drug delivery systems (DDS) to be discov-
ered and approved by the FDA. Liposomes are lipid-based nanocarriers of phospho-
lipid spherical bilayer vesicles exhibiting a similar structure as that of the human
cell membrane and are biodegradable and biocompatible [4, 25]. Li et al. developed
flexible liposomes of galanthamine hydrobromide (GH) to check the pharmacoki-
netic characteristic of acetylcholinesterase inhibition via i.n. administration.
Negative-stained TEM micrographs of flexible liposomes demonstrated that it is
spherical and consists of multilamellar vesicles. The size distribution range of
112 ± 8 nm confirmed the homogeneity of the preparation. It exhibited a zeta poten-
tial of about −49.2 ± 0.7 mv, which indicates the stability of liposome by electro-
static repulsion which inhibits the aggregation. It showed excellent entrapment
efficiency of 83.6 ± 1.8%, which is attributed due to the high lamellarity of vesicles.
The cell viability on PC-12 cells confirmed the non-toxicity of the cultured cells.
The acetylcholinesterase activity was determined in male SD rats using a reagent
kit. The animals were administered with the liposome via i.n. at a GH dose of 3 mg/
kg. The animal which was given with GH-loaded formulation showed good perme-
ability in the rat nasal mucosa and better anticholinesterase activity. The cerebral
microdialysis technique was used to assess the pharmacokinetic properties of for-
mulation in the brain. In this technique, analytes will move through a semi-permeable
membrane due to concentration gradient, and the perfusion fluid will be analyzed to
check drug concentration by analytical methods like HPLC or microsensors.
GH-loaded liposomes showed Cmax of 13.98 μg/ml and AUC of 55.42 μg h/ml,
which is notably more than other animal groups. These results suggest the deep
penetration of liposomes into the nasal mucosa and efficient delivery of GH [26, 27].
Yang and his co-workers developed cell-penetrating peptide (CPP) liposomes of
rivastigmine (RS) to enhance its distribution in the brain and minimize the side
effects. RS transport across the BBB was time-dependent and the results suggested
that the liposomes can potentiate the transmembrane effect. Bio-distribution and
pharmacokinetic studies were performed in male SD rats. A higher concentration of
RS was observed in the brain region by i.n. delivery compared to i.v. This study
demonstrated that the intranasal delivery of liposome increased the rivastigmine
distribution. The developed CPP liposome Pharmacodynamic study demonstrated
that cilia movement and hemolytic effect were the same as that of physiological
saline, indicating non-toxicity of the formulation [28]. Upadhyay et al. developed
nanoliposomes of quetiapine for the improved and direct delivery to the brain for
treating schizophrenia. A thin film hydration technique was used to prepare lipo-
somes with the particle size and surface charge of 139.6 nm and −32.1 mV, respec-
tively. The maximum entrapment efficiency was observed in the 1:3 ratio of egg
phosphatidylcholine (EPC):cholesterol (CH), which is attributed due to the neces-
sity of the large amount of EPC to make the lamellar structure of liposomes and
cholesterol to increase the stability. An increase in ratio resulted in the leakage of
liposomes due to disruption of the liposomal membrane by CH. Scintigraphy study
demonstrated that the liposome can efficiently deliver quetiapine compared to sim-
ple solution and dispersion. These results suggest the use of this platform for target-
ing the brain [29].
Solid-lipid NPs (SLNs) are nanocarrier systems manufactured with solid lipids
and are efficient in loading high amounts of the drug that can load both lipophilic
and hydrophilic agents. As the SLNs exhibit smaller particle size and lipophilicity,
108 N. Dhas et al.
this delivery system can make use for brain drug delivery [30]. In an investigation,
Bhatt et al. formulated SLNs of astaxanthin (AST) using lecithin, poloxamer 188,
and citric acid to improve the brain targeting via i.n. delivery. The double emulsion
solvent displacement technique was employed to prepare AST-loaded SLNs. The
SLN was optimized by response surface methodology. The PDI of the formulation
was +0.349 and the average particle size was about 205.85 nm, which indicates the
narrow size distribution. The developed SLN showed excellent entrapment effi-
ciency (77.42% ± 1.15%) and better loading capacity (47.63 ± 1.07%). SEM and
TEM results indicated that the SLNs are in nanosize range and spherical. The
in vitro drug release study showed the drug release in a controlled and sustained
manner (81.40% release in 24 h). The optimized formulation was then radiolabeled
with technetium-99 m (99 mTc) and the labeling efficiency was confirmed by thin-
layer chromatography (TLC). A bio-distribution study was performed in male
albino Wistar rats, and administered the SLNs via i.n. and i.v. route. The drug con-
centration in the brain was higher and lesser in blood followed by i.n. administra-
tion. Intravenous administration of SLNs showed higher radioactivity in the spleen,
kidney, and lungs. The brain accumulation of SLNs on i.n. (1.70 ± 0.13) showed
200% more than i.v. (0.844 ± 0.12). Gamma scintigraphy was performed to visual-
ize the brain uptake followed by i.n. and i.v. administration, which confirmed the
internalization of the drug in the brain. The above results strongly suggest that the
AST-SLNs can be effectively used for brain targeting [31]. Patel et al. formulated
risperidone (RES)-loaded SLNs for targeted delivery to the brain via the intranasal
route to cross BBB. The RES-SLNs were prepared by solvent emulsification-
evaporation technique. It exhibited better entrapment efficiency of 59.65 ± 1.18%,
the particle size of 148.05 ± 0.85 nm, and PI of 0.148 ± 0.028, which indicates the
physical stability of the formulation. The zeta potential was found to be
−25.35 ± 0.45 mV, negative charge is due to the surfactant and lipid used. The
in vitro drug release of the formulation was 25.74% ± 0.65% and 48.90 ± 1.01%
after 7 h and 24 h respectively, which demonstrates the controlled release of the
drug. The pharmacodynamic study was carried out by using paw test with Perspex
platform, which exhibited prolonged hind limb reaction time (HRT) on RES-SLNs
administration than RES solution. These findings indicated that the SLNs delivery
via nose-to-brain is an excellent technique to drug delivery [32].
Nanostructured lipid carriers (NLCs) are a novel type of nano-mediated carrier
system which integrates both liquid and solid lipids. Compared with SLNs, NCLs
exhibit enhanced stability and better drug loading [33]. Singh et al. formulated gly-
col chitosan functionalized lipid carrier (GC-NLC) for intranasal delivery of ase-
napine (AS). The optimized formulation of AS-NLC showed a particle size of
167.30 ± 7.52 nm, excellent entrapment efficiency of 83.50 ± 3.48%, and zeta
potential of −4.34 ± 1.37 mV. GC-coated AS-NLC showed a significant increase in
particle size, zeta potential, and entrapment efficiency. Increased particle size may
be attributed due to the deposition of GC on the surface of AS-NLC and positive
zeta potential indicates the cationic nature of GC. The dialysis bag technique was
performed to evaluate in vitro drug release, where 90% of ASM was released in
12 h. Cell viability of GC-AS-NLC was checked in A549 cells by MTT assay. They
observed a negligible difference in the viability between ASM and GC-AS-NLC. In

vivo pharmacokinetic evaluation was carried out in Charles foster rats. The systemic
absolute bioavailability of GC-AS-NLC was 141.50%, which was 2.3-folds higher
than ASM (59.02%) followed by i.n. delivery. To confirm the brain targeting of the
formulation, the AS concentration from GC-AS-NLC was compared with pure AS
on i.v. and i.n. which demonstrated the high concentration of AS from GC-AS-NLC
at all the time points. The systemic bioavailability of AS in the brain, from GC-AS-
NLC, was 407.89% and ASM was 103.31% via i.n. route. The overall result indi-
cates that GC-AS-NLC through i.n. the route can efficiently target the brain with
increased bioavailability [34]. Rajput et al. formulated resveratrol (RES)-loaded
NLCs in situ gel for i.n. administration. The emulsification-probe sonication tech-
nique was used to prepare RES-loaded NLCs. The particle size distribution of NLCs
was from 70 ± 6 nm to 189 ± 109 nm. As the NLCs showed PDI less than 0.3, it is
considered as uniform particle size. TEM showed that NLCs are spherical and
exhibited narrow size distribution. Pharmacokinetic and biodistribution parameters
were evaluated in Sprague-Dawley rats. The biodistribution study showed that a
higher concentration of RES was found in the brain on NLC in situ gel administra-
tion than RES suspension, due to direct brain delivery of NLCs via olfactory lob and
its enhanced lipophilicity. A pharmacokinetic study also confirmed the sufficient
amount of RES in the brain. The developed formulation could be a better strategy
for efficient drug delivery in Alzheimer’s disease [33].
2.3 Metallic Nanocarriers
Metallic nanocarriers exhibit their outstanding application in the area of biomedical

sciences and engineering. It possesses a wide variety of applications like image-
guided therapy and targeted drug and gene delivery. The majority of imaging tech-
niques like MRI, SERS, and ultrasound techniques need a contrast agent for proper
functioning. This is the basis of the development of metallic NPs like silver, mag-
netic iron oxide, and gold [35–37].
Gold NPs (AuNPs) are the most used metallic nanocarrier for targeted delivery
of the drug and bio-imaging. Sukumar et al. developed gold-iron oxide NPs conju-
gated with microRNAs (miRNA) for multimodal imaging and presensitization of
glioblastoma (GBM). As biological barriers like BBB are the major challenging
factors for GBM treatment, the approach was to bypass BBB via a direct nose-to-
brain pathway. This formulation is expected to presensitize the GBM cells to deliver
chemotherapeutical agent temozolomide (TMZ) and for image-guided treatment.
Followed by the synthesis of gold iron oxide NPs (GIONPs), it is coated with a
β-cyclodextrin-chitosan polymer and conjugated with miRNA. Then it was surface
modified with PEG-T7 peptide via CD-adamantane host-guest chemistry. A partial
negative potential on the GIOn surface was attributed due to the coating with
cyclodextrin-chitosan (CD-CS) polymer. The in vivo i.n. delivery of formulation
was performed in an orthoptic xenograft mice model (U87-MG GBM derived). The
110 N. Dhas et al.
MR imaging and optical fluorescence studies suggested the better localization of

Cy5-loaded miRNAs in mice administered with GIONPs. The survival rate of mice
treated with the formulation was higher compared to other groups. This theranostic
platform has good capability to improve the therapeutic effects of TMZ in GBM
patients [38, 39]. Salem et al. developed nanoemulsion conjugated with AuNPs for
the brain delivery of RES via i.n. delivery for the therapy of Alzheimer’s disease.
AuNPs were synthesized by a simple reduction technique and conjugated with
RES-loaded transferosomess (RES-Tr-AuNPs). The TEM images of AuNPs indi-
cated they are uniformly distributed and spherical, having a diameter and PDI of
10.30 ± 2.4 and 0.130 ± 0.05, respectively. Micrographs showed that RES-Tr-
AuNPs are uniform and spherical with a measured size of 94.93 ± 5.6 nm. The zeta
potential of the formulation was about −28.7 ± 4.7, which confirms the uniform
dispersion and enhanced stability of the system. PDI was less than 0.3, which indi-
cates the uniform and narrow size distribution. The ex vivo penetration study dem-
onstrated the deep penetration of RES-Tr-AuNPs via nasal mucosal layers,
indicating uniform distribution and showing higher fluorescence intensity. The his-
topathology studies showed enhanced accumulation of AuNPs in the cytoplasm and
nucleus of brain tissues. Thus the developed formulation can show an efficient
brain-targeting effect and can be used for CNS diseases [40]. Betzer et al. developed
non-invasive glucose-coated AuNP (G-AuNPs) system for tracking and neuroimag-
ing of exosomes in brain structures. The G-AuNPs were loaded into exosomes via
GLUT-1 glucose transporters. The optimal particle size of the formulation was
about 5 nm and the administration route of i.n. intranasal administration resulted in
excellent in vivo brain accumulation and efficient in vivo bioimaging. The mouse
model was used to track AuNPs, which also demonstrated the specific localization
of the system. These results suggested the use of this system as a potential diagnos-
tic tool [41].
Silver nanoparticles (AgNPs) show cytotoxicity in normal lung, skin, and fibro-
blast cells [42]. AgNPs are broadly used in nutraceutical and consumer products due
to their better antibacterial and therapeutic characteristics. It has an excellent capa-
bility to cross BBB via intranasal administration and to concentrate in the brain.
Jonathan and his co-workers compare the transport and biological characteristics of
i.n. administration of AgNPs with silver ion. First, they compared the antimicrobial
activity of AgNPs and silver ions on the bacteria responsible for clinical rhino sinus-
itis. AgNPs exhibited reduced antimicrobial activity (minimum bactericidal concen-
tration (MBC) = 15 ppm) compared to silver ions (MBC = 5 ppm). After that, they
evaluated the residence time of silver in the sinus cavity followed by i.n. administra-
tion of both AgNPs and silver ions to mice, and checked the sinonasal mucosal
distribution of silver. The uptake level of AgNPs via respiratory epithelium was
minimal in the olfactory bulb and brain. The reduced retention and biodistribution
of silver on i.n. administration suggesting the safety of AgNPs [43]. Lung et al.
loaded poorly soluble phytochemicals like chrysin and curcumin into mesoporous
silica NPs (MSNs) for nose-to-brain phytochemical delivery. The formulated MSNs
exhibited a spherical shape along with an average particle size of 220 nm. DSC,
TGA, and FTIR techniques were used to confirm the efficient loading of
phytochemicals into the MSNs. In vitro drug evaluation indicated the pH-dependent
release of drugs at lower pH (5.5) at a better release rate of 53.2 ± 2.2% for cur-
cumin and 9.4 ± 0.6% for chrysin over 24 h. The cellular toxicity of MSNs was
evaluated in OBGF400 cells for 24 h, and MSNs were found to be non-toxic. The
confocal microscopy study demonstrated that FITC-labelled MSNs showed
membrane-specific and cytoplasmic accumulation on 2 h of post-incubation. Thus
the MSNs are capable of targeting and delivering drugs to the brain by bypassing
BBB [44].
Magnetic nanoparticles (MNPs) are nanocarriers that show magnetic properties.
It exhibits a unique property called magnet operation, in which MNPs produce
pores in the cell membranes including BBB for a shorter duration of time, which
enhance the targeting efficiency and delivery of drugs [45]. MNPs have potential
significance as magnetic contrast agents and magnetic vectors [46]. Abbas et al.
formulated superparamagnetic iron oxide (SPION) loaded with nano lipid carrier
(SLN and NLC) for the targeted clonazepam (CZ) delivery to the brain via intrana-
sal olfactory mucosa. To enhance the efficiency of CZ delivery, lipid carriers were
conjugated with mucoadhesive in situ gel. NLC formulation showed better entrap-
ment efficiency than SLNs. All the formulations showed zeta potential above 20 mV
and negative surface charge, which proved that the nanosuspensions possessed good
stability and were well dispersed. The lipid nanocarriers showed a PDI below 0.5,
indicating the narrow size distribution. NLC formulation showed better entrapment
efficiency (59.3 ± 1.68% – 65.7 ± 1.81%) than SLNs (49.2 ± 1.55% – 52.6 ± 1.35%).
NLC showed most satisfactory properties than SLNs and are conjugated with
SPION to form NLC-SPION. Both NLC and SPION were then loaded into an in
situ system containing sodium alginate (0.75%) and pluronic 127 (15%). The anti-
convulsant activity of NLC/in situ and NLC-SPION/in situ was evaluated in Albino
mice with chemically induced convulsion. The study resulted in a pronged onset of
convulsion and significantly protect from its death [47, 48].
3 Characterization of Nanocarriers for Nasal Delivery
Many studies proved the efficient delivery of drugs to the brain followed by the i.n.
administration of the drug-loaded nanoformulation is better than free drug formula-
tions. As the nanocarrier system offers non-invasive drug delivery, it is highly
patient compliant. Surface functionalization improves the targeted delivery of drugs
via i.n. route. It is better to keep particle size below 100 nm to evade opsonization
for a prolonged duration. The nanocarrier systems should be biodegradable and
biocompatible to keep away from an immune reaction, should show controlled
release, no leakage of drugs, be able to load peptides, proteins, and drugs, cost-
effective and reduce drug-excipient incompatibility [49].
Different mechanisms have been suggested for drug delivery to the brain. The
first one is by the release of a chemotherapeutic agent into the mucus-epithelial
interface or at mucus membrane by the interaction of drug-loaded nanocarrier with
112 N. Dhas et al.
the mucus layer. The second one is by uptake of drug-loaded nanocarriers by the
neurons after crossing the mucosal barrier, and then it will be moved into trigeminal
or olfactory nerve axons and release the drugs. The third one is explained by the
crossing of drug-loaded nanocarrier through the neuroepithelium and respiratory
epitheliums that will uptake these carriers. Here the drug will be released and enter
the CNS through perineural space. The above-mentioned mechanisms indicate that
the delivery of drug molecules and the efficacy of the drug delivery system depend
on the physicochemical properties of nanocarriers. So it is evident that surface
hydrophobicity/hydrophilicity, surface charge, size distribution, composition, and
surface charge will affect the activity of nanocarriers in the biological environment.
These characteristics can influence the brain delivery of drugs by affecting the
drug’s release kinetic profile, neuroepithelial/epithelial-mediated uptake of nano-
carrier, and mucus interaction. These aspects on the properties of nanocarriers
reveal that the physicochemical properties are important and to be considered while
making the formulation for the effective, targeted, and safe delivery of a drug into
the brain [50].
To demonstrate the influence of these physicochemical characters on the brain
delivery of the drug, many researchers have evaluated in vitro as well as in vivo
transport of dug-loaded nanocarriers. In recent work, Gartziandia and his co-
workers evaluated the permeability characteristics of the nanocarriers (NLCs) as per
the change in physicochemical properties. DiR (1-1′dioctadecyl-3,3,3′,3′-tetranethy
lindotricarbocyanine), a fluorescent probe was conjugated with the nanocarriers to
trace the drug delivery. Compared to PLGA NPs, chitosan-coated NLCs showed
higher permeation. The coating of chitosan with NLCs showed a change in surface
charge from negative to positive charge and enhanced transcellular transport, which
is threefold higher than uncoated NLCs. Surface functionalization of nanocarriers
with cell-permeating peptides such as penetration resulted in enhanced permeabil-
ity [51].
Ahmad et al. evaluated the transportation of nanoemulsions (NEs) by fluorescent
bioimaging techniques. They compared the residence duration of NEs in the nasal
cavity of rats by administering 100 μL of NEs of different size ranges as 80, 200,
500, and 900 nm from 0.5 to 16 h. NEs with smaller droplet sizes exhibited pro-
longed retention time compared with larger ones. NEs having a >200 nm showed
mucociliary clearance after 4 h of NE application. The remaining evaluation studies
were carried out with three optimized formulations NE of particle size 80 nm
(NE-80), 900 nm (NE-900, uncoated NEs) and chitosan-coated 108 nm (NEs-108,
chitosan coated). The order of retention time after 1 h of nasal instillation of NEs is
108 nm chitosan-coated NEs > NE80 > NE900-uncoated NE [52].
Gabal and his co-workers evaluated the influence of surface charge of nanocarri-
ers for the efficient delivery of a hydrophilic therapeutic agent, ropinirole hydro-
chloride (RPHCL) to the brain via i.n. route. They formulated both anionic and
cationic NLCs optimized based on their zeta potency and particle size. Both cat-
ionic and anionic NLCs showed a particle size below 200 nm and a zeta potential of
about 34 mV. These optimized formulations were loaded into poloxamer in situ gel
and determined the efficacy of the formulation after administering it to rats by
checking its behavior in the brain and plasma to measure pharmacokinetic vari-
ables. A toxicity study was performed in rats for 14 days for both cationic and
anionic NLCs and gel-dispersed nanocarriers. Toxicity study showed nasal mucosal
lining destruction in rats treated with cationic NLCs and nasal epithelial reversible
inflammation in anionic NLCs-treated rats. Gel-loaded NLCs have not shown any
histopathological changes in the treated animal. The absolute bioavailability showed
by the anionic NLCs (44%) and cationic NLCs (77.3%) in situ gels were signifi-
cantly larger than i.n. solution of RPHCL. The maximum drug concentration (Cmax)
shown by the cationic NLCs-in situ gel was more than the anionic NLCs-in situ gel.
Anionic NLCs showed a better targeting effect (158.5) which is 1.2-folds better than
cationic NLCs. The poloxamer 188 showed an efficient protective mechanism to
safeguard against inflammation and oxidative stress. The reduced toxicity could be
attributed due to the restricted movement of NPs via gel network and reduced con-
tact between epithelium and NPs [53].
The relation between brain distribution and peptide-based nanocarriers was eval-
uated by Kanazawa and his co-workers. Arginine-rich oligopeptide, which pos-
sesses excellent transmissibility and adhesiveness, was loaded with stearic acid
(hydrophobic nature) or PEG-PCL block copolymer (hydrophilic nature) to make
micellar formulations. The particle size of PEG-PCL-peptide (P-P-Pep) and
stearate-peptide (St-Pep) was 50 and 100 nm and zeta potential of about +15 and
+20 mV, respectively. The formulations were conjugated with Alexa-dextran (AD)
conjugate of molecular weight 10,000 D, a fluorescent probe for assessing biodistri-
bution. Both St-Pep and P-P-Pep carriers showed better uptake in nasal mucosa than
AD alone. The St-Pep complex showed intense fluorescence than P-P-Pep in the
nasal mucosa, but St-Pep showed less fluorescence than P-P-Pep in the trigeminal
nerve. The St-Pep exhibited intense fluorescence in the forebrain, but no fluores-
cence was observed in the hindbrain. Intranasal administration of P-P-Pep showed
fluorescence in the entire brain, which confirmed the distribution of formulation on
the entire brain. The study results suggest that the delivery pathways of drugs can be
modified based on the characteristics of different nanocarriers [54].
Functionalization of nanocarriers with appropriate polymers could enhance the
efficiency of nasal drug delivery. Several natural (gelatine and alginates), semisyn-
thetic (cellulose derivatives), and synthetic polymers (crospovidone and polyacry-
lates) potentiated the nasal drug delivery [55]. The nasal formulations containing
pectins and chitosan showed prolonged residence time in the olfactory area; besides
that, sodium hyaluronate enhanced the brain delivery of 4 kDa dextran conjugated
with fluorescein after i.n. administration to rats [56, 57].
4 Biomedical Applications of Nanocarrier in Nasal Delivery
Several nanocarriers are used for drug delivery: chitosan nanoparticles, liposomes,
solid lipid nanoparticles, dendrimers, polymeric nanoparticles, micelles, and nano-
emulsions. The objective of nanocarrier in drug entrapment is to increase the
114 N. Dhas et al.
effectiveness of the drug to the target cell and to decrease the toxicity of the drug to
other organs. This will be advantageous in cancer treatment. An anticancer drug like
paclitaxel has been investigated for the nanoparticles. It has been shown that the
nanoparticles entrapped by the paclitaxel had increased sustainable therapeutic
effect and also increased cytotoxic effect on cancerous cells.
Due to nano-size, the nanoparticles have several advantages like they can effec-
tively bind to the proteins and can penetrate the cell membrane. They can save from
the lysosome after entering into the cell by endocytosis [58]. Along with the drug
delivery to the brain, a nasal route can also be utilized for the delivery of stem cells.
Several studies have been confirmed that mesenchymal stem cells, neural stem
cells, and pluripotent stem cells localized in the brain after administered through the
nasal route. Studies were also done on delivery of the mesenchymal stem cells to the
brain which successfully treated various brain-related diseases like Parkinson’s dis-
ease, Alzheimer’s disease, Huntington’s disease, and also stroke. Radiation therapy
is successfully used in the treatment of brain tumors. But the disadvantage of radia-
tion therapy is it causes damage to the surrounding healthy tissue. To overcome this
disadvantage mesenchymal stem cells were administered intranasal which showed
improved neurological function [2].
Liposomes Liposomes for drug delivery have been investigated. As liposomes can
incorporate hydrophilic as well as lipophilic substances, it has extensive use in the
delivery of biomedicines. The following drugs have been investigated as liposomal
drug delivery system: anticancer drugs like doxorubicin, daunorubicin, cisplatin,
amikacin, amphotericin B, liposomal lidocaine, annamycin, nystatin, and retinoic
acid [59, 60]. Carboxymethyl chitosan nanoparticles have been prepared for the
carbamazepine drug. It has been observed that the concentrations of CBZ remained
higher in the brain than the plasma over 240 min. Also, sodium alginate nanoparti-
cles were formulated for the venlafaxine to treat the depression and administered
intranasally. The concentration of the drug in the brain was compared with the ven-
lafaxine solution administered through IV and the venlafaxine solution adminis-
tered intranasally. It was found that the blood/brain ratio of the VLF concentration
was higher in the nanoparticles than in other formulations. These prove the promi-
nence of the nanoparticle which directly transports the drug to the brain [4].
Solid lipid nanoparticles (SLN) Few examples where SLN systems have been
utilized for the delivery of anticancer drugs are docetaxel, doxorubicin, paclitaxel,
methotrexate, and 5-fluorouracil (5-FU).
Dendrimers Doxorubicin, cisplatin, and 5 fluorouracil have been investigated in

the form of dendrimers which have greater effectiveness as compared to the drug
used in free form.
Virus-based nanoparticles It was investigated to deliver drugs, in gene therapy, vac-

cination, targeted drug delivery system. Various viruses like bacteriophage, insect
virus, cowpea chlorotic mottle virus, cowpea mosaic virus, red clover necrotic mosaic
virus, tobacco mosaic virus poliovirus, and adenovirus are employed in developent of
virus-based nanoparticles. Advantages of using virus-based nanoparticles are: They

are morphologically uniform in size, biocompatible, available in various sizes and
shapes, Chemical and genetic alteration can be possible, greater drug entrapment effi-
ciency. If the surface of the virus-based nanoparticle is PEGylated then the circulation
time in the host cell can also increase. Carbon nanotube has been investigated for the
delivery of various anticancer drugs like methotrexate, paclitaxel, doxorubicin, cispla-
tin, carboplatin, and mitomycin C [61]. Along with the drug delivery to the brain, the
nasal route can also be utilized for the delivery of the stem cells. Several studies have
confirmed that mesenchymal stem cells, neural stem cells, and pluripotent stem cells
localized in the brain after administration through the nasal route. Studies were also
done on delivery of the mesenchymal stem cells to the brain which successfully
treated various brain-related diseases like Parkinson’s disease, Alzheimer’s disease,
Huntington’s disease, and also stroke. Radiation therapy is successfully used in the
treatment of brain tumors. The main disadvantage of radiation therapy is it causes
damage to the surrounding healthy tissue. To overcome this disadvantage, mesenchy-
mal stem cells were administered intranasally which showed improved neurological
function [2] versus applications as shown in Table 7.1.
4.1 Targeted Delivery
Parkinson’s disease, Alzheimer’s disease, glioblastoma, epilepsy, and multiple scle-

rosis are neurological disorders. The central nervous system (CNS) still lacks ade-
quate medication delivery in therapeutic doses [62]. Mainly through the nasal
cavity, the target organ is the brain. Nanocarriers are investigated for the delivery of
the drug to the brain through the nasal cavity (nose-to-brain drug delivery system).
Figure 7.4 illustrates the drug transport from nose-to-brain and systemic circulation.
It is a non-invasive method to directly approach the brain by bypassing the blood-
brain barrier. Thereby it decreases the systemic side effects. It is potentially investi-
gated as an alternative route for administration for the delivery of the drugs which
are used in CNS diseases. They also provide the controlled drug release and thereby
decrease the administration frequency. NBDDS has been investigated for the treat-
ment of glioblastoma. It is the most lethal form of brain tumor. The absorption of the
drug through the nasal route depends on the nature of the drug (hydrophilic or lipo-
philic) and the molecular weight of the drug. Bioavailability is very less for the
drug, which has M.W. greater than 1 kDa. The drug with lyophilic nature and hav-
ing M.W. less than 1 kDa may have a bioavailability near to 100%.
Nanoparticles are investigated for the treatment of brain-related diseases through
the nasal cavity. The drug may be administered in the nanoparticle in the form of a
matrix or the drug may be encapsulated into the polymer coat. Due to their small
size nanoparticles are more effectively transported to the brain transcellular. The
disadvantages of nasal administration are the drug is poorly penetrated through the
nasal mucosa, fast mucociliary clearance, and degradation of the drug by the
enzyme. Encapsulating the drug into nanoparticle will overcome these disadvan-
tages. Besides, NP may offer improved drug delivery to the brain, since they can
Table. 7.1 Application of polymeric/lipidic nanocarrier in various diseases through nasal route
In-vivo
Type of Target Therapeutic Method of Particle Encapsulation animal Ex-vivo
nanocarrier diseases molecule preparation Carrier/polymers size PDI Zeta potential efficiency model cell line Reference
Liquid Epilepsy Rosu Hydrotrope- Glyceryl monooleate, 219.15 0.24 −26.2 mv 70.30 ± 1.84 Swiss – [81]
crystalline vastatin based method phenytoin, PTZ, PEG ± 8.14 nm ± 0.03 albino
nanoparticles 400, and poloxamer male
407 mice
Liposome Cancer Ovalbumin Thin film Dimethyl dioctadecyl 265.9 (± – 56.5 (± 11.9) 70.30 ± 1.84% Female L-132 [82]
hydration ammonium 51.9) nm C57/ Human
technique BL6 lung
mice epithelial
cells
Liposomes Cancer Ovalbumin Reverse pH Chitosan Carbopol® 200– – −5.33 mV 73.15 ± 0.21% – Calu-3 [83]
974P 250 nm ± 2.019 cells
In situ Pain- Opiorphin Thin layer Chitosan, 141 ± 0.184 −0.62 ± 0.18 59 ± 3 – Nasal [84]
mucoadhesive- killing evaporation hydroxypropylmethyl 4 nm ± 0.024 porcine
thermosensitive method cellulose, Poloxamer, mucosa
liposomal gel Carbopol
Polymer–lipid Systemic Tenofovir Melt Acconon CO-7 239 nm −44.16 ± 2.51 87.14% – – [85]
nanocarrier circulation disoproxil emulsification- (PEG-7 glyceryl
fumarate probe cocoate), Carbopol
sonication 934 P
technique
In-vivo
Type of Target Therapeutic Method of Particle Encapsulation animal Ex-vivo
nanocarrier diseases molecule preparation Carrier/polymers size PDI Zeta potential efficiency model cell line Reference
Nano-micelles Nose-to- Camp Water-in-oil Poly(ethylene 70–80 nm 0.4 5.98 ± 1.32 85.7 ± 6.11 Tumor- Gastrin- [86]
brain tothecin (w/o) emulsion glycol)- bearing releasing
delivery polycaprolactone rats peptide
block polymer, receptor
ε-caprolactone (GRPR)
positive
cells
Micelle Nose-to- Sumatriptan Micellization Transcutol P®, 23.1 – – Male – [87]
brain Pluronic® ± 0.4 nm Wistar
delivery F127 (PF127; rats
poly[ethylene oxide]–
Poly[propylene
oxide] block
copolymer)
118 N. Dhas et al.
Fig. 7.4 Illustration of drug transport from nose-to-brain and systemic circulation
Table 7.2 Study on the use of various nanocarrier in nasal delivery for the treatment of
glioblastoma
Name of drug Nanocarrier Reference
Bevacizumab Polymeric NPs (PLGA) [63]
Melatonin Polymeric NPs (PCL) [64]
Temozolomide Polymeric NPs (PLGA) [65]
Nano-emulsion [66]
Curcumin Nanostructure lipid carrier [67]
Methotrexate Polymeric nanodispersion (PLA) [68]
Carboplatin Polymeric NPs (PCL) [69]
prevent extracellular transport by P-glycoprotein (P-gp) efflux proteins localized in

the olfactory epithelium and the endothelial cells that surround the olfactory bulb.
Table 7.2 shows the summary of various nanocarriers, and have been investigated
for the treatment of glioblastoma.
Alzheimer’s disease (AD) has been managed by meloxicam (MEL). Scientists for-
mulated poly (lactic-co-glycolic acid) nanoparticles (PLGA NPs) and solid lipid
nanoparticles (SLNs) and SLN coated by the chitosan (c-SLNs). SLNs showed higher
encapsulation efficacy (EE) and drug loading (DL) than PLGA NPs. When compared
to a native drug, nanocarriers (c-SLNs,>SLNs> PLGA NPs) had a greater sustained
release profile, permeation characteristics, and adhesion. As a result, encapsulating
MEL in C-SLNs and delivering it via the intranasal route could improve its brain
bioavailability [70]. Another study has been done to deliver the drug to the brain by
formulating dexamethasone (DXM)-loaded, mixed polymeric micelle-based drug
delivery system. Micelles formed had high water solubility due to high surface polar-
ity and low z average. So it shows a high in vitro permeability value on polar brain
(porcine) lipid extract .diffusion study showed the rapid diffusion through the nasal
mucosa. The formulation had also sufficient mucoadhesive property [71–76].
4.2 Gene Therapy
Apolipoprotein E (ApoE) gene therapy is a promising disease-modifying method

for Alzheimer’s disease. ApoE can control Aβ clearance so it reduces its level and
thereby prevent pathogenic plaque development. So it protects neurons. Keeping
this concept scientists formulated transferrin-penetrating modified (dual functional-
ized) liposomes containing ApoE encoding a plasmid DNA (pDNA). The pDNA
was complexed with chitosan to increase the transfection’s vitality. They confirmed
that the dual-ligand liposome nanoparticles could preserve pDNA from enzymatic
nuclease destruction while delivering the therapeutic gene and enhancing ApoE
expression. Greater BBB penetration has been observed in the dual-functionalized
liposomes than single modified (either transferrin or penetration) formulations. The
authors believe that this liposomal-based gene delivery technology has a lot of
promise for preventing and treating Alzheimer’s disease [77].
4.3 Vaccine Delivery
Apart from the advantages of the nasal drug delivery system there are several disad-
vantages like permeability of the nasal mucosa for the hydrophilic drugs is less so
we get less bioavailability, the dose we can apply is very low, degradation of the
drug due to enzymes present in the nasal mucosa and mucociliary clearance is very
high. To solve these disadvantages there were many strategies developed. Among
that one of the strategies is the formulation of a nanocarrier drug delivery system.
Along with the nanocarrier permeation enhancers also be used to increase the pen-
etration of the drug into the nasal mucosa. To prevent mucociliary clearance bioad-
hesive polymers have been investigated. The principal location for inducing nasal
immunity against given antigens is the nasal-associated lymphoid tissue (NALT).
NALT consists of B cells, T cells, and a dense network of antigen-presenting cells
conveniently located to improve the nasal. Another promising strategy to improve
the antigen uptake at nasal mucosa is targeting the formulation for DCs or M cell
capture using specific ligands or antibodies. Attenuated influenza vaccine (LAIV)
FluMist® (or Fluenz® at Europe): it is the most successful nasal vaccine to date. It
is used for more than 10 years in children and adults. The vaccine contains three
types of LAIV. The vaccine is in suspension form administered in a specific dose
with the help of an intranasal sprayer [78]. The scientist also studied the monovalent
influenza subunit vaccine for nasal delivery in form of N trimethyl chitosan nanopar-
ticles as a carrier. The vaccine was formulated by combining TMC and monovalent
influenza A subunit H3N2 with a tripolyphosphate (TPP) solution. Incubation of the
120 N. Dhas et al.
particles in phosphate buffer solution showed that greater than 75% of the protein
remain connected with the nanoparticles. The vaccine was administered intranasally
as well as intramuscularly. The strong hemagglutination inhibition and total IgG
responses have been created in both the routes but response in intranasal (IN)
administration was significantly higher than in intramuscular administration.
Nanoparticles prepared from trimethyl chitosan induce greater immune responses
than the other intranasal antigen formulations. It can be concluded that TMC
nanoparticles are a powerful novel delivery system for influenza antigens [79].
4.4 Theranostic Application
Theranostics is a combination of two terms therapeutics and diagnostics. It is a new

discipline that has the potential to help with medicine delivery to the brain.
Theranostics combine radiological science with the delivery of medicinal agents. As
a result, customized medicine’s goals are likely to be advanced. The main principle
of theranostics is it includes a nanocarrier that contains medicinal agents and imag-
ing labels. So they functionalized as a therapeutic carrier as well as molecular imag-
ing. It also enables real-time tracking of the drug molecule.
The nanoparticles were formulated using the polylactide (PLA)–1,2-distearoylp
hosphatidylethanolamine (DSPE) coated with the polyethylene glycol. The aerosol-
ized formulation has been administered to the rat and measured the quantity of the
nanoparticles that reached the brain. The novel nano-theranostics was evaluated for
quantitative temporal and spatial testing. The nanoparticles were also administered
intravenously. After being administered intranasally via nasal tubing and intrave-
nously imaging was done by PET/CT. Imaging was done for 2 h and the animals
were sacrificed and different parts of the brain were isolated to compare the activity
in each brain region with the corresponding PET/CT region. Greater activity in the
brain was found in intranasal administration as compared to intravenous administra-
tion. This result was correlated with the ex vivo gamma counting [80].
4.5 Diagnostics Application
The nanocarriers containing active moiety are labeled with the radioactive sub-
stances. Nanocarriers are administered through the nasal route. The amount of the
drug accumulated at the site of action can be determined by various imaging tech-
niques like PET, CT, SPECT, etc. A scientist has developed the chitosan nanoparti-
cles of the zolmitriptan (ZMT) which was labeled with the technetium-99 (99mTc).
The nanoparticle was prepared by the modified ionic gelation of anionic sodium
triphosphate and cationic chitosan. Three formulations were investigated for the
activity in the brain. Those were ZMTNP, ZMT pure drug solution administered
intranasal, and ZMTNP administered intravenously all three formulations were
radiolabeled with the 99mTc. The accumulation of nanocarrier into the brain was
visualized by the single-photon emission computerized tomography (SPECT). The

percentage of radioactivity in the brain was found greatest in ZMTNP administered
intranasally. Therefore, nanocarrier with nasal route is the convincing drug deliv-
ery system.
5 Conclusions and Future Perspectives
Drug delivery with a nanocarrier system is a viable approach for the treatment of
various diseases compared to the conventional formulation. The research done so far
had majorly focused on the targeting to the brain via the nasal route. Several nano-
carriers were tried in research and showed efficacy. However, still there is a strong
need to consider the safety and efficacy aspects in designing nano-carrier-based
formulations. The polymeric and lipidic nanocarriers are most preferred for deliver
drug to brain via nasal route. Furthermore, there is a need to examine the targeting
pathway from the nose-to-brain region to understand its mechanism and thereby
designing robust formulation. The pharmacodynamics and pharmacokinetic studies
in higher animals and clinical proof in humans would be required for judging the
nanocarriers for the therapeutic purpose. It is projected that the effective use of new
tools and imaging techniques of diagnosis might help for the same in the future.
Acknowledgments The authors would like to thank Nirma University, India for providing finan-
cial assistance in the form of Nirma University fellowship-SRF to Atul Garkal (NU/Ph.D./IP/
GAD/19-20/1496).
Conflicts of Interest The authors declare that there are no conflicts of interest for this
publication.
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Chapter 8
Delivery of Vaccines via the Nasal Route
Seth Kwabena Amponsah and Emmanuel Boadi Amoafo
Abstract Several methods, including novel formulations and production systems,

have been proposed as ways to improve drug delivery. Nasal delivery of drugs is
traditionally employed when local effects (allergies, congestion, and respiratory ill-
nesses) and/or systemic effects (pain treatment) are required. Over the last couple of
years, the nasal route has been sought as a site for vaccine delivery. Data suggests
that when vaccines are administered via the nasal route, they elicit powerful immune
system responses. There are a number of vaccines that have been developed for
nasal administration, some of which include Nasalflu, FluMist®, and Coronavac,
among others. The nasal route for delivering vaccines has many merits; however, a
few drawbacks limit the use of this route. Nonetheless, scientists are still trying to
exploit this route as a potential for vaccine administration.
Keywords Absorption · Liposomes · Mucociliary · Nasal delivery · Vaccine
1 Introduction
Historic data suggests that smallpox killed about 375 million people throughout the
twentieth century alone. However, after a successful vaccination (eradication) cam-
paign in the 1970s, few deaths from smallpox have been recorded. Currently, more
than 70 vaccines, against nearly 30 microorganisms, exist [1, 2]. Indeed, vaccina-
tion has reduced morbidity and mortality associated with infections. The history of
vaccination began when Thucydide (430 BC) realized that people who survived
fatal infectious diseases were not likely to contract that same disease again [3].
S. K. Amponsah (*)
Department of Medical Pharmacology, University of Ghana Medical School, Accra, Ghana
e-mail: skamponsah@ug.edu.gh
E. B. Amoafo
Department of Pharmaceutical Sciences, North Dakota State University, Fargo, ND, USA
e-mail: emmanuel.amoafo@ndus.edu
https://doi.org/10.1007/978-3-031-23112-4_8
128 S. K. Amponsah and E. B. Amoafo
Later on in the fifteenth century, people in China started to practice variolation, a

method of exposing healthy individuals to air-dried pustules of smallpox. While
effective, the procedure was extremely dangerous. Contemporary medicine made
great advancements in vaccination when Jenner discovered that it was worth inocu-
lating people with pustulae derived from cowpox [4, 5]. Later on, Pasteur pioneered
the development of vaccines by exposing people to dead or attenuated microorgan-
isms that mimicked the infectious agents [6, 7].
Vaccination leads to induction of an immune response from the host and this is
to confer protection against infection or disease upon subsequent exposure to a
pathogen. To achieve this, the vaccine should have either antigens from the caus-
ative microorganism or synthetic antigens that are components of causative micro-
organism [8]. Currently, vaccine design takes advantage of the principle of eliciting
protective immunity against diseases by simulating immune response against a
pathogen that causes the disease, without inducing disease. Additionally, it is impor-
tant to consider factors that influence interaction between the host and the infectious
agent at the population, individual, cell, and, more recently, genetic level [9].
Generally, licensed vaccines are administered intramuscularly (IM), however,
some are also available for subcutaneous or intradermal (SC or ID) use [10].
Administration of vaccines via mucosal membranes is becoming a promising ave-
nue to drug delivery. There are currently about 5 approved vaccines (i.e., Dukoral®
given orally for cholera, Biopolio™ B1/3 given orally for poliomyelitis, Rotarix®
given orally for rotavirus, Vivotif® given orally for typhoid and FluMist™ given
intranasally for influenza), that are administered via mucosal membranes. There are
several advantages of using mucosal routes in vaccination. For instance, mucosal
tissues cover large surface area (on average 400 m2), and this makes them a point of
entry for many pathogens [11]. Likewise, nearly 80% of human immunocytes are
found on mucosal surfaces; therefore, vaccination via this route helps achieve both
systemic and mucosal immunity, unlike parenteral vaccination which merely stimu-
lates systemic immunity [12, 13]. Furthermore, administration via mucosal routes
has other advantages over parenteral administration, and these include lower infec-
tion risks, increased patient compliance (particularly for pediatric populations), and
minimal need of skilled personnel [12, 13].
It is noteworthy, however, that mucosal tissues also have their limitations. They
have mucus or cell barriers and enzymes that can affect vaccine function.
Nonetheless, there is a lot of effort being made to develop vaccines that can be
administered to overcome these limitations [14].
2 Mucosal Delivery of Vaccines
Infections of viral or bacterial origin can start at mucosal surfaces [15, 16]. Immune
responses mediated by mucosal vaccination are relevant because antibodies can be
released into the mucus, thereby, eliminating causative microorganisms. This
approach serves as first line of protection on mucosal surfaces. Indeed, this is a big
8 Delivery of Vaccines via the Nasal Route 129
step in vaccine development [17]. As a needle-free method of administering vac-

cines, the mucosal route would address the challenge of needle reuse. Around 16
billion injections were given worldwide in 2000, with reused needles resulting in
approximately 266,000 new cases of human immunodeficiency virus (HIV), two
million new cases of hepatitis C, and 21 million new cases of hepatitis B [18].
Although not all injections were associated with the process of vaccination, the use
of the mucosal route can help minimize needle reuse. Additionally, mucosal vacci-
nation can reduce sharp wastes and prevent needle-stick injuries [17].
3 Nasal Route
3.1 Anatomy of the Nose
Compared to the skin, the nasal mucosa does not have a highly keratinized stratum
corneum; rather, it is composed of numerous microvilli and a rich vascular network
[19]. Traditionally, the nasal route has been employed in the administration of drugs
for local and systemic effects. In the past decade, systemic-acting drugs and vac-
cines have been formulated to be delivered via the nasal route [14].
The nose is divided into two symmetrical halves by the median septum; both
halves open toward the face via the nostrils and reach the nasal cavity posteriorly
[20]. The nasal cavity is protected by the membranous viscerocranium. The atrium
occupies an intermediate area between the respiratory region of the nose and the
vestibule. The respiratory region of the nasal cavity, the nasal turbinates, has three
sub-sections: inferior, superior, and middle turbinates [21]. Characteristics of vari-
ous portions of the nose are summarized in Table 8.1.
3.2 Physiology of the Nose
The nose is involved in an array of functions, some of which include olfactory (the
sense of smell), as well as filtering, humidifying, and regulating the temperature of
air entering the respiratory system. The nose is also a point of entry for pathogens.
Hair at the entrance of the nostrils provides the first barrier to foreign bodies, since
it effectively keeps out large particles [26].
A mucus layer covers the entire nasal cavity, trapping smaller particles. The
mucus produced by this layer is viscoelastic and made from mucins that are secreted
by mucus sub-glands and goblet cells [27–29]. Through mucociliary clearance, cilia
transport mucus blankets packed with pathogens to the back of the throat at a rate of
5–6 mm per minute in either direction, either to destroy the pathogens in the stom-
ach or expel them via sneezing and/or coughing [30].
Table 8.1 Characteristics of the different portions of the human nose

Surface
Nasal area
region Cells present (function) (cm3) Vascularization Permeability Reference
Vestibule Nasal hair, stratified ≈ 0.6 Low Poor [22]
squamous and
keratinized epithelial
cells (support and
protection).
Atrium Stratified squamous cells Not Low Reduced [23]
(support). found
Pseudostratified cells
(support).
Respiratory Columnar non-ciliated ≈ 130 Very high Good [24]
region cells (support).
Columnar ciliated cells
(support and muciliary
clearance).
Globet cells (mucus
secretion).
Basal cells (progenitors
of other cell types).
Olfactory Sustentacular cells ≈ 15 High Direct access to [25]
region (support). central nervous
Olfactory receptor cells system
(olfaction perception).
Basal cells (progenitors
of other cell types).
4 The Mucosal Immune System
Mucosal surfaces have physical, chemical, and immunological defense mechanisms

against infection [31]. In the mucosa of the nose are lymphoid tissues that are
responsible for immunity. They are often referred to as mucosa-associated lym-
phoid tissue (MALT). MALT can also be found in the gastrointestinal tract, lungs,
vagina, and rectum [32]. To effectively fight infection, MALT suppresses pathogen-
specific immune responses and releases immunoglobulin A (IgA) at mucosal sur-
faces [31, 33]. MALT can be subdivided based on their location. One associated
with the nasal route is nasopharyngeal-associated lymphoid tissue (NALT) [34].
5 Nasopharyngeal-Associated Lymphoid Tissue (NALT)
Induction of nasal immunity against administered antigens via vaccines takes place
primarily at NALT [35]. In simple terms, the NALT is an integrated immune system
in the nasal mucosa consisting of lymphoid tissue, B cells, T cells, and
antigen-presenting cells (APCs) which are covered by epithelia containing memory

cells [36]. The Waldeyer’s ring is formed by the physical arrangement of multiple
lymphoid tissues in humans, including two palatine tonsils, two tubal tonsils, an
adenoid, and a lingual tonsil [37]. These lymphoid tissues sample antigens from
food, water, and air and contribute to host immunity [37, 38].
Pathogens are usually identified by APCs (macrophages and dendritic cells) after
going through nasal epithelium. Antigens are processed by these APCs and immu-
nogenic features of the antigens are presented to T cells in the lymph node. As a
result, the immune response cascade is activated [39]. Additionally, the NALT
drains into lymph nodes, where it undergoes further antigen processing [40].
6 Drug Uptake in the Nose
Once a drug dissolves in the nasal mucus, extremely vascularized surfaces and low
enzymatic activity facilitate absorption [41]. Absorption of drugs from the nasal
mucosa can be via transcellular and/or paracellular processes (Fig. 8.1). The muco-
sal lining over the turbinates or conchae is the most efficient location for absorption
in the nasal cavity because it is highly vascularized [21]. By bypassing the liver,
drugs administered via the nasal route avoid first-pass hepatic processing, making
the nose an ideal target for low orally bioavailable drugs. There is evidence, how-
ever, that the nasal mucosa can metabolize substances such as cocaine and proges-
terone [42, 43].
Fig. 8.1 Drug uptake mechanism in the nasal cavity

7 Absorption Through Nasal Route
Nasal absorption of drugs can happen rapidly; with concentration versus time for
nicotine and butorphanol being almost similar to that of intravenous route [44]. A
nasally administered drug has to cross a mucus layer and epithelial membrane
before reaching the bloodstream [45]. Vaccine administration through the nasal
route can be affected by a number of factors. Two of the most important factors are
nasal physiology and the physiochemical properties of the vaccine.
7.1 Nasal Physiological Factors
7.1.1 Blood Flow
The nasal mucosa is well supplied with blood and has a large surface area, making
it ideal for drug absorption. The sphenopalatine artery supplies the nasal septum
from behind, the anterior ethmoid artery from above, and the superior labial branch
of the facial artery from below. Furthermore, the palatine and ethmoid arteries sup-
ply the inferior and lateral walls of the nasal cavity, respectively, while the spheno-
palatine artery supplies the remainder of the blood flow to the nasal cavity [46].
Since the absorption of drugs occurs by diffusion, blood flow plays an important
role in the concentration gradient at the absorption site [47].
7.1.2 Mucociliary Clearance
Mucociliary clearance system prevents unwanted particulate material from entering

the lower airways. Mucociliary clearance reduces the time during which drugs are
exposed to the mucosa and thereby affects the absorption of active principles of
drugs. Inhaled agents are usually eliminated by this mechanism within 15–30 min
at the mucus layer. Transit times longer than 30 min may indicate mucociliary clear-
ance abnormality [22, 23]. Mucociliary clearance is often compared to a “conveyor
belt,” with cilia acting as the propeller and mucus, a sticky fluid that discards foreign
particles [23].
7.1.3 Degradation and Excretion of Nasally Administered Drugs
Due to the availability of a number of metabolic enzymes in nasal tissues, drugs

may be metabolized in the nasal cavity or during transit over the nasal epithelial
barrier [47]. Endopeptidases and/or carboxypeptidases are present in the nasal epi-
thelium. These enzymes are involved in the metabolism of drugs as well as the
degradation of native molecules [41]. Isoenzymes of the cytochrome P450 family
have been identified as metabolizers of pharmaceuticals (progesterone, cocaine,

nicotine, and decongestants) via the nasal route [48, 49]. Aminopeptidases and pro-
teases can affect the absorption of peptide drugs (insulin, calcitonin, and desmo-
pressin) via the nasal route. These aforementioned enzymes may alter the
pharmacokinetics and pharmacodynamics of nasally administered drugs [50, 51].
7.2 Physicochemical Properties of Drugs
7.2.1 Lipophilicity/Hydrophilicity, Molecular Weight, and Degree

of Ionization
Hydrophilic molecules must employ the paracellular pathway to cross the epithe-
lium, whereas lipophilic molecules can freely diffuse across. For paracellular transit
across tight junctions, high molecular weight of a drug may be a limiting factor.
Nasal absorption is fast when drugs have a molecular weight below 300 Da, whereas
molecules with a weight above 1 kDa are absorbed relatively slowly [22, 45, 52,
53]. Ionization is also an important factor for diffusion; hence drug absorption [53].
Physicochemical factors that can affect the absorption of the active principles of a
drug are summarized in Fig. 8.2.
Fig. 8.2 Factors that affect the absorption of xenobiotics across the nasal epithelium
7.2.2 Solubility
Drug absorption is often dependent on the dissolution of the drug at the absorption
site. Only molecularly dispersed forms of the drug can pass through biomembranes.
For nasal absorption to occur, the drug must dissolve in the watery fluids of the
nasal cavity. Thus, it is important that the drug dissolves adequately in the nasal
environment to allow adequate contact with the nasal mucosa [54].
8 Types of Nasal Vaccines
Studies over the years have tried to develop intranasal drug delivery systems for
vaccines. When administered via the nasal route, vaccines may elicit powerful
immune system responses. In order to improve the absorption of vaccines, delivery
systems can comprise enzyme inhibitors, nasal absorption enhancers, or mucoadhe-
sive polymers [47]. Examples of vaccines administered via the nasal route are sum-
marized in Table 8.2.
Some of the formulations of vaccines may include liposomes, microspheres, and
nanoparticles.
8.1 Liposomes
Liposomes, also known as phospholipid bilayer vesicles, have been well investi-
gated as drug delivery vehicles [60]. Hydrophilic drugs can be entrapped in the
aqueous interior of liposomes, and lipid-soluble pharmaceuticals can be integrated
into the hydrophobic core of the phospholipid bilayer [61]. Liposomes are small,
amphiphilic, and biocompatible, hence, offer promising delivery systems.
Liposomes are identified by APCs (macrophages and dendritic cells) and then
Table 8.2 Available vaccines that are administered via the nasal route
Potential
Vaccine name Type use Company References
Nasalflu Inactivated virosomal-subunit Influenza Berna Biotech [55]
FluMist® Live attenuated influenza Influenza AstraZeneca [56]
vaccine
Comirnaty mRNA vaccine (nucleoside SARS- Pfizer, BioNtech, [57]
(BNT162b2) modified) CoV-2 Fosun Pharma
Ad26.COV2.S Adv serotype 26(Ad26) COVID-19 Johnson & [58]
vector-based DNA vaccine Johnson
(non-replicating viral vector)
Coronavac Inactivated vaccine COVID-19 Sinovac Biotech [59]
presented to other lymphoid cells for immunological responses [62]. Liposomal

drug delivery systems provide several benefits, including the ability to encapsulate
drug molecules with different solubility and pKa values [63]. The ability of lipo-
somes to change the pharmacokinetics of their associated medications is a key fea-
ture that makes them useful as drug delivery systems [64, 65].
Liposomes have been found to increase membrane penetration of peptides such
as insulin and calcitonin by increasing their nasal absorption [66, 67]. The Swiss
Serum Institute recently launched a nasal influenza vaccine based on a liposome
(virosomal) formulation of influenza virus components [68].
8.2 Microspheres
Microsphere technology has been widely used in the development of nasal medica-
tions. Microspheres are small spherical particles that range from 0.1 to 200 μm in
diameter and are made of biodegradable and non-biodegradable materials [69].
Microsphere-based systems have the potential to extend the half-life of active con-
stituents, while also regulating the release of bioactive compounds [70].
Degradable starch microspheres (Spherex®) are the most utilized microsphere
technology for nasal drug delivery. Usually, an emulsion polymerization approach
(starch cross-linked with epichlorohydrine) is used to make these microspheres
[71]. Microspheres can protect vaccines from degradation by enzymes and improve
their efficacy [72].
8.3 Nanoparticles
Drug delivery using nanoparticulate systems is known to improve intranasal drug

delivery. Nanoparticles are natural or artificial polymers ranging in size from 1 nm
to 1000 nm. Nano-biotechnology in drug delivery is to improve precision of medi-
cine, reduce toxicity, and enhance therapeutic effect [73].
In vaccines, nanoparticles can be adjuvants in which the active agent can be dis-
solved, entrapped, encapsulated, adsorbed, or chemically attached [74].
Nanoparticles can protect vaccines against the hostile environment of the nasal
mucosa and also aid in the activation of the immune system response [21].
9 Challenges with Nasal Vaccines
Vaccines administered via the nasal route have many merits but also have some
limitations. Water soluble molecules with low membrane permeability (especially
those with high molecular mass), mucociliary clearance, mucus barriers, and
enzymatic environment can adversely affect vaccine administration via the nasal
route [75]. Additionally, the poor deposition of vaccines in nasal mucosa poses a
challenge to the use of this route [76].
To overcome these challenges, absorption boosters and/or protease inhibitors can
be added to vaccines. Cell-stabilizing and cell-penetrating agents can also be added
to vaccines. The use of nanoparticulate drug delivery systems also holds promise
[40, 77].
10 Conclusion
Administration of vaccines via the nasal route has promise based on the research
available. By using the nasal route, we could possibly solve many unmet medical
needs and improve vaccination of masses due to better compliance (compared to
parenteral vaccines). Furthermore, employing the nasal route avoids the risk of dis-
ease transmission via needle reuse. Also, the nasal route for vaccination can trigger
powerful mucosal and systemic immune responses.
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Chapter 9
An Overview on Nanocarriers for Nasal
Delivery
Sunita Dahiya and Rajiv Dahiya
Abstract Nasal delivery has come up as a promising approach to deliver diverse

therapeutic agents including small drug molecules to biomacromolecules like pep-
tides, proteins and genes to treat various disorders of central nervous system includ-
ing depression, epilepsy, migraine, schizophrenia, Parkinson’s disease, Alzheimer’s
disease, and brain tumor. Nasal route unveils the possibility for delivering the drug
directly from nose to brain, circumventing the challenging blood-brain barrier
enabling their effective brain delivery. The past couple of decades have witnessed
tremendous enthusiasm about exploiting nanotechnology-based approaches in
the drug delivery area, specifically due to the exponential growth in research efforts
employing the nanocarriers’ delivery via different administration routes including
the nasal route. Nanocarrier-based nasal delivery of drugs has progressed over the
years with an assumption that the use of nanocarriers would enable the drug to
access tissues and organs that could otherwise not be accessed effectively by con-
ventional nasal delivery. The present chapter sets out to discuss applications of dif-
ferent nanocarriers in nasal delivery along with their transportation mechanisms and
toxicity concerns.
Keywords Nano carrier · Intranasal administration · Nasal delivery · Nasal

toxicity · Nasal transport
S. Dahiya (*)
Department of Pharmaceutical Sciences, School of Pharmacy,
University of Puerto Rico – Medical Sciences Campus, San Juan, PR, USA
e-mail: sunita.dahiya@upr.edu
R. Dahiya
School of Pharmacy, Faculty of Medical Sciences, The University of the West Indies,
St. Augustine, Trinidad and Tobago
e-mail: Rajiv.Dahiya@sta.uwi.edu
https://doi.org/10.1007/978-3-031-23112-4_9
142 S. Dahiya and R. Dahiya
1 Introduction
The global market for nasal drug delivery is forecasted to reach US$59.2 billion by
2027 at a CAGR of 4.1% from the estimates of US$44.7 billion in 2020 [1].
Traditionally, nasal delivery is limited to topical delivery of drugs in combating
nasal problems like nasal congestions, allergic rhinitis, and common cold. In recent
years, an increased interest has emerged to utilize nasal route as non-invasive alter-
native to achieve systemic effects of drugs and vaccines over the invasive parenteral
route based on its efficient and cost-effective delivery. In particular, nasal mucosa is
highly vascularized and offers benefits including fast onset of action, improved bio-
availability, enhanced patient acceptance, and higher immune response to favor vac-
cine delivery [2]. Some studies indicated that a drug delivered via intranasal route
provides quicker therapeutic response than an oral mixture or tablet and the response
could be achieved as fast as intravenous injection [3]. In addition, intranasal vacci-
nation contributes to local immune protection [4]. Further, the nose is a promising
route for delivery of macromolecules and biotechnology-derived multifarious pro-
teins. Combining the nasal formulation advancements with the contemporary nano-
technology concepts may render vital benefits for the delivery of drugs and vaccines
[5–7]. More efficient dosage forms not only offer patient compliance but can also
get long patent periods upon their successful commercialization that help to sustain
market share and revenue of pharmaceutical company under rationally limited
investment. Marketed nasal formulations containing biologic macromolecular drugs
and small molecule drugs are summarized in Tables 9.1 and 9.2 respectively [8, 9].
Table 9.1 Marketed nasal formulations containing biologics

Therapeutic agent Molecule Brand
(Dosage form) (MW) Indication name(s)
Calcitonin Peptide Osteoporosis Fortical®
(Nasal spray) (3432 Da) Miacalcin®
Desmopressin Peptide Diabetes insipidus, Hemophilia A, Minirin®
(Nasal spray) (1183 Da) Nocturnal polyuria Stimate®
Noctiva™
Oxytocin Peptide Start or strengthen uterine Pitocin®
(Nasal spray) (1007 Da) contractions during labor Syntocinon®
Nafarelin Peptide As part of a fertilityprogram, Synarel®
(Nasal spray) (1321 Da) endometriosis
Cyanocobalamin Peptide Deficiency of vitamin B12 Nascobal®
(Nasal spray) (1355 Da)
Live attenuated influenza Virus-based Influenza FluMist®
vaccine vaccine
(Nasal spray)
Human live attenuated H1N1 Swine flu NASOVAC™
influenza vaccine
(Nasal spray)
9 An Overview on Nanocarriers for Nasal Delivery 143
Table 9.2 Nasal delivery of small molecule drugs for systemic effects
MW
Drug (log P) Indication Brand name(s)
Butorphenol 327.5 (3.7) Migraine; pain management Stadol NS®
Estradiol 296.4 (4.3) Hormone replacement therapy Aerodiol®
Fentanyl 336.5 (4.1) Breakthrough cancer pain Instanyl®
PecFent®
Naloxone 327.4 (0.6) Opioid overdose Narcan®
Nicotine 162.2 (1.09) Smoking cessation Nicotrol NS®
Sumatriptan 295.4 (0.8) Migraine and cluster headaches Imitrex®
Zolmitriptan 287.3 (1.6) Migraine Zomig®
2 Key Anatomical Regions and Functions of Nasal Cavity
Three major segments of nasal cavity are nasal vestibule, respiratory region, and
olfactory region, each one performs distinct functions [10]. The nasal vestibule, also
known as nostril, is the area surrounding anterior external opening to the nasal cav-
ity. Nostrils contain nose hairs that behave like baffle and performs filtration of
inhaled air. The olfactory region is situated at the peak of the nasal cavity. Olfactory
region is lined by olfactory cells with olfactory receptors. From this region, odorant
particles are transported to olfactory epithelium and further solubilized by binding
with the odorant-binding proteins, attach to olfactory receptors, and allow higher
processing before entry into the brain. The respiratory region is lined by a ciliated
pseudostratified epithelium and spread along mucus-secreting goblet cells. The
respiratory region is the most significant segment performing the functions to
humidify, warm, filter, protect, and remove debris [11]. The olfactory epithelium is
located in the upper part of nasal cavity and occupies about 10% of the nose area.
The olfactory region is the only site in the human body that allows direct contact of
the CNS with the external environment. This region is of prime importance for the
drug delivery because nasal administration can directly transport the drugs to this
region which can further diffuse through the olfactory mucosa to reach the CNS
through the epithelial pathway [12].
3 Nasal Delivery: Rationale and Design
The delivery of drugs via nasal route has logical reasons in terms of the easy access
of nasal cavity, rich vascularity that help to achieve effective drug concentrations in
blood even if after topical administration, which is specifically convenient com-
pared to the use of catheter required for the invasive intravenous route as a means of
direct drug delivery to the blood [13]. Higher blood concentrations can be achieved
via nasal route by employing the drug solution as fine droplets or mist rather than as
larger droplets of drug solution, as larger droplets may run off and fail to be absorbed
[14]. The rich vascularity of nasal cavity enables entry of highly permeable drug
directly into the blood stream. Further, this direct systemic absorption bypasses the
first pass hepatic metabolism of the drug by live enzymes which is common for
several drugs administered via oral route. Thus, for the drugs undergoing extensive
first-pass metabolism, nasal route offers a non-invasive way of enhancing bioavail-
ability over oral route [15]. In addition, nasal administration of many drugs shows
comparable plasma concentrations to those achieved by their intravenous route or
typically better plasma concentrations that are achieved by intramuscular or subcu-
taneous routes [16, 17]. Nasal administration is non-invasive, painless, and readily
self-administered by the patient, in contrast to the invasive intravenous administra-
tion that requires medical aid for their administration. Another advantage of nasal
route is its nearby location to the brain, which may achieve higher drug concentra-
tions in cerebrospinal fluid (CSF) and spinal cord as compared to that of the plasma
[18]. Typically, small and lipophilic drug molecules permeate through the nasal
mucosal membranes easily at near physiological pH [19]. However, the important
concern in nasal drug delivery system is that the adequate drug concentration be
present in volume as low as 0.25–3 mL which is the volume that the nasal cavity can
accommodate per nostril. For intranasal administration, a drug must be concen-
trated to a degree that a dose can be administered in <1 mL; however, only limited
number of therapeutic agents meet this criteria [20, 21].
Besides several advantages, the special interest in nasal drug delivery also lies in
its ability of delivering a vast variety of compounds, which includes peptides and
proteins for systemic effects [22]. The nasal delivery provides a user-friendly way
of self-administration and improves patient compliance in comparison to injections
[16–18]. The favorable nasal anatomy in terms of large surface area and rich blood
supply in nasal mucosa helps rapid distribution and absorption of drugs into the
abundant blood vessels, allowing direct access to systemic circulation. This is spe-
cifically useful for drugs that undergo hepatic first-pass metabolism and compro-
mise their bioavailability when delivered by oral route [23]. The improved drug
pharmacokinetics following the intranasal drug absorptions was found comparable
with intravenous injection. Further, it permits a better control of systemic drug con-
centrations in patients, thereby avoiding the risk of disease-related chronic compli-
cations [24]. In certain diseases where nasal drug delivery alone is not appropriate,
it can be used as a supplement to parenteral therapy. The peptide and protein drugs
are assumed to be effectively delivered via nasal route; however, several factors
limit their absorption from nasal mucosa resulting in poor bioavailability and limit-
ing their systemic delivery via intranasal administration as compared to parenteral
administration. The poor intranasal bioavailability is one of the major delivery hur-
dles for development of peptides and proteins for intranasal administration.
Although the absorption enhancers are employed in intranasal preparations of pep-
tide and protein drugs, they are generally not adequate to overcome the barriers
posed by typical physiology of human nose [2]. The major barriers that contribute
to limited bioavailability of peptides and proteins include poor permeability of the
nasal epithelium, mucociliary clearance caused by physical removal of the dosage
content in the nasal cavity, and enzymatic insufficiency in the mucus and epithelium
[25]. Hence, achievement of expected results from nasal delivery of peptide and
protein; a thorough knowledge of anatomical and physiological aspects of nasal
cavity along with its barriers that limit drug absorption is essential. Such knowledge
is utilized to develop strategies that overcome the potential barrier functions of the
nasal and improves the nasal delivery outcomes [19].
4 Applications of Nasal Delivery
4.1 Vaccine Delivery
The entry of air into the body starts from the nasal passage, which eventually
initiates the respiratory airflow of inhaled antigens. The inhaled air becomes warm
and moist, and then filtered before reaching to lungs. There is an active ciliary
movement which continuously impels the mucus layer toward the nasopharynx,
causing clearance of the inhaled particles entrapped in the mucus, by expelling them
out from the nasal passage [26]. However, certain factors like air pollution, low
temperature, and presence of diseased conditions like common cold or inflammation
alter the physicochemical properties of mucus layer or interfere with ciliated cells’
movement; both of which lead to reduced mucociliary clearance of the trapped par-
ticles. The epithelial barrier divides mucosal surface of nasal passage from external
environment, thereby protecting mucosal surface via mucosal secretion, mechanical
cleaning, or mucosa-associated lymphoid tissue (MALT), which is a specific immu-
nological function of the lymphoid tissues [27].
Nanocarriers find wide applications as adjuvants or carriers for the delivery of
vaccines via nasal route. The purpose of incorporating adjuvant in vaccine formu-
lation is to increase the magnitude of an antigen-specific immune response by
accelerating the immune response against even the highest infective form for any
infection or malignancy [28, 29]. Moreover, vaccine adjuvants shall be capable of
providing increased immunization efficacy from weak antigens, increased T cell
responses of required types, and creation of versatile immune responses while
maintaining the safety. Nanocarriers can be interestingly utilized to produce novel
adjuvants for nasal vaccine of desired efficacy and safety based on their several
unique characteristics [30, 31]. Nanocarrier adjuvants are capable of increasing
the antigen amounts that finally reach the immune system with a controlled anti-
gen delivery for a prolonged period, enabling the antigen-induced immune
response for prolonged periods [32]. Besides, nanocarriers-enabled antigen deliv-
ery can be combined with immunomodulation and/or immunostimulation [33].
Lipid-based nanocarriers and polymeric nanocarriers have been researched vastly
for the nasal vaccine delivery [34–36]. Besides anatomical and physiological
favor of human nose, nasal route is economic, patient-friendly, easily accessible,
and needle-free. Due to these benefits, nasal route is capable of achieving
vaccination goals of massive population cohorts as a cheaper and safer vaccine

[37]. Currently, prophylactic nasal vaccine products are available for clinical use
for influenza conditions and available as nasal spray (see Table 9.1). These nasal
vaccines are FluMist/Fluenz™ by MedImmune, MD, USA and the Nasovac™
live attenuated A(H1N1) influenza by the Serum Institute of India, for protection
against swine flu [38]. With these results, recent research focuses in developing
therapeutic nasal vaccine via nasal immunotherapy, where biomaterial immunol-
ogy can be employed in treating different cancers, diabetes mellitus, atherosclero-
sis, multiple sclerosis, rheumatoid arthritis, and lupus, Alzheimer’s disease, and
Crohn’s disease [39].
4.2 Topical/Local Delivery
About 75% of nasal products market is occupied by topical decongestants and

topical steroids for patients suffering from chronic allergic conditions or non-
allergic mucosal inflammatory conditions like rhinitis and sinusitis [40]. Since
persistent rhinosinusitis and nasal polyps can lead to asthma, these chronic condi-
tions require lifelong treatment as nearly 80% of patients with asthma report some
form of rhinitis [41]. However, the topical steroids are poorly distributed in the
sinuses and nose, limiting therapeutic outcomes. This fact prompted the research-
ers to search superior treatment methods like nanotechnology-enabled approaches
for treating the chronic rhinitis and chronic sinusitis with an aim to enhance bio-
availability and patient compliance overcoming the shortcomings of conventional
topical nasal delivery. Mucoadhesive polymers have been widely attempted to
increase the product’s retention time on mucosal surface and reduce interpatient
variations in their administration process, for example, the head position of the
patient [42]. Nanocarriers of different categories such as lipid nanoparticles
including liposomes, solid lipid nanoparticles, and nanoemulsions, or polymeric
nanoparticles including nanogel, nanofibers, and dendrimers are developed to
enhance solubility and stability of a drug or to achieve controlled release with
improved brain delivery via nasal route.
Triamcinolone acetonide (TA) was encapsulated in different lipid nanocarriers
such as polymeric nanocapsules with oily core, nanostructured lipid carriers, and
nanoemulsion to study its in vivo nasal deposition, mucin interaction, and mucosal
permeation and retention. Among three types of TA-loaded nanocarriers, nanocap-
sules exhibited the highest nasal drug deposition, best mucin interaction parameters,
and 46.14% ± 0.048% of initial TA dose indicating best mucosal drug retention after
24 h. As a result, the systemic absorption of nasal TA treatment showed enhanced
efficacy in rhinitis treatment with reduced systemic absorption of TA as compared
to its suspension and marketed product. In addition, nanocapsules maintained better
stability and exhibited lower mucosal irritation than nanoemulsion system, indicat-
ing the potential of polymeric oil-core nanocapsules in developing superior nasal
delivery system for localized nasal delivery than the existing commercial nasal
spray product containing TA suspension [43].
4.3 Systemic Delivery
Due to the fact that the human nose functions to filter the air and also possesses
extensively vascularized large mucosa, the nasal route can be exceptionally vital for
absorption of difficult-to-deliver therapeutic agents requiring parenteral administra-
tion with conventional routes [2, 21]. Extensive research efforts have been per-
formed to exploit the non-invasive nasal route as a potential alternative to the
invasive parenteral route to deliver the challenging pharmacological molecules
including protein and peptide biomacromolecules which undergo degradation in
stomach and metabolic first-pass degradation in liver as the nasal route provides
protection against these degradations [25]. However, small molecule drugs usually
exhibit relatively high bioavailability whereas large molecule drugs like peptides
and proteins show relatively low nasal bioavailability as compared to that of the
parenteral route [8].
Nasal delivery is a non-invasive, smart, and needle-free method that allows easy
access for self-medication and thereby higher patient compliance while providing
fast absorption and rapid onset for management of acute conditions such as heart
attacks, epileptic seizures, low sugar levels, nausea, and vomiting. Systemic nasal
delivery of most drugs can be achieved through nasal sprays or aerosols; however,
such devices are meant to prolong the mucosal drug retention time. Salmon calcito-
nin for osteoporosis, desmopressin for diabetes insipidus, and many non-steroidal
anti-inflammatory drugs are commercially available as nasal dosage forms (see
Table 9.1), whereas many more are in the clinical development [2]. Although the
clinical success of conventional dosage forms for systemic nasal delivery is evident
from their commercial availability, intranasal formulation containing nanocarrier is
not yet available in market despite ample research efforts in this area. The interest
and hope in developing successful nanocarrier-based systemic nasal delivery of
already approved large molecule drugs is based on their intended delivery via non-
parenteral route, which is justified considering their clinical need and develop-
ment cost.
4.4 Nose-to-Brain Delivery
Nose-to-brain delivery offers a prospective option for delivery of drugs from nose
to the central nervous system (CNS) by avoiding blood-brain barrier (BBB),
enabling direct access of drugs to CNS through olfactory region or trigeminal nerves
[12, 18]. When nasal formulations are designed for targeting the CNS, the drug
absorbed into the blood, the cerebrospinal fluid (CSF), and the lymphatic systems
are considered to be delivered via intracellular or extracellular transport mecha-

nisms [3, 17] unlike the conventional systemic route in which the drugs can reach
the lungs and blood circulation before it comes to the brain. Due to this, systemic
administration of drugs requires more time to cross the BBB and reach the CNS,
leading to increased time for attaining desired therapeutic effect and restricting ade-
quate amount of drug to reach to brain [45]. Also, the actual amount of drug reach-
ing the brain after nasal delivery shows interpatient variations. The drug is eliminated
by renal and hepatic mechanisms after intranasal administration [46]. Whether the
drug will go to the brain by direct route or systemic route mainly rely on drug’s
properties like lipophilicity. After intranasal administration, a lipophilic drug enters
the brain via systemic pathway [47]. However, since the systemic pathway limits the
brain delivery, CNS disorders like epilepsy, schizophrenia, depression, migraine,
brain tumor, etc., are ill-treated via this pathway. Inadequacy in accessing the CNS
is mainly related to barrier that separates the brain from blood circulation, that is the
BBB [48]. However, nanocarrier-based nasal delivery systems can breach the bar-
rier through olfactory region of the brain which is placed in upward divisions of
nasal channels remote to the BBB [49].
Intranasal delivery of peptides and proteins enables them to permeate into the
brain via olfactory bulb and trigeminal pathways [22]. Different drug molecules
with conformational similarities with hormones can cause hormonal side effects
upon reaching blood circulation. Nanocarrier-based nasal delivery can markedly
decrease the amount of drug reaching to blood and targeting to the brain due to the
olfactory region. Thus, olfactory region or trigeminal nerves are involved in the
direct brain transfer of a drug leading to its enhanced bioavailability [44]. Nasal
administration of a drug can enter into the general blood circulation either by
directly permeating the brain or by following any of these pathways for its transpor-
tation: (i) drug directly enters from nasal mucosa to blood circulation, (ii) drug
directly enters from nasal mucosa to brain via (a) olfactory region or (b) trigeminal
nerves, another pathway to allow entry of a drug to the brain could be via olfactory
bulb through the neurons by axonal transport [13]. The olfactory pathway shows
brain targeting of a drug within 0.33 h as compared to about 1.7 h required by the
trigeminal pathway [50].
4.4.1 Nasal Transport Mechanisms from Nose-to-Brain
Olfactory pathway in the olfactory mucosa is one of the routes that allow direct
access of drug from nose-to-brain. In this pathway, the drug goes to the olfactory
bulb and passes through the olfactory nerves via intraneuronal and extraneuronal
transport to reach to the olfactory cortex, and further passes to the cerebrum and
cerebellum in the CNS [51].
In case of intraneuronal transport, the drug has to pass through the axons. This is
a slow process which takes hours to days to reach to olfactory bulb and then to reach
the CNS. Contrarily, extraneuronal transport is quicker as it undergoes paracellular
and transcellular transport, which are faster, and the drug can access olfactory bulb
and CNS in only a few minutes [52]. In addition, the olfactory mucosa can be
accessed via supporting cells by transcellular transport or along supporting cells by
paracellular transport through lamina propria [11]. In paracellular transport, drug
passes through tight junctions like occludin, claudin, and zonula occludens present
between the cells. In general, paracellular transport is used by hydrophilic drugs.
Lipophilic drugs usually adopt transcellular transport via passive diffusion or
receptor-mediated endocytosis. Thus, the drug can be transported from olfactory
region to CNS within the nerves by intraneuronal or outside the nerves by extraneu-
ronal transport. However, combination of both routes is more likely to be used for
drug transport instead of a single route [53].
Trigeminal nerve pathway connects to the tail part of the brain such as spinal
cord, the medulla, and the pons and the drug is transported from nose via trigeminal
nerve pathway using intracellular transport (axonal transport) or by endocytosis
[54]. The trigeminal nerve is the fifth and largest cranial nerve that divides into three
branches namely mandibular, ophthalmic, and maxillary nerves, that merges in the
trigeminal ganglion, enters in the brain, and ends in the brainstem. Maxillary and
ophthalmic branches of the trigeminal nerve connect the nasal cavity to the brain,
therefore are more significant in nose-to-brain delivery. Trigeminal nerve causes
nerve impulses to stimulate the olfactory and respiratory epithelium [55].
The prediction of drug absorption via direct nose-to-brain transport of a drug
after nasal delivery is challenging due to characteristics anatomical and physiologi-
cal features of nasal cavity and presence of other barriers. Moreover, the size of
nasal cavity limits the volume of formulation to be administered, restricting the drug
dose to reach the brain. In addition, the residence time of the drug in the nasal
mucosa is shortened by mucociliary clearance and presence of enzymes [56, 57]. To
address these challenges, formulation and manufacturing factors such as molecular
weight of drug, solubility of drug, the design and fabrication of nasal delivery device
are critically considered for improved drug deposition in the nasal cavity [2]. For
example, correct nasal delivery device helps right administration of drug formula-
tion to facilitate drug targeting to the upward area of nasal cavity to reach the olfac-
tory bulb and trigeminal nerve for facilitating drug passage toward the brain. In
addition, researchers have suggested other nose-to-brain transportation pathways
such as a pathway through the CSF and nasal lymphatics in which the drugs are
transported from the nasal passage to the CSF to the brain with no significance entry
into the blood, but it is influenced by the drug’s properties such as degree of ioniza-
tion, lipophilicity, and molecular weight. However, more in vivo studies can provide
deeper insights to confirm direct drug transportation pathway for nose-to-brain
delivery following intranasal administration [58]. Major transportation pathways
from nose-to-brain are illustrated in Fig. 9.1.
Fig. 9.1 Major transportation pathways from nose-to-brain
5 Nanocarriers for Nasal Delivery
Nanotechnology has diverse applications in drug delivery area with unique

attributes. Particularly, different kinds of nanocarriers are excellent vehicles for
carrying variety of cargos and deliver them to the desired target site in the body. The
nanocarriers loaded with different drugs can be delivered via intranasal delivery.
Additives such as permeation enhancer, enzymatic inhibitor, or mucoadhesive
polymers are included in the formulations of nanocarriers to enhance nasal
permeation and retention time of formulations to achieve improved bioavailability
or to improve the stability of nanocarrier systems. Researchers have employed
different lipid-based and polymer-based nanocarrier systems to target brain via
nasal delivery route (Fig. 9.2).
5.1 Liposomes
Liposomes comprise of aqueous core surrounded by phospholipids bilayers.

Liposomes are one of the oldest and proven nanocarrier used for the targeted drug
delivery and several liposomal formulations have gained clinical success in the
treatment of different carriers. Liposomes can be engineered by modifying their
sizes and/or phospholipid composition to suit drug properties. Liposomes offer an
advantage regarding effective encapsulation of both small and large therapeutic
molecules having varied hydrophilicity due to the preferred entrapment of more
hydrophilic drugs within an aqueous core or lipophilic drugs within the hydropho-
bic shell [59]. Liposomes can enhance nasal absorption of therapeutic agents by
enhancing membrane penetrability or by prolonging the retention period of peptides
or by protecting the encapsulated peptides from enzymatic degradation or disturbing
Fig. 9.2 Different
nanocarriers used for nasal
delivery
the mucosal membrane [19]. In fact, liposome-based formulations can lead to

protective immunity irrespective of the nature of the antigen like protein, peptide,
DNA, or its mode of loading such as encapsulation, coupling, or membrane embed-
ding, when delivered through the nasal route [60]. Liposome surfaces can be func-
tionalized to increase the delivery efficiency of wide range of therapeutic agents. In
addition, liposome formulation can aid improving the bioavailability of lipophilic
drugs that undergo hepatic metabolism as well as permeation denial through the
brain barriers [61]. For instance, an anti-schizophrenic drug quetiapine fumarate
was loaded in nanoliposome for nasal administration. Nanoliposome formulations
displayed enhanced diffusion rate and higher amounts of drug distribution within
the brain were found using gamma scintigraphy in rats as compared to that of sim-
ple drug dispersion [62].
Functionalization improves liposome permeation through the brain barriers. For
example, in one such study, cationized liposomes were produced by surface coating
with stearylamine and 1,2-Distearoyl-sn-Glycero-3-Phosphoethanolamine –
methoxyl poly(ethylene glycol) (DSPE-mPEG). This polymer is used for function-
alization as it is a linear phospholipid PEG conjugate with both hydrophilicity and
hydrophobicity. The functionalized liposomes showed improved entrapment of an
anti-schizophrenic drug risperidone with preferential in vivo nose-to-brain transport
along with higher bioavailability and lesser clearance rate than non-functionalized
liposomes [63]. In another study, liposomal formulation was developed for an anti-
Alzheimer drug donepezil. Liposomes were formulated by thin film hydration tech-
nique employing cholesterol, polyethylene glycol, and
1,2-distearyl-sn-glycero-3-phosphocholine (DSPC) via intranasal administration in
rats. The prepared liposomes were uniform in shape and size having 102 ± 3.3 nm
diameter, and 84.91% ± 3.31% encapsulation efficiency. The liposomal formulations
followed sustained-release behavior showing significantly increased bioavailability

of donepezil in plasma and brain without toxic effects [64]. Liposomes can facilitate
the direct drug absorption through the nasal mucosa. A study reported acyclovir-
loaded liposomes incorporated in mucoadhesive gel to investigate its effects on drug
absorption. The liposome-incorporated gel showed 60.72% bioavailability compared
with intravenous route indicating their potential in promoting the contact time and
permeability resulting in direct absorption of drug through nasal mucosa [65].
5.2 Solid Lipid Nanoparticles (SLNs)
Solid lipid is the main building material of the SLNs along with the use of surfactants
to decrease surface tension between water and lipid to stabilize SLNs. The SLNs’
preparation involves dispersion of melted solid lipid(s) in water containing a
surfactant. The SLNs are formed by aqueous dispersions of nanosized particles of
sizes 50–1000 nm via high-pressure homogenization or microemulsification. The
drug molecules, mainly lipophilic compounds, are incorporated in solid lipid and
can undergo controlled release upon administration [66, 67]. Commonly employed
solid lipids for SLNs preparation include mono-, di- and triglycerides, partial glyc-
erides, sterols, free fatty acids, fatty alcohols, and waxes. These lipids are selected
based on their ability to form an extremely nonpolar lipid matrix in which the drug
can either dissolve or disperse. Thus, the selection of lipid or lipid blend and surfac-
tant is critical in the formation of desired SLNs [68]. Besides the lipid being physi-
ologically compatible and stable at room temperature, the selection of single lipid
or blend of lipids also accounts for their ability to produce nanosize particles; bio-
degradable properties; drug-loading efficiency and stability during storage. In addi-
tion, the matrix structure in SLNs can be ordered, less ordered, or disordered in
SLNs. The type and concentration of surfactant selection in SLNs are based on
selected lipid(s) and the intended administration route [69]. Research efforts using
SLNs for brain delivery [70–77] are summarized in Table 9.3.
5.3 Nanoemulsions
Nanoemulsion can be either oil-in-water or water-in-oil biphasic dispersion

containing surfactant or blend of appropriate surfactants for stabilization of two
immiscible liquids. The multicomponent nanoemulsion is thermodynamically
stable, transparent, or translucent emulsions comprising of nanosize dispersed
phase droplets of sizes 20–200 nm [78]. The nanoemulsions show enhanced physical
stability based on the nanosize of the dispersed phase which prevent sedimentation
or creaming due to Brownian motion of colloidal particles of the dispersed phase.
The colloidal size also prevents other destabilization phenomena such as coalescence,
flocculation, and subsequent phase separation which contribute to long-term
Table 9.3 Significant research efforts in nasal delivery of SLNs

Therapeutic agent
(category) Key research outcomes References
Agomelatine High brain targeting efficiency of drug by intranasal route [70]
(antidepressant) compared to intravenous route
Displayed a good direct transport percentage indicating an
achievement of significant extent of direct nose-to-brain
pathway in the brain drug delivery
Ondansetron HCl Rapid localization of the drug in the brain [71]
(anti-nauseating and No adverse response of SLNs on sheep nasal mucosa
antivomit)
Carvedilol Significantly high amounts of carvedilol permeated through [72]
(Antihypertensive) the nasal mucosa
The absolute bioavailability of the intranasal SLNs was
significantly higher than oral formulation
Risperidone Superior pharmacokinetics and biodistribution of intranasal [73]
(Antipsychotic) risperidone compared to intravenous route
Localized risperidone in brain indicating existence of
nose-to-brain delivery
Haloperidol High brain/blood ratio indicating direct nose-to-brain [74]
(Anti-schizophrenic) transport, bypassing the blood-brain barrier
Significantly higher Cmax in brain from intranasal
administration with the highest drug-targeting efficiency and
direct transport percentage as compared to other
formulations, indicating improved brain targeting efficiency
as compared to both intranasal and intravenous solution
Piribedil SLNs were loaded in thermoresponsive in situ gel to delay [75]
(anti-Parkinson’s) mucociliary clearance
Intranasal administration at the olfactory region showed
about 4-folds increased absorption and 2.3- fold reduced
plasma concentration compared to plain intranasal
suspension and showed efficient direct nose-to-brain uptake
as compared to intranasal suspension
Paeonol SLN-in situ gel accumulated effectively in the olfactory [76]
(Neuroprotective) brain area after intranasal administration with observed
fluorescence response in brain regions like olfactory bulb,
cerebellum and striatum
Naloxone Improved pharmacokinetics and biodistribution as compared [77]
(opioid receptor to solution with no toxicity even after administering
antagonist) three-folds of normal dose
physical stability of nanoemulsion systems. Another advantage of nanoemulsion is

that it can solve problems of low drug solubility along with improving chemical
drug stability problems like oxidation, pH, hydrolysis, or biological stability prob-
lems like enzymatic degradation at the mucosal level under physiological condi-
tions [79]. The oily phase of the nanoemulsion can dissolve hydrophobic drugs
followed by its release from the nanoemulsion system due to nanoprecipitation
upon contacting the surrounding aqueous environment, resulting in formation of
particles with an immensely greater surface area with exceptionally high
improvement in drug dissolution based on Noyes–Whitney equation [80].

Nanoemulsions can mask the bitter or unpleasant taste of medicaments and can be
used as nanocarriers to load natural compounds. Nanoemulsion preparation uses
high-energy methods or low-energy methods. High-energy methods such as ultra-
sonication, microfluidics, and high pressure homogenization employ mechanical
devices that consume high amount of energy to generate force that ruptures oil and
water phases into tiny globules. Low energy methods use specific physicochemical
processes like phase inversion temperature and emulsion inversion points that occur
as a response toward change in composition or temperature of emulsion system to
form tiny droplets of oil and water phase consuming only low amounts of energy [81].
Several studies claimed the potential of nanoemulsions in brain delivery for the
treatment of various CNS conditions. In one of the recent studies, nanoemulsion has
been explored as a theranostic system in brain diseases for which a nanoemulsion
rich in cholesterol content was developed to imitate typical lipoprotein molecules
for delivery of a photosensitizer compound, aluminum phthalocyanine chloride.
The nanoemulsion showed higher efficiency particularly in glioblastoma as com-
pared to classical liposomes using the U-87 MG cell line. The nanoemulsion formu-
lation was devoid of dark cytotoxicity at the light dose of 1.0 J·cm−2 on U-87 MG
cells due to photodynamic therapy-induced apoptosis. These results strongly sup-
ported the use of cholesterol-rich nanoemulsion as a new theranostic system [82]. In
another study, nanoemulsion of a potential agent to treat resveratrol neurodegenera-
tive disorders, namely resveratrol, was formulated with coconut oil, Pluronic-P107,
and Cremophor EL by ultrasonication process. Intranasal administration of resvera-
trol nanoemulsion at 2 mg/kg dose in rats demonstrated high brain targeting efficacy
indicating it as a promising option for treatment of Alzheimer’s disease [83].
Chitosan can be used to modify nanoemulsion properties to serve as an excellent
carrier for nasal vaccine, as it can enhance immune efficacy of nasal vaccine by
prolonging its residence time in the nasal cavity and improving the cellular uptake
efficiency via inducing higher systemic and mucosal antibody levels. In one such
study, a chitosan-modified cationic nanoemulsion was developed and evaluated for
antigen encapsulation efficiency, stability, uptake efficiency, and cytotoxicity of
vaccine in the nasal cavity in mice. The cationic nanoemulsion-based nasal vaccine
demonstrated relatively high cellular uptake of antigens on DC2.4 and MDCK cells
along with displaying the enhanced residence time of the antigen, with a good quan-
tity of vaccine residing within nasal cavity following 60 min of its administration.
The systemic antibody levels in the serum and mucosal antibody levels in nasal
lavage fluids of the immunized mice were found to be significantly higher in chito-
san modified nasal vaccine as compared to nanoemulsion without chitosan modifi-
cation as well as free antigen, demonstrating the potential of chitosan modification
of nanoemulsion for nasal vaccine administration [84].
5.4 Nanogels
Nanogels are highly viscous hydrogel materials with a three-dimensional structure

of crosslinked and swellable polymers that have high water-holding capacity. A
variety of synthetic polymers, biopolymers, or combination can be used for nanogel
preparation. The internal polymer crosslinking is a very important phenomenon in
nanogel preparation as it influences the stability and drug-loading capacities of
nanogels. The swelling capacity of nanogel is affected by the crosslinking capacity
of nanogel is influenced by chemical composition of polymer matrix, charge den-
sity, and degree of crosslinking [85]. In case of stimuli-responsive nanogel, external
triggers determine the swelling capacity. The nanogel can be tailored to achieve
desired physicochemical properties for which their chemical composition can be
modified for amphiphilicity, surface charges, size, and porosity. The swelling and
collapsing capacity of nanogels determines the drug loading and release behavior
from nanogel. This is because upon swelling, the solvent penetration occurs occu-
pying the free spaces within nanogel causing an increased volume either in a rapid
or in a slow manner depending on the degree of crosslinking [86]. In addition, the
release of a drug from a nanogel system is affected by gel crosslinking, gel network
degradation rate, drug-polymer chain interactions, and molecular weights of the
polymer. The small size of nanogels enables their enhanced cellular uptake at
desired target sites [87]. The major advantage of nanogels lies in its reduced muco-
ciliary clearance based on high viscosity, which leads to decreased taste effect based
on low post-nasal drip approaching nasopharynx. However, nanogel deposition
inside of nasal cavity is significantly affected by administration mode because the
high viscosity of nanogels limits their spreadability, necessitating well-designed
applicators. To address this problem, in-situ gel-forming agents can be incorporated
in the nanogel formulation so as to keep the nanogel in liquid form during storage
but convert into gels at the site of application. Such a liquid-to-gel conversion is
triggered by pH, temperature, and presence of ionic or biological compounds [88].
Nanogels have been explored for nasal delivery of drugs for several brain
conditions as they can deliver both hydrophilic and hydrophobic and vaccine
delivery also. Nanogels are very useful systems in treating brain diseased based on
their capability of delivering both hydrophilic and hydrophobic drugs to bypass the
BBB after intranasal administration [89]. In an important study, poly(N-vinyl
pyrrolidone) nanogels were synthesized with free insulin and insulin covalently
attached was synthesized by the e-beam irradiation method. The insulin covalently
attached to polymer exhibited high biocompatibility, lack of immunogenicity by
nasal mucosa, higher biodistribution in brain regions, and clearance from the blood
and urine within 24 h in comparison to free insulin in the mouse model. The results
displayed higher potential of synthesized nanogel in brain insulin delivery via intra-
nasal route to treat neurodegenerative diseases like Alzheimer’s disease [90].
Another recent study employed teriflunomide, loaded in a lipid-based in situ nano-
gel composed of carbopol as mucoadhesive agents, gellan gum as gelling agent, and
gelucire 44/14 as a surfactant in glyceryl di-behenate and glyceryl mono-linoleate
lipid blend for intranasal delivery in the treatment of glioma. The gellan gum with
surfactant system exhibited superior permeation enhancement (IC50 7.0 μg/mL) as
compared to gellan gum with carbopol system (IC50 4.8 μg/mL) in human U-87MG
glioma cell line. Further, a twofold increased brain Cmax was observed for techne-
tium (99mTC) labeled intranasal gellan gum-surfactant system than gellan gum-
carbopol system by both intranasal and intravenous routes, which confirmed that the
surfactant with natural gelling polymer are promising systems for improved drug
permeability and brain delivery via nasal route [91]. Nanogel formulations have
been researched for protein-based antigen delivery of mucosal infections like
Clostridium botulinum [92], and lifestyle diseases like obesity and hypertension
[93, 94].
5.5 Polymeric Nanoparticles
Polymeric nanoparticles are solid nanocarriers made up of synthetic, natural, and/or

biodegradable polymers within 1–1000 nm size range and can be structurally clas-
sified as matrix type (nanospheres) and reservoir type (nanocapsules). The drug is
dissolved or dispersed in nanospheres’ polymer matrix whereas it dissolves or dis-
persed in liquid oily or aqueous core nanocapsules, the drug is either dissolved or
dispersed in oily/aqueous core enclosed by a polymeric membrane or can be
adsorbed or chemically conjugated to the nanoparticle surface forming a vesicular
core-shell system [95]. The preparation of polymeric nanoparticles uses dispersion
of natural/synthetic polymers via solvent evaporation, salting out, nanoprecipita-
tion, dialysis, and supercritical fluid technology, or by using direct polymerization
of monomers via emulsification polymerization, interfacial polymerization, and
controlled/living radical polymerization [96]. Polymeric nanoparticles proffer
improved drug stability due to avoidance of systemic degradation, and their surfaces
can be readily functionalized to tailor their properties [97]. However, complicated
purification and storage and irregular drug-release patterns are limitations in their
full exploration as intranasal delivery vehicles. Polymeric nanoparticles have been
explored as nanocarrier to entrap, encapsulate, dissolve, adsorb, or chemically
attach different drug molecules for the brain-targeting applications. In addition, the
macromolecular polymer materials used in their preparation enable these nanocar-
riers to be utilized as adjuvants in vaccines. Polymeric nanoparticles with the small-
est size only can penetrate the mucosal membrane in restricted quantities via the
paracellular route [98]. The research reports regarding the beneficial use of poly-
meric nanoparticles in intranasal delivery are ambiguous [99, 100].
The poly-lactic-co-glycolic acid (PLGA) nanoparticles of an antidepressant drug
agomelatine were developed for direct nose-to-brain delivery. The nanoparticles
showed stability for longer period of time and exhibited low particle size (<200 nm),
high entrapment efficiency, sustained release, and higher ex vivo permeation
through goat nasal mucosa [101]. PEGylation is an effective strategy in overcoming
the mucociliary clearance, low epithelial permeation, and local enzymatic
degradation of nanocarriers. Polymeric nanoparticles of polycaprolactone (PCL)

were used to encapsulate a neuroprotective compound, bexarotene. Among
PEGylation with 1, 3, 5, and 10% PCL-PEG, only 5 and 10% of PCL-PEG coating
were able to ensure NP stability and homogeneity in mucus whereas other formula-
tions with 1 and 3% PCL-PEG were ineffective to particle size or morphology.
Upon incubation with artificial nasal mucus, the PCL-PEG5% and 10% demon-
strated fast mucous penetrability without affecting their uptake by RMPI 2650 cells.
The PCL-PEG5% demonstrated 3- and twofold higher concentration, indicating
that about 4% of dose was directly delivered to the brain as compared to non-
PEGylated nanoparticles [97]. Chitosan is an attractive natural alkaline polysaccha-
ride with good biocompatibility and biodegradability to be employed for nanoparticle
preparation as carrier for drug encapsulation and as a coating material to achieve
surface modifications of nanoparticles suitable for brain delivery. Chitosan affects
tight junctions and enhances drug permeability across the BBB. Further, chitosan
bears positive charge and can be absorbed to the negatively charged cell membrane,
thereby increasing residence time of nanoparticles on nasal mucosa to benefit nose-
to-brain delivery. A wide range of derivatives have been synthesized due to the pres-
ence of free amino groups on the chitosan particle surface which allow specific
chemical modification [102, 103].
5.6 Polymeric Nanomicelles
Polymeric nanomicelles comprise of hydrophobic central core enclosed by

hydrophilic shell. The hydrophobic core encapsulates the hydrophobic drugs while
the hydrophilic shell stabilizes and protects them from cellular interactions [104,
105]. Polymeric nanomicelles in the range of 10–100 nm are formed by self-
assemblies of block or copolymers in specific solvent. Due to the small particle
sizes, low polydispersity index, and high surface polarity, polymeric nanomicelles
can achieve significantly increased water solubility for improved dissolution in
nasal and axonal conditions which is beneficial during nasal pathological conditions
[106]. Polymeric nanomicelles can be a promising way to treat neuroinflammation
using non- steroidal anti-inflammatory drugs via intranasal delivery [107].
Soluplus®-based polymeric micelles were developed for intranasal administration
of a poorly water-soluble drug meloxicam. The nanomicelles exhibited more than
20-fold faster meloxicam dissolution rate and fivefold higher nasal permeability
compared to pure meloxicam and demonstrated excellent potential for higher
in vivo brain distribution via nose-to-brain delivery [108].
CNS neurodegenerative diseases due to oxidative stress and inflammation can be
benefitted from polymeric nanomicelle formulations by enhancing solubility and
nasal bioavailability, which in fact shows poor solubility and low oral bioavailabil-
ity. A poorly soluble drug baicalein was encapsulated into PEG-PLGA micelles for
nasal inhalation to achieve enhanced brain distribution of drug in C57BL/6 mice.
PEG-PLGA nanomicelles reduced drug toxicity in SH-SY5Y and BV-2 cells with
no toxicity at concentrations of 50 μM or less. Further, the intranasal administration

of polymeric nanomicelles showed substantially reduced levels of inflammatory
factor TNF-α at 5 μM and IL-6 at 20 μM with 1.50-fold higher absorption in com-
parison to oral administration [109]. Moreover, polymeric nanomicelles have been
surface-functionalized with cell-penetrating peptides to achieve significantly higher
brain distribution and cellular uptake of drugs. Methoxypolyethylene glycol-
polycaprolactone co-polymers were used to form nanosized micelles of 100–600 nm.
It was revealed that the polymer nanomicelles with a diameter of around 100 nm
could promote nasal absorption and increase the delivery of drug into the CNS with
lower concentrations in non-targeted tissues to facilitate the direct intranasal brain
delivery as compared to intravenous administration [110].
5.7 Nanofibers
Electrospun nanofibers are fibers in nanometer range having different physical

properties based on polymers used including poly(vinyl alcohol), poly(ethylene
oxide), poly(ε-caprolactone), poly(acrylic acid), ethyl cellulose, cellulose acetate,
hydroxypropylmethyl cellulose, poly(acrylonitrile), cellulose acetate phthalate, and
poly(urethane) for different biomedical applications [111]. Nanofibers naturally
possess very high surface-to-volume ratio that can result in enhanced cell attach-
ment, higher drug loading, and improved mass transfer properties. Adhesive nano-
fibers show fast dissolution and delay or controlled drug release properties, making
them optimistic nanocarrier for drug delivery research [112]. Intranasal delivery of
self-assembling peptide nanofiber vaccines is a promising method of a needle-free
and adjuvant-free option to induce protective immunity against viral infections. In
one recent study, intranasally delivered peptide nanofibers significantly increased
the number of persisting antigen-specific tissue-resident memory CD8+ T cells in
the lung, allowing for a more rapid response to infection at 6 weeks post-vaccination
as compared to subcutaneously delivered nanofibers [113].
5.8 Dendrimers
Dendrimers are nanosized, hyper-branched macromolecules with highly defined

chemical structures. Typically, dendrimers consist of a central core, repeating units
attached to the central core known as “generations”, and terminal functional groups
present on the dendrimer surface. All these structural features account for den-
drimer’s unique physicochemical and biological properties [114]. A generation 5
(G5) polyamidoamine (PAMAM) dendrimer nanocomposites functionalized with
PEG methyl ether (mPEG) was developed as in situ ionic-sensitive gel of gellan
gum and compared with the PAMAM nanocomposite solution. Following nasal
administration of the in situ gel, maximum accumulation was observed at 12 h

against only a small amount of accumulation by nanocomposite solution at 2 h,
indicating delivery potential of dendrimer-based functionalized nanocomposites as
in situ gel for significantly improving the nasal brain transport efficiency [115].
6 Toxicity Concerns
When the material is reduced from its original size to a nanoscale, it shows entirely
new or modified physicochemical and pharmacokinetic properties that may pose
health hazard to a patient. For instance, certain drug molecules unable to cross the
biological membrane can do so when converted as nanoparticle or they can enter
other cellular compartments posing potential risk of adverse effects [116].
Nanocarriers are composed of polymers, lipids, and other adjuvants such as surfac-
tants. Mostly, the use of biodegradable lipids or polymer matrix materials is consid-
ered safe due to their degradability within the body followed by their elimination from
the body. This is the reason for the preferential use of lipid or polymeric nanocarri-
ers over metallic nanocarriers. For example, polymeric nanoparticles developed
using biodegradable material are metabolically converted into biodegradable prod-
ucts like lactic acid, butanol, and 6-hydroxycaproic acid that are regarded as safe by
USFDA [117]. However, crosslinking of biodegradable polymers via ionic or cova-
lent binding may or may not retain their biodegradability [118]. A thermorespon-
sive, naringenin-loaded-nanoemulsion-in situ-gel formulated with chitosan did not
show any toxicological response in terms of morphological changes in the micro-
structure of brain as well as nasal mucosa tissues in animals which accounts for the
minimal interaction of nanoparticles with the nasal mucosa without showing any
visual signs of inflammatory or necrosis [119].
Nanocarrier-based nasal drug delivery systems contain surfactants to enhance
nasal permeation, but the main concern is the associated toxicity such as nasal irrita-
tion and ciliotoxicity. Poloxamer 188 is one of the most widely employed surfactant
in nasal nanoformulations. Like Poloxamer 188, Cremophor EL and laurate sucrose
ester have been found safe non-ionic surfactants for nasal delivery [120].
Concentration-dependent toxicity and irritation were evident for Tween-80 at a con-
centration higher than 10.0% w/w in thermoresponsive in situ nanoemulgel formu-
lation. Another important additive used to increase contact time of drug with nasal
mucosa is the use of mucoadhesive polymer which provides adhesive properties and
therefore, increased exposure of drug to nasal tissues should be rationally accounted
for the safety of the nasal nanocarrier product [121].
Despite limited research data on nanoformulations’ toxicities, literature supports
nanocarriers’ superiority over conventional nasal dosage forms like solutions or
gels as these require greater amounts of possibly toxic excipients including perme-
ation enhancers, stabilizers, and enzyme inhibitors. Also, the use of nanocarriers for
nasal delivery can reduce the drug’s side effects by using its decreased required
dose. However, the final decision about using the nanocarriers for nasal delivery
should consider attentive choice of excipients along with their concentrations,

absorption from nasal mucosa and transportation mechanism, biodegradability, and
long-term toxicity before moving forward with their clinical application.
7 Conclusion
The conventional drug delivery systems for treatment of CNS diseases are limited
due to different biological conditions including first-pass effect, enzymatic degrada-
tion, presence of BBB, and systemic clearance, that end up in poor bioavailability
and inadequate brain therapy. Nasal delivery of drugs could address these chal-
lenges up to a significant extent and is forecasted as a promising approach for non-
invasive and effective drug delivery options. Nanomedicine comprising of
nanocarriers can be employed for effective delivery of small and large therapeutic
molecules including genes to deliver to the brain in a controlled and targeted man-
ner which also reduces the potential peripheral side effects. Nanocarriers can be
tailored via different functionalization strategies to enhance targetability and to
address the clinical needs of the therapy. In spite of extensive research efforts in the
area of nasal delivery of nanocarriers, increased efforts are required to understand
the transport mechanisms for nose-to-brain delivery in order to produce clinically
successful nanomedicine to be administered via nasal route for enhanced brain
delivery. Currently, a substantial number of therapeutic agents including drugs, pep-
tides, proteins, and genes have shown promising results in animal models and are
under clinical development. If their preclinical studies support clinical data, nasal
route could be a boon in the treatment of brain diseases in the coming years.
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Chapter 10
Nose-to-Brain Delivery of Peptides
and Proteins
Meltem Ezgi Durgun, Gamze Çamlık, İsmail Tuncer Değim,

and Yıldız Özsoy
Abstract The fact that BBB is an important obstacle in drug delivery to the brain
has revealed the need o try alternative drug administration routes. One of these
alternative ways, also called non-invasive administrations, is intranasal drug admin-
istration. Due to the high molecular weight of proteins/peptides, their bioavailabil-
ity cannot be high after administration with conventional dosage forms. In addition,
the hydrophilic part in their structures makes it difficult for them to pass through the
BBB and cannot accumulate in the brain in sufficient quantities. For this reason,
intranasal administration stands out as an important alternative for protein/peptides.
However, the properties that limit the bioavailability of protein/peptides prevent
their nasal absorption from being adequate.
Carrier systems are used to modify drugs to gain different structural properties.
Thus, drugs can be easily delivered to the desired target region, and their bioavail-
ability can be increased. Nanocarriers also offer great advantages for targeting pro-
teins and peptides. Proteins/peptides produced in nanoscales and given a certain
level of lipophilic character can easily pass through the BBB and reach the brain
after intranasal administration.
In this chapter, rate-limiting steps in nose-to-brain targeting of proteins/peptides,
possible pathways through which transmission occurs, points to be considered
while developing nanocarriers, and examples of working with some peptides, which
are frequently the subject of studies, are examined. In addition, the potential for
commercialization and future perspective of nanocarriers developed for
protein/peptides are discussed.
M. E. Durgun · Y. Özsoy (*)

Department of Pharmaceutical Technology, Faculty of Pharmacy, Istanbul University,
Istanbul, Turkey
e-mail: mezgi.kilic@istanbul.edu.tr; yozsoy@istanbul.edu.tr
G. Çamlık · İ. T. Değim
Department of Pharmaceutical Technology, Faculty of Pharmacy, Biruni University,
Istanbul, Turkey
e-mail: gcamlik@biruni.edu.tr; tdegim@biruni.edu.tr
https://doi.org/10.1007/978-3-031-23112-4_10
170 M. E. Durgun et al.
Keywords Nose-to-brain delivery · Nanocarriers · Intranasal drug administration ·

Protein · Peptide · Olfactory
1 Introduction
Proteins and peptides are macromolecules that can control various biological
events in the body. These structures attract the attention of scientists due to their
target-specific binding properties and low toxicity. Protein and peptide therapeu-
tics have undergone rapid development since the synthesis of the first bioactive
peptide proposed by Robert Bruce Merrifield in 1953 [1]. Peptides and proteins
are biomolecules that can be potentially used in the treatment of cancer, diabetes,
cardiovascular diseases, autoimmune diseases as well as central nervous system
(CNS) diseases. However, there are hurdles that must be overcome before these
complex biomolecules can reach to the brain in a therapeutically meaningful
amount [1, 2].
The brain is protected by restrictive barriers such as the blood-brain barrier
(BBB) and the blood-cerebrospinal fluid barrier (BCSFB), which are essential
for maintaining CNS homeostasis. This defense mechanism also prevents the
transport of therapeutics to the brain. In addition, the high metabolic activity of
these barriers can cause the degradation of drug molecules. Various approaches
such as targeting and increasing permeability have emerged to improve the dis-
tribution of therapeutics across the BBB by overcoming the aforementioned
barriers [2]. However, the systemic side effects, low bioavailability, and toxicity
of the excipients used should be carefully evaluated. Increasing evidence from
research reports indicate that intranasal (IN) drug delivery allows both small
and large molecules to bypass the BBB via the olfactory and trigeminal
nerves [2, 3].
In this chapter, the nasal route of administration, its advantages and disadvan-
tages, the routes for the penetration of protein and peptide therapeutics through the
nose to the brain, and the drug delivery systems used are reviewed.
2 Brain Drug Delivery
Brain is a delicate organ and it manages many vital functions. It protects itself
with different barrier mechanisms against inflammation and microhemorrhages,
which can be caused by environmental factors such as air pollution or exposure to
chemicals. The mechanisms unfortunately work as a negative for desired mole-
cules that brain needs. In other words, these barriers limit the desired molecules’
10 Nose-to-Brain Delivery of Peptides and Proteins 171
entries. In the case of drugs therapy, the active molecules cannot pass the men-
tioned barriers easily and it becomes very difficult for drug molecules to reach the
CNS. The strategies developed to deliver drugs (therapeutics) to the brain can be
classified into two major categories as invasive and non-invasive strategies/tech-
niques. Invasive methods include the disruption of the BBB, intracerebral
implants, and intrathecal applications but these procedures are not preferred due
to their short life spans, not being safe, or some difficulties of the application
process. When these disadvantages are present, the intranasal pathway appears to
be a promising, non-invasive approach, and it has been gaining attention over the
years [4, 5].
Delivering active agents through the nose to the brain needs to solve the issues
by bypassing the BBB, improves the bioavailability of protein and peptides, and
provides a more convenient drug administration way than the parenteral route [5].
Also, different uncommon approaches, such as using nano-carrier systems, target-
ing active sites, surface modifying, and using permeation enhancers, are used by the
researchers to enable the cellular permeation and absorption of macromolecules
from nose-to-brain drug transport [5, 6].
3 Barriers to Nose-to-Brain Drug Delivery
Delivering drugs through the nasal route has gained more and more attention over
the years due to the easy formulation and process development [7]. As it is true for
all drug administration routes, IN delivery has its own physiological, physicochemi-
cal, and mechanical challenges obscuring the absorption of the active agent [7].
These challenges include the following:
–– Mucociliary clearance
–– Diffusion through mucus layer and transportation across the epithelial membrane
–– Active agent physicochemical challenges
–– Potential efflux drug transporters
–– Nasal structure and site of drug release
–– Potential degradation by extracellular and intracellular enzymes [7]
4 Nasal Route
The air we breathe is cleaned up through two phases. In the first stage, particles
bigger than 3 μm are blocked from getting in the nose by the hairs on the nasal
vestibular region. In the next stage, particles bigger 0.5–3 μm get stacked at the
nasal mucosa. Particles with lower diameter than 0.5 μm can pass through nose
to the lower airways [8]. The thickness of the nasal mucosa is around 5 μm and
the pH is 5.5–6.5 in adults and 5.0–6.7 in children. The surface covered with
mucous layers and its form is gel or sol. The gel layer is the outermost layer and
it is more viscous and thicker, the sol layer fills up the gap between the gel layer
and the epithelial cell cilias are present there and they can even be able to move
[9, 10].
At the cilia’s tips, the sol layer is present and in contact with the gel layer.
Particles are pushed through to rhino-pharynx area by the movement of cilias and
this phenomenon is called mucociliary clearance (MCC) [8]. The nasal mucosa is
responsible for both moisturizing the air and protecting the mucosa from environ-
mental factors such as unwanted gases, bigger particles, and temperature changes.
The mucosa consists of 95% water and 2% glycoproteins. The liquid produced by
mucosa every day is 1.5–2.0 liters on average. Also, in the mucosa, eosinophils,
neutrophils, mast cells, and immunologically active compounds are secreted mainly
by the goblet cells. The MCC mechanism is responsible for swiftly getting the for-
mulation into the nasal cavity to the rhino-pharynx [8, 11]. This mechanism is one
of the factors negatively affecting the absorption of the therapeutics which are
applied through the intranasal way. To overcome MCC, different bioadhesive excip-
ients should be used in formulations [4, 12].
4.2 Transporter and Efflux System
At the moment, alteration of transporters for the multidrug resistance have already
been identified in human nasal respiratory and olfactory mucosa, which may be
involved in the transport of a wide variety of hydrophobic and amphiphilic drugs.
P-glycoprotein (P-gp) is one of the efflux transporter that exists in the apical area of
ciliated epithelial cells and in the submucosal vessels of the human olfactory region
[4]. Several studies demonstrated that P-gp has also an important role in actively
preventing the influx of drugs from the nasal membrane [4].
4.3 Enzymatic Degradation
Enzymatic degradation is another factor causing a low transport, especially for pep-
tides and proteins across the nasal membrane. Enzymatic degradation of the sub-
stance can occur either within the nasal cavity or around nasal cavity when passing
across the epithelial barrier. Both sites contain exopeptidases that can divide
peptides and endopeptidases to disrupt internal peptide bonds [13]. The use of
enzyme inhibitors and/or saturation of enzymes can be considered to overcome this
barrier [14].
5 Pathways for the Intranasal (IN) Drug Delivery
A substance can pass olfactory epithelium (OE) by three different pathways:

–– Intracellular pathway, in which the drug is carried to olfactory bulb (OB) by
intracellular axons
–– Paracellular pathway, in which the active substance is carried to the lamina pro-
pria by the tight junctions between the cells of OE and olfactory nerves
–– Transcellular pathway, in which the active substance is carried to the lamina
propria with sustentacular cells (SUS) by receptor-mediated endocytosis or pas-
sive diffusion (Fig. 10.1) [15, 16]
The substances reaching to the blood circulation can be absorbed by the blood ves-
sels (BV) and lymph vessels (LV) or drained to the deep cervical lymph nodules on
the neck. Also, the active substances may be delivered by the perineural cavities
between olfactory cells and olfactory nerve fibroblasts [15]. After passing the crib-
riform plate, theoretically, the active substance can reach cerebrospinal fluid and
diffuse to different brain parts [15].
6 Systemic Pathways
The systemic pathway is an indirect delivery route. The drug is absorbed from the
nasal epithelium and being transported to the systemic circulation. The drug should
first enter the nasal cavity and avoid being eliminated by enzymes and MCC [17].
Then the drug is transported to the systemic circulation to pass the BBB to reach
brain parenchyma. This pathway is mainly considered for lipophilic substances with
low molecular weights [18]. More hydrophilic molecules generally follow the para-
cellular route. Depending on their molecular weight, the bioavailability of more
hydrophilic molecules is higher with lower molecular weights [19]. The structure
and shape of the molecule and net positive charge are the other factors affecting the
permeation of the drugs [20]. The drawbacks of the systemic pathway also include
increased systemic exposure, hepatic and renal drug metabolism, and accumulation
in other tissues resulting in some toxic effects [21].
Fig. 10.1 Pathways of the passage of substances through the olfactory epithelium [15]
This pathway remains the least explored and studies have been carried out only
in animal models. Drug access to the systemic circulation does not guarantee its
direction to the BBB, and whether it can cross the BBB or not is also very impor-
tant. Transfer of the drug from the blood to the brain can also occur across the cho-
roid plexus. The drug will enter the cerebrospinal fluid (CSF) depending on its
molecular weight and may diffuse into the brain tissue [22]. The CSF is drained
totally to the peripheral bloodstream every 5 hours, causing the transport from blood
to the CSF to be in one direction [19].
7 Mechanism of Drug Absorption Via Nose
At the first step of the absorption, the drug from the nasal cavity can pass through
the mucosa. Although neutral, lipophilic, and small particles can pass through this
layer easily, the passage is more difficult for charged, hydrophilic, and larger parti-
cles [16]. Musin, a major protein that can bind the substances which are soluble in
the mucosa, prevents the diffusion of the drugs. Also, changes in the mucosa
structure are possible by environmental changes such as pH and temperature [15,

16]. Although the mechanism of nasal delivery of drugs is not fully understood, it is
reported that CSF is responsible for transporting therapeutic substances from the
nasal cavity to the brain by combining the vascular system and lymphatic pathways
[16]. The drug accumulated on the respiratory epithelium passes first to the sys-
temic circulation and is then transmitted to the CNS. On the other hand, the drug
accumulated on the olfactory epithelium is transmitted to the CNS through intracel-
lular, paracellular, and transcellular pathways by olfactory neurons and olfactory
epithelium cells [15, 16, 23]. Another pathway to deliver drug intranasally is tri-
geminal nerves. Olfactory and trigeminal nerves bind the brain to the nasal cavity
and make possible drugs to reach the brain without passing through the BBB [23].
7.1 Paracellular Route of Transport
The first route is paracellular route, which is associated with the intercellular spaces
and tight junctions. Paracellular transport has an important route particularly for the
absorption of peptides and proteins, so it has been reported that the paracellular
route should be reversibly opened and enhances the nasal absorption of peptides.
Mucosal absorption increases due to the hydrophilic characteristic of drugs [24].
7.2 Transcellular Route of Transport
The second route is the transcellular route which is achieved with passive diffusion
or active transport mechanism. It is important for the absorption of lipophilic mol-
ecules. Molecules can also be recognized by the membrane (active carrier trans-
port) [24].
8 Advantages and Disadvantages of IN Drug Delivery
Compared to other drug delivery pathways, nasal pathway has many advantages
[23, 25]:
–– Can reach to brain easily and provides non-invasive application.
–– Being a noninvasive application, it has low infection risk.
–– Lets direct drug delivery to the CNS without having to pass through the BBB.
–– Provides direct delivery to both the CNS and systemic circulation.
–– Easy self-application process provides high patient compliance.
–– Prevents metabolization of the drugs in the gastrointestinal pathway.
–– Prevents hepatic first-pass effect.
–– Porous and thin membrane structure helps with the fast absorption and fast onset
of action.
–– Provides high bioavailability with a low dosage.
–– It has a wide absorption area (160 m2 in humans) and a wide olfactory epithelium
area (12.5 cm2 in humans).
Despite having many advantages, nasal pathway also has some disadvantages
[23, 25]:
–– Complete dosage should be given in 25–200 μL in volume (low volume).
–– It may not be feasible for all drugs.
–– Mucociliary clearance of the delivery system causes drugs to be rapidly cleaned
out from nasal cavity.
–– Nasal cytochrome P450/peptidases/proteases may cause some enzymatic
degradation.
–– When compared to the gastrointestinal system, the absorption area is much
smaller.
–– Nasal blockage and nasal irritation may disrupt the drug absorbance.
–– High doses should be used for hydrophilic drugs. If absorption enhancers are not
used with hydrophilic drugs, high dosages cause some problems.
–– Shows individualities.
–– Epithelium pH is low, so it may not be suitable for all drugs.
9 Factors Influencing Nasal Drug Absorption
9.1 Factors Related to Drugs
9.1.1 Solubility and Dissolution Rate
Solubility is a prerequisite for the absorption of a drug. Absorption of a drug that

remains in the form of particles without dissolving cannot be expected. For nasal
absorption of a drug to occur, the drug must first be dissolved in the aqueous fluids
of the nasal cavity [26]. Therefore, the absorption profile of less soluble drugs in
water and require high doses can be improved by increasing their solubility in
water [27].
The chemical form of the drugs can also affect the absorption as it affects the
solubility of that drug. For this reason, the use of salt or ester forms of drugs is a
frequently preferred method. In a study, Huang et al. searched the effect of chemical
structure on nasal absorption of L-Tyrosine. The results showed that the nasal
absorption of the carboxylic acid ester of L-Tyrosine was much higher than that of
L-Tyrosine [28].
Polymorphism directly affects the absorption mechanism as it affects the disso-
lution rate and solubility of drugs. For this reason, it is recommended to examine the
polymorphic stability and purity of the active pharmaceutical ingredient (API) used
in pre-formulation studies. This general rule also applies to the R&D processes of
nasal drugs.
9.1.2 Lipophilicity
The cell membrane structure in the nose is one of the most effective factors in drug
absorption. Cell membranes with a lipid bilayer structure facilitate the absorption of
drugs with high lipophilic character. The nasal mucosa is a second parameter that
facilitates nasal absorption of lipophilic drugs. In fact, the nasal mucosa also has a
hydrophilic character. However, the lipophilic nature of this mucosa is more domi-
nant while fulfilling the natural barrier function [29]. For this reason, as the lyo-
philic character of drugs increases, their nasal absorption is expected to increase
[26]. In studies with antihypertensive beta-blockers such as metoprolol, alprenolol,
and propranolol, nasal absorption of metoprolol, which has a hydrophilic character,
was found to be lower. Since alprenolol and propranolol have lipophilic character,
their nasal absorption was found to be high [30].
However, it should not be forgotten that it must be dissolved in the aqueous fluids
of the nasal cavity for drug absorption. Therefore it must have certain water solubil-
ity. Although lipophilicity will increase absorption, drugs with very high lipophilic
character may not be sufficiently absorbed.
9.1.3 Molecular Weight
The relationship between molecular weight (MW) and nasal absorption of a drug
varies according to the lipophilic or hydrophilic character of the drug. For lipophilic
drugs, there is a direct relationship between MW and absorption. Lipophilic drugs
with an MW of less than 1 kDa are almost completely absorbed from the nasal cav-
ity via transcellular mechanisms, while nasal absorption of lipophilic drugs greater
than 1 kDa is significantly reduced [31]. In hydrophilic drugs, while the absorption
of drugs with MW value <300 Da is not affected by physicochemical properties, the
same properties become important parameters when MW value is ≥300 Da [26].
For this reason, while the bioavailability of some small hydrophilic molecules is
expected to be around 10%, this value drops to 1% for large molecules such as pro-
teins [32].
9.1.4 Partition Coefficient and pKa
It is known that the non-ionized form of drugs is more easily absorbed than the ion-
ized form [26]. For this reason, the partition coefficient and pKa values of drugs
affect their absorption. Jiang et al. determined diltiazem hydrochloride and
paracetamol as model drugs and examined their nasal absorptions. The results
showed a quantitative relationship between the partition coefficient and the nasal
absorption constant [33]. Similarly, in a study with aminopyrine, it was found that
nasal absorption was associated with the partition coefficient and the ionization of
the drug [34]. However, studies with salicylic acid and benzoic acid showed that
both drugs were absorbed even in ionized form [34, 35]. For this reason, it is thought
that the partition coefficient and pKa are the main factors affecting nasal absorption,
but other transport routes may also be important for hydrophilic drugs.
9.2 Factors Related to Formulation
9.2.1 pH and Mucosal Irritancy
The pH of the formulation affects nasal drug absorption by two different mecha-
nisms. The first mechanism is directly related to the compatibility between the pH
conditions of the nasal surface and the pH of the formulation. The pH of nasal for-
mulations should be in the range of 4.5–6.5 to avoid any nasal irritation. Because the
lysozyme enzyme located in the nasal mucosa, which is a part of the immune sys-
tem, is active at acidic pHs. At alkaline pH values, the lysozyme enzyme becomes
inactive. Thus, the nose and then the whole body becomes open to infections. In a
possible infection, drug absorption is affected as physiological conditions will
change [36].
The second mechanism arises from the relationship between the pH of the for-
mulation and the pKa value of the drugs. We mentioned that drugs are absorbed at
higher rates in the non-ionized form in the drug-related factors affecting the absorp-
tion section. For this reason, it is important to develop a formulation with a pH range
where the drug is in stable and non-ionized form [26].
9.2.2 Osmolarity
Hypertonic drugs administered by the nasal route cause damage to the structure of
the nasal epithelium. Nasal tissues and mucous membranes cannot fully fulfill their
duties due to this damage and their permeability increases. Thus, the absorption of
drugs increases. However, hypertonic preparation of formulations is not a preferred
method to increase the absorption of drugs. Because prolonged stimulation of the
nasal epithelium and mucosa with a hypertonic agent may cause serious toxicity in
the tissues [26].
9.2.3 Viscosity
Increasing viscosity of the formulation increases the absorption of drugs through

multiple mechanisms. With the increase in viscosity, the mucociliary clearance of
drugs decreases; thus, the contact time of the drug with the nasal tissues is pro-
longed and the absorption value increases [26].
9.2.4 Dosage Form of the Formulation
The dosage form of a drug directly affects the nasal absorption of that drug [26].
Nasal drugs are divided into conventional dosage forms and drug delivery systems.
Although nasal drops, which are one of the conventional dosage forms, are one of
the simplest dosage forms, they have disadvantages such as dripping during applica-
tion and not being able to control the dose easily [37]. Nasal powders can be used in
cases where drug stability cannot be achieved. However, it is an important disadvan-
tage that they cause nasal toxicity [37]. Gels, which are included in conventional
drugs, offer the advantage of increasing the contact time and absorption of the drug
with nasal tissues [26]. Drug delivery systems have been developed to overcome the
disadvantages of conventional dosage forms such as dosing frequency, difficulty in
dose adjustment, low bioavailability, and high systemic side effects.
9.3 Nasal Effect
9.3.1 Membrane Permeability
Membrane permeability is an important factor since nasal absorption of drugs

occurs through the nasal membrane. The nasal membrane, which has a lipophilic
character, acts as an important barrier for hydrophilic drugs and high molecular
weight drugs such as proteins/peptides [38].
9.3.2 Environmental pH
Nasal pH, like formulation pH, is effective in the presence of drugs in ionized and
non-ionized forms depending on the pKa value [39].
9.3.3 Mucociliary Clearance
The nasal mucosa and mucociliary clearance, which are part of the immune system,
prevent the lungs from microbiological agents, allergens, and toxins. However, at
the same time, drugs applied to the nasal cavity also adhere to this mucosa and
decrease the amount absorbed [39].
9.3.4 Cold and Rhinitis
In the case of colds and rhinitis, changes in nasal secretion and vascular width affect
the absorption of drugs [39].
10 Protein/Peptide Drug Delivery: Nose-to-Brain
High molecular weight and hydrophilic drugs such as proteins and peptides cannot
easily cross the BBB, and drug absorption cannot occur at an ethereal level. For this
reason, non-invasive methods have been developed to enable drugs to reach the
CNS by bypassing the BBB. Intranasal drug delivery systems are one of these non-
invasive methods. The most important advantage of nose-to-brain drug delivery is
that the administered drug can be transmitted directly to the brain transsynaptically
[40]. However, the surface area of the nasal epithelium, where drug permeability
occurs, is approximately 150 cm2, which may limit drug absorption. Higher bio-
availability values are obtained if the epithelial permeability can be increased with
a suitable agent to be included in the formulation. This effect can be achieved by
adding an excipient that will increase mucoadhesion into conventional drugs [41].
However, the surface area still constitutes a significant disadvantage for conven-
tional drugs. Carrier systems provide significant advantages in the nose-to-brain
transport of proteins/peptides by increasing the solubility profile of drugs to the
desired range, increasing permeation and penetration into tissues.
10.1 The Importance of Nanocarriers
The advantages offered by carrier systems are many in the transmission of a

protein/peptide to the brain by passing through the BBB after intranasal administra-
tion. The hydrophilic groups in their structures and high molecular weights are the
parameters that reduce the nasal absorption of proteins/peptides. By masking these
properties, drug delivery systems can increase the permeation and penetration of
proteins/peptides into the nasal mucosa, change their size, and facilitate their pas-
sive or active transport [42].
Nose-to-brain drug targeting of protein/peptides from the 2000s to the present is
an issue that pharmaceutical technologists work intensively on and still maintain its
importance. Although many different carrier system types have been tried, mostly
polymeric nanoparticles, liposomes as lipid-based carriers, or hydrogels that
increase mucoadhesion are preferred. Undoubtedly, this situation is related to gain-
ing desired properties to proteins/peptides with nanoparticles, liposomes, and
hydrogels more easily.
Drug delivery systems can be micro- and nano-sized. Particle size is a parameter
that directly affects the ability of the carrier system to pass through the membranes
and be delivered to the target area. For this reason, they can be micro- or nano-sized
according to the target tissue. In studies on the olfactory epithelium morphology, it
has been observed that the membrane pore size in humans is between 100 and
700 nm [43–45]. Still, the nasal membrane pore size differs between species. The
results obtained in studies with drug delivery systems and the data of morphology
studies show compatibility. It has been shown that high permeation and penetration
into the nasal epithelium and mucosa are higher in nano-sized carriers. Nanocarriers
up to 200 nm in size have been found to be effective in different species of in vivo
animal models [42, 45, 46], and nanocarriers up to 100 nm have been reported to be
transported along the olfactory or trigeminal pathway [47]. When the morphologi-
cal, in vitro, and in vivo studies conducted to date are evaluated, it has been con-
cluded that if a drug administered intranasally has a size of 70–150 nm, it can easily
pass through the nasal membranes and be transmitted to the brain [42].
The effect of zeta potential, which is one of the characteristic features of nano-
carriers, on nasal absorption is controversial. Since the nasal mucosa is negatively
charged, it is thought that the penetration and absorption of positively charged nano-
carriers will increase in the nasal mucosa. However, studies on this topic are not
clear enough [42, 48, 49].
It is known that the polymers or lipids used during the preparation of the nano-
carrier also affect nasal absorption. Many studies have shown that nanocarriers pro-
duced using chitosan and cyclodextrins increase the affinity of proteins/peptides to
the nasal mucosa [26]. Similarly, nasal absorption of proteins/peptides increased in
nanocarriers using biodegradable polymers PLA, PLGA, and PCL [46, 50–52]. It
has been proven that this increase reaches even higher values, especially with copo-
lymers obtained from these polymers due to PEGylation also the bioavailability of
peptides increases the in the brain [42]. Lipids and surfactants used in the prepara-
tion of non-polymeric nanocarriers increase the permeability of the epithelial layer
and the absorption of proteins/peptides [53, 54]. Polymers used in the preparation
of nasal hydrogels are known to increase mucoadhesion [26].
10.2 Proteins and Peptides
Proteins and peptides that are frequently used for intranasal administration of drug
targeting to the brain are detailed below.
10.2.1 Insulin
Insulin is generally known as a hormone responsible for regulating glucose metabo-

lism in the body. However, it also supports memory enhancement, synaptogenesis,
and synaptic remodeling [55]. Different drug delivery systems have been developed
to use insulin in the treatment of Alzheimer’s disease (AD), especially since the
changes in the insulin mechanism have been associated with AD [56]. Born et al. in
2002 demonstrated that Insulin and other melanocortin-like peptides were success-
fully transferred to the CSF after intranasal administration in 36 healthy human
volunteers [57]. Although serum levels of proteins/peptides could not be deter-
mined, this work was a breakthrough in 1995 and 1997 when Frey et al. first dem-
onstrated the passage of peptides/proteins through OE [58, 59]. Today, Insulin has
the feature of being the most studied hormone in nose-to-brain drug targeting.
Clinical trials on intranasal administration of Insulin in the treatment of many dif-

ferent neurological and psychiatric diseases such as schizophrenia, multiple sclero-
sis, delirium, different types of dementia, and bipolar disease have been
completed [60].
In one of these clinical trials completed and published in 2019, the effect of intra-
nasal insulin in Parkinson’s disease was evaluated against a placebo. After treat-
ment, the IN Insulin group had a higher overall verbal fluency score than the placebo
group. In addition, IN Insulin was found to improve Unified Parkinson’s Disease
Scale and modified Hoehn-Yahr scale scores. The hypoglycemic attack did not
develop in any of the patients, and it was shown that IN Insulin treatment was well
tolerated. It was concluded that IN Insulin is safe and can provide clinically signifi-
cant functional improvement in patients with Parkinson’s disease and multiple sys-
tem atrophy [61].
In a different study with in vivo animal experiments, Dwivedi et al. developed
Insulin-loaded PEGylated PLGA nanoparticles. The results showed that the
nanoparticles successfully crossed the BBB, delivered Insulin to the brain, and did
not cause hypoglycemia in the peripheral region. The developed nanoparticles are
promising for the intranasal administration of Insulin in the treatment of Alzheimer’s
disease [62]. Picone et al., developed nanogels of Insulin. In in vivo studies, it was
determined that the nanogels did not cause any damage to the nasal mucosa after
application and that Insulin transmission and activity increased in different parts of
the brain [63].
10.2.2 Albumin
Albumin is the most common protein in the body; is synthesized in the liver; and is
found on muscles, skin, sweat, tear film, gastric fluids, bile, and approximately 60%
of the blood. It is responsible for transporting drugs in the body by binding to drugs
through different mechanisms. For this reason, it is frequently used as a targeting
agent in drug delivery systems [64]. In studies to see the effect of intranasal admin-
istration of albumin on drug targeting to the brain, albumin was detected in 11 dif-
ferent brain regions in 5–60 minutes after intranasal administration. In addition, it
has been observed that the transfer of albumin to the brain and blood circulation is
dose-dependent [65].
Katona et al. selected albumin as a carrier agent for targeting meloxicam to the
brain by intranasal administration. It was found that the produced nanoparticles
improved the in vitro dissolution and permeation of meloxicam threefold and five-
fold, respectively. Also, in this study, when Tween80 was used in addition to albu-
min in the preparation of nanoparticles, it was observed that there was a synergistic
effect between both excipients. Thanks to this synergistic effect, fourfold and six-
fold increases in in vitro dissolution and permeation were observed, respectively. In
addition, in vivo studies have demonstrated nanoparticles’ trans-epithelial and axo-
nal transport [66]. In another study, Piazzini et al. produced a model carrier system
by coating albumin with chitosan. In this study, in which the mucoadhesion
properties of nanoparticles were evaluated by turbidimetric and indirect methods, it

was shown that nanoparticles have the advantage of opening tight junctions between
hCMEC/D3 cells by reducing ZO-1 expression levels [67]. In studies with
R-flurbiprofen (R-FP), which is used to treat Alzheimer’s disease (AD), it has been
observed that R-FP-loaded albumin nanoparticles significantly improve basal and
maximal mitochondrial respiration [68].
10.2.3 Oxytocin
Oxytocin is a peptide with known psychological effects. The effectiveness of intra-

nasal oxytocin administration for the treatment of post-traumatic stress disorder has
been proven [69]. It is also the second most nose-to-brain peptide used after insulin
[42]. In in vivo studies, it was found that the nanoparticle structure increased the
passage of oxytocin to the BBB. In addition, it was observed that oxytocin-loaded
nanoparticles administered both acutely and continuously intranasally for 3 days
increased the social interaction of mice [70]. Another study was performed on mice
derived from SCN1A mutations thought to be responsible for social behavior and
epileptic seizures. In this study, it has been shown that oxytocin-loaded nanoparti-
cles provide robust and continuous protection against seizures and restore more
normal social behavior [71]. The results obtained in both studies showed that intra-
nasal administration of oxytocin drug delivery systems has therapeutic potential in
epilepsy and other neurological disorders.
10.2.4 Leptin
Leptin is an endogenous hormone that regulates appetite, energy metabolism, and

glucose uptake in the CNS [56]. It has been shown that leptin administered intrana-
sally easily crosses the BBB and accumulates in the frontal lobe. It also suppresses
appetite and body weight gain and reduces plasma triglyceride levels in rats in
repeated applications. Due to these effects, it is seen as a potential therapeutic agent
for obesity [72]. Duan et al. produced a conjugate of Leptin using Pluronic P85 in
their study with Leptin. It has been observed that this intranasal administered Leptin
conjugate accumulates more in the hypothalamus and hippocampus than natural
Leptin administered intranasally [73].
11 Future Perspectives and Conclusion
Targeting drugs to the brain is very difficult because of the BBB. For this reason,
alternative routes have been developed for drug targeting to the brain. Intranasal
administration is one of the most preferred methods in this field. In fact, the struc-
ture of the nasal mucous membrane does not allow the absorption of every drug in
high amounts. Especially drugs with high molecular weight and hydrophilic groups
in their structure create an obstacle for the nasal absorption of proteins/peptides.
However, the development of suitable drug delivery systems, thanks to nanotech-
nology, creates an important advantage in increasing the nose-to-brain drug delivery
of proteins and peptides and bioavailability in the brain.
With the full discovery of the pathways that proteins and peptides follow after
nasal administration, studies on intranasal drug delivery systems targeted to the
brain have intensified. Since the beginning of the 2000s, many studies in this field
have been registered in the literature. These developments did not only remain in
in vitro or in silico models, but also in vivo animal experiments were carried out.
Some nanocarriers were transferred to phase studies after successful results. These
studies are still ongoing in different phase stages from phase I to phase IV. On the
other hand, there are also completed phase studies. However, when the published
results of the completed studies are examined, it is seen that all of them emphasize
the need for larger clinical studies. On the other hand, there is no brain-targeted
intranasal drug delivery system approved by a health authority such as FDA or
EMA yet. This shows that there is still a long way to go in this field.
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Chapter 11
Novel Mucoadhesive Polymers for Nasal
Drug Delivery
Ljiljana Djekic
Abstract Nasal drug administration is attractive noninvasive approach to rapidly

achieve local or systemic effects of small molecular drugs, proteins, and peptides,
as well as for nasal immunization and nose-to-brain targeted delivery. However,
conventional nasal dosage forms such as nasal drops, sprays, and nasal powders, are
often suboptimal to provide bioavailability higher than 10% of drug molecules with
high lipophilicity, hydrophilicity, or molecular mass, due to the nasal mucociliary
clearance mechanism. The most extensively investigated formulation strategy for
improving nasal bioavailability is the development of mucoadhesive drug delivery
systems. Novel nasal liquid preparations with in situ gelling polymers have been
proposed as a promising concept suitable for convinient instillation, precise dosing,
good spreadability, enhanced nasal retention, and achievement of controlled drug
release. Furthermore, mucoadhesive polymeric nanoparticles (up to 300 nm) have
been evaluated as carriers for increased nose-to-brain bioavailability. This chapter
reviews widely used mucoadhesive polymers and their derivatives considered so far
as components of nasal drug delivery systems, including: natural polymers (chito-
san, cellulose, starch, xanthan gum, gellan gum, pectins, alginates, gelatin) and their
derivatives, synthetic polymers (polyacrylates, polycarbophils), copolymers and
polymer blends (physical mixtures, polyelectrolyte complexes, cross-linked poly-
mers). Many of novel polymers enable enhanced permeation enhancement capacity,
in situ gelling and drug delivery control in response to nasal temperature, pH, ions,
or enzymes, thus providing significant improvements in bioavailability of both
small molecules and macromolecules. The versatility of the concept grows continu-
ously by combining such responsive (smart) polymers or by functionalization of
polymers by targeting ligands. Future prospects in development of mucoadhesive
polymers as pharmaceutical excipients for nasal application are also briefly
overviewed.
L. Djekic (*)
University of Belgrade-Faculty of Pharmacy, Department of Pharmaceutical Technology and
Cosmetology, Belgrade, Serbia
e-mail: ljiljana.djekic@pharmacy.bg.ac.rs
https://doi.org/10.1007/978-3-031-23112-4_11
190 L. Djekic
Keywords Nasal · Mucoadhesive · Polymers · Spreadability · Natural
1 Introduction
Nasal administration of drugs is painless and easily feasible by the patients. The
nasal route is implied when it is necessary to achieve local effects in the treatment
of rhinitis of various origins. Also, increasing interest is put on the development on
nasal vaccines in order to take advantage of the nasal mucosa as a natural entrance
of many pathogens and induce both systemic and local immune response at the
beginning of infection. Moreover, nasal drug delivery has been recognized as a
promising approach for achievement of systemic effects or to target the structures of
the central nervous system (CNS) (i.e., the nose-to-brain delivery). Important
advantages of the nasal mucosa as a site of absorption of drug substances, compared
to the other mucosal barriers (e.g., buccal and different regions of the gastrointesti-
nal tract (GIT)), are lower rates of enzymatic degradation and greater permeability.
Its relatively large surface (150–200 cm2) is highly vascularized, so the drugs can be
absorbed into the systemic circulation relatively fast while avoiding the degradation
in variable pH conditions in the GIT, slow/variable absorption, and first pass metab-
olism. The onset of therapeutic action can be rapid and for some small lipophilic
drugs (e.g., fentanyl, propanolol, progesterone, and pentazocine) occurs in a few
minutes, similar to that obtained after an intravenous injection [1–5]. Also, nasal
mucosa has been intensively explored as a noninvasive alternative route for sys-
temic administration of drugs with poor oral bioavailability such as proteins, pep-
tides, and steroids [6–10]. During the last decade, the delivery of drugs from the
nasal cavity directly to the CNS has been assessed as a promising strategy for cir-
cumventing the blood–brain barrier (BBB). The nose-to-brain delivery takes place
via the olfactory epithelium and olfactory and trigeminal nerve cells situated on the
olfactory region at the loft of the nasal cavity. It can be fast and selective so that the
therapeutic effect is achieved within 3–5 min and the drugs can be delivered into
specific regions of the CNS (e.g., the olfactory bulb, the hippocampus, cerebellum
and brain stem, piriform cortex, amygdale, and hypothalamus). The nose-to-brain
delivery is especially of interest for fast treatment of epileptic conditions and pain
associated with migraine and headaches as well as for targeting of brain tumors and
long-lasting treatments of schizophrenia and chronic neurodegenerative disorders
such as Alzheimer’s disease, Parkinson’s disease, and Huntington’s disease [11–
16]. Several highly potent drugs have been marketed so far in the form of nasal
sprays, as for example zolmitriptan (Zomig®), buserelin (Suprecur®), calcitonin
(Fortical®, Miacalcin®), desmopressin (Desmospray®), ergotamine mesylate
(Migranal®), butorphanol tartarate (Stadol NS®), nafarelin (Synarel®), and oxytocin
(Syntosinon®), as well as nasal powders of sumatriptan (Onzetra Xsail®), beclo-
methasone dipropionate (Teijin Rhinocort®), and dexamethasone cipecilate
(Erizas®).
11 Novel Mucoadhesive Polymers for Nasal Drug Delivery 191
Despite high expectations of nasal delivery to the systemic circulation and the
CNS, studies to date also have shown that the bioavailability of drug substances
through the nasal mucosa is variable and generally lower than 10%, or even less
than 1% in the case of highly lipophilic or hydrophilic drugs, macromolecules, pep-
tides, proteins, and genes [2, 7, 8, 11, 17]. The main factor that reduces the bioavail-
ability of nasally administered drugs is the mucociliary clearance mechanism
(MCCM) [11, 18, 19]. This physiological mechanism enables rapid clearance of
any foreign material from the nasal mucosa into the nasopharynx and further toward
the esophagus. Therefore, MCCM decreases the contact time between the drug sub-
stance and the absorption area. A half-life of nasal clearance on the human respira-
tory epithelium is about 12–30 min, which is not enough for complete absorption of
the majority of drug substances. Methods for improving nasal absorption are pri-
marily focused on overcoming mucociliary clearance. The most extensively inves-
tigated formulation strategy is the development of mucoadhesive nasal drug delivery
systems with optimal spreadability (for liquids and semisolids), flowability (for
powders), or wettability (for powders and nasal inserts) [20]. Common formulations
of nasal preparations are solutions and suspensions for application as nasal drops
and nasal sprays [21]. Their overall retention in the nasal cavity is considerably
shorter compared to semisolids and dry powders. Nasal liquids of low viscosity
quickly leak out from the anterior part of the nasal cavity or clear down to the naso-
pharynx and oropharynx [11]. Increasing the viscosity of the liquid formulation
may prolong the retention at the absorption site and may adversely affect the ease of
instillation. Semisolid formulations are usually hydrogels with suitable rheological
properties to remain in the nasal cavity and increase the contact time with the nasal
mucosa. However, the spreadability of such viscous systems is limited and they are
usually not suitable for spraying. Novel formulation strategy of in situ gelling poly-
mer solutions combines the convenient instillation and precise dosing of a liquid
preparation with the enhanced retention of a viscous hydrogel formed by the sol-gel
transition upon contact of the solution with the nasal mucosa. Such in situ gelling
nasal preparations have certain potential for achievement of controlled-release drug
delivery [22, 23]. Nasal sprays spread on the nasal mucosa finely; however, the
droplet size should range from 30 to 50 μm to avoid deposition at the front of the
nose or at lungs. Olafsson and Gizurarson [24] reported that an increased viscosity
of the nasal solution decreased the spray cone angle and thus allowed drug delivery
to the olfactory region; however, the dose reaching the target site of absorption was
small and the reproducibility was poor. Furthermore, the deposition pattern of the
nasal sprays is significantly affected by intersubject variability regarding the nasal
airway anatomy, particularly in the posterior region which is relevant for the nose-
to-brain drug delivery [25]. The individual nasal resistance to airflow and particle
diameter is of critical importance for deposition of nasal powders. For dry powders,
the optimal size range of microparticles is considered to be 10–30 μm, whereas
smaller particles can escape to lungs, while the larger ones will be retained in the
anterior segment of the nasal cavity. Polymer nanoparticles with particle size
between 10 and 300 nm are promising for nose-to-brain delivery via olfactory path-
way. Although the correlation between the properties of the formulation and the
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therapeutic efficacy of the nasal preparation has not yet been fully elucidated, it has
been demonstrated that the retention time of the nasally administered drugs on the
mucosal surface and nasal bioavailability can be significantly increased by the intro-
duction of polymer excipients that have mucoadhesive properties [11, 21, 25–33].
Widely used macromolecular pharmaceutical excipients considered so far for
nasal drug delivery are natural polymers (e.g., chitosan, alginate, gelatin, or starch);
derivatives of cellulose such as microcrystalline cellulose and cellulose ethers; and
synthetic polymers (e.g., polyacrylates, poloxamers). Mucoadhesive polymers
interact with the mucus layer covering the nasal epithelium. The following are the
main components of the nasal mucus: water (95%), mucin (2.5–3%), electrolytes
(sodium, calcium, and potassium ions) (2%), proteins, lipids, enzymes (mainly
P450 enzyme, rhodanase, glutathione S-transferases, and carboxylesterases), and
antibodies (mainly IG-G and Ig-A). The heterogeneous mucin glycoproteins with
numerous oligosaccharide chains are responsible for the viscoelastic (gel-like)
properties and negative charge of mucus which enable noncovalent interaction
between mucin and hydrated polymer chains [8, 34]. Mucoadhesion sustains the
ciliary movement and thereby increases the residence time of the drug delivery sys-
tem at the absorption site. Moreover, it allows more intimate contact between the
drug delivery system and nasal mucosa and higher local concentration gradient of
the drug substance toward the absorptive membrane [8, 34]. The mechanisms of
mucoadhesion fundamentally involve: (i) wetting and swelling of the polymer; (ii)
intimate contact between the polymer and the nasal mucosa; (iii) penetration of the
polymer into the tissue crevices; and (iv) the interpenetration between the polymer
chains and mucin chains and non-covalent bonding involving hydrogen bonding,
van der Waals interactions, and hydrophobic and electrostatic forces [35]. The eval-
uation of mucoadhesion has shown the differences between the polymers. For
example, mucoadhesiveness is rated as excellent in some cellulose ethers (carmel-
lose sodium, hydroxypropylmethyl cellulose), carbomers, polycarbophils, traga-
canth, and sodium alginate, very good mucoadhesive properties have gelatin, karaya
gum and guar gum, while the mucoadhesiveness of pectin, acacia, chitosan, and
hydroxypropyl cellulose was characterized as good [36]. Mucoadhesive polymers
should possess surface energy properties favorable for spreading onto mucus, high
molecular weight and chain flexibility, positively charged groups, and/or ability to
form hydrogen bonds. It should be considered that the physiological pH-values of
mucin are slightly acidic (5.5–6.5), and its chemical buffer capacity is small, thus
the mucoadhesiveness based on ionic and electrostatic interactions depends also on
the pKa value of the polymer. It is well known that hydration and swelling of par-
ticulate mucoadhesive drug delivery systems may diminish the risk for enzymatic
degradation of the drug substance and provide controlled (sustained) drug release.
In addition, some mucoadhesive polymers (e.g., chitosan, alginates, and cellulose)
cause the dehydration of the epithelial cells and temporal widening of intercellular
tight junctions that improve the paracellular absorption of macromolecules, genes,
and vaccines, but usually the effect of the polymer as a nasal absorption enhancer
offers a limited ability for improvement of absorption of hydrophilic drugs and
macromolecules [30, 31, 34, 37–39]. The variability of physiological pH, ionic
strength, and nasal fluid volume can significantly affect drug delivery performance
of nasal preparations [23]. In this research area, over the last years, attention is paid
to the introduction of novel formulation strategies and polymers with improved
mucoadhesiveness and enhanced permeation enhancement capacity. The main strat-
egies for modulation of the polymer performances include chemical derivatization
of natural and synthetic polymers, synthesis of the copolymers, preparation of inter-
polymer complexes, and physical blends, which are more mucoadhesive and/or sen-
sitive to stimuli in nasal cavity, in comparison with the starting polymer(s). Many of
novel polymers enable in situ gelling and drug delivery in controlled manner in
response to nasal temperature (32 ± 2 °C) (temperature-responsive polymers), pH
(pH-responsive polymer), ions (ion-responsive polymers), or enzymes (enzyme-
responsive polymers) present in mucus [21, 40, 41].
This chapter reviews novel mucoadhesive polymers, including stimuli-responsive
(smart) polymers, that have been used in the development of various nasal drug
delivery systems, their physicochemical properties, mucoadhesion potential, and
in vitro and in vivo nasal drug delivery performances. The differences in nasal drug
delivery potential between the carriers based on novel polymers and polymer com-
binations compared to the starting (parent) polymers are highlighted.
2 Natural Mucoadhesive Polymers and Their Derivatives
Natural polymers such as polysaccharides (chitosan, starch, xanthan gum, gellan

gum, alginates, and pectins) and proteins (albumin, gelatin) have been frequently
applied as pharmaceutical excipients in nasal drug delivery. They have been consid-
ered abundant, relatively inexpensive, non-toxic, biocompatible, and nonirritating
for nasal mucosa. In addition to the adhesion capacity for the nasal mucosa, many
of them show thermo-responsive character. Polyelectrolytes such as cationic chito-
san and anionic sodium alginate, albumin, and hyaluronic acid are pH-responsive
excipients. The suitability of such polymers for in situ gelation and interaction with
the nasal mucus has been tailored by synthesis of various chemical derivatives.
Table 11.1 summarizes examples of natural polymers and their derivatives with
relevance for nasal drug delivery.
2.1 Chitosan and Its Derivatives
Chitosan is a natural linear polysaccharide (50–2000 kDa) obtained by partial

(40–98%) alkaline deacetylation of chitin mainly originating from crustacean shells.
Chitosan molecule consists of monosaccharide units of glucosamine (2-amino-2-
deoxy-β-D-glycopyranose) and N-acetylglucosamine (2-acetamide-2-deoxy-β-D-
glucopyranose) linked by β-(1-4) glycosidic bonds which could be digested by
lysozyme [36, 40, 41, 42]. The value of pKa of the amino groups on the polymeric
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Table 11.1 Examples of natural polymers derivatives evaluated for nasal drug delivery
Mucoadhesive
polymer Derivative Benefit References
Chitosan ChiSys® (chitosan Nasal absorption enhancer [54]
glutamate with mean
molecular weight
~200 kDa and degree of
deacetylation 80–90%)
Chitosan-thiobutylamidine Concentration-dependent moderate [80]
to severe cilio-inhibitory effect; in
situ pH-dependent gelation
Thiolated chitosans Enhanced mucoadhesiveness; [77–79,
permeation enhancement ability; 82–84]
enzyme inhibitory activity; enhanced
nasal absorption of small molecule
drugs and peptides
N-trimethyl chitosan Enhanced nasal absorption at neutral [75,
and alkaline pH 85–89]
N-trimethyl chitosan Formation of nanoparticles suitable [90]
for encapsulation of peptides,
transport to submucosal layers and
enhanced nose-to-brain delivery
N-trimethyl chitosan Coating of liposomes for [91]
achievement of better protein
entrapment efficiency and
mucoadhesive strength and enhanced
penetration through the nasal mucosa
Diethylaminomethyl- Higher drug loading, encapsulation [93]
(diethyldimethylene efficiency, in vitro drug release
ammonium)n profiles and mucoadhesiveness
methylchitosan
Methylpyrrolidinone Better mucoadhesiveness of [94]
chitosan microparticles
N-succinyl chitosan Improved aqueous solubility, nasal [95]
absorption enhancement effect and
increased absolute bioavailability of
isosorbide in rats
Deoxycholate-chitosan- Self-assembly into pH-sensitive [40]
hydroxybutyl spherical nanoparticles which swell
at physiological pH, expand in the
presence of lysozyme, and shrink due
to thermo-response
Cellulose CMC, MCC, HPC Improved nasal availability of small [2, 51,
hydrophobic drugs and hydrophilic 98–102]
macromolecules
Microcrystalline cellulose Enhanced induction of systemic and [103]
(Ceolus® PH grade) mucosal immune responses with
ovalbumin in monkeys
(continued)
Mucoadhesive
Starch Cross-linked starches Control drug release carriers [110, 111]
(microparticles, nanoparticles)
Hydrolyzed potato starch The nasal absorption enhancement of [112]
semisynthetic sodium-human insulin
from microspheres
Xanthan gum The entirely preactivated Improved mucoadhesion and [125]
xanthan thiomer conjugate enhanced stability toward oxidation
Gellan gum Thiolated gellan gum Superior in situ gelling properties in [131]
the presence of physiological cation
concentrations
Aminated gellan gum Enforced interactions to mucus and [23]
temporarily inhibited ciliary beating
Ionic-triggered in situ Increase in adhesiveness and [132]
gelling deacetylated gellan viscosity upon gelling; enhanced
gum solution resveratrol bioavailability via direct
nose-to-brain pathway
Pectins Low methylated pectins Rapid gelation and prolonged [11, 133,
retention in sheep nasal cavity 134]
Alginates Thiomer alginate-cysteine Concentration-dependent cilio- [79]
inhibitory effect
Gelatin Aminated gelatin Increased mucoadhesiveness, nasal [143, 144]
absorption-enhancing effect for
insulin and fluorescein
isothiocyanate-dextran (4.4 kDa) in
rats; sustained drug release
Sperminated gelatin Enhanced nasal absorption of insulin [145]
in rats by modulation of the epithelial
tight junctions
backbone is about 6.5, thus chitosan is insoluble in water at neutral and alkaline pH
values, but soluble and positively charged in weakly acidic solutions in presence of
acetic acid, glutamic acid, citric acid, hydrochloric acid, or lactic acid [41, 43]. The
solubility and charge density of chitosan depend also on the molecular weight, the
degree of deacetylation, and ionic strength of a solution. The higher the ionic
strength the lower the solubility and the positive charges [34]. The positively
charged amino groups of chitosan are available for ionic interactions with the nega-
tively charged residues of the sialic acid of mucus, thus enabling mucoadhesion and
extended residence time of a drug delivery system at the site of drug absorption
[37]. Soluble, protonated chitosan enhances the epithelial permeability by the tran-
sient opening of the tight junctions [21]. These effects significantly promote para-
cellular absorption and transnasal bioavailability of small molecular weight drugs
(e.g., verapamil [44], carvedilol [45], morphine [46, 47], alniditan [48, 49], carbam-
azepine [50], hormones [estradiol] [51]), as well as peptides and proteins (e.g., insu-
lin [52], LHRH-analogue [goserelin] [53]). Glutamate salt of chitosan with a mean
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molecular weight of around 200 kDa and a degree of deacetylation of 80–90%

(ChiSys®) is used as nasal absorption enhancer in Rylomine™ nasal delivery sys-
tem, currently in phase 3 clinical development for rapid delivery of morphine mesyl-
ate to control moderate to severe pain [54]. The permeation enhancement capacity
of chitosan clearly depends on the surrounding pH [55].
Chitosan of different molecular weights and degree of deacetylation has been
used in various mucoadhesive nasal drug delivery systems. The high flexibility of
the chitosan linear chain is favorable for achieving mucoadhesion [56]. The medium
molecular weight chitosan exhibited the strongest mucoadhesion [37]. It is a suit-
able excipient for the preparation of both liquid and particulate carriers for nasal
mucosal delivery. Chitosan may cause a strong flavor in the mouth due to relatively
rapid drainage of nasal solutions into the nasopharynx [57]. The pH-dependent sol-
ubility of chitosan is associated with a risk of reduced polymer solubility at physi-
ological pH of the nasal epithelium. The pH of nasal fluid is slightly acidic (5.5–6.5);
however, it reaches neutral values in inflammation [58]. Such variations in pH value
could diminish solubility and charge of the chitosan molecules. Nazar et al. [59]
have reported that chitosan solutions show the ability of thermoreversible gelling at
pH 6.5–6.9, but the transition from sol to gel at body temperature was slow. Chung
et al. [60] suggested cross-linking of chitosan for formulation of thermo-sensitive
nasal gels for sustained release of insulin. Although no precise conclusions are yet
available, it was demonstrated that chitosan-based particulate drug delivery systems
could improve nasal drug adsorption [61–65]. Chitosan-based microparticles and
nanoparticles are usually applied as nasal dry powder sprays. Particulate formula-
tions in nasal cavity swell and form mucoadhesive gels or polymer matrices that
control the release rate of the incorporated drug (e.g., zolmitriptan [66] and ondas-
etron [67]). Recent study of Akel et al. [68] demonstrated that the use of chitosan as
a coating agent for both poly (lactic-coglycolic acid) and solid lipid nanoparticles
enabled the superior sustained release of meloxicam, mucoadhesive properties, and
in vivo brain distribution of the nanoparticles. Comparative studies have shown that
nasal administration of drugs encapsulated in chitosan-based microparticle carriers
achieved greater bioavailability compared to the corresponding solutions and
nanoparticles, probably due to longer maintenance in the nasal cavity [69]. Chitosan
has been studied also for the preparation of the antigen-loaded microparticles and
nanoparticles for nasal vaccine formulations [70–72]. It shows an adjuvant effect
busting the systemic and mucosal immune reactions in nasal vaccines, for example,
against Bordetella pertussis filamentous hemagglutinin, recombinant pertussis
toxin, and influenza virus vaccines [73, 74]. In addition, micro- and nanoencapsula-
tion of antigens may protect them from enzymatic degradation, while they may be
released from the carrier in the nasal cavity, or the insoluble particles with a diam-
eter less than 5 μm enable their transport across the nasal mucosa and toward the
nose-associated lymphoid tissue (NALT) cells to achieve the local immune response
[75]. Upon cellular uptake, particles may get into the compartments with lower pH
which would induce the carrier dissolution and release of the antigen [27].
Aqueous solubility, charge density, mucoadhesion, nasal absorption enhancement,
and thermosensitivity of chitosan can be improved by the synthesis of different
derivatives via chemical functionalization of the amine and hydroxyl groups [34,
36]. Representative examples described in available literature are: thiolated
chitosans, quaternized chitosans, methylpyrrolidinone chitosan, N-succinyl
chitosan, deoxycholate-chitosan-hydroxybutyl, and methylchitosans.
Thiolated chitosans (thiomers) are conjugates formed by establishing a chemical
bond between the amino group of chitosan and a thiolating reagent (e.g., thiogly-
colic acid, 2-iminothiolane). Such derivatives of chitosan were developed for the
first time and intensively investigated for the nasal drug delivery by the group of
Bernkop-Schnürch [76]. Generally, thiomers are recognized as excipients with in
situ gelling ability as well as superior permeation enhancing effect and mucoadhe-
siveness. The thiolated polymers interpenetrate the mucus gel layer resulting in an
intimate contact with the nasal mucosa, a higher drug concentration on the mucosa
and thus facilitating nasal absorption [76, 77]. Mucoadhesive properties and resi-
dence time on the nasal mucosa of thiolated chitosans are superior over the unmodi-
fied polymer due to a thiol/disulphide exchange reaction between the thiomers and
cysteine-rich subdomains of mucus glycoproteins [78]. Palmbereger et al. [79]
observed that gels with 1–2% of chitosan-thiobutylamidine, which was synthesized
by attaching iminothiolane to chitosan, had the concentration-dependent moderate
to severe cilio-inhibitory effect, followed by a partial recovery in ciliary beating.
Thiolated chitosans are pH-sensitive polymers. In situ pH-dependent gelation based
on the conversion of thiol groups of the polymer to thiolated ions available for for-
mation of intermolecular and intramolecular disulphide bonds at physiological pH
values, is typical for the chitosan thiomers. The gels based on thiolated chitosan are
viscoelastic and their elastic properties at pH 5.5 were found to increase signifi-
cantly with the degree of thiolation [80]. Furthermore, thiolated chitosans exhibit
permeation enhancement ability and enzyme inhibitory activity and hence signifi-
cantly enhance the nasal absorption of small molecules as well as peptide drugs
[77–79]. In a study, Patel et al. [81] evaluated the effect of thiolated chitosan
nanoparticles on the nose-to-brain distribution of tizanidine from the nasal cavity of
mice in comparison to drug solutions and drug-in-chitosan nanoparticles. The high-
est brain uptake and the most pronounced antinociceptive effect was found for the
thiolated chitosan nanoparticles, likely due to the better mucoadhesiveness of this
chitosan derivative. In the later study, the same group demonstrated enhanced anti-
hyperalgesia of cyclobenzaprine hydrochloride encapsulated in thiolated chitosan
nanoparticles in comparison with the control. Shahnaz et al. [82] demonstrated the
higher absolute bioavailability of leuprolide from the nasal cavity of rats when the
drug was encapsulated into the thiolated chitosan-based nanoparticles (18.5%) in
comparison with the nanoparticles prepared from unmodified chitosan (4.3%) and
the leuprolide solution (2.6%). Nazar et al. [83] observed the higher bioavailability
of insulin from dry microparticles (1.4–80 μm) consisted of chitosan-
thiobutylamidine (6.9 ± 1.5%) as compared to the dry chitosan microparticles
(4.2 ± 1.8%), after nasal administration to rats. Insulin loaded chitosan-
thiobutylamidine microparticles swelled 4.39 ± 0.52-fold and showed controlled
release of insulin over 6 h. It was suggested that the improved absorption enhancing
effect was due to the higher mucoadhesiveness of the thiolated chitosan
198 L. Djekic
formulation. N-acetyl-L-cysteine grafted chitosan (chitosan-NAC) is a thiomer used

for formulation of positively charged nanoparticles (140–210 nm) for intranasal
delivery of insulin [84]. The drug loading capacity of the nanoparticles ranged from
13% to 42%. Mucoadhesiveness of the chitosan-NAC nanoparticles was 1.8 fold
higher in comparison with the unmodified chitosan. This was explained by the
building of disulphide bonds between the sulfhydryl (thiol) groups of chitosan-NAC
and nasal mucosa components, which consequently “anchor” the system to the
mucosa. This likely enabled an increased level of insulin absorption and a larger
decrease of glycemia (40%) as compared to unmodified chitosan nanoparticles
in vivo (rats).
N-trimethyl chitosan (TMC) is the most thoroughly investigated quaternized
derivative of chitosan with a capacity to enhance nasal absorption even at neutral pH
[85]. The degree of quaternization of TMC correlates with the amount of positive
charges and thereby influences the extent of nasal absorption improvement [86].
Hydrosoluble TMC hydrochloride, with high (61.2%) and low (12.3%) degrees of
quaternization at two polymer concentrations (0.25% w/v and 0.5% w/v), were con-
sidered for the enhancement of nasal delivery of insulin in rats. The study high-
lighted that at pH 4.4 all investigated polymers increased insulin absorption, while
at pH 7.4 only the more quaternized TMC was able to enhance the absorption [75,
87]. Junginger et al. pointed that TMC with the quaternization degree higher than
25% has a pH-independent solubility in water, comparable absorption enhancement
efficiency in Caco2 cells to unmodified chitosan at acidic pH, and, in contrast to
unmodified chitosan, also performs absorption enhancing effect at pH higher than 7
[75, 88, 89]. Amidi et al. reported the use of TMC with 25% degree of quaterniza-
tion to obtain positively charged nanoparticles (about 350 nm) loaded with ovalbu-
min. The nanoparticles did not exhibit in vitro cytotoxicity in Calu-3 cell lines. In
vivo studies in rats showed that albumin-loaded nanoparticles can cross the mucosal
layer and be taken up by nasal epithelia and NALT cells, and transported to sub-
mucosal layers. Encapsulation of the analgesic peptide leucine-enkephalin in TMC-
based nanoparticles enables enhanced nose-to-brain delivery and higher brain
concentration in mice in comparison with nasally administered marker solution.
The observed enhancement in brain uptake was followed by improvement in
observed antinociceptive effect of leucine-enkephalin [90]. Chen et al. [91] devel-
oped spray-dried bovine serum albumin (BSA)-loaded liposomes coated with TMC,
for nasal administration as a dry powder. BSA was used as a model antigen.
Uncoated liposomes are easily removed from nasal mucosa by MCCM. Coating
with TMC gave better results for entrapment efficiency, glass transition (Tg), and
mucoadhesive strength in comparison with liposomes coated with alginate or chito-
san as well as uncoated liposomes. In spite of the reduction of the encapsulation
efficiency by coating, the encapsulated BSA levels were maintained at 60–69%, and
its structural integrity and release profiles were consistent. The TMC-coated lipo-
somes also showed enhanced penetration through the nasal mucosal tissue over
uncoated liposomes.
Mei et al. [92] assessed and compared permeation enhancement of chitosan,
thiolated chitosan, and TMC for the nasal absorption of tetramethylpyrazine
phosphate in rats. Both chitosan derivatives are soluble at basic pH values. Chitosan
of around 100 kDa was the most promising absorption enhancer and thiolation did
not increase the absorption of the drug above that of unmodified chitosan, whereas
TMC did possess a stronger enhancing efficiency at pH 6. Rassu et al. [93] described
the quaternary ammonium chitosan derivative diethylaminomethyl-
(diethyldimethylene ammonium)n methylchitosan for nasal formulation of micro-
spheres loaded with the macrolide rokitamycin intended for nose-to-brain delivery
in treatment of granulomatous amoebic encephalitis. The study showed the higher
drug loading (up to 22.7 ± 1.4 w/w) and encapsulation efficiency (up to 94.5 ± 3.7
w/w) of the derivative in comparison to the unmodified chitosan. In vitro drug
release profiles and mucoadhesiveness of the derivative-based microspheres were
similar or better than those based on unmodified chitosan. The investigated methyl-
chitosan derivatives did not cause any cytotoxicity effect compared to human umbil-
ical vein endothelial cells.
Gavini et al. [94] used hydrosoluble derivative methylpyrrolidinone chitosan
(MPC) for development of the microspheres (about 10 μm) loaded with metoclo-
pramide hydrochloride. In in vitro release test, the drug was completely released
after 30 min from both MPC and unmodified chitosan microspheres. Although
swelling capacity of MPC microparticles was lower, they showed better mucoadhe-
siveness. The swelling capacity was significantly affected by the drug content for
both types of microparticles. The ex vivo drug permeation through sheep nasal
mucosa was slower, achieving 40% for MPC microparticles, 80% for the pure drug,
and 60% for unmodified chitosan based microparticles, after 3 h.
N-succinyl chitosan is a hydrosoluble derivative of chitosan obtained by inclusion
of succinyl groups on the N-terminal glucosamine units of the parent polymer. Na
et al. [95] showed improved aqueous solubility and nasal absorption enhancement
effect of isosorbide in rats, in presence of this derivative versus unmodified chitosan.
The absorption enhancing efficiency of the investigated nasal solutions increased
with increase in degree of chitosan substitution within the investigated range.
Although Cmax was higher for the unmodified chitosan, the absolute bioavailability
achieved in the presence of the derivative was 69.9% while the one achieved in
presence of the unmodified chitosan was 55.4%.
Sun et al. [40] synthesized amphiphilic derivative deoxycholate-chitosan-
hydroxybutyl (DCCH) from chitosan with deacetylation degree of 85% and average
molecular weight of 700 kDa by carbodiimide reaction. DCCH of various degree of
substitution of deoxycholate and cetirizine groups was obtained by conjugating
these groups to the amino groups of previously prepared hydroxybutyl-chitosan.
Amphiphilic DCCH could self-assemble into spherical nanoparticles which swell at
physiological pH due to the pH-sensitivity of the polymer, expand in the presence
of lysozyme, and shrink due to thermo-response at 33 °C and pH 5.5. DCCH with
covalently grafted hydrophobic H1-antihistamine cetirizine was used to prepare
nanoparticles. The drug causes irritation of the nasal mucosa, so the application in
the nasal cavity is limited. The cetirizine grafted DCCH was used to obtain stimuli-
responsive nanoparticles with free cetirizine encapsulated. Positively charged
nanoparticles (~120 nm) were prepared and their lower critical solution temperature
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(CST) was variable (29, 33, and 37 °C). Under physiological conditions (pH 5.5,
33 °C) the diameters of the nanoparticles increased slightly (~129 nm to ~134 nm)
because of the pH-responsive expansion of the polymer. The free cetirizine release
was burst (desirable for quick relief of allergic symptoms) and the mechanism of the
process was thermo-sensitive drug squeeze out of the nanocarrier. The drug release
amount was significantly different among the nanoparticles with different CST val-
ues. Furthermore, incubation of nanoparticles with lysozyme (30 μg/ml) caused
swelling with ~two-fold increase of sizes, while covalently grafted fraction of the
drug (~5%) was released additionally due to the digestion of chitosan backbone,
which could prolong the anti-allergic onset time.
2.2 Cellulose Derivatives
Cellulose is a most abundant natural polysaccharide containing 8000–10,000

glucose units linked by β-1,4 glucosidic bonds. Chemical modification of the
hydroxyl groups on the glucose residues yielded a number of nontoxic derivatives
which are extensively used as pharmaceutical excipients for different administration
routes including nasal route. In nasal formulations, the use of water-soluble
derivatives such as hydroxypropylmethyl cellulose (HPMC) (also known as
hypromellose), carboxymethyl cellulose (CMC) and its sodium salt (NaCMC),
hydroxypropyl cellulose (HPC), methyl cellulose (MC), hydroxyethyl cellulose
(HEC), as well as poorly water-soluble derivatives such as microcrystalline cellulose
and ethylcellulose (EC) has been reported. These cellulose derivatives show
mucoadhesion and prolong the residence time of the drugs in the nasal cavity. Their
mucoadhesion depend on the type of derivative and the molecular weight. Although
some authors point out that the mucoadhesion of hydrosoluble cellulose derivatives
is of limited importance [21, 34, 51], they have gel-forming properties and due to
high viscosity of such gels the drug release is usually sustained. NaCMC is an
example of polyanionic derivative of cellulose whose aqueous solutions of high and
medium viscosity show thixotropic behavior [96]. HPMC is a thermosensitive
polymer whose solutions form gels at the critical temperature. HPMC gels show
mucoadhesive character due to strong hydrogen bonding with the mucin and provide
sustained drug release [36]. El-Gizawy et al. [96] showed that mucoadhesive nasal
gels of HPC (1%) or MC (1.5%) can enhance the rate of absorption of oxybutynin
chloride. HPC gel showed the best release profile, optimum viscosity, and best
mucoadhesive characteristics. The mucoadhesive properties of HPC and MC gels
were similar and likely based on the hydrogen bond formation between the mucous
membrane and free hydroxyl groups of the cellulose derivatives. A correlation
between the viscosities of the nasal gels and their mucoadhesiveness was
insignificant, indicating that the prolonged retention could be attributed to their
mucoadhesiveness rather than their viscosities [97].
The potential of cellulose derivatives to improve nasal availability has been
demonstrated in small hydrophobic drugs and hydrophilic macromolecules [51, 98,
99]. In vivo study in rabbits showed that the relative availability of apomorphine,
administered nasally with CMC, was 102% compared with subcutaneous injection
[100]. Also, administration of ketorolac tromethamine with MCC enabled absolute
bioavailability up to 90.77% [101]. Several studies have described improvements in
the nasal bioavailability of insulin, calcitonin, and leuprolide from microspheres
based on microcrystalline cellulose, alone or with HPC [2, 102]. In the study of
Suzuki and Makino [99] leuprolide, salmon calcitonin and FITC-dextran (FD4
peptide), when administered nasally in rabbits with MCC/HPC, reached an absolute
bioavailability of 34.9%, 16.4%, and 35.5%, respectively. Torikai et al. [103]
investigated a proprietary nasal powder carrier with a mean particle size of 50 μm
based on microcrystalline cellulose (Ceolus® PH grade) combined with ovalbumin
for applicability in monkeys to induce systemic and mucosal immune responses.
The nasal ovalbumin powder showed comparable and higher antigen-specific IgG
antibody titer to an injection and nasal liquid formulation, respectively, while the
antigen-specific IgA antibody response was detected only for the nasal ovalbumin
powder. Therefore, the technology based on microcrystalline cellulose has been
considered promising for nasal vaccines enabling both a mucosal immunity response
as the first line of defense and systemic immunity response as a second line of
defense against infection. Some authors have pointed to the limited capacity of
cellulose derivatives such as MC, HEC, EC, and HPC, to improve nasal bioavailability
and the need to include permeation enhancers such as a mucolytic agent, N-acetyl-
L-cysteine (NAC), polysorbate 80 or Azone®, for absorption enhancement of human
parathyroid hormone [104], insulin [105], ciprofloxacin [106], and dopamine [107].
Hence, the data published so far on the achieved improvement in the bioavailability
of nasally administered drugs are inconsistent for defining final conclusions on the
potential of carriers based on cellulose derivatives.
2.3 Starch and Its Derivatives
Starch is a polysaccharide comprising glucose monomers which are linked so as to

form a linear polymer amylose and a branched polymer amylopectin. Amylopectin
consists of a linear chain and a number of side chains usually in sequences of
25 units, which may be branched further. Glucose units are linked in a linear way
via α(1 → 4) glycosidic bonds, while the branching is accomplished by the estab-
lishment of α(1 → 6) and likely α(1 → 3) bonds. The branching in amylopectin
makes its molecule soluble and susceptible to enzymatic degradation. Starches from
different natural resources are widely used in pharmaceutical products, and among
them corn starch is the most preferred [108]. By modifying the hydroxyl groups,
processed starches, such as drum-dried waxy corn starch, carboxy methyl starch,
sodium starch glycolate, soluble starch, and degradable starch, were obtained.
Generally, starch polymer chains only interact weakly with mucin. The drum-dried
waxy corn starch showed better bioadhesion properties compared to starch pro-
cessed by other methods [109].
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Cross-linked starches swell in aqueous media (including nasal mucus) and form
gel-like systems which control the drug release. Yadav and Mote [110] prepared
domperidone-loaded cross-linked starch microspheres for intranasal administration.
The microspheres (22.8–102.63 μm) were prepared by the emulsification cross-
linking technique using epichlorhydrine as a cross-linking agent. The microparti-
cles showed good mucoadhesive property and swelling behavior. The strength of
mucoadhesion depended on the degree of cross-linking. The formulation variables
(drug concentration and polymer concentration) affect in vitro drug diffusion
through the sheep nasal mucosa. The release amount of domperidone ranged from
73.11% to 86.21%. Domperidone was released from the microspheres in a sustained
release manner, and it was directly proportional to the concentration of the drug.
Jain et al. [111] prepared cross-linked starch nanoparticles loaded with insulin. The
nanoparticles were prepared with different cross-linkers and using emulsion method
and gel method. Emulsion cross-linked nanoparticles (351 nm) were smaller in size
compared to those prepared by gel method (997 nm), and also size was further
reduced by utilization of epichlorohydrin (194 nm) as a cross-linking agent com-
pared to phosphoryl trichloride (810 nm). A size-dependent first-order diffusion-
controlled insulin release with an initial burst effect was found. In vitro release rate
was higher for nanoparticles of smaller size and least cross-linking. Small size of
the nanoparticles increased the surface area, the insulin release rate, and the concen-
tration gradient across the nasal epithelium. The absorption was increased to the
greatest extent in the nanoparticles with sodium glycocholate which produced 70%
reduction of plasma glucose and hypoglycemic effect over 6 h post-administration.
Illum et al. [112] prepared degradable starch microspheres (48 μm) by emulsion
polymerization of hydrolyzed potato starch loaded with semisynthetic sodium-
human insulin and compared the effect of different bile salts (lysophosphatidylcho-
line, glycodeoxycholate, and sodium taurodihydroxyfusidate) as absorption
enhancers in sheep. Dependent on the potency of the enhancer system, the absorp-
tion enhancement was shown to be from 1.4 to 5 times higher than that obtained for
the corresponding solution of absorption enhancer. The highest nasal bioavailability
of insulin was observed from the microspheres administered along with glycode-
oxycholate. The starch microspheres were shown to synergistically increase the
effect of the absorption enhancers on insulin transport across the nasal membrane.
2.4 Xanthan Gum and Xanthan Thiomers
Xanthan gum is a natural extracellular polysaccharide (exopolysaccharide) produced

by a Gram-negative bacteria genus Xanthomonas. This branched polysaccharide
consists of a main linear chain of β- (1,4)-D-glucose (that is similar to cellulose
backbone) and trisaccharide side chains consisting of two mannose residues with a
glucuronic acid residue between them (in a 2:2:1 molar ratio). The mannose-(β-1,4)
glucuronic acid-(β-1,2)-mannose side chains are attached to alternate glucose
residues in the main chain by α-1,3 linkages [113–117]. D-mannose unit linked to
the main chain contains an acetyl group at position O-6, while approximately one-
half of the terminal D-mannose contains a pyruvic acid residue linked via keto
group to the 4 and 6 positions, with an unknown distribution. The molecular weight
distribution of xanthan gum ranges from 2 × 106 to 20 × 106 g/mol. It is stable at
different temperatures and under both acidic and alkaline conditions, swells in
water, and dissolves slowly [114, 116–119]. In aqueous solutions at pH > 4.5 it has
polyanionic characteristics [118, 120]. Although aqueous solutions show weak gel-
like properties originating from relatively weakly bound polymer molecules by
hydrogen bonds [118, 121], however, a large number of hydroxyl and carboxylic
groups of xanthan gum molecule are available for chemical cross-linking, conjuga-
tion with other polymers (polysaccharides), and chemical grafting. The concept of
thiolated polymers provides excipients with improved adhesiveness for nasal
mucosa by disulfide bond formation, but they are prone to intramolecular oxidation,
particularly in solutions at pH > 5 [76, 122, 123]. Oxidation limits stability and
sprayability of liquid nasal dosage forms. The strategy of entirely preactivated thi-
omers, which do not undergo intramolecular oxidation reactions while retaining
gel-forming properties and adhesiveness toward mucosal thiol groups, has been
introduced by Hintzen et al. [124]. The entirely preactivated xanthan conjugate
(PXC) was synthesized by carbodiimide-mediated amide bond formation between
primary amino groups of the ligand 2-((2-amino-2-carboxyethyl)disulfanyl)nico-
tinic acid (i.e., 2-mercaptonicotinic acid coupled to L-cysteine by disulfide exchange
reaction) and the carboxylic acid group of xanthan gum backbone. Afterwards, car-
boxylic acid moieties were activated by adding N-(3-dimethylaminopropyl)-N0-
ethylcarbodiimide hydrochloride. The reversibility of the nasal ciliary beat
frequency of porcine nasal epithelial cells pointed sufficient nasal safety of
PXC. The liquid nasal formulation with PXC showed improved mucoadhesion as
well as enhanced stability toward oxidation. Rheological investigations of polymer
mucus mixtures revealed a 1.7-fold and 2.5-fold enhanced mucoadhesion of PXC
compared to thiolated xanthan and unmodified xanthan. Tensile force evaluation
reported a 2.87-fold and 5.11-fold higher total work of adhesion and a 1.63-fold and
2.41-fold higher maximum detachment force of PXC compared to thiolated xanthan
and unmodified xanthan. The viscosity and in situ gelling behavior of the PXC-
based liquid formulation can be furthermore customized by the addition of polyva-
lent cations [125].
2.5 Gellan Gum and Its Derivatives
Gellan gum is a linear exopolysaccharide produced by Sphingomonas elodea. It is

composed of 1,3-β-D-glucose, 1,4-β-D-glucoronic acid, 1,4-β-D-glucose, and
1,4-α-L-rhamnose repeating units in molar ratios of 1:2:1 [23]. This anionic poly-
mer has the characteristic property of temperature-dependent and ion-induced in
situ gelation. A three-dimensional network responsible for gelation forms upon
complexation with cations, as being present in body fluids like nasal fluid, and
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formation of double helical junction zones due to hydrogen bonding with water
[126, 127]. The study of Cao et al. [128] demonstrated decreased symptoms of
motion sickness in rats by nasal delivery of scopolamine hydrobromide from in situ
gelling formulation with gellan gum, in comparison with subcutaneous and oral
administration, due to prolonged retention of gel in nasal cavity. However, the
mucoadhesive properties of gellan gum gels are based on weak hydrogen bonds and
van der Waals forces, which may limit the extent of retention. Moreover, the vol-
ume, ionic composition, and pH of nasal fluid are critical for drug delivery perfor-
mances of the gellan gum-based systems [129, 130].
Krauland et al. [131] developed a thiolated derivative of gellan gum by linking
covalently l-cysteine to deacetylated gellan gum via carbodiimide chemistry.
Rheological characterization showed that the deacetylated gellan gum-cysteine con-
jugate was capable of forming inter- and/or intramolecular disulfide bonds in aque-
ous solution (1.5% m/m) at pH 7, in the presence of physiological cation
concentrations displaying superior in situ gelling properties compared to unmodi-
fied polymer.
Jelkman et al. [23] proposed derivatization of gellan gum by reductive amination
in order to intensify mucoadhesiveness and prolong the residence time. Positively
charged aminated gellan gum derivatives were obtained by introducing amino
groups to the polymeric backbone (Fig. 11.1). In a previous step, oxidation under
addition of periodate was performed, which evokes ring opening and thus enables
higher chain flexibility contributing to improved adhesive interactions. The synthe-
sized gellan gum derivatives revealed coupling rates of amino groups up to
1259.50 ± 75.98 μmol/g polymer, accelerated hydration and superior mucoadhesion
Fig. 11.1 Pathway for synthesis of aminated gellan gum derivative by introduction of primary
amino groups via oxidative cleavage and subsequent reductive amination with ammonia. (With
permission from Ref. [23])
in presence of divalent cations, compared with the unmodified polymer, including a

32-fold increase in viscosity of polymer/mucus mixtures, a 14-fold extended muco-
sal adhesion time, a 9-fold higher total work of adhesion, and a 3.75-fold elevated
maximum detachment force. Synthesized derivatives exhibited enforced interac-
tions to mucus enabled by the attached amino groups. Ciliary beating of excised
porcine ciliated epithelial cells in presence of the gel was temporarily inhibited and
completely recovered after wash-off. Although the gellan gum derivative showed
sufficient nasal safety along with prolonged mucoadhesion, prior pharmaceutical
use should be considered a proper balance between amination and polymer degra-
dation, as side reactions of periodate oxidation.
Hao et al. [132] formulated a composite in situ gelling formulation comprising
nanoparticles of resveratrol in an ionic-triggered deacetylated gellan gum (DGG)
solution for intranasal delivery. DGG formed ionic-sensitive solution which trans-
forms into a gel matrix in the nasal cavity. Optimal gelling ability and viscosity
were observed at 0.6% w/v of DGG. In situ DGG-based gels showed an increase in
adhesiveness and viscosity compared to the DGG solution. Bioavailability in the
brain achieved by intranasal resveratrol-loaded in situ gel formulation was 2.88-
times enhanced, likely via direct nose-to-brain pathway, bypassing the BBB.
2.6 Pectins and Methylated Pectins
Pectins are ionic polysaccharides comprising a backbone of galactosyluronic acids

units linked by α-1,4 bonds with neutral sugars such as galactose, xylose, rhamnose,
and arabinose, either in the backbone or as side chains. A linear chain of 1,4-linked
α-D-galactosyluronic residues is homogalacturonan. It contains large neutral
regions. Some of the carboxyl groups of homogalacturonan are methyl esterified
and they may also be O-acetylated at the C-2 and C-3 positions. Methyl esterifica-
tion is common in native pectins and a random distribution of the methyl esters
groups over the galacturonan backbone was found. In commercially extracted pec-
tins the distribution depends on the raw material and the extraction and de-
esterification conditions. Low methylated pectins (LMP) have a degree of
esterification (DE) ≤50%, while high methylated pectins (HMP) have DE >50%.
Gelling of pectins is significantly affected by degree of esterification. LMP can form
gel in the presence of divalent cations, such as calcium, following a similar mecha-
nism of gellan gum due to the formation of intermolecular junction zones by side by
side association of homogalacturonic smooth regions of different chains. HMP form
gel with sugar and acid. LMP gelled rapidly after administration in the nasal cavity
of sheep and retained for an extended period of time [133, 134]. The example of
commercially available ion responsive in situ gelling nasal drug delivery is PecFent®
(fentanyl citrate) nasal spray which uses Archimedes Pharma’s patented drug deliv-
ery system PecSys™. It is the LMP-based drug delivery system designed for admin-
istration as fine mist spray which forms gel upon contact with the nasal mucosa.
This system enables fast onset of action [135]. The gelling of the product prolongs
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the delivery of fentanyl in a controlled manner as well as absorption of the drug, but
does not increase the bioavailability [37]. PecFent® exhibited better tolerability pro-
file when compared with fentanyl, chitosan and fentanyl in chitosan-poloxamer 188
system, as well as superior pharmacokinetic profile compared to transmucosal oral
fentanyl citrate [136]. Pectin-based liquid nasal formulations are rated as controlled
drug release vehicles; however, these polymers showed low effect on drug mole-
cules transport in cell monolayers, as demonstrated for the model substances man-
nitol and propranolol [11].
LMP-based liquid nasal formulations have been shown to be promising for
achieving mucoadhesion in the olfactory region. Charlton et al. [11] investigated
nasal drug delivery systems prepared with LMP. It was found that all formulations
were able to reach the olfactory region in the nasal cavity of human volunteers when
delivered using a simple nasal drop device whereby the volunteer is in the supine
position with the head tilted back. Moreover, the formulations displayed a signifi-
cantly increased residence time on the olfactory epithelial surface, potentially
enhancing the delivery of drugs to the brain, in contrast to a non-bioadhesive control
administered with the same device. Also, the pectin formulation administered with
a nasal spray system did not show an increase in residence time in the olfac-
tory region.
2.7 Alginates
Alginates are salts of anionic linear polysaccharides consisting of varying ratios of

glucuronic and manuronic acid units [137]. Sodium alginate is the purified product
obtained by alkaline extraction from brown seaweed. It dissolves in water, slowly
forming a viscous, colloidal solution. The solution of this hydrosoluble monovalent
salt undergoes sol-gel transition upon addition of divalent ions (e.g., calcium) [138].
Alginate gels with high glucuronic acid contents are more rigid and porous in com-
parison with gels comprising alginates with low glucuronic content, which are more
randomly packed and less porous [139]. Mucoadhesiveness of alginates is relatively
poor because it is based on formation of hydrogen bonds with mucin-type glycopro-
teins through carboxyl–hydroxyl interactions [140, 141]. It can be improved by
covalent attachment of L-cysteine to the polymeric backbone. Thiomer alginate-
cysteine exhibited a concentration-dependent cilio-inhibitory effect. The observed
effect was moderate and followed by a partial recovery of ciliary beat frequency at
the polymer concentration of 1%, while at 2% cilio-inhibition was severe and par-
tially reversible [79]. In spite of promising observations, alginate thiomers have
been scarcely investigated for the nasal drug delivery.
2.8 Gelatin and Its Derivatives
Gelatin is a fibrillar protein derived from collagen. The product of acidic collagen
hydrolysis is gelatin A, while the product of alkaline treatment of collagen is gelatin
B. The molecular weight is about 75 kDa. Gelatin dissolves in water forming ther-
mally reversible and pH-sensitive gels. The gelation temperature is about 35 °C and
may vary with the pH of the solution [34]. Morimoto et al. [142] have prepared gela-
tin microspheres (10.9 μm) with negative charge and microspheres (3.4–71.5 μm)
with positive charge using basic gelatin [isoelectric point (IEP) value of 5.0] and
acidic gelatin (IEP = 9.0), respectively. The adhesion to the nasal mucosa of posi-
tively charged gelatin (PCG) microspheres was significantly higher than that of
negatively charged gelatin (NCG) microspheres. The nasal absorption of salmon
calcitonin in rats was enhanced by both types of gelatin microspheres; however, the
hypocalcemic effect after administration of PCG microspheres of 11.2 μm was sig-
nificantly greater than that of NCG microspheres of the same size. The hypocalce-
mic effect of the NCM microspheres tended to appear more slowly and last longer
compared to that of PCG microspheres.
In order to enhance the nasal drug delivery potential of gelatin, aminated
derivatives were prepared and investigated. Wang et al. [143] prepared aminated
gelatin by cationization of gelatin with ethylene diamine. Positively charged
aminated gelatin can interact with negatively charged mucin, thus upgrading the
mucoadhesiveness. Moreover, it showed the absorption-enhancing effect, in both
liquid and powder dosage forms, for insulin and fluorescein isothiocyanate-dextran
(4.4 kDa) in rats. The hypoglycemic effect was affected by the pH of the formulations
and the concentration of aminated gelatin. Aminated gelatin showed a concentration-
dependent (0.1–0.4%) but relatively small effect on the lactate dehydrogenase
leaching in an in situ perfusion rat nasal epithelial membrane model. In the continu-
ation of the research, aminated gelatin microspheres (AGM) were prepared and
investigated as a nasal delivery carrier for fluorescein-labeled insulin and dextran
(4.4 kDa) [144]. Insulin release from the microparticles was significantly slower
than from native gelatin microspheres (GMS); however, the release of dextran from
both AGM and GMS was fast, likely due to the different electrostatic interactions
between the model drugs and the microspheres. The plasma glucose levels of
healthy rats following intranasal administration of insulin-loaded AGM showed that
AGM could significantly increase the absorption of the drug when administered in
a dry powder formulation, but no significant hypoglycemic effect was observed
when given in suspensions. The absorption enhancement effect was ascribed to the
hydration of AGM with water from nasal mucosa resulting in a temporary dehydra-
tion of the epithelium membrane and opening of the tight junctions. Indirectly,
mucoadhesiveness contributed to the absorption enhancing effect. In a similar study,
sperminated gelatin (SG) also enhanced the nasal absorption of insulin in rats by
modulation of the epithelial tight junctions [145]. SG was prepared by the addition
of spermine, which is a small organic polyamine. In Calu-3 cell monolayer perme-
ation experiments, SG showed significant enhancing effects on
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Table 11.2 Examples of synthetic polymers and their derivatives evaluated for nasal drug delivery
Mucoadhesive
Polyacrylates Carbomers pH-responsive/sensitive polymers; [21, 26, 36,
swelling upon hydration; suitable for 100, 147–151]
achievement of controlled
(sustained) drug release; enhanced
nasal bioavailability of drugs
Polycarbophil Sustained drug release and increased [152–154]
thiomers drug uptake by nasal mucosa
Miscellaneous Polyvinylacetal pH-responsive and thermo- [155]
synthetic diethylamino acetate responsive gelling on the rat nasal
polymers mucosa to control the drug release
Polyamidoamine Enhancement of aqueous drug [156–162]
(PAMAM) solubility; transmucosal drug
dendrimers delivery; nose-to-brain drug delivery
and cellular uptake
5(6)-carboxyfluorescein (CF), FITC-dextran (MW 4400, FD4), and insulin.

Although the cylindrical pore radius was not changed, the pore occupancy/length
ratio of the permeation pathways for water-soluble molecules in the tight junctions
increases indicating that SG increase the pore number, while retaining the sieving
property of the epithelial membranes.
3 Synthetic Mucoadhesive Polymers
The synthetic mucoadhesive polymers offer advantages such as stability, mechanical

strength, and easy production, but there are only few types that show adhesion to the
nasal mucosa and sensitivity to conditions in the biological environment. Examples
are given in Table 11.2. So far, in nasal drug delivery carbomers and polycarbophils
have been considered the most often [36]. Their mucoadhesion is based on formation
of hydrogen bonds between the functional groups of the polymer and mucosal layer.
The interaction with the nasal mucus can be altered by chemical derivatization of
the parent polymer. Thiomers have been synthetized with the free thiol groups
which form disulfide bonds with the cysteine-rich residues of mucin [146].
3.1 Polyacrylates
Polyacrylates are polymers of acrylic acid very frequently used in liquid, semisolid,
and solid dosage forms for different drug administration routes. As mucoadhesive
polymers, polyacrylates attach to nasal mucosa and ensure intimate and prolonged
contact between the drug delivery system and the membrane surface at the site of
absorption. In the nasal drug delivery systems, carbomers and polycarbophils are
mostly considered. Carbomers are polymers of acrylic acid that have been cross-
linked with either allyl sucrose or allyl ethers of penta erythritol. They are hydro-
soluble polyelectrolytes with pendant carboxyl side chains which are ionized at
neutral pH. Such acidic carboxylic groups get deprotonated at the basic pH and
acquire negative charge. The similar charge causes electrostatic repulsion between
the polymer chains, and the material expands in dimensions. At acidic pH, carboxyl
groups lose their charge hence the repulsion is eliminated, and the material regains
its original shape. Therefore, at acidic pH carbomers are in solution, while they
form soft gel at pH 6–8, so they can be considered as pH-responsive/sensitive poly-
mers toward nasal route of administration. Also, carbomers are powders which
readily absorb water, become hydrated, and swell. The cross-linked structure and
swelling behavior make them suitable for achievement of controlled (sustained)
drug release mechanisms [36, 147].
Carbopol® 974 (1%, 1.5%, and 2%) was used for preparation of nasal gels for
progesterone delivery in rabbits. The drug-loaded gels were prepared by dispersing
polymer in distilled water followed by addition of progesterone/progesterone-beta
cyclodextrin complex dissolved in propylene glycol and neutralization. The abso-
lute bioavailability of progesterone from nasal gels was significantly increased in
comparison to intravenous injection. The nasal absorption of progesterone was pro-
moted by the beta cyclodextrin complex; however, the significance of mucoadhe-
sion has not been assessed [148]. Carbopol® 934P gel produced a significant
hypoglycemic response in rabbits, whereas no response was seen following nasal
administration of the insulin solution formulation. The bioavailability of insulin
from the nasal gel formulation was 20.6% higher in comparison with the intrave-
nous injection. Although there have been no attempts to assess its mucoadhesive-
ness, it was concluded that the Carbopol® 934P 0.4% gel promoted the nasal
absorption of insulin due to its sprayability with commercially available spray
pumps [149]. Ugwoke et al. [100] investigated the residence time of apomorphine
administered in rabbit nasal cavity in the form of a mucoadhesive preparation based
on the oral grade carbomer (carboxypolymethylene (Carbopol® 971P)) or carboxy-
methyl cellulose. The drug clearance from the nasal cavity to the stomach and intes-
tine was evaluated and compared by gamma scintigraphy. Although apomorphine
itself inhibited nasal mucociliary clearance, the lowest drug clearance percentages
(3% and 12% at 30 min and 3 h post insufflation, respectively) were observed in the
presence of Carbopol® 971P. The use of Carbopol® 971P increased the nasal resi-
dence time and provided sustained nasal delivery of apomorphine and achievement
of its peak plasma concentration while the formulation was still within the nasal
cavity. Another study of Ugwoke et al. showed that the Tmax of apomorphine deliv-
ered by the Carbopol® 971P-based formulation was 52.21 min, that is, 5-fold higher
in comparison with that of the lactose-containing formulation. Simultaneously, the
Cmax of the Carbopol® 971P-containing formulation was lower than that of the
lactose-containing formulation [150]. Nandgude et al. [151] demonstrated pH-
responsive gelation caused by deprotonation of Carbopol® 934 at nasal pH. This in
situ gelling formulation comprising 0.4–0.5% w/v of carbomer enabled sustained
210 L. Djekic
release and enhanced bioavailability of salbutamol sulphate. Sustained release of

drugs results in a more stable blood concentration-time curve [21]. The study of
Abd El Haamed and Kellaway [26] showed that Carbopol® 934P-based micro-
spheres had superior mucoadhesiveness compared to those made from chitosan,
polyvinyl alcohol, and HPMC.
3.2 Polycarbophils
Polycarbophils are polyacrylic acid-based polymers cross-linked with divinyl

chloride. Thiolated polycarbophils were used to study the nasal delivery enhancement
potential of leu-enkaphlin and human growth hormone [152, 153]. The results
showed that the thiomers of polycarbophils provide sustained release and diminish
the risk for the degradation of leu-enkaphlin, while the drug uptake by nasal mucosa
was increased by 80-fold [152]. Gel based on thiolated polycarbophil effectively
improved the nasal delivery and plasma level of human growth hormone [153].
Greimel et al. [154] used polycarbophil thiomer (polycarbophil-cysteine) for devel-
opment of microparticles (up to 100 μm) for nasal administration. The thiomer was
synthesized by the covalent attachment of L-cysteine to neutralized polycarbophil
via formation of amide bonds between the primary amino group of cysteine and the
carboxylic acid group of the polymer. The microparticles provide controlled release
of the model compounds sodium fluorescein and fluorescein isothiocyanate-dextran,
while their transport through the freshly excised bovine nasal mucosa was increased
1.70-fold and 2.64-fold, respectively, in the presence of permeation mediator gluta-
thione. Evaluation of ciliary beat frequency of human nasal epithelial cells excluded
the risk of ciliotoxicity caused by thiolated polycarbophil.
3.3 Miscellaneous Synthetic Polymers
Aikawa et al. [155] developed pH-responsive polyvinylacetal diethylamino acetate

(PDA) hydrogel and evaluated its potential for nasal delivery of chlorpheniramine
maleate and tetrahydrozoline hydrochloride. PDA forms transparent solution at
pH 4, while abrupt formation of porous hydrogel occurred in vitro, in phosphate
buffer at pH 7.4 at 37 °C. In vivo formation of the hydrogel on the rat nasal mucosa
was visually confirmed. The gelling phenomenon included shrinkage of the pores
due to increase in the temperature from 25 to 37 °C at pH 7.4, which was related
with the hydrogel potential to control the drug release. The higher the PDA concen-
tration, the lower drug release rate was observed. The in vitro drug release profiles
from dialysis tubes showed a rapid and a slow phase, with an inflection point, at
which hydrogel formation occurred.
Dendrimers are three dimensional, highly ordered, branched polymeric molecules
with unique structural architecture. Polyamidoamine (PAMAM) dendrimers have
gained attention for nasal drug delivery due to their capacity for enhancement of aqueous
Fig. 11.2 (a) Synthesis schematics for the PAMAM dendrimer nanocomposites. (a) Synthesis of
pegylated PAMAM dendrimer (mPEG-PAMAM G5.NH2); (b) synthesis of mPEG-PAMAM G5.
NHAc by the addition reaction between anhydride and the amino group; (c) paeonol (PAE) loading
in the cavities of mPEG-PAMAM G5.NHAc; (d) the FITC tracer was used to label PAMAM to
observe the nasal brain transport of PAMAM dendrimer nanocomposite in vivo; (b) Transmission
electron microscopy (TEM) images of nanocomposites. (a) PAMAM G5.NH2; (b) PAE/mPEG-
PAMAM G5.NHAc; (c) high-resolution image of a single PAE/mPEG-PAMAM G5.NHAc; (d)
the image of PAE/mPEG-PAMAM G5.NHAc based DGG (deacetylated gellan gum) in situ
gel [158]
212 L. Djekic
Fig. 11.2 (continued)
drug solubility, transmucosal drug delivery, and cellular uptake as well as the nose-to-
brain drug delivery [156, 157]. PAMAM dendrimers are biocompatible, biodegradable,
and low-immunogenic spherical molecules with internal cavities and peripheral func-
tional groups available for modification to encapsulate drugs. The core of PAMAM
dendrimers is usually ethylenediamine, to which methyl acrylate and ethylenediamine
are repeatedly added according to the desired number of generations G0, G1, G2, G3,
G4, and also generation called G5 obtained by terminating the reaction sequence after
addition of methyl acrylate that leads to terminal carboxylate groups (Fig. 11.2).
Superficial branches of PAMAM molecule could be terminated by amino-, hydroxyl-,
aldehyde-, methoxycarbonyl-, tert-butyloxycarbonyl-, methyl-, or -COONa groups. The
amino group is typically employed in intracellular delivery of genetic material
[158–160].
PAMAM dendrimers were evaluated for enhancement of solubility and nose-to-
brain delivery of haloperidol. Nasal administration of PAMAM-based formulation
enabled achievement of the same behavioral response by 6.8-fold lower doses as the
one induced by the intraperitoneal injection [161]. Kamei et al. [162] attached
siRNA onto PAMAM dendrimers to target against high mobility group box 1
(HMGB1). Upon intranasal administration, siRNA was distributed in different
regions of brain and depleted the target gene in peripheral cortex and striatum.
PAMAM dendrimers also had potential diagnostic applications on neurological
biomarkers in mouse brain by nasal route, although it is not clear whether the brain
delivery realized through systemic circulation or via olfactory pathway [163].
4 Mucoadhesive Copolymers
Synthetic copolymers as well as copolymers obtained by covalent conjugation of

natural polymers with synthetic polymers find application in the design of mucoad-
hesive nasal drug delivery systems with sensitivity to temperature and/or pH in the
nasal cavity. Copolymer formation allows the expansion of excipients performances,
such as increased water solubility, amphiphilicity, increased molecular weight and
mucoadhesiveness, and/or extended drug release [22, 164]. Examples of copolymers
considered so far for nasal drug delivery are: poloxamers, poly(N-
isopropylacrylamide) (PNiPAAm), PNiPAAm-co-polyacrylamide, PEGylated
chitosan, alginate polyethylene glycol acrylate, poly lactic-co-glycolic acid (PLGA),
PEGylated PLGA, lectin conjugates of PLGA, and PEG-PLGA (Table 11.3).
Poloxamers are nonionic poly(ethylene oxide)-poly(propylene oxide)-
poly(ethylene oxide) (PEO-PPO-PEO) triblock copolymers. The hydrophobic
polypropylene oxide segment connects two hydrophilic polyethylene oxide
segments. Poloxamers are commonly named with the letter “P” (for poloxamer)
followed by three digits. The first two digits × 300 give the approximate molecular
mass of the polyoxypropylene core, and the last digit × 10 gives the percentage
polyoxyethylene content [41, 165, 166]. In aqueous solutions poloxamers show
amphiphilic character and form micelles at concentrations above the critical micellar
concentration (CMC). An increase in the polymer concentration leads to arranging
of the micelles in various liquid crystalline phases such as lamellar, cubic, and
hexagonal. An increase in temperature causes desolvation (i.e., the rupture of the
hydrogen bonds present between the hydrophilic polyoxyethylene chains and
solvent) as well as the enhancement of the hydrophobic interactions among the
polyoxypropylene chains that induce formation of cubic structure and formation of
gel [167, 168]. Poloxamers are thermo-responsive polymers whose aqueous
solutions exhibit in situ gelling (sol-gel transition) upon exposure to the nasal
temperature. Their thermoreversible gelling properties depend on the copolymer
concentration, molecular weight, and ratio of molecular weight of hydrophilic core
to molecular weight of hydrophobic core [169, 170]. Poloxamer 407 is frequently
used in drug delivery systems and its lower critical solution temperature (LCST)
varies between 25 °C and 37 °C as a function of the copolymer concentration.
Aqueous solutions of poloxamer 407 at the polymer concentration of 16–18%
exhibited thermoresponsive gelling at the temperature of 32 ± 2 °C, which is close
to the nasal temperature [171, 172]. It was demonstrated that poloxamers in
concentrations below CMC may increase the transnasal drug delivery, but this was
not observed at concentrations above CMC [173]. The study of Na et al. [95]
reported better permeation enhancing effect of poloxamer 188 in comparison with
hydroxypropyl-beta-cyclodextrin and chitosans of different molecular weight, for
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Table 11.3 Examples of mucoadhesive copolymers and achieved benefits for nasal drug delivery
Mucoadhesive copolymers Benefit References
Poloxamers In situ gelling upon exposure to the nasal [169–172]
temperature
Permeation enhancing effect for intranasal [95]
absorption of drugs
Inhibition of drug efflux transporters in [174]
olfactory region and at the BBB
Poloxamer/carbomer Thermogelling property and enhanced [175, 176]
graft-copolymers mucoadhesion
Poly(N-isopropylacrylamide) Thermosensitive gelling in nasal cavity [177, 178]
(PNiPAAm) and
(PNiPAAm)-co-polyacrylamide
Acrylated Eudragit® E PO Enhanced mucoadhesiveness due to [179]
covalent linkages between acrylated groups
and mucins
PEGylated chitosan Aqueous solubility at alkaline pH; [180–182]
pH-dependent permeability enhancing
effect; enhanced nasal absorption
Alginate polyethylene glycol Gelling properties with strong [184, 185]
acrylate mucoadhesion; controlled drug release rate
Poly lactic-co-glycolic acid Design of nanoparticles for brain targeted [186, 187]
(PLGA) delivery via olfactory neurons
Wheat germ agglutinin-conjugated Nanoparticles for improved nose-to-brain [189]
PLA-PEG absorption
Solanum tuberosum lectin- Nanoparticles suitable for rapid absorption [190]
conjugated PLGA and higher brain targeting efficiency
Solanum tuberosum lectin- Nanoparticles for achievement of enhanced [194, 195]
conjugated PEG-PLGA brain bioavailability of nasally administered
nanoparticles peptides; protection of peptide and
protein-based drugs from peptidase
degradation in nasal cavity
intranasal absorption of isosorbide dinitrate in rats. In addition to thermo-sensitivity

and permeation enhancement ability, poloxamers have been attributed the ability to
inhibit the drug efflux transporters, such as Pglycoprotein, that are present throughout
the nasal cavity, in olfactory region, and at the BBB [174]. Despite these favorable
characteristics, uncharged poloxamer molecules are considered weak mucoadhesive
agents. Mucoadhesion can be enhanced by graft-copolymerization of the
hydrophobic segment of poloxamers with a suitable polyelectrolyte segment having
ionizable groups. The example is poloxamer 407/carbomer copolymer consisting of
a thermosensitive polymer and a pH-sensitive mucoadhesive polymer, respectively.
The copolymer has shown thermogelling property and enhanced mucoadhesion at
low concentrations due to the presence of carboxylic groups in carbomer. Also, the
carboxylic groups get deprotonated at the basic pH and acquire negative charge that
causes repulsion between the copolymer molecules leading to their expansion and
gelling of the system. Relevant studies reported the enhancement of the viscosity for
10–103 fold for 1–5% w/v solution of the poloxamer/carbomer (1:1) graft-
copolymers and increase in the residence time for fluorescent labels in rat nasal
cavity, while the physical mixture and/or random copolymer of the thermosensitive
and the pH- sensitive polymer loses its thermosensitivity and shows only
pH-sensitivity [175, 176].
Poly(N-isopropylacrylamide) (PNiPAAm) and (PNiPAAm)-co-polyacrylamide
are examples of copolymers which aqueous solubility significantly changed around
their LCST due to rapid hydrophobic and hydrophilic interactions between the
polymeric chains and the aqueous media. At the temperatures lower than the LCST
the copolymer is soluble in water and hydrophilic interactions between the copoly-
mer and water are dominant, but at the temperatures higher than the LCST, hydro-
phobic interactions between the copolymer chains become stronger [177]. The
LCST of PNiPAAm and (PNiPAAm)-co-polyacrylamide is 30–32 °C, so they can
enable localization of the drug delivery system in the nasal cavity. Thermosensitive
gel based on (PNiPAAm)-co-polyacrylamide was evaluated for nasal delivery of
insulin in rats [178]. The linear cationic terpolymer obtained by copolymerization
of N,N-dimethylaminoethyl methacrylate with methylmethacrylate and butylmeth-
acrylate is a commercially available pharmaceutical excipient Eudragit® E PO
(EPO), which is freely soluble in water only under acidic conditions (Evonik techni-
cal notes, 2018). EPO’s molecule is convenient for chemical modification using
acryloyl chloride, which results in the formation of acrylated polymers with
enhanced mucoadhesive properties due to the formation of covalent linkages
between the acrylated groups and thiols present in mucins. Biocompatibility of the
prepared EPO’s acrylated derivatives was demonstrated by the slug mucosal irrita-
tion test. Liquid formulations were prepared using EPO and its acrylated deriva-
tives. The formulations were loaded with sodium fluorescein as a model compound.
The formulations prepared with acrylated derivatives exhibited superior mucoadhe-
sive effect and greater retention of sodium fluorescein on freshly excised sheep
nasal mucosa in comparison with those prepared with EPO [179].
The formation of a positively charged copolymer of chitosan and macrogol
(PEGylated chitosan) provided solubility at alkaline pH values, while permeability-
enhancing effect was pH-dependent at pH 6 and higher in comparison with unmodi-
fied chitosan [180, 181]. Zhang et al. [182] reported that PEGylated chitosan
nanoparticles (150–300 nm), with insulin loading efficiency of 20–39%, enabled
the plasma drug concentration maximum of 350 μU/ml after 30 min and subsequent
reduction of the initial glycemia by 60% after 90 min in rabbits. The nasal absorp-
tion of insulin from the nanoparticles was better in comparison with the physical
mixture of insulin and the PEGylated chitosan.
Alginate polyethylene glycol acrylate (alginate-PEGAc) is an anionic copolymer
comprising an alginate backbone with acrylated polyethylenglycol groups attached
to it. This copolymer combines strength and gelling properties of alginates with
strong mucoadhesion which is the result of the penetration of polyethylene glycol
into the mucus surface and binding with sugar moieties on glycosylated proteins by
hydrogen bonds, while the acrylate group binds with the sulphide group of
glycoproteins present in the mucus and β-d-mannuronic acid moiety binds to the
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glycoproteins in the mucus through carboxyl–hydroxyl interactions. The introduc-

tion of acrylic acid compensated the erosion of alginate in neutral pH, but also
controlled the release rate of drugs and improved its adhesive properties. Addition
of PEG increased the viscosity of the resulting copolymer, retarding its disintegra-
tion and removal from the mucosal surface [183].
Poly lactic-co-glycolic acid (PLGA) is a biocompatible and biodegradable
synthetic copolymer of polylactic acid and polyglycolic acid [184, 185]. It has been
used for preparation of nanoparticles to study their uptake on olfactory ensheathing
cells. Higher level of rhodamine-incorporated PLGA nanoparticles was observed in
olfactory cells than that of nanoparticles prepared from polylactic acid or chitosan.
PLGA nanoparticles are considered more promising for brain-targeted delivery via
olfactory neurons [186]. For example, the brain uptake efficacy of olanzapine loaded
in PLGA nanoparticles increased by 10.86-fold after intranasal administration and
6.35-fold for intravenous administration compared with free drug solution. Md
et al. [187] reported enhanced brain uptake of PLGA nanoparticles for nose-to-brain
delivery of donepezil. Coupling PEG and/or lectins with PLGA or poly(lactic acid)
(PLA) is a useful strategy to upgrade the polymer performance to target the brain
via nasal mucosa. Lectins are structurally diverse proteins that are usually extracted
from gorse, soybean, peanut, and lentil. They are promising for use in nasal delivery
due to low ciliary irritation risk and high permeability. They have shown specific
and reversible binding to the mucosa due to their capability to identify and bind to
specific sugar moieties like those present in the mucus layer. Depending on the
number of carbohydrate-recognizing domains (CRD), lectins are classified into:
merolectins (1 CRD), hololectins (≥2 crd), and chimerolectins (with additional
unrelated domains). Conjugates of lectins with synthetic polymers are considered
promising excipients for development of nose-to-brain drug delivery systems.
Wheat germ agglutinin is a biorecognitive ligand-lectin which was used for conju-
gation with PLA-PEG nanoparticles in order to improve their nose-to-brain absorp-
tion by specifically binding to N-acetyl-D-glucosamine and sialic acid moieties,
both of which were abundantly observed in the nasal cavity particularly in the olfac-
tory mucosa [188–192]. The lectin-conjugated nanoparticles were prepared by
incorporating maleimide in the PLA-PEG molecule so that its thiol group binds
2-iminothialane thiolated wheat germ agglutinin. The nasal toxicity of the nanopar-
ticles was negligible. The brain uptake of wheat germ agglutinin-functionized
nanoparticles was about two-fold higher in different brain tissues compared with the
unmodified ones [189]. Solanum tuberosum lectin (STL) was used for conjugation
of PLGA nanoparticles (STL-nanoparticles). The in vitro uptake study in Calu-3
cells showed markedly enhanced endocytosis of STL-nanoparticles compared to
unmodified PLGA nanoparticles and significant inhibition of uptake in the presence
of inhibitor sugar (chitin hydrolysate). Following nasal administration, a marker
coumarin-6 carried by STL-nanoparticles was rapidly absorbed into blood and brain
segments (olfactory bulb, cerebrum, brainstem, and cerebellum) achieving
1.89–2.45 times higher brain targeting efficiency than unmodified NP. Mild
cytotoxicity and negligible cilia irritation demonstrated safety of STL-nanoparticle
[190]. Moreover, Zhang et al. [193] developed SLT-conjugated PEG-PLGA
nanoparticles with entrapped fibroblast growth factor (I-bFGF). After intranasal

administration nanoparticles exhibited significantly higher area under the concen-
tration time curve (AUC) in different parts of the brain than intravenously adminis-
tered STL-conjugated nanoparticles and nasal drug solution. Hence, lectin
conjugated nanoparticles may protect peptide- and protein-based drugs from pepti-
dase degradation in nasal cavity. Conjugation of PEG-PLGA nanoparticles by lac-
toferrin enabled targeting of lactoferrin receptors which are present on the respiratory
epithelium cells and neurons in nasal cavity [194]. It was reported by Yan et al. that
lactoferrin-modified nanoparticles delivered higher concentration of the antiparkin-
sonian drug rotigotine in olfactory bulb, striatum, and other brain regions compared
to unmodified nanoparticles [195].
5 Mucoadhesive Polymer Blends
Many published studies have focused on combining two or more mucoadhesive

polymers with different physicochemical properties in order to integrate their prop-
erties relevant for nasal drug delivery and possibly achieve synergy that would
expand functionality (mucoadhesion, thermo-sensitivity, pH-sensitivity, ion-
sensitivity, and/or solubility) and enhance drug delivery potential of the nasal for-
mulation. Polymer combinations are mostly physical mixtures when there is no
chemical bond between the polymers. The possibility of physical and chemical
cross-linking of polymers in order to create a carrier with improved mucoadhesive
properties and drug release kinetics, in comparison with the starting molecules, has
been also considered.
5.1 Physical Blends
In physical polymer, blends often use poloxamers to provide thermoreversibility of

the formulation, as well as chitosan and carbomers to emphasize mucoadhesion
and/or to introduce pH-sensitivity. Selected examples, which are commented on in
this subchapter, illustrate the potential of this strategy.
Poloxamers are responsible for in situ gelling; however, their mucoadhesiveness
is weak due to their nonionic character and relatively low molecular weight. The
combination of poloxamers with other mucoadhesive polymers, such as carbomers,
polycarbophils, and PEGs, has been allowed for the tuning of the residence time in
the nasal cavity and sol-gel transition temperature as well as to control the in vitro
drug release. Thermoresponsive nasal gel of Nardostachys jatamansi with polox-
amer 407 and PEG 4000 [196] showed an increase in the gelling temperature as the
concentration of the PEG 4000 increases. PEG 4000 is a nonionic, hydrophilic
compound, which may establish intermolecular hydrogen bonding with poloxamer
chains and water and thus delay gelation time and increase the gelation temperature,
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during which these hydrogen bonds weaken and hydrophobic interaction between
poloxamer chains becomes dominant leading to gel formation. Addition of mucoad-
hesive polymers such as carbomers, chitosan, and cellulose derivatives reduces the
gelling temperature of the poloxamers [41, 197]. Srivastava et al. prepared thermor-
eversible and mucoadhesive in situ gels by combining xanthan gum, HPMC, or
carbomer, in different concentrations (0.5, 1, or 1.5% w/v), with poloxamer 407
(10% w/v), with an aim to improve the antioxidative and anti-inflammatory effect of
polyherbal (Moringa olifera and Embelia ribes) extract in the treatment of allergic
rhinitis. After administration into the nasal cavity, formulations rapidly transformed
into viscous hydrogels. The combining of poloxamer 407 with the mucoadhesive
polymers increased the gel strength, so the formulations prepared with high concen-
trations of xanthan gum, HPMC, or carbomer exhibited better mucoadhesiveness
than poloxamer 407 itself. The pH values of the formulations ranged between 5.2
and 5.9 and their spreading ability was from 7.6 ± 0.21 to 11.7 ± 0.65 cm. Investigated
in situ nasal herbal gels did not exhibit nasal redness, edema, inflammation, and
irritation in mice. Poloxamer 407-based thermoreversible formulation for intranasal
delivery of fexofenadine included 0.1–0.3% (w/v) of chitosan [167]. Besides the
increased nasal residence time, the presence of chitosan increased the nasal perme-
ability and drug bioavailability in rabbits of about 18-folds. The nasal permeation
activity of chitosan was confirmed also in vivo, where the Cmax increased from
52.96 ± 9.43 ng/ml to 78.25 ± 21.25 ng/ml, employing 0.1% and 0.3% of this poly-
mer, respectively. Shelke et al. [198] developed thermoreversible intranasal gel with
zolmitriptan-loaded nanoethosomes (171.67 nm) with entrapment efficiency of
66%. Thermoreversible gel vehicle with phase transition temperature at 32–33 °C
was prepared by using varying concentrations of poloxamer 407, Carbopol® 934,
and HPMC. In vitro drug release from the optimized formulations followed
Korsemeyer-Peppas model indicating non-Fickian release profiles. The gels were
non-toxic on columnar epithelial cells. The study of Perez et al. [199] demonstrated
enhancement of nose-to-brain delivery of siRNA dendriplexes from in situ mucoad-
hesive gels comprising a blend of poloxamer 407 with mucoadhesive chitosan or
carbopol 974P NF. In situ gel maintained the stability of dendriplexes and enhanced
their uptake by olfactory neurons in comparison with intravenously administered
dendriplexes and intranasally administered naked siRNA within gel vehicle.
Chitosan has been used often in combination with other synthetic and natural
polymers due to its mucoadhesiveness and permeation enhancer effect. There is an
interest for thermosensitive chitosan-based formulations since they may be suitable
for application in nasal cavity as liquids which transform into gel of increased
mucoadhesiveness resulting in prolonged residence as well as with potential for
increased drug absorption. Agrawal et al. [200] incorporated insulin in thermosensi-
tive gel formulation based on chitosan (3%) and polyvinyl alcohol (PVA) (2%).
Formulation showed thermoresponsive gelling, high swelling index, and the poten-
tial of controlling the blood sugar level for 6 h. PVA is a water-soluble polyhydroxy
polymer. At room temperature, the intermolecular hydrogen bonds exist between
hydroxyl and amino groups of chitosan and hydroxyl groups of PVA as well as
between water and PVA. These hydrophilic interactions lead to dissolution of
chitosan chains. At nasal temperature, intermolecular hydrogen bonding gets

ruptured, and thus chitosan chains’ mobility enhances, surrounding water molecules
are removed, and hydrophobic chitosan chains associate with each other to form
gel. Thermosensitivity of the formulation was dependent on the polymers mass
ratio. Temperature sensitive in situ gelling property does not occur if PVA/
chitosan ratio exceeded 10:1 [201]. Al-Ghananeem et al. [202] promote nasal
absorption of Δ9 -tetrahydrocannabinol (THC) by formulation of a mucoadhesive
gel spray based on chitosan (2% w/v) and PEG 400 (10% w/v). Although no
significant difference between the absolute bioavailabilities of the THC solution
(13.3 ± 7.8%) and the chitosan-based gel (15.4 ± 6.5%) was observed, the THC
delivered with the chitosan-based gel reached the higher Cmax (31 ± 4 ng/ml) in
contrast to the drug solution formulation (20 ± 3 ng/ml). Jose et al. [203] reported
the sustained release of lorazepam over 24 h from mucoadhesive chitosan
microspheres loaded in a thermosensitive poloxamer-based gel formulated for nose-
to-brain delivery. For comparison, lorazepam-loaded microspheres in phosphate
buffer solution pH 6.2 and lorazepam powder loaded in poloxamer gels showed a
release profile over 9 h and 15 h, respectively.
Zafar et al. [204] aimed to formulate the microspheres by using chitosan and
ethyl cellulose for the systemic delivery of domperidone via the nasal route. In vivo
study for optimized DOM-Microsphs (F1) formulation was performed on Wistar
rats for the evaluation of bioavailability and pharmacokinetic parameters after nasal
administration and compared to nasally administered DOM solution (DOM-Sol),
and orally administered DOM-Sol and commercially available tablet formulation
(ComTab). The optimized microspheres were spherical with 21.12 ± 0.51 μm par-
ticle size, 84.79 ± 1.39% entrapment efficiency, 50.68 ± 0.96% drug loading, and
81.2 ± 6.75% drug release in 8 h. Domperidone-loaded microspheres demonstrated
2-folds ex vivo permeation higher than commercial tablet formulation. The relative
bioavailability of optimized microsphere formulation administered nasally was
superior 3.75-fold, 2.61-fold, and 1.77-fold in comparison to the drug solution and
tablets administered orally, and solution administered nasally, respectively. Gavini
et al. [205] prepared zolmitriptan-loaded microspheres from chitosan and HPMC of
different molecular weights. Microparticles in the form of powder were adminis-
tered nasally to rats. The nasal bioavailability of zolmitriptan was around 35%;
however, the concentrations of the drug in the CSF after 1 h were similar for the
intravenous injection and the nasal chitosan microsphere formulation, albeit they
had shown different plasma concentrations. Nasal gel based on chitosan and HPMC
blend provide 8.5 times higher concentration in brain of a dopamine D2 agonist
ropinirole, than nasal drug solution [38]. Jose et al. [203] formulated poloxamer-
based thermosensitive gel loaded with mucoadhesive microspheres prepared from
chitosan. This drug delivery system, upon nasal administration in rats, provided
sustained release of lorazepam over 24 h. The microspheres in phosphate buffered
saline (pH 6.2) and lorazepam-loaded poloxamer gels released drug over 9 h and
15 h, respectively.
The chitosan-based gels undergo a slow sol-gel transition at physiological pH
[25], while chitosan derivative TMC retains the key characteristics of the parent
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polymer, but presents improved aqueous solubility and thermosensitivity, enhanced

mucoadhesiveness, and a significant absorption enhancing effect over a wide pH
range [206, 207]. In addition, combinations of TMC with other mucoadhesive poly-
mers were considered. Nazar et al. [59] synthesized thermosensitive in situ nasal gel
from TMC with PEG 4000 which provided additional sites for hydrogen bonding
and allows the formation of more extensive gel network. TMC, synthesized from
chitosans of three different average molecular weights, have been co-formulated
with PEG 4000 into stable thermosensitive liquid formulations suitable for admin-
istration via nasal spray or drops. The hydrogels derived from TMC with a low
degree of quaternization and high or medium average molecular weight exhibited
relatively short sol-gel transition time at physiologically relevant temperatures,
good water-holding capacity, and strong mucoadhesive potential. TMC of medium
average molecular weight and low degree of quaternization (3.6% w/v) with 5.8%
w/v PEG 4000 and 2.5% w/v glycerophosphate undergo thermal gelation at 32.5 °C
within 7 min [208]. In a study of Wu et al. [206], thermosensitive hydrogel was
formulated using N-[(2-hydroxy-3-trimethylammonium)propyl] chitosan chloride
(HTCC) and PEG 4000 with a small amount of α, β-glycerophosphate. Negatively
charged moieties of glycerophosphate may interact with various bioactive compo-
nents [200]. HTCC is a water soluble, mucoadhesive derivative of chitosan which
retains absorption enhancement effect on nasal mucosa. After being administered
into the nasal cavity, the solution transformed into viscous hydrogel at body tem-
perature, which decreased nasal mucociliary clearance rate and drug release rate.
The hydrogel enabled insulin retention and absorption, while blood glucose concen-
tration was drastically decreased (almost 40–50% of initial glycemia for at least
4–5 h) after administration in male Sprague–Dawley rats [206]. Introduction of
alginate into chitosan/tripolyphosphate nanoparticles was used to design a hybrid
carrier for nasal delivery of insulin (5 IU/kg) in rats [209]. This hybrid nanocarrier
had a high positive surface charge (ζ N + 41 mV). Alginate molecular weight was
related with the duration of the hypoglycemic response. Although the chitosan/tri-
polyphosphate system showed higher reducing capacity of the glucose level (35%)
compared to the hybrid system (30%) after 1 h, the glucose level reduction (20%)
from the hybrid nanocarrier was prolonged up to 5 h. Blend of pH-responsive chito-
san and thermo-responsive pNIPAAM has been used for the formulation of dual
responsive hydrogels for effective nasal delivery of pilocarpine hydrochloride [210].
Intranasal gels comprising PEGs and carbomer (Carbopol® 934) were evaluated
for insulin in vitro release, and its bioavailability was compared with an intranasal
solution and subcutaneous injection in rabbits. Increasing the molecular weight of
PEG and Carbopol® 934 concentration increased the gel viscosity and affected the
drug release mechanism. A lower viscosity gel was prepared by applying heat and
it was suitable to achieve a higher insulin release. A stronger and longer hypoglyce-
mic effect with 1.7-fold and 3.1-fold higher maximum decrease in glycemia and
AUC, respectively, were provided by the gel, when compared with the subcutaneous
injection [211].
5.2 Polyelectrolyte Complexes
Polyelectrolyte complexes are formed by establishing noncovalent interactions

between anionic and cationic polymers at pH in the vicinity of pKa interval of the
two polymers. In this way, various polyelectrolyte complexes of chitosan with poly-
anions such as xanthan gum, pectin, and hyaluronic acid were obtained and consid-
ered for use in nasal drug delivery.
The chitosan/xanthan polyelectrolyte complex was assessed by Dehghan and
Kazi [212] for design of the mucoadhesive nasal insert for treatment of motion sick-
ness by promethazine hydrochloride. The complexation of the oppositely charged
polyelectrolytes enabled the formation of a three-dimensional network with a
capacity to control the release of the incorporated drug. The study focused on evalu-
ation of the effect of concentration of the parent polymers on viscosity of polyelec-
trolyte complex solution, water uptake of nasal inserts (at pH 2, 5.5, 7.4), bioadhesion
potential, and in-vitro drug release at Q6h. The higher content of xanthan gum in
polyelectrolyte complexes retarded in vitro drug release. The polycation/polyanion
concentration as well as pH of the medium strongly influenced the water uptake
behavior of nasal insert and formation of a mucoadhesive gelled system. Luppi et al.
[213] suggested the preparation of chitosan/hyaluronic acid polyelectrolyte com-
plexes with mucoadhesive properties for nasal delivery of vancomycin and insulin
in the form of nasal inserts. In vitro swelling, mucoadhesion, and drug release were
evaluated. The selection of suitable conditions for preparation of the polyelectrolyte
complexes was of significant importance for modulation of swelling behavior and
prolonged drug release from the inserts during 6 h. The same group investigated
capacity of chitosan/pectin polyelectrolyte complexes in the form of nasal inserts
for improvement of bioavailability of an antipsychotic drug chlorpromazine hydro-
chloride [214]. Chitosan/pectin polyelectrolyte complexes (Fig. 11.3a) were pre-
pared at pH 5.0 from citrus peel pectin (Mr 30,000–100,000; esterification degree
60%; pKa 4.0) and chitosan (Mr 150,000; deacetylation degree 97%; pKa 6.3). The
selection of suitable chitosan/pectin molar ratio during complex preparation allowed
the modulation of insert water uptake behavior and drug release and permeation
across sheep nasal mucosa. The higher amount of pectin in the complexes, with
respect to amount of chitosan, produced a more porous nasal inserts, improving
water uptake ability (Fig. 11.3b) and mucoadhesion capacity (Fig. 11.3c). Also,
pectin interacted with the drug inducing the formation of less hydratable inserts and
sustained drug release and permeation.
5.3 Cross-Linked Polymers
The use of cross-linked polymers in the development of nasal drug delivery systems
of various types has not been intensively considered so far. A representative exam-
ple is the thermo-responsive gels formulated by using chitosan, cross-linked by
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Fig. 11.3 (a) Scanning electron micrographs of the different chitosan/pectin complexes; (b) water
uptake ability of nasal inserts in different pH conditions (n = 5, the SD did not exceed the 5%); (c)
mucoadhesive capacity (expressed as detachment force, mean ± SD, n = 3) of chitosan hydrochlo-
ride, pectin, and chitosan/pectin complexes at pH 5.5. (With permission from Ref. [214])
glutaraldehyde, and poloxamer which interpenetrated gels. These gels were evalu-
ated as carriers for nasal delivery of insulin in diabetic rats. The in vitro release of
insulin from the gels followed a Fickian diffusion model, and it was for about six
times longer than release from poloxamer gels. Also, hyperglycemic effect was pro-
longed and pharmacological efficiency significantly improved in vivo [60]. Another
example is the study of Deutel et al. [215] which was focused on utilization of
interpolymer complex formed between poly(vinyl pyrrolidone) (PVP) and
poly(acrylic acid) (PAA) or poly(acrylic acid)-cysteine (PAA-Cys) for preparation
of microparticles for nasal delivery of insulin. The mean particle size of PAA/PVP/
insulin and PAA-Cys/PVP/ insulin microparticles was 2.6 ± 1.6 μm and 2.8 ± 1.7 μm,
respectively. A strong network based on formation of intra- and intermolecular
disulphide bonds between thiomer/polymer and insulin was established. Nasal
safety of formulation and native compounds was confirmed in in vitro ciliary beat
frequency test. The release of insulin from the microparticles at pH 7.4 occurred
within the first 60 min likely due to swelling process, the maximum was reached
after 120 min, while the release amounts detected after 6 h were 52.3 ± 18.9% and
37.0 ± 10.7% regarding formulations containing the thiomer and the polymer,
respectively. There was no significant difference between the microparticles; how-
ever, thiomer-based microparticles then showed a lower release likely due to a
greater stabilization as a result of disulphide bond formation within the thiomeric
network. The release of insulin was hindered by electrostatic and hydrophobic
drug/polymer interactions as well as steric effect and hydrogen bonds.
6 Conclusion and Future Perspectives
Nasal drug delivery strategies based on the mucoadhesive polymers have been
intensively researched for achievement of local and systemic immune response, sys-
temic drug delivery, and nose-to-brain drug delivery. The absorption of polar drugs
and macromolecules through the nasal mucosa is a permanent challenge, thus the
enhancement of intranasal retention time and trans-nasal bioavailability could be
particularly exciting prospects. Contemporary chemical engineering techniques
enable the synthesis of derivatives of natural polymers, copolymers, polyelectrolyte
complexes, and cross-linked polymers with improved solubility, mucoadhesiveness,
and drug release kinetics in response to unique physiological conditions (tempera-
ture, pH, ions, or enzyme) in the nasal cavity, thus providing significant improve-
ments in the bioavailability of both small molecules and macromolecular active
substances. Novel stimuli responsive polymers can be optimized in accordance with
the nasal temperature and pH value. Also, by combining thermo-responsive and pH-
responsive polymers, dual (thermo-/pH)-responsive nasal drug delivery systems can
be formulated. Functionalization of polymers by special molecules, such as lectins,
enables the targeted delivery to specific receptors in the olfactory region, which is
promising for improving generally low direct transport of drugs into the CNS. The
increasing number of more complex and fragile molecular drugs, which can be
224 L. Djekic
applied exclusively by injection, will expand the future development of the nasal
drug delivery systems and the role of the mucoadhesive polymers. However, there
are a number of limitations and challenges that need to be overcome. Natural poly-
mers are available from renewable sources; however, to obtain safe excipients of
satisfactory functional characteristics, a high degree of purity and/or chemical treat-
ment are required. Further work is needed to characterize the various types of the
polymers available on the market, including a variety of molecular weights, degree
of chemical treatment, and/or chemical derivatization. The data published so far on
the strength of mucoadhesion of different polymers, both intrinsic and in nasal for-
mulations, are inconsistent and sometimes contradictory. Safety aspects of mucoad-
hesive polymers, especially neurotoxicity, as well as their potential for enhancing
the intranasal absorption of macromolecular drugs, mechanisms and dynamics of
the polymer phase transitions in the nasal environment, the polymer-mucus interac-
tions, the drug release, and trans-nasal delivery, have not been sufficiently investi-
gated. Of particular importance could be studying these aspects of nasal drug
delivery under disease conditions which can significantly affect mucus production
and/or ciliary movement.
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Chapter 12
Novel Approaches in Nasal In Situ Gel
Drug Delivery
Cinzia Pagano, Luana Perioli, and Maurizio Ricci
Abstract The nasal cavity represents a suitable administration route both for local
and systemic treatments. Conventional nasal formulations (liquid, solid, semisolid)
show a limited residence time responsible for an incomplete drug absorption with
consequent impaired therapeutic efficacy. In situ nasal gels represent a suitable for-
mulative strategy able to overcome this problem. These formulations are liquid at
room temperature, making an easy administration, and become gel once in the nasal
cavity. This is possible thanks to the use of polymers able to form a viscous gel
under specific stimuli as temperature, pH, ions in the nasal fluid. The chapter illus-
trates the evolution from conventional nasal formulations to innovative in situ gel
delivery systems and the advantages of such formulations. It presents also recent
approaches, based on the combination of in situ gels with nanocarriers, useful to
protect the drug either improve the biopharmaceutical properties and promote a
controlled release. The safety aspects have been examined as well.
Keywords Nasal formulations · Residence time · In-situ gelification · Polymers ·

Nanocarriers · Safety
1 Introduction
According to the official pharmacopoeias, “nasal preparations” are classified as

“liquid (solutions, suspensions and emulsions), semi-solid or solids (powders)
intended to be administered in the nasal cavity to obtain a systemic or local affect”
[1, 2]. The nasal cavity is a useful administration route especially for drugs sub-
jected to first-pass metabolism as the absorption through the nasal mucosa allows to
C. Pagano (*) · L. Perioli · M. Ricci

Department of Pharmaceutical Sciences, University of Perugia, Perugia, Italy
e-mail: cinzia.pagano@unipg.it
https://doi.org/10.1007/978-3-031-23112-4_12
236 C. Pagano et al.
reach the systemic circulation bypassing the liver [3]. This route is also interesting
for the administration of biological drugs and biotherapeutic products, such as
recombinant proteins, peptides, and vaccines as antibodies, which generally have a
high molecular weight and a complex structure. Biological molecules are extremely
sensitive to the physical and chemical conditions of the gastrointestinal environ-
ment and their permeability across the intestinal mucosa is extremely poor [4]. For
these reasons they are currently administered parenterally and, more recently, by
intranasal route as a valuable alternative to perform systemic treatments.
Moreover, nasal formulations represent a useful tool for the treatment of “spe-
cial” patients such as newborns, infants, elderly, and unconscious patients that often
are non-cooperative and/or presenting swallowing difficulties with risks of suffoca-
tion in case of oral therapy. Nasal formulations are noninvasive, safe, easy to admin-
ister, and characterized by a short onset time, useful to obtain a rapid effect included
the case of emergency treatments [5] as a valid alternative to intravenous route when
it is not possible to obtain venous access. The drugs mainly administered intrana-
sally in emergency rooms are fentanyl, morphine, naloxone, midazolam, flumaze-
nil, ketamine, lidocaine, glucagon, haloperidol, and dexmedetomidin [3, 5–7].
Conventional nasal formulations are mainly represented by instillations (solu-
tions, suspensions), applied in the nasal cavity by means of proper dispensers as
dropper, and ointments (semisolid) spread on the nasal mucosa by the finger or
using cotton swabs. Recently the tubes have been provided by special cannula-tips
in order to perform a better and precise dosing in the nasal cavity.
The main limitations of this kind of formulations are the small dose that can be
administered and the low residence time responsible for an incomplete absorption
of the administered drug. Generally, most of the drug delivered in the nasal cavity
are physically removed in less than 80 min [8, 9].
In addition, the application of drops or ointments produces nasal discharge with
loss of drug, discomfort, and the necessity to perform many daily administrations
for an efficacious treatment [3]. The nasal discharge represents the main factor
responsible for therapy failure. It depends on two main factors: physiological,
mainly due to patient orthostatic posture and nasal mucociliary clearance (mecha-
nisms having the objective to protect the respiratory system from damage by inhaled
substances) [9], and formulative. The development of formulations compatible with
nasal cavity anatomy and physiology could allow to perform a safe, standardized,
and effective therapy [3].
The nasal products available on the market are conventional formulations
intended both for local and systemic treatments (Table 12.1). The local treatments
are generally performed by solutions of small and water-soluble drugs as deconges-
tants, antihistaminics, and fluidifying agents and suspensions, generally used for the
nasal administration of poorly water-soluble corticosteroids. In regard to systemic
therapies, commercially available products are solutions administered by sprays
containing small molecules that can be easily absorbed through the nasal mucosa
such as benzodiazepine (e.g., diazepam) for the treatment of seizures in epilepsy,
alkaloids such as dihydroergotamine mesylate and zolmitriptan for migraine treat-
ment, hormones as estradiol hemihydrate for menopausal syndrome treatment,
12 Novel Approaches in Nasal In Situ Gel Drug Delivery 237
opioids as fentanyl for severe pain treatment, and nicotine for smoking cessation. In
addition, few other intranasal formulations are available on the market as powders
(Table 12.1).
In order to better exploit the nasal route, new nasal drug delivery systems are
expected to be on the market in the near future considering that the success of a
nasal therapy mainly depends both on the suitable formulation and device. The for-
mulation should offer advantages in terms of high drug bioavailability with conse-
quent reduced daily administrations and thus decrease of the total amount of daily
administered drug (reduced side effects). At the same time, increased patient com-
pliance and improved adherence to therapy can be achieved. A suitable device offers
advantages in terms of easy, precise, and reproducible dosing and thus therapeutic
efficacy [10].
Table 12.1 Examples of conventional nasal dosage forms for drug administration available on
the market
Formulation Drug Treatment
Solutions Decongestants: For example, oxymetazoline hydrochloride (e.g., Topical
Oxywell® nasal drops, Otrivin®oxy nasal spray), xylometazoline
hydrochloride (e.g. Otrivin® nasal drops, Otrivin® nasal spray),
naphazoline (e.g. Naprisol® nasal drops, Privine® nasal spray),
phenylephrine (e.g. NTR® nasal drops, Equate™ nasal spray), silver
(Argotone® nasal drops, Rinosilver® nasal spray)
Antihistaminics: azelastine (Rinazina® nasal spray), tramazoline
hydrochloride (Rinogutt® nasal spray), levocabastine (Levoreact®
nasal spray)
Fluidifying agents: For example, acetylcysteine (Rinofluimucil® nasal
spray)
Solutions Benzodiazepine (diazepam – Valtoco®) Systemic
Opioids (naloxone-hydrochloride – Narcan®, fentanyl-PecFent® and
Instanyl®, butorphanol tartrate – Stadol®)
Antidepressants (esketamine – Spravato®)
Antiemetic (metoclopramide – Gimoti®)
Alkaloids (dihydroergotamine mesylate – Trudhesa™)
5HT1-receptor
agonists (zolmitriptan – Zomig®, sumatriptan – Imitrex®)
Hormones (estradiol hemihydrate – Aerodiol®)
Polypeptides (calcitonin – Miacalcin™)
Vaccines (virus strains: an A/H1N1 strain, an A/H3N2 strain and two
B strains – FluMist®)
Suspensions Corticosteroids: For example, fluticasone propionate (Flixonase® Topical
nasal spray), mometason furoate (Nasorex® nasal spray),
beclomethasone dipropionate (Rinoclenil® nasal spray), triamcinolone
acetonide (Nasacort® nasal spray), budesonide (Benacort® nasal
spray)
Semisolids Antimicrobial agents (mupirocin – Bactroban® ointment; Topical
chlorhexadine + neomycin – Naseptin® cream)
Powders 5HT1-receptor agonists: sumatriptan (ONZETRA™ Xsail) Systemic
Peptidic hormone: For example, glucagon (Baqsimi™)
Steroids: For example, beclomethasone dipropionate (QNASL™)
2
In Situ Nasal Gel
The hydrophilic semisolid nasal formulations (gels) available on the market are
medical devices, intended only for nasal mucosa hydration, mainly represented by
hydrogels based on saline solutions, aloe vera, and hyaluronic acid, while no hydro-
gels intended for drug administration can be found. In fact, the gel administration in
the nasal cavity could be difficult (applicator blockage due to gel drying) with low
dosing precision and discomfort for the patient.
Nasal in situ gel forming drug delivery systems represent the formulative answer
to the limitations of conventional gels and, in general, to the problem of the low resi-
dence time of conventional nasal dosage forms. The rationale for the development
of such kind of formulations is based on the idea to obtain a formulation liquid at
room temperature able to increase its viscosity once in the nasal cavity where it
forms a gel. The liquid form at room temperature allows an easy administration/
dosing (e.g., by spray), being very practical to use and with high patient compliance.
In situ gel formation takes place thanks to polymers that, once in the nasal cavity,
gel after specific stimuli, such as temperature, pH, ions in the nasal fluid [11–18]
(Table 12.2).
2.1 Temperature-Induced In Situ Gel System
Temperature is the most commonly used physical stimulus and is based on the fact
that at room temperature (20–25 °C) the formulation is in a liquid state while under-
goes a rapid sol-gel phase transition when in contact with body fluids (35–37 °C)
Table 12.2 Examples of in situ nasal gels

Stimulus for
Drug In situ gelling agent gelification Disease Reference
Lamotrigine Gellan and xanthan Nasal fluid ions Epilepsy [13]
gum
Timosaponin BII Poloxamer 407/ Temperature/ Alzheimer [14]
deacetylated gellan nasal fluid ions
gum
Fluticasone Gellan gum Nasal fluid ions Nasal inflammatory [15]
disorders
Docetaxel Pluronic F127 Temperature Brain tumor [16]
Piribedil Methyl cellulose Temperature/ Parkinson’s [17]
nasal fluid ions
Ropinirole Pluronic F-127 Temperature Parkinson’s [23]
Influenza virus Pluronic F127 and Temperature Influenza [26]
nucleoprotein F68
Breviscapine Gellan gum Nasal fluid ions Cerebrovascular and [27]
cardiovascular
Rufinamide Xyloglucan Temperature Epilepsy [28]
Fig. 12.1 Schematic representation of temperature-induced gelification of poloxamer
(Fig. 12.1). This type of behavior is characteristic of polymers, which can be syn-

thetic such as poloxamers; semi-synthetic such as methylcellulose (MC), hydroxy-
propyl methyl cellulose (HPMC), and ethyl (hydroxyethyl) cellulose (EHEC); or
natural such as xyloglucan and gellan gum.
Among them, poloxamers are widely used in pharmaceutics as thermo-responsive
gelling agents (as well as non-ionic surfactant) [19–23]. They are synthetic, A-B-A
type, linear, tri-block co-polymers made of hydrophilic end-groups of poly(ethylene
oxide) (PEO) and poly(propylene oxide) (PPO) hydrophobic core group [(PEOx-
PPOy-PEOx)]. They are commercially known with the trade names Pluronics®,
Lutrol®, Kolliphor®, Antarox®, and Synperonics®.
Although the exact gelation mechanism is not yet fully elucidated, it seems to be
correlated to the following steps. In water, the copolymers, due to their amphiphilic
character, self-aggregate into micelles with an inner core formed by PPO hydropho-
bic blocks and an outer shell constituted by PEO hydrophilic units. With the rising
of temperature, a packing and entanglement of the micelles, with concomitant dehy-
dration of the PPO block and the expulsion of water from the micelle core, is
observed [24] resulting in gelation of the system.
2.2 pH-Triggered Systems
The gelation process triggered by changes in pH is typical of polymers containing

pendant acidic or basic groups that either accept or release protons in response to
changes in environmental pH. These polyelectrolytes behave differently toward pH,
depending on whether they are anionic or cationic polymers. In the case of anionic
polymers, gelation occurs with the increase of the external pH, while the opposite is
observed with cationic polymers.
The polyacrylic acid (Carbopol®) is the most representative of the pH-responsive
polymers. The Carbopol®-based formulations are initially maintained at a pH value
Fig. 12.2 Schematic representation of pH-induced gelification of polyacrylic acid. Neutralization

with a base creates negative charges along the backbone of the polymer. These repulsive forces
uncoil the polymer into an extended, highly swollen structure
between 4 and 5 and when in contact with nasal cavity, where the pH is approxi-
mately 6.2, the polymer changes conformation resulting in sol-gel transformation
(Fig. 12.2). At low pH values, approximately 3.5, the carboxylic acid groups are
protonated and thus no effective charge is present in the molecule. This results in
limited polymer-solvent interactions, leading to a collapsed conformation of the
polymer with low hydrodynamic volume [25]. As the pH increases the carboxylic
groups are deprotonated, exhibiting a negative charge along the backbone. These
repulsive forces uncoil the polymer into an extended, highly swollen structure, with
a significant increase in viscosity of the aqueous solutions.
2.3 Ionic Gelation
Some polymers are ion-sensitive, undergoing a phase change in the presence of

various ions such as K+, Ca2+, and Na+. These ions are found in the nasal fluid rep-
resenting another stimulus useful to induce the gelification of ion-sensitive polymers.
For example, in the case of the carrageenan (polysaccharide formed by galactose
and 3,6-anhydro-D-galactose units), water-soluble biopolymers obtained from red
algae, the type k-carrageenan is able to form rigid and brittle gels mainly in pres-
ence of K+ ions while iota(ι)-carrageenan forms elastic gels in the presence of
Ca2+ ions.
The anionic polysaccharide gellan gum, obtained from cultures of Sphingomonas
elodea, gels with both mono- and divalent cations, including Ca2+, Mg2+, K+, and
Na+. The linear polysaccharide sodium alginate, obtained from brown algae, under-
goes ionic gelation in the presence of multivalent counterions Ca2+ and Mg2+, due to
cross-linking reaction between bivalent ions and guluronic acid block in alginate
chains forming hydrogels with a characteristic structure called “egg-box” (Fig. 12.3).
All the above-mentioned polymers are often combined with others having a bio-
adhesive capacity (e.g., cellulose derivatives, polyvinylpyrrolidone), that is, the
-OOC OH -OOC
OH
O O
OH OH
-OOC HO O -OOC HO
O O O
HO O O HO O
O OH OH OH
HO HO O O O O HO
O -OOC
-OOC OH OH OH OH
-OOC -OOC
Ca++ Ca++
-OOC -OOC
-OOC OH OH OH -OOC OH
O HO O O
HO O O HO OH
O OH OH
HO O O HO O
-OOC HO O
G O O -OOC O
HO
O OH OH
M
O
M OH -OOC G OH -OOC
M M
Ca2+
G: 1,4-α-L--Guluronic acid
M:1,4-β-D-Mannuronic acid
Fig. 12.3 Gelification mechanism of alginate in the presence of ions
ability to interact with mucin chains on the nasal mucosa, thus prolonging dosage
form residence time. This can allow a greater absorption of the drug and longer last-
ing protective effect of the nasal mucosa, if needed.
Bedford et al. [26] developed an in situ gel for vaccine nasal administration using
a mixture of pluronic F127 and F68 combined to the mucoadhesive polymer chito-
san loaded with the model antigen ovalbumin. In vivo studies performed on mice
highlighted that a prolonged retention and a better absorption of the antigen in the
nasal upper respiratory tract are observed using the in situ bioadhesive gel com-
pared to the same formulation without the thermosensitive polymers (pluronic).
Moreover, the number of antigen-positive cells resulted 6.7-fold more as well.
These results confirm the advantages derived from the use of thermo-responsive
polymers in the formulation. The same authors observed a better protection against
a respiratory virus infection loading the in situ gel with influenza virus nucleopro-
tein. Mice, previously infected by a heterologous influenza A virus, treated with the
in situ gel vaccine formulation showed a reduction of virus growth 100-fold in com-
parison to mice treated with the same formulation without thermo-responsive poly-
mers. Moreover, the virus was not detected in the lung of 80% of mice [26].
As an alternative to pluronic, polymers from natural sources have been employed
to obtain the in situ gelification. For example, Chen et al. [23] prepared a nanosus-
pension of the poor soluble molecule breviscapine in 0.5% of gellan gum (m/v), a
polysaccharide produced by the bacterium Sphingomonas elodea, which forms an
in situ gel, thanks to the interaction with the cations present in the nasal fluid. In
vivo studies performed on rats showed that, after intranasal administration, the for-
mulation increases the drug retention time in the nasal cavity resulting in an
improved concentration in cerebrum, cerebellum, and olfactory bulb tissues com-
pared to i.v. administration. For example, the AUC0–8h measured in the cerebrum was
3194.833 ± 501.745 μg/L·h vs 139.603 ± 33.691 μg/L·h for intranasal and i.v.
administration, respectively. Moreover, evaluating the intranasal administration of
breviscapine in comparison to the oral administration, a higher absolute bioavail-
ability was measured resulting 57.12% vs 0.40%, respectively. The tmax of intranan-
sal administration resulted low (0.167 ± 0.000 h), indicating the rapid drug
absorption from nasal mucosa [27].
Dalvi et al. [28] used xyloglucan as an in situ gelling agent. It is a tamarind seed
polysaccharide in which galactose residues were removed to obtain the thermo-
reversible form.
The in situ gel, loaded with the anti-epileptic drug rufinamide, allowed to obtain
a preferential accumulation of the drug in the brain compared to the aqueous sus-
pension of the drug administered intranasally. In fact, the bioavailability resulted
enhanced as testified by the AUC values (AUC0 → tlast) measured resulting
201.8 min·μg/g vs 104.28 min·μg/g for in situ gel and drug suspension, respec-
tively [28].
Recently, the nasal route is under investigation for a new nasal gel for COVID-19
respiratory infections treatment. In fact, on April 2021 a clinical trial started (still
ongoing) to evaluate the efficacy of a nasal gel loaded with the antiviral drug
LTX-109 for the treatment of severe acute respiratory syndrome SARS-CoV-2 [29].
3 Recent Approaches
Despite the use of film-forming and bioadhesive polymers solves the problem of the
low residence time, however the nasal absorption of molecules showing poor bio-
pharmaceutical properties (solubility and/or permeability) and/or stability problems
(e.g., chemical and enzymatic) is still a challenge. Therefore, new approaches
involve the incorporation of nano- and microcarriers into the in situ nasal gel in
order to protect the drug, improve the biopharmaceutical properties, and control the
release (if needed) [30–32] (Table 12.3).
This technological approach was investigated both for local and systemic thera-
pies [33]. In particular it represents a more valuable platform for a nose-to-brain
delivery promoting the drug absorption via the olfactory neuroepithelium in order
to improve the bioavailability of drugs used in the treatment of central nervous sys-
tem (CNS) diseases [34, 35] as well as alternative for brain cancer therapy [36].
Among the carriers used for this application, liposomes [37, 38], niosomes, nano-
structured lipid carrier (NLCs) [39], dendrimers [40], and β-cyclodextrin [41], prop-
erly customized, are the most useful.
Liposomes are versatile carriers that, due to their structure, can entrap both
hydrophilic and lipophilic drugs, improving their absorption through the nasal
mucosa [37].
Mura et al. [42] used PEGylated liposomes for nasal administration of the anal-
gesic opiorphin that, due to its peptidic nature, is susceptible to degradation.
Table 12.3 Examples of in situ nasal gel combined to nanocarriers

In situ Stimulus for
Drug gelling agent gelification Nanocarrier Disease Reference
Voriconazole Gellan gum Nasal fluid Nanotransferosomes Nasal fungal [33]
ions infection
Carbamazepine Poloxamer Temperature NLC Epilepsy [35]
407 (P407)
and
poloxamer
188 (P188)
Ondansetron Poloxamer Temperature Niosomes Cancer [36]
407
Donepezil Poloxamer Temperature Liposomes Alzheimer’s [38]
407 and
poloxamer
188
Apixaban Poloxamer Temperature Ethosomes Thromboembolic [43]
407 and disorders
poloxamer
188
Sumatriptan Poloxamer Temperature Nanotransferosomes Migraine and [45]
succinate 407 and cluster headaches
poloxamer
188
Buspirone Carbopol pH Niosomes Anxiety [46]
hydrochloride 974P and disorders
poloxamer
407
Teriflunomide Carbopol- pH-nasal NLC Glioma [51]
gellan gum fluid ions
Resveratrol Gellan gum Nasal fluid NLC Alzheimer’s [53]
ions
Disulfiram Deacetylated Nasal fluid Nanoemulsion Glioblastoma [54]
gellan gum ions
Duloxetine Pluronic Temperature Cubosomes Depressive [55]
F127 and disorders
PF68
Paeonol Gellan gum Nasal fluid PAMAM dendrimer Parkinson’s [56]
ions
Clonazepam Poloxamer Temperature Cyclodextrins Seizures, panic [58]
407 disorder, and
akathisia
Liposomes were loaded and then dispersed in a mixture of poloxamer 407 (as
thermo-responsive polymer) and carbopol 934P (as bioadhesive polymer) able to
form a gel at 34 °C with a rapid gelation time (10 s). Ex vivo studies performed on
nasal porcine mucosa showed a considerable enhancement of opiorphin permeation
from the thermosensitive hydrogel containing liposomes (Papp 14.3·10−4 cm/min)
compared to the same formulation containing the drug in free form (Papp
2.5·10−4 cm/min).
El-Shenawy et al. [43] developed a formulation based on a thermosensitive-
bioadhesive gel associated to ethosomes loaded with the anticoagulant drug apixa-
ban, a molecule characterized by low permeability (class III of the Biopharmaceutics
Classification System, BCS). Ethosomes were prepared using lecithin, cholesterol,
and ethanol and dispersed in a mixture of poloxamer 407/ poloxamer 188 (thermo-
sensitive polymers) and carbopol 934 (bioadhesive polymer). In vivo studies on
rabbits showed that this formulation is able to produce high plasmatic concentra-
tions improving drug bioavailability (Cmax 0.618 ± 0.073 μg/ml, AUC0–6h
2.078 ± 0.084 μg h/ml) compared to the nasal solution of the drug (Cmax
0.298 ± 0.04 μg/ml, AUC0–6h 0.710 ± 0.04 μg h/ml). A further study showed that the
in situ gel exhibits a bioavailability 5.5 times more than the oral solution of apixa-
ban [43].
Hosny et al. [44] prepared a nasal formulation for fungal sinusitis treatment con-
taining the antimicotic agent amphotericin loaded in nanotransferosomes prepared
using soybean lecithin and clove oil, which were dispersed in a gellan gum base.
Amphotericin is a molecule that, due to its physico-chemical properties, is not able
to cross the membranes. The use of nanotrasferosomes showed an improved pene-
tration as testified by ex vivo studies performed on goat nasal mucosa (permeability
coefficient 3.101 × 10−3 cm/min vs 0.411 × 10−3 cm/min of aqueous suspension).
Omar et al. [45] developed an in situ gel, for migraine and cluster headaches
treatment, based on thermosensitive polymers (mixture of poloxamer 407 and
poloxamer 188) and the bioadhesive carrageenan containing nanotransferosomes
loaded with sumatriptan succinate. As this drug molecule has a low oral bioavail-
ability (15%), the choice of intranasal administration route appeared to be advanta-
geous. In vivo studies performed on two rabbit groups, one treated with an oral
solution of the drug and the other one treated intranasally with the developed in situ
gel, showed differences in the bioavailability. The AUC0–12 resulted low for the rab-
bits treated orally, 186.81 ng·h/mL plasma; 158.95 ng·h/mL in the brain vs.
723.65 ng·h/mL plasma and 742.37 ng·h/mL brain for the group treated with the
developed in situ nasal gel.
Abdelnabi et al. [46] developed an in situ gel for the intranasal administration of
buspirone hydrochloride, an anxiolytic agent, characterized by limited oral bio-
availability (4%) as subject to first-pass metabolism and short half-life (2–3 h). The
drug was loaded into niosomes then formulated in carbopol 974P solution able to
form a gel in situ at the nasal fluid pH value. In vivo studies highlighted that the
intranasal administration of drug in situ gel is more advantageous than the oral one.
In fact, the bioavailability measured as AUC0–24 resulted higher for the in situ gel
(141.86 ± 13.15 ng·ml−1·h) in comparison to oral administration
(67.54 ± 7.12 ng·ml−1·h). The study performed on animals treated with the devel-
oped nasal in situ gel highlighted a further improvement of AUC0–24 value
(462.95 ± 10.15 ng·ml−1 h), suggesting the advantage in the use of a formulation
able to remain in the application site for a prolonged time allowing better drug
absorption.
In situ nasal gels were developed in combination to lipid carriers mainly

employed to treat diseases affecting the central nervous system (CNS) as Alzheimer’s
[39, 47, 48], Parkinson’s [49], and glioblastoma [50]. In fact, due to their nature, the
lipid nanocarriers are able to cross the blood-brain barrier, allowing a preferential
drug accumulation in the brain [39, 51].
Rajput and Butani [52] formulated the poor soluble and unstable (susceptible to
enzymatic degradation) molecule resveratrol in nanostructured lipid carrier (NLCs)
dispersed in a mixture of the polymer gellan gum combined to xanthan gum (poly-
saccharide gum produced by the microorganism Xanthomonas campestris) [53]. In
vivo experiments were performed on rats divided into two groups: the first one
submitted to intranasal administration of the in situ gel loaded with NLC-resveratrol
and the second one treated with a resveratrol suspension (control). The plasmatic
curves obtained showed a rapid (Tmax 0.5 h) and higher localization of resveratrol in
the brain in comparison to the control. In fact, the AUC values (AUC0 → 8) measured
in the brain were 2572 ± 338 ng·h/ml vs. 1809 ± 206 ng·h/ml obtained from a nasal
suspension.
Qu et al. [54] investigated an ion-sensitive nanoemulsion in situ gel formulation
for disulfiram nasal administration to perform a more effective treatment of glio-
blastoma. Disulfiram, characterized by low solubility and high instability, was
loaded in the lipophilic phase of a nanoemulsion. In vivo studies performed on
glioma-bearing rats demonstrated that the intranasal administration of the ion-
sensitive nanoemulsion is able to promote the localization of the drug in the tumor
cells compared to a saline solution administered intranasally used as control. The
measured median survival time resulted 1.6 folds higher than the control group.
Recently “cubo-gels”, a combination of the cubosomes and a thermosensitive
hydrogel, were prepared for the nasal delivery of duloxetine, a drug used for the
treatment of depressive disorders [55]. In vivo experiments were performed on
Swiss albino rats divided into three groups. The first two were administered intrana-
sally with in situ “cubo-gel” and drug solution, respectively. The third one was
administered intravenously. The plasmatic AUC0–72 measured for the three different
groups highlighted that the “cubo-gel” was able to produce higher values
(457.96 ± 4.53 ng·h/mL) compared to the other two control formulations
(243.14 ± 8.16 ng·h/mL and 301.04 ± 9.03 ng·h/mL for intranasal solution and i.v.,
respectively). The brain bioavailability was evaluated as well, and the obtained
results confirmed the best results for rats treated with “cubo-gels” measuring higher
AUC values (179.62 ± 6.30 ng·h/mL) compared to intranasal solution
(109.27 ± 5.3 ng·h/mL) and i.v. administration (106.32 ± 3.7 ng·h/mL).
Another kind of carrier, combined to thermosensitive polymer, is represented by
polyamidoamine (PAMAM) dendrimers.
Xie et al. encapsulated paeonol, a neuroprotective agent useful for Parkinson’s
disease treatment, in the hydrophobic cavities of polyamidoamine (PAMAM)-
modified dendrimer [56]. The obtained complex was dispersed in a mixture of
deacetylated gellan gum (0.45% w/w) and HPMC (0.30% w/w) able to form a gel
in the presence of ions (Na+, K+, and Ca2+). In vitro studies demonstrated that the
molecule is released by a sustained mechanism within 12 h. The developed
formulation (containing a fluorescent molecule) was administered in the nasal cav-

ity of 25 rats in order to evaluate the nasal brain transport. A comparison was carried
out with the loaded dendrimer alone. The fluorescence images collected at estab-
lished times (0 h, 2 h, 6 h, 12 h, and 24 h) showed that the administration of the
complex by the in situ gel allows to obtain a better and higher localization of the
drug in the brain tissue than the loaded dendrimer alone. This result has been
explained considering that the latter is easily cleared from the nasal cavity while the
in situ gel is able to prolong the contact time with the nasal mucosa allowing a better
release and absorption.
Ahmed et al. [57] prepared an in situ gel based on gellan gum in which were
dispersed micelles loaded with the poor soluble molecule raloxifene hydrochloride
intended for the treatment and prevention of osteoporosis. In vivo studies performed
on rats showed a higher bioavailability (AUC) of the drug obtained from both the in
situ gel containing the micelles (11.26 ± 2.53 μg·h/ml) and the in situ gel containing
the drug in free form (2.34 ± 1.93 μg·h/ml) compared to oral tablets of raloxifene
hydrochloride (0.83 ± 0.47 μg·h/ml), suggesting the advantages in the use of
nasal route.
Cirri et al. developed a nasal thermosensitive and mucoadhesive gel to perform
clonazepam (anti-epileptic drug) intranasal delivery in order to overcome the prob-
lems associated with both oral and parenteral administration of this drug [58].
Poloxamer was employed as thermosensitive polymer and chitosan glutamate and
sodium hyaluronate as mucoadhesive and permeation enhancer, respectively. In
addition, a randomly methylated β-cyclodextrin was used to improve diazepam
solubility. In consideration of all the variables of the formulation, including the
drug-cyclodextrin ratios, the authors used a screening DoE for a systematic evalua-
tion of the formulation composition on gelation temperature and gelation time and
prepared many loaded gels at different clonazepam-cyclodextrin concentrations. All
selected formulations showed properties suitable for an in situ mucoadhesive ther-
mosensitive gel formulation, and they demonstrated that the gel formulations were
significantly more effective in the improvement of clonazepam permeation than the
drug solution. In vitro permeation studies showed that the complex clonazepam-
methylated β-cyclodextrin formulated as an in situ gel allows to obtain an improved
permeability of clonazepam (Papp ~ 9 × 10−5 cm/s) in comparison to clonazepam
solution (Papp ~ 2 × 10−5 cm/s).
4 Safety Aspects
Recently nasal drug delivery has widely increased, especially for peptides and pro-
teins, mainly due to rapid absorption to the systemic circulation and good bioavail-
ability of drugs, advantages offered by the particular anatomical structures of the
nasal cavity. In fact, the site of absorption offers a high contact area presenting only
two cell layers separating the nasal lumen from the dense blood vessel network in
the lamina propria, which is probably the main reason for the rapid absorption of
drugs via this route. On the other hand, it is very important to take into account that
the administration of nasal formulations could cause damage to this thin and deli-
cate epithelium [59].
For this reason, safety is a key issue when an innovative and effective drug for-
mulation, as nasal in situ gel, is planned. Total safety and not only that of the active
ingredient must be considered when developing a new product. In fact, in this kind
of formulation the excipients play a very important role and particular attention
must be paid to the penetration enhancers, eventually used, and to the polymers
(mucoadhesive and film-forming) which prolong the contact time with the nasal
mucosa. Their action could in fact significantly reduce the safety of the final medici-
nal product [60].
With regard to local effects on the nasal mucosa, the tolerability of the medicinal
product depends on many factors and differs, obviously, from individual to indi-
vidual. In fact, environmental aspects (temperature, local humidity), presence of
infections or inflammations, and previous illnesses or allergies can influence the
interactions between the nasal mucosa and the applied formulation.
The blood flow of the nasal mucosa is the factor most responsible for the regula-
tion of temperature and humidity of the inhaled air, and some nasal administered
drugs can both decrease blood flow (e.g., vasoconstrictors) and increase it (hista-
mine, albuterol, isoproterenol, and fenoterol). These variations, together with the
presence of polymers able to bind the mucin chains present on the mucosa, can
cause irritation, especially in case of continuous use. Rare side effects such as nose-
bleeds and the onset of perforation of the nasal septum have only rarely been
observed [60].
Particular attention must be paid to the penetration enhancers, largely used in
intranasal gels, generally able to improve the transport of drug molecules through
the nasal epithelium and mucosa due to the opening of the tight junctions, the altera-
tion of the mucus layer, and the inhibition of proteolytic enzymes. In turn, those
functions can have a disruptive character and thus lead to adverse side effects,
thereby significantly reducing the safety of the final pharmaceutical product. In fact,
this approach is very useful for drugs that possess low permeability, but the penetra-
tion enhancers alter the natural nasal barrier function damaging epithelial cells,
mucus, or the cilia. Thus, the barrier function may be lost.
The boundary between the irritant (dryness, irritation, sneezing, itching) and the
toxic (rhinitis medicamentosa, congestion, nasal lesion) activity depends on the
mechanism of action exerted but above all on the local exposure time. In this per-
spective it is possible to believe that these substances and compounds act as irritants
to the nasal mucosa but are non-damaging [59, 60].
Most nasal formulations, as hydrogels, are formulated as multidose preparations
requiring a preservative to prevent the growth of microorganisms upon repeated use.
The effects of various preservatives on mucociliary transport rate have been in vitro
studied in order to investigate their influence on the safety characteristics of the final
product. Methyl-p-hydroxybenzoate (0.02% and 0.15%), propyl-p-hydroxybenzoate
(0.02%), and chlorbutol (0.5%) halted mucociliary transport. The ciliostatic effect
of methyl-p-hydroxybenzoate and propyl-p-hydroxybenzoate was reversed after
rinsing with normal saline solution, whereas in the case of chlorbutol (0.5%) and
methyl-p-hydro-xybenzoate (>0.15%) the reverse of the inhibiting effect was vari-
able [59]. Benzalkonium chloride exhibits surfactant properties that can destroy the
ciliary membrane and thus might be expected to be toxic to cilia. In contrast, nasal
administration of 0.01% bezalkonium chloride in humans was well tolerated and
did not change nasal clearance after a single administration. Long-term administra-
tion of 0.02% benzalkonium chloride (6 weeks) in humans did not change the nasal
clearance rate without changes in mucosal morphology [59].
Ethylenediaminetetraacetic acid (EDTA) halted mucociliary transport irrevers-
ibly by disruption of ciliated epithelium. Due to its chelating ability, EDTA causes
expansion of the intercellular spaces and therefore could permit an increase in the
permeability of the tissue to various molecules [59].
The human nasal mucosa has an average physiologic pH of 6.3 (slightly acidic),
and the maintenance of the pH value in the mucus ensures the function of the ciliary
clearance. Therefore, the pH of nasal hydrogels should be within a pH range from
4.5 to 6.5 to avoid nasal irritation [60]. Not only the pH but also the osmolarity has
an influence on the mucus rheological properties and on the ciliary beat contributing
to local toxicological considerations [61, 62].
These aspects must be taken into consideration in the case of nasal hydrogels
based on filming and mucoadhesive polymers whose action derives from the ability
to establish numerous bonds, mainly hydrogen bond and van der Waals forces, with
water and biological structures [63].
Although the residence time in the cavity is considerably increased for polymeric
formulations that absorb water, when in contact with the mucosa to form a film, it is
not the only factor in the increased absorption of drugs. It has been suggested that
the gels swell by taking water from the mucus layer and the underlying epithelial
cells, resulting in a temporary widening of the tight junctions.
Also the osmotic pressure of the nasal hydrogels is an important parameter to
keep under control because hypotonic or hypertonic gels can have different effects
on the nasal mucus rheology and clearance [64]. Rossi et al. [65] demonstrated that
hypertonic formulations cause a weak interaction with the biological structures.
Most of the studies carried out to evaluate the toxicity of nasal formulations
focused mainly on acute drug-induced or short-term effects.
In vitro techniques often provide a good screening method for identifying sub-
stances with potential deleterious effects on the nasal mucosal structure although a
good correlation with in vivo use is not observed. Moreover, to make accurate and
reliable evaluations of the potential side effects of nasal gels, researchers have to
determine the effects of its long-term use in animals and humans [59, 60].
Despite the fact that in situ nasal gels represent a promising therapeutic tool,
pharmacovigilance data will be useful to perform a deep analysis of the safety
aspects.
Finally, patient compliance must be considered for an optimal nasal formulation.
In this respect, gel strength should be considered as an important physical parameter
in order to improve patient adherence to therapy. In fact, a gel strength in the range
25–50 sec is recommended [66]. A less strong gel hardly maintains its integrity
while a stiff gel could be responsible for irritation and consequent discomfort.
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Chapter 13
Nasal Delivery of High Molecular Weight
Drugs: Recent Trends and Clinical
Evidence
Emine Kahraman, Sevgi Güngör, and Yıldız Özsoy
Abstract In recent years, the nasal route has been a promising administration for
systemic drug delivery. However, the nasal epithelial barrier hinders high molecular
weight medications, especially hydrophilic drugs, while low molecular weight
drugs are rapidly absorbed through the nasal mucosa. Additionally, enzymatic deg-
radation and mucociliary clearance in the nasal cavity negatively affect the bioavail-
ability of these drugs in the peptide-protein structure. As a result of the limitations,
the nasal bioavailability of these drugs is generally less than 1%. Thereby, the recent
studies have been focused to increase the nasal bioavailability of high molecular
weight drugs, using various strategies such as particulate drug delivery systems,
mucoadhesive/thermosensitive polymers, absorption enhancers, and enzyme inhibi-
tors. In this chapter, the recent advances that developed to improve nasal bioavail-
ability of high molecular weight drugs have been reviewed in the light of literature
studies and preclinical and clinical trials.
Keywords High molecular weight drugs · Macromolecular drugs · Nasal delivery

· Peptide-protein drugs · Absorption enhancers
1 Introduction
The nasal drug delivery has been used for therapeutic reasons for centuries.
Nowadays, it is commonly utilized for the treatment of local inflammation, common
rhinitis, and allergic rhinitis, with active ingredients such as glucocorticoids and
antihistamines [1]. In recent years, the nasal delivery of the drugs into the systemic
E. Kahraman · S. Güngör · Y. Özsoy (*)

Department of Pharmaceutical Technology, Faculty of Pharmacy,
Istanbul University, Istanbul, Türkiye
e-mail: yozsoy@istanbul.edu.tr
https://doi.org/10.1007/978-3-031-23112-4_13
254 E. Kahraman et al.
circulation has gained a great importance as an alternative approach to oral and

parenteral drug administration because of its many advantages such as rapid drug
absorption resulting in a quick onset of action, prevention of the hepatic first-pass
metabolism, reduced adverse effect, increased patient compliance, and low costs
[2]. Besides these advantages, the nasal delivery of high molecular weight drugs has
some limitations including enzymatic barriers for peptide-protein drugs and low
permeability of the nasal epithelia. Thus, the nasal bioavailability of high molecular
weight drugs is low and variability is high, while the bioavailability of low molecu-
lar weight drugs is relatively high and variability is low in comparison with injec-
tions [3].
Thereby, the recent studies have been focused to increase the nasal bioavailability
of high molecular weight drugs, especially peptide-protein drugs, by using various
strategies such as particulate drug delivery systems, mucoadhesive/thermosensitive
polymers, absorption enhancers, and enzyme inhibitors. In this chapter, the recent
advances that developed to improve nasal bioavailability of high molecular weight
drugs have been reviewed in the light of literature studies and preclinical and
clinical trials.
2 High Molecular Weight Drugs
The high molecular weight drugs (also referred as macromolecules) are defined as

compounds having a number of average molecular weight that are greater than or
equal to 1000 Da. These molecules having mostly peptide-protein structures have
rapidly grown with development of biotechnology over the past 20 years [4]. Herein,
nasal delivery of peptide-protein drugs which are commercially in another dosage
forms (e.g., injection) on the market is intensively highlighted in this section.
2.1 Insulin
Insulin is a peptide hormone composed of 51 amino acids with a molecular weight

of 5778 Da. Since its discovery in 1922, it has been used in the management of
diabetes mellitus, which is a chronic disease characterized by elevated levels of
blood glucose [5, 6]. Nowadays, insulin has been subcutaneously administered with
side effects such as possibility of hypoglycemia episodes, weight gain, and inade-
quate post-meal glucose control [7]. Kupila et al. [8] reported that short-term use of
intranasal insulin was well tolerated with minimal hypoglycemia risk and no local
irritation. Moreover, Schmid et al. [9] reviewed safety of intranasal insulin using
articles published between 1999 and 2017. This retrospective review on the intrana-
sal insulin presented that no hypoglycemia episode or severe adverse effect were
reported after administration of intranasal insulin. However, further data are needed
to ensure long-term safety of chronic insulin administration. In addition to the man-
agement of diabetes mellitus, many studies including preclinical and clinical
13 Nasal Delivery of High Molecular Weight Drugs: Recent Trends and Clinical… 255
indicate that intranasal insulin might improve cognition and memory in the patients
with age-related cognitive impairments such as Alzheimer’s and Parkinson’s dis-
ease [3, 10–12].
Over the years, numerous formulations have been developed to improve
absorption of intranasal insulin. However, no approved formulation of intranasal
insulin is currently available on the market, with being the most studied molecule in
the formulation development [3].
2.2 Desmopressin
Desmopressin is a synthetic analogue of the natural antidiuretic hormone arginine

vasopressin, which has a molecular weight of 1069 Da [6]. It has been clinically
used to treat nocturnal enuresis, central diabetes insipidus, and congenital bleeding
disorders including hemophilia A and von Willebrand diseases over 40 years [13–
15]. On the market, it is available in several dosage forms including injection, oral
tablet, fast-dissolving tablet, sublingual tablet, nasal spray, and nasal drops [5, 16,
17]. Bypassing the intestinal first-pass effect, nasal spray has the higher bioavail-
ability (5–10%) than sublingual and oral tablets, but it exhibits high variation in its
bioavailability, which could lead to severe adverse effects such as hyponatre-
mia [18].
2.3 Salmon Calcitonin
Calcitonin is a peptide hormone composed of 32 amino acids with a molecular

weight of 3432 Da [6], which acts in the calcium homeostasis, regulating intestinal
calcium absorption and renal calcium reabsorption and inhibiting osteoclast activity
[19]. Calcitonin, which is commercially available as salmon calcitonin, has been
used in the treatment of osteoporosis and Paget’s disease. In 1995, its nasal spray
(Miacalcin®, Novartis) was approved by the Food and Drug Administration (FDA),
but its nasal absorption was very low (3%) [5].
2.4 Oxytocin
Oxytocin is a peptide hormone with a molecular weight of 1007 Da [6], which

normally plays a role in social bonding, labor, and post-partum period in the human
body. On the market, it is available in two pharmaceutical forms: injection and nasal
spray. Its parenteral form is intravenously used for labor induction, abortions, and
control of post-partum bleeding as nasal spray is used for stimulation of post-partum
milk ejection [3]. Additionally, some studies have indicated in the recent years that
intranasal oxytocin has created positive results on psychiatric disorders, social

behavior, autism, and obesity [20–22].
2.5 Glucagon
Glucagon is a peptide hormone composed of 29 amino acids with a molecular

weight of 3483 Da [6], which is used in the treatment of severe hypoglycemia that
could be life threatening. Since 1960, there have been only injectable preparations
on the market despite lack of ability to administer in the emergency conditions [23].
In 2019, nasal powder glucagon (Baqsimi®, Eli Lilly) was approved by FDA, to
meet the unmet medical need for an easily administrated glucagon [24].
2.6 Human Growth Hormone
The major isoform of human growth hormone is a protein composed of 191 amino
acids with a molecular weight of approximately 22 kDa [6]. It is used in the treat-
ment of growth hormone deficiency, Turner syndrome, Prader–Willi syndrome,
chronic kidney failure, and idiopathic short stature in children [25]. Its nasal deliv-
ery can greater simulate the normal endogenous pulsatile human growth hormone
secretion pattern when compared to a subcutaneous injection, but there has been
only its injection preparation on the market.
2.7 Teriparatide (Recombinant Human Parathyroid Hormone)
Teriparatide is a form of parathyroid hormone consisting of the first (N-terminus) 34

amino acids, which is the bioactive part of the hormone [26]. The drug, which is a
peptide with a molecular weight of 4118 Da [6], was approved by the FDA for par-
enteral treatment of osteoporosis in postmenopausal women and in men with idio-
pathic or hypogonadal osteoporosis that are at high risk for fractures [27]. There has
been still commercially no nasal form, despite several formulation are being under
development.
2.8 The Miscellaneous
Beside aforementioned macromolecules, there have been some molecules

(interferon-beta, basic fibroblast growth factor, glial-derived neurotrophic factor,
etc.) which are only under preclinical and clinical studies for the nasal delivery.
Table 13.1 The molecular weights and indications of high molecular weight drugs
Molecular
weight Structure
Molecule (Da)a (Peptide/protein) Indication Reference
Insulin 5778 Peptide The management of diabetes [28]
mellitus
Desmopressin 1069 Peptide The treatment of nocturnal [13–15]
enuresis, central diabetes
insipidus, and congenital
bleeding disorders
Salmon 3432 Peptide The treatment of osteoporosis [19]
calcitonin and Paget’s disease
Nafarelin 1323 Peptide The treatment of endometriosis [29]
Buserelin 1239 Peptide and fertility [30]
Leuprolide 1209 Peptide The treatment of endometriosis, [31]
prostate cancer, and central
precocious puberty
Goserelin 1269 Peptide The treatment of breast and [32]
prostate cancer
Oxytocin 1007 Peptide The stimulation of post-partum [33]
milk ejection
Glucagon 3483 Peptide The treatment of severe [24]
hypoglycemia
Teriparatide 4118 Peptide The treatment of postmenopausal [27]
and idiopathic osteoporosis
Interferon-beta 1663 Peptide The management of multiple [34]
sclerosis
Octreotide 1019 Peptide The management of acromegaly, [35]
diarrhea, and flushing caused by
carcinoid tumors and vasoactive
intestinal peptide-secreting
adenomas
Insulin-like 7649 Peptide The treatment of growth failure [36]
growth factor-I and short stature in children with
(IGF-I) severe primary IGF-I deficiency
Orexin A 3561 Peptide The treatment of narcolepsy [37]
Exenatide 4186 Peptide The adjunctive treatment of [38]
diabetes type 2
Hirudin-2 6892 Peptide The prophylaxis and treatment of [39]
heparin-induced
thrombocytopenia, venous and
arterial thrombosis, and shunt
thrombosis or treatment of
disseminated intravascular
coagulation
(continued)
Molecular
weight Structure
Molecule (Da)a (Peptide/protein) Indication Reference
Human growth ca 22,000 Protein The treatment of growth [25]
hormone hormone deficiency, Turner
syndrome, Prader–Willi
syndrome, chronic kidney
failure, and idiopathic short
stature in children
Erythropoietin ca 34,000 Protein The treatment of anemic patients [40]
with insufficient erythropoietin
production
Basic fibroblast In range of Protein The treatment of brain trauma, [41]
growth factor 16,000 and ischemic stroke, and
18,500 neurodegenerative diseasesb
Single-domain ca 15,000 Protein The treatment of severe [42]
antibody pneumoniab
Glial-derived ca 36,000 Protein The treatment of several [43]
neurotrophic neurodegenerative disorders
factor including Parkinson’s diseaseb
Nerve growth ca 140,000 Protein The treatment of [44, 45]
factor (NGF) spermatogenesisb
www.pubchem.com
a
Non-approved by no regulatory authority, only under preclinical and clinical trials

b
Further, only few studies have been to improve the nasal absorption in the literature.
Hence, information on these molecules is summarized in Table 13.1.
3 Superiorities and Limitations of Nasal Administration

for High Molecular Weight Drugs
3.1 Nasal Blood Flow
When compared to other biological membranes, the nasal mucosa is a relatively

porous, thin, and highly vascularized epithelial membrane. Also, it comprises a
large absorption area (150 cm2) with microvilli in epithelial cells. As a result of
these characteristics, the nasal mucous membrane is well supplied with blood. The
greater blood circulation, the easier it is for the drugs to be absorbed and distributed
in the system. Hence, the nasal administration ensures fast absorption of drugs,
rapid onset of action, and low risk of overdose [46].
3.2 Enzymatic Activity
Enzymes such as carboxylesterases, epoxide esterases, glutathione S-transferases,

epoxide hydrolases, aminopeptidases, endopeptidases, and exopeptidases exist
in the nasal cavity. They negatively impact stability of nasally applied peptide-
protein drugs. These drugs might be degraded by peptidase and proteases. The
endopeptidases and exopeptidases can cleave internal peptide bonds and peptides
at their N, C termini, respectively. This results in decreased nasal bioavail-
ability [2].
The nasal mucosa is covered with 5 μm of thick mucus which has a viscous gel on
the upper part and an aqueous sol layer on the lower part. The nasal mucus renews
itself approximately every 10–15 min at a speed of 5–6 mm/min. This occurs with
ciliary activity to clean the nose from foreign particles and pathogens. All foreign
bodies in the air inhaled are enclosed by the mucous layer and pushed from the nasal
cavity to nasopharynx to be thrown into the gastrointestinal tract. This movement of
mucus is named “mucociliary clearance”, which is the main defense mechanism of
the body against foreign particles and pathogens. Therefore, the retention time of
drugs is limited in the nasal cavity. Particularly, hydrophilic drugs are readily solu-
ble in the mucus and quickly removed from the nasal cavity by mucociliary clear-
ance, resulting in poor nasal absorption. Also, the mucociliary clearance being
affected by external factors (air pollution, smoking, lung diseases, etc.) might cause
variation in the nasal absorption [5, 47].
3.4 Nasal Absorption
Nasal absorption can be described as diffusion of a drug into the circulation via the
nasal mucosa. The physicochemical characteristics (ionization, lipophilicity, etc.),
surface charge, and especially the molecular size of drug affect its nasal absorption,
and then bioavailability. Low molecular weight drugs are well absorbed through the
nasal mucosa. However, the nasal mucosa is an obstacle for absorption of high
molecular weight drugs, especially for more than 1000 Da in size. Thus, the nasal
bioavailability of particularly hydrophilic peptide-protein drugs is mostly less
than 1% [5].
3.5 Physical Condition of Nose
The constant nasal administration is a challenge in the cold season because the
physiological change that is based on a disease in the nasal mucosa affects nasal
drug absorption. In the case of a disease, the mucous membrane swells after inflam-
mation or irritation, leading to decreased drug absorption. Moreover, itching and
sneezing during the disease intensify this effect [48].
4 Recent Trends in Nasal Delivery of High Molecular

Weight Drugs
Intranasal drug delivery is not complicated, hence all types of preparations (solution,
powder, spray, gel, suspension, ointment, insert, etc.) could be administrated for the
nasal delivery of high molecular weight drugs as well as small molecular weight
drugs. However, it has some limitations for high molecular weight drugs, as
explained in Sect. 3. Thereby, several strategies have been used to enhance the nasal
bioavailability of high molecular weight drugs, including improvement in the half-
life of drugs, higher mucoadhesion and retention time, protection from degradation
of enzymes in the nasal cavity such as aminopeptidases and endopeptidases, and use
of absorption agents that enable delivery through tight junctions of nasal epithelium.
4.1 Particulate Drug Delivery Systems
4.1.1 Microparticulate Systems
Microparticulate systems including microparticles and microspheres are matrix

carriers where the drug is dispersed in a polymeric material, with diameter in the
range of 1 μm and 1000 μm. These systems are fabricated by different encapsulation
methods including mostly spray-drying, emulsification solvent evaporation, and
phase separation [5]. The microparticulate systems which are produced using muco-
adhesive polymers such as chitosan and alginate could protect the peptide-protein
drug from degradation and ensure prolonged drug release via nasal mucosa into the
systemic circulation [49]. Mostly, these nasally applied systems are insoluble in
water, but absorb water into the matrix, resulting in swelling of the microparticles/
microspheres and formation of gel [50]. Chitosan is one of the most used polymers
to prepare these microparticulate systems. As a result of a combination of chitosan
and microparticle properties, the microparticles provide controlled drug delivery
while the chitosan increases the residence time in the nasal cavity and enhances
drug absorption by opening the tight junctions between the epithelial cells. Hence,
drug absorption through the nasal mucosa has shown a great extent as also reported
by Varshosaz et al. [51]. Further, the bioavailability of thiolated chitosan micro-
spheres (7.24 ± 0.76%) was 3–4 times higher than that of chitosan microspheres
(2.04 ± 1.33%) [52]. When thiolated chitosan microspheres are utilized, they
increase the bonds with the mucin protein because of their enhanced interactions
with the cysteine residues of the mucus glycoprotein, resulting in a longer retention
time in the nasal cavity [53]. Similarly, increased positive charge of aminated gela-
tin microspheres and thiomer polycarbophil-cysteine microparticles contributed to
insulin and human growth hormone absorption through the nasal mucosa, respec-
tively [54, 55]. Nema et al. [56] revealed that thiolated microspheres loading insulin
showed greater reduction in the blood glucose level than that of non-thiolated
microspheres in the streptozotocin-induced diabetic rabbits. Additionally, some
studies are available about thiolated microparticles and microspheres, as seen in
Table 13.2.
Differently, Balduci et al. [92] formulated desmopressin microparticles as
chimera agglomerates to improve nasal drug bioavailability. In vitro permeation
studies indicated that permeation of desmopressin in the novel formulation through
nasal mucosa was significantly higher than that of commercial liquid nasal spray. In
rats, it induced a significant reduction in urine production. In another study, Serim
et al. [59] produced spray freeze-dried lyospheres® with diameter in the range of
190 μm and 250 μm for nasal administration. Because of their low density, 90% or
greater of lyospheres deposited in the nasal cavity, resulting in the nasal bioavail-
ability of insulin of 7.0 ± 2.8%.
4.1.2 Nanoparticulate Systems
Nanoparticulate systems with diameter in the range of 20 nm and 200 nm have
become attractive as a promising administration for nasal delivery in recent years.
Despite complicated preparation process and paradoxical effectiveness reports, they
improve drug permeation through the nasal mucosa, protect peptide-protein drugs
from enzymatic degradation in the nasal cavity, increase delivery of vaccines to the
lymphoid tissue in the nasal cavity with an adjuvant activity, and offer a way for
peptide-protein drug delivery particularly into the brain and systemic circulation.
Moreover, these systems could be targeted for nose-to brain delivery of drugs and
decrease adverse effects of the drugs [49, 50, 93]. Generally, polymeric nanocarriers
(nanoparticles, nanocomplexes, dendrimers, nanogels, etc.) and lipid-based parti-
cles (liposomes, solid lipid nanoparticles, nanostructured lipid carriers, etc.) have
been reported to be used for nasal delivery of high molecular weight drugs
(Table 13.2). Chitosan and chitosan derivates have been widely used in the prepara-
tion of the nanoparticles for the nasal delivery, because of their mucoadhesive and
absorption-enhancing characters, being safety polymers. Zhang et al. [61] revealed
that intranasal administration of PEG-g-chitosan nanoparticles in the rabbits
resulted in a greater insulin absorption through the nasal mucosa in comparison
with insulin-PEG-g-chitosan suspension and control insulin solution. In other stud-
ies, insulin-loaded PEGylated trimethyl chitosan nanocomplexes and chitosan/
Table 13.2 The examples of particulate drug delivery system, mucoadhesive/thermosensitive

polymer, and absorption enhancer used to improve the nasal delivery of high molecular drugs
Polymer/others Molecule Delivery system Absorption enhancer Reference
Thiomer Human growth Microparticles – [55]
polycarbophil- hormone
cysteinea
Poly (acrylic Exenatide Microparticles – [57]
acid)-cysteinea
Poly (acrylic Insulin Microparticles – [58]
acid)-cysteine-poly
(vinyl pyrrolidone)a
Thiolated carbopol Insulin Microspheres – [56]
cysteinea
Polyvinylpyrrolidone Insulin Lyospheres Sodium taurocholate or [59]
cyclodextrins
Poly (L-aspartic acid) Insulin Submicron – [60]
and chitosan capsules
Polyethylene glycol Insulin Nanoparticles – [61]
(PEG)-grafted
chitosan
Chitosan/sodium Insulin Nanoparticles – [62]
alginate
Starch Insulin Nanoparticles Na glycocholate or [63]
Lysophosphatidylcholine
Chitosan-N-acetyl-L- Insulin Nanoparticles – [64]
cysteinea
Chitosan/ Insulin Nanoparticles – [65]
cyclodextrin
derivativesb
Lectins modified Basic Nanoparticles – [66]
PEG-polylactide- fibroblast
polyglycolide growth factor
Phenylboronic-acid- Insulin Nanoparticles – [67]
functionalized
dextran
Chitosan–ZnO Insulin Nanocomposites – [68]
PEGylated trimethyl Insulin Nanocomplexes – [69]
chitosan
Amine-modified poly Insulin Nanocomplexes – [70]
(vinyl
alcohol)-graft-
poly(L-lactide)a
(continued)
Chitosan, glyceryl Glial Chitosan-coated – [71]
distearate, caprylic/ cell–derived nanostructured
capric triglyceride, neurotrophic lipid carriers
poloxamer 188 factor with surface
Tween 80b modified
transactivator of
transcription
(TAT) peptide
Gelatin, poloxamer Basic Nanostructured – [72]
188, Tween 80b fibroblast lipid carriers
growth factor
Chitosan, cholesterol, Leuprolide Liposomes with – [73]
dicetyl phosphate, acetate chitosan
stearyl amine,
hydrogenated soya
phosphatidylcholineb
Soybean Salmon Ultraflexible – [74]
phosphatidylcholine, calcitonin liposomes
cholesterol, sodium
deoxycholateb
Alginate, propylene Insulin, Phospholipid – [75]
glycol, magnesium epidermal vesicles
salt, phospholipidb growth factor, (Phospholipid
oxytocin Magnesome)
Lauroyl proline Human growth Protein- – [40]
hormone, lipoamino acid
erythropoietin, complexes
and salmon
calcitonin
Polyamidoamine Insulin, Dendrimer – [76]
(PAMAM) calcitonin
Poly(N-vinyl Insulin Nanogel – [77]
pyrrolidone)
Chitosan Insulin Mucoadhesive Saponin, sodium [78]
gel deoxycholate,
ethylendiamine
tetra-acetic acid (EDTA),
lecithin
Carbopol Insulin Mucoadhesive – [79]
gel
Carbopol/ Insulin Mucoadhesive – [80]
hydroxypropyl gel
methylcellulose
Gelatin Basic Mucoadhesive – [81]
fibroblast gel
growth factor
(continued)
Chitosan/hyaluronate Insulin Insert – [82]
polyelectrolyte
complexes
Hydroxypropyl Insulin Insert – [83]
methylcellulose
Carbopol/Starch Salmon Powder – [84]
calcitonin,
human growth
hormone
N-[(2-hydroxy-3- Insulin Thermosensitive – [85]
trimethylammonium) gel
propyl] chitosan
chloride, poly
(ethylene glycol),
a-b-glycerophosphate
Chitosan, poly vinyl Insulin Thermosensitive – [86]
alcohol gel
Glyceryl Insulin Solution – [87]
monocaprylate-
modified chitosana
Sperminated gelatina Insulin Solution – [88]
Aminated gelatina Insulin Solution – [89]
Chitosan Insulin Solution EDTA, Tween 80, [90]
cyclodextrins
Chitosan Hirudin-2 Solution Glycyrrhizic acid [91]
monoammonium salt,
azone, hydroxylpropyl-
beta-cyclodextrin,
lecithin, EDTA, sodium
dodecylsulfate, Brij35,
Tween 80, menthol
Cationized polymers to increase the mucoadhesion
a
At least one component of the delivery system is an absorption enhancer

b
cyclodextrin nanoparticles showed 34–47% and ˃35% reduction in the blood

glucose concentration, respectively [65, 94]. In contrast to these results, Dyer et al.
[95] reported that chitosan powder for the nasal insulin delivery (bioavailability of
17.0%) was more effective in reducing the blood glucose concentration than chito-
san nanoparticles and chitosan solution (bioavailability of 1.3% and 3.6%, respec-
tively). Besides chitosan derivates, phenylboronic acid-functionalized glycopolymers
(3-acrylamidophenylboronic acid-r-N-acetyl glucosamine, 2-lactobionamidoethyl
methacrylate-random-3-acrylamidophenylboronic acid, phenylboronic-acid-
functionalized dextran, etc.) have recently attracted attention because of their muco-
adhesive and enzyme-inhibitory properties [67, 96, 97].
Lipid-based particles have been mostly applied by coating/incorporating a
mucoadhesive polymer into the nasal mucosa. In recent years, glial cell–derived
neurotrophic factor was encapsulated into chitosan-coated nanostructured lipid

carriers with surface-modified transactivator of transcription (TAT) peptide for
treatment of Parkinson’s disease in the study of Hermando et al. [71]. Similarly,
Akel et al. [98] indicated that chitosan-coated solid lipid nanoparticles had
superiority in comparison with native insulin for management of Alzheimer’s
disease.
4.2 Polymers
4.2.1 Mucoadhesive Systems
The mucoadhesive polymer approach has been developed to enhance nasal drug
absorption, extending formulation’s intimate contact time with the nasal mucosa by
attaching to the mucus layer’s surface. This is achieved by several drug delivery
systems including microspheres, powders, gels, inserts, using mucoadhesive poly-
mers such as chitosan, poly (acrylic acid) (Carbopol), hydroxypropyl cellulose,
hydroxypropyl methylcellulose, starch, and gelatin [54, 80, 99, 100]. This approach
has low toxicity, and the most promising method, especially when combined with
absorption enhancers [50]. Chitosan is the most used polymer of mucoadhesive
systems for nasal delivery of high molecular weight drugs. It has positive charge at
slightly acidic pH due to its glucosamine residues. Because of its ionization, chito-
san is soluble in water at pH˂6, and it becomes water insoluble at more than 6 of
pH. As a result of this, it could be utilized as a viscosity agent in the acidic medium
[101]. More importantly, it exhibits a strong adhesion ability to mucosal tissues
consisting of anionic sialic acid groups in the nasal cavity, owing to its cationic
amino groups. Additionally, solvent-drag mechanism of chitosan leads to widening
the tight junctions, and then high molecular weight drugs transports through the
nasal mucosa into the blood capillary, enhancing nasal absorption [50]. Further, it
has a high loading capacity of drug molecules, owing to interactions with peptide-
protein drugs of hydroxyl and amino functional groups in the chitosan [102]. It is a
promising mucoadhesive polymer for nasal delivery of high molecular weight
drugs. Hinchcliffe et al. [103] reported that relative bioavailability of salmon calci-
tonin from the chitosan solution was increased twofold in comparison with mar-
keted nasal product in animal studies. Additionally, most of the studies demonstrated
that nasal bioavailability of peptide-protein drugs increased greater than that of only
chitosan formulations when chitosan and chitosan derivates were combined with
absorption enhancers [78, 90, 91]. Apart from chitosan, carbopol, hydroxypropyl
methylcellulose, starch, gelatin, and their combinations with/without absorption
enhancers were widely studied to improve nasal absorption of several peptide-
protein drugs by numerous of research groups (Table 13.2).
4.2.2 Thermosensitive Systems
The thermosensitive systems which would be herein called as in-situ gelling systems
form gel from solution state in specified temperature conditions. In the solution
state, they may be readily dropped or sprayed into the nasal cavity and spread on the
nasal mucosa. Following the formulation into the nasal cavity, the solution could
transform into viscous gel at body temperature. As a result, the rate of nasal
mucociliary clearance reduces, and drug release gets slower. Poloxamers are widely
used for the preparation of thermosensitive gels [50, 104]. In recent years,
Bahmanpour et al. [105] have developed a novel thermosensitive hydrogel consist-
ing of chitosan, chitosan quaternary ammonium salt, and gelatin for intranasal insu-
lin administration. This novel formulation exhibited low gelation time, uniform
pore structure, and desirable swelling rate, which resulted in adequate encapsulation
and prolonged release of insulin over 24 h. Similarly, Wu et al. [85] and Agrawal
et al. [86] had separately optimized thermosensitive gels consisting of N-[(2-
hydroxy-3-trimethylammonium) propyl] chitosan chloride, poly (ethylene glycol),
a-b-glycerophosphate and chitosan, poly vinyl alcohol to improve nasal insulin
absorption, respectively.
4.3 Absorption Enhancers
The use of absorption enhancers is the most common and effective strategy to
enhance absorption of high molecular weight drugs through the nasal mucosa. The
absorption enhancers increase the permeability of epithelial cell layer based on vari-
ous mechanisms, by improving drug solubility or stability, inhibiting enzyme activ-
ity, reducing mucus viscosity or elasticity, decreasing mucociliary clearance, and
opening tight junctions between the cells. Ideally, they are compatible with drugs in
the formulation, reversibly rapid-acting on the absorptive properties without sys-
temic absorption. They provide predictable and reproducible absorption enhance-
ment degree. Although they are mostly safe, some of the mechanisms can lead to
severe irritation and damage to the nasal mucosa at concentrations required to effec-
tively promote nasal absorption, especially in case of chronic nasal administration
[50, 104]. Generally, absorption enhancers are mainly subdivided into six groups:
(i) bile salt and its derivatives, (ii) surfactants also composing of fatty acid and its
derivatives, (iii) chelators such as ethylendiamine tetra-acetic acid (EDTA), (iv) cat-
ionized polymers, (v) cyclodextrins, and (vi) cell-penetrating peptides.
The bile salts (sodium deoxycholate, sodium taurodihydrofusidate, etc.) are
conjugates of bile acids with taurine or glycine residues. They are utilized in the
formulation alone [106] or in combination with other absorption-enhancing methods
such as mucoadhesive polymers and particulate drug delivery systems [59, 63, 78].
The surfactants (lysophosphatidylcholine, dodecylmaltoside, tetradecylmaltoside,
tetradecylsucrose, dodecanoylsucrose, Laureth-9, sucrose cocoate, soybean-derived
sterol, sterol glucoside, etc.) can exhibit detrimental effect on the nasal membrane
at concentrations required to effectively promote nasal absorption, especially for

chronic nasal administration. Thereby, they have been evaluated over the years in
animal models [50, 107, 108]. The esters of hydroxystearic acid have recently been
of great interest because of its low systemic toxicity and negligible local toxic effect
on the nasal mucosa [109–112]. Williams et al. [110] indicated in rat models that
Cmax (13.7 ± 1.6 ng/mL) of human parathyroid hormone with polyethylene glycol
(15)-hydroxystearate (Solutol® HS15) was equivalent to Cmax (14.8 ± 8 ng/mL) in
subcutaneous injections. Additionally, Wang et al. [109] reported that polyethoxylated
12-hydroxystearic acid (Kolliphor® HS15) was effective and biosafe as a permeation
enhancer in the intranasal formulation of human parathyroid hormone.
The cationized polymers (thiolated chitosan, sperminated gelatin, aminated
gelatin, etc.) (Table 13.2) are the most effective and safest materials to increase
nasal absorption of high molecular weight drugs. Besides strong mucoadhesion
properties, as explained in Sect. 4.2.1, they lead to an ionic interaction with the
luminal surface of nasal mucosa, and then induce signals which open tight junctions,
resulting in intercellular permeation [50].
The cyclodextrins are oligosaccharides which is widely used in the pharmaceutical
area. In the nasal formulations, they could effectively utilize as absorption enhancer
and solubilizer [113]. Among cyclodextrin derivatives, dimethyl-ß-cyclodextrin is
the most effective absorption enhancer, especially for the powder formulations. The
absolute bioavailability of insulin with dimethyl-ß-cyclodextrin powder was
13 ± 4% in rabbits [114]. However, hydroxypropyl-ß-cyclodextrin and randomly
methylated-ß-cyclodextrin exhibit lower toxicity than dimethyl-ß- cyclodextrin
[115]. Principally, they increase the nasal absorption, by decreasing affinity of
peptide-protein drugs to the physical and/or metabolic barriers. Hence, (i) they
protect the peptide-protein drugs against enzymatic as well as chemical degradation.
(ii) The hydrophilic cyclodextrins could remove some lipids through nasal mucosa
via the formation of rapid and reversible inclusion complexes. (iii) They could
interact with the hydrophobic chain of peptide-protein drugs, altering its intrinsic
aggregation or permeability through the phospholipid bilayer. (iv) They might
change distribution of tight junction proteins, thereby opening the tight junctions
between epithelial cells [50].
The cell-penetrating peptides (D, L-penetratin, D, L-octaarginine, transactivator
of transcription (TAT), etc.), which are also known as “protein transduction
domains,” are novel high-capacity delivery vectors for various hydrophilic macro-
molecules. They effectively internalize various molecular cargoes of the drugs
through plasma membranes, without altering their activities [116–118]. L-Penetratin,
which is characterized by a high density of basic amino acids (Arg and Lys) and by
the presence of hydrophobic residues (importantly Trp), is one of the most promis-
ing cell-penetrating peptides as a universal vector for various high molecular weight
drugs including insulin, glucagon-like peptide-1 (GLP-1), exendin-4, and inter-
feron-ß [119–121]. L-Penetratin ensured that nasal bioavailability of insulin was
50.7% relative to subcutaneous administration, without causing detectable damage
to the integrity of cells in the nasal respiratory mucosa [119].
Besides aforementioned absorption enhancers, tight junction modulating

peptides (zonula occludens toxin, clostridium perfringens enterotoxin, etc.), tight
junction modulating lipids (glycosylated sphingosines, oxidized lipids and ether
lipids, etc.), nitric oxide donors (S-nitroso-N-acetyl-DL-penicillamine, etc.), and
N-acetyl-L-cysteine at a high concentration (20%) have been also studied to improve
nasal absorption of high molecular weight drugs [50, 122–124].
4.4 Enzyme Inhibitors
The enzyme inhibitors (soybean trypsin, bacitracin, nafamostat mesilate,

phosphoramidon, aprotinin, bestatin, etc.) protect the peptide-protein drugs from
enzymatic degradation in the nasal cavity. Namely, the enzyme inhibitors cannot
dramatically improve the nasal bioavailability, but they improve stability of peptide-
protein drugs at the absorption area by restricting enzymatic activity and reducing
degradation rate of the drugs [104]. For example, camostat mesilate, an
aminopeptidase and trypsin inhibitor, improved the nasal delivery of vasopressin
and desmopressin because of protecting the drugs from enzymatic activity in the
nasal cavity [125]. However, they are not very effective to improve nasal absorption
of peptide-protein drugs. Additionally, they might affect the normal metabolism of
body, and then lead to severe adverse effects [50].
4.5 The Miscellaneous
The other strategies are chemical modification of primary peptide structure or

preparation of pro-drugs and use of deep eutectic solvents to improve absorption of
peptide-protein drugs.
The chemical modification protects peptide-protein drugs from enzymatic
degradation or improves absorption characteristics of the drugs. In the first condition,
chemical modifiers such as for polyethylene glycol (PEG), poly styrene-maleic acid
copolymer, albumin, and dextran are used to protect the peptide-protein drugs.
However, this might not be feasible for the nasal delivery of hydrophilic large
peptide-protein drugs because of making it more hydrophilic and larger, which
reduced nasal absorption of peptide-protein drugs. In the second condition, the
drug’s absorption across the nasal mucosa improves owing to the increased lipophi-
licity, because the preparation of pro-drug makes the drug more lipophilic. However,
it can cause to decrease pharmacological activities of parent peptides [50, 104]. Due
to these limitations, there are no examples of chemical modification for nasal
administration of peptide-protein drugs.
The use of deep eutectic solvents is a novel strategy, which is rarely used to
improve nasal absorption of macromolecules. The deep eutectic solvents are mix-
tures of compounds having lower melting points than melting points of the
compounds individually. They exhibit as greater solubilization as higher viscosity

properties. Additionally, they could readily interact by hydrogen bonds, electro-
static forces, and van der Waals forces. These could lead to increased nasal absorp-
tion of the macromolecules [104]. Li et al. [126] demonstrated that choline chloride
and maleic acid eutectic mixture increased solubility and release rate of insulin.
Also, increased viscosity of the formulation improved the residence time of insulin
in the nasal mucosa, and then its nasal absorption.
5 Clinical Evidence on Nasal Delivery of High Molecular

Weight Drugs
In recent years, the number of high molecular weight drugs administered nasally
has progressively increased on the market, despite temporarily or permanently stop-
ping the production of some molecules. Additionally, clinical studies of novel nasal
formulations of high molecular weight drugs are ongoing. The information about
marketing status and clinical studies of these molecules are presented in Table 13.3.
Presently, a range of nasal peptide formulations on the market exhibits very low
bioavailability (less than 1%). However, these drugs are intended for non-parenteral
administration considering clinical superiorities and development costs [5]. The
marketed formulations generally contain none of efficient nasal absorption strate-
gies, because of the poor nasal tolerability of the most known absorption enhancers
[112]. However, recently a commercial product (Baqsimi®, Eli Lilly) and a few
clinical studies have emerged. Baqsimi® dry powder formulation (commercial
product) which contains synthetic glucagon utilizes phospholipid dodecylphospho-
choline as surfactant and absorption enhancer, and beta-cyclodextrin (β-CD) as
filler/bulking agent and absorption enhancer [127]. Additionally, dodecyl maltoside
(Intravail™ Technology, Aegis Therapeutics) and cyclopenta decalactone (azone,
CPE-215) have been in development as the nasal delivery systems of teriparatide
and insulin (Nasulin, CPEX Pharm), respectively [93].
6 Future Directions and Conclusion
The nasal route is a promising administration for drug delivery into the systemic
circulation because it offers large absorption area, relatively porous, thin, and highly
vascularized epithelial membrane. However, the nasal epithelial barrier constitutes
an obstacle for drugs greater than 1000 Da. Thereby, high molecular weight drugs,
particularly those which are hydrophilic (e.g., peptide-protein drugs), cannot trans-
port through the nasal mucosa. Further, enzymes in the nasal cavity are a hurdle for
stability of high molecular weight drugs with peptide-protein structure. Numerous
strategies have been developed to overcome these issues. Among these, absorption
Table 13.3 The high molecular weight drugs administered via intranasal delivery on the market
and under clinical trials
FDA
Marketing Dosage Commercial approval
Molecule status form Strength name Company date
Desmopressin Prescriptiona Spray, 0.01 mg/ Minirin Ferring 2002
acetate metered spray
Discontinuedb Spray, 0.01 DDAVP Ferring 1978
metered mg/spray
Discontinuedb Spray, 0.15 mg/ Stimate Ferring 1994
metered spray
Discontinuedb Spray, 0.15 mg/ Octostimc Ferring 2018
metered spray
Buserelin Prescription Spray, 0.1 mg/ Suprefactc Sanofi-Aventis –
Nafarelin Prescription Spray, 0.2 mg/ Synarel Pfizer 1990
Interferon Prescription Spray, 500 IU/ Genferond Biocad 2011
alfa-2b metered spray
Glucagon Prescription Powder 3 mg/ Baqsimi Eli Lilly 2019
dose
Oxytocin Discontinuedb Solution 40 USP Syntocinon Novartis 1960
units/mL
Salmon Discontinuedb Spray, 200 IU/ Miacalcin Mylan 1995
calcitonin metered spray
Recombinant Discontinuedb Spray, 200 IU/ Fortical Upsher Smith 2005
salmon metered spray Labs
calcitonin
Insulin Phase 2 Spray, 100 IU/ Nasulin CPEX Pharm –
metered dose
Phase 2 Solution 40 IU/ – Beth Israel –
dose
Oxytocin Phase 2 Solution 24 IU/ – Massachusetta –
dose General
Hospital
Phase 2 Solution 24 IU/ – TriGemina –
dose
Teriparitide Phase 1 Solution 90 mcg/ – Nottingham –
dose University
Hospital
Somatotropin Phase 1 Solution n.d. – Critical Pharm –
a
Cold storage formulation
b
If generic versions of this product have been approved by the FDA, there may be generic equiva-
lents available
c
Approved by Canadian and EMA authorities
d
Approved by Russian authority
enhancer method is the most common and effective strategy to enhance absorption
of high molecular weight drugs through the nasal mucosa, which is used in a com-
mercial product (Baqsimi®, Eli Lilly). In light of recent studies, ongoing preclinical
and clinical trials, it is asserted that novel nasal formulations containing high molec-
ular weight drugs and absorption enhancers will become gradually available on the
market in the next decades.
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Chapter 14
Niosomes-Based Drug Delivery
in Targeting the Brain Tumors Via Nasal
Delivery
Mahmoud Gharbavi, Sepideh Parvanian, Milad Parvinzad Leilan,

Shabnam Tavangar, Maedeh Parchianlou, and Ali Sharafi
Abstract Targeting tumors has always been a herculean task. Moreover, the

presence of blood brain barrier (BBB) acts as a physical barrier and restricts the
transportation of therapeutic molecules across the brain. Targeted delivery of the
therapeutic payload across the blood brain barrier has gained widespread attention
over the past few years. Intranasal route offers delivery to the brain via the trigemi-
nal and olfactory route surpassing BBB. It also offers various other advantages such
as surpassing biotransformation, and systemic absorption increasing the efficacy.
Over the last few decades, several novel drug delivery systems such as liposomes
and other lipid nanoparticles targeting brain, have gained widespread attention. The
Niosomes are vesicular nanoparticle flatforms comprised of non-ionic surfactants,
which are biodegradable, more stable than liposomes. This current review discusses
the potential use of niosomes as a delivery vehicle for targeting brain tumors via the
nasal route.
Keywords Niosomes · Nasal route · Blood brain barrier · Nanoparticles · Tumours
M. Gharbavi
Nanotechnology Research Center, Ahvaz Jundishapur University of Medical Sciences,
Ahvaz, Iran
S. Parvanian
Faculty of Science and Engineering, Åbo Akademi University & Turku Bioscience Center,
Turku, Finland
M. P. Leilan · S. Tavangar
Department of Medical Biotechnology, School of Medicine, Zanjan University of Medical
Sciences, Zanjan, Iran
M. Parchianlou · A. Sharafi (*)
Zanjan Pharmaceutical Biotechnology Research Center, Zanjan University of Medical
Sciences, Zanjan, Iran
e-mail: alisharafi@zums.ac.ir
https://doi.org/10.1007/978-3-031-23112-4_14
280 M. Gharbavi et al.
1 Introduction
Chemotherapy, radiation therapy, and targeted drug therapy play a significant role in
the treatment of glioblastoma and central nervous system (CNS) diseases to decrease
the mortality rate [1]. The main problem is the inability of drugs to cross the blood-
brain barrier (BBB) to reach the brain tissue in sufficient quantities to achieve thera-
peutic levels. It is estimated that the BBB prevents intrusion of approximately 98%
of low molecular weight drugs and about 100% of macromolecules drugs leads to
poor bioavailability for drug delivery to the CNS [2]. Different strategies including
intracerebroventricular or intraparenchymal injections, mini-pump-assisted intra-
cranial delivery, catheter infusions, accurate ultrasound methods, or electromag-
netic force-field are used to deliver active drug agents. However, these methods are
aggressive and dangerous for patients [3]. BBB is the first limiting factor to the
delivery of drugs to the brain through systemic circulation [1]. Several studies are
underway to cross the BBB through the nose-to-brain approach. Intranasal delivery
is a promising alternative approach compared to the invasive methods mentioned
above for drug delivery to the brain, because the nasal cavity has so many arteries
that provide a high absorption level for the prescribed drug. It also allows this path-
way to bypass the BBB and provide fast and direct drug delivery to the brain [4].
Also, this route (delivery through the nose) limits unnecessary drug systemic expo-
sure and reduces systemic toxicity [5].
As the olfactory nasal segment cavity extends to the cranial cavity, nasal drug
delivery can provide direct access to the brain [3]. At present, nanotechnology-
based drug delivery systems provide a great opportunity for intranasal drug delivery
to the brain. Nano-drug delivery systems have been widely studied in the last
decades as a new strategy to solve the problem of poor bioavailability of various
drugs [6]. Successful delivery of the drug to the brain through liposomes, den-
drimers, microspheres, nanoemulsions, carbon-based nanoformulations, micro-
spheres, and dendrimers has been reported in different studies [7].
The major goal of vesicular structure development is to change distribution
profiles, control drug release over time, and deliver drugs to target sites. Vesicular
systems can handle high amounts of drugs and generate an appropriate surface for
targeting. It enables the drugs to carry both hydrophilic and lipophilic components.
The non-ionic surfactant vesicles (Niosomes), systems with the advantages of lipo-
somes and the permeability of membranes, are created in the aqueous phase from
non-ionic surfactants. The integrity of niosomes in biological fluids is a critical
requirement for their function as a medication carrier. Niosomes are to circulate in
the body while simultaneously protecting the medicine for a certain period, connect
with the target site, and convey their contents into the target cells as a carrier.
Niosomes are preferred in comparison to other bilayer structures, because of chemi-
cal stability, biodegradability, biocompatibility, low production cost, low toxicity,
and easy storage and handling. Niosomes have been used by different delivery
routes, such as oral, intramuscular, intravenous, transdermal, and so on. In this
chapter, we will discuss and suggest the niosomes as versatile nasal formulations for
brain targeting of drugs.
14 Niosomes-Based Drug Delivery in Targeting the Brain Tumors Via Nasal Delivery 281
2 Nasal Drug Delivery Route
2.1 The Blood-Brain Barrier (BBB) and Targeted Drug

Delivery to the Brain
Despite advances in the treatment of brain diseases, the blood-brain barrier (BBB)
is a major barrier to the delivery of drugs to the central nervous system (CNS).
Crossing BBB barrier is a challenging problem for most of the effective drugs on
central nervous system diseases such as neuropeptides, proteins, chemotherapeutic
agents, monoclonal antibodies, recombinant proteins, and antisense or gene therapy
agents. The BBB is made up of tight junctions between the brain capillaries endo-
thelial cells with low endocytic activity. This structure leads to a capillary wall that,
like the lipid bilayer of the cell membrane, prevents the passage of polar and insol-
uble substances across the BBB. With significant advances in nanotechnology, sev-
eral strategies have been developed for drug delivery to the CNS. Some strategies,
such as modifying the drug itself, binding it to the transcytosis vector, and using
appropriate carriers, increase the capacity of therapeutic agents to cross the
BBB. One of the current challenges is to develop a targeted drug delivery system
that can effectively cross the BBB barrier while the drug agent remains intact [8, 9].
2.2 Transmitting to the Brain Through Nasal Passages
Understanding the anatomy and physiology of the nasal cavity is essential to the
success of nasal drug delivery systems. The nasal cavity can be divided into three
areas: the olfactory area, respiratory area, and vestibule. The respiratory area is rich
in blood vessels; thus, it can provide systemic absorption of the drug after intranasal
administration and subsequent indirect delivery of the drug to the brain. The vesti-
bule is a small area and the drug absorbed through this area is very low [10, 11]. The
respiratory area is suitable for the delivery of the vaccines by the intranasal route.
The olfactory area also plays an important role in the direct delivery of drugs to the
brain and cerebrospinal fluid (CSF) [1, 12]. The main purpose of these drug delivery
routes is to deliver the desired drug concentration to the drug activity site. Due to the
permeability of the nasal epithelium, high overall flow, porous endothelial mem-
brane, large surface area, and evading of the first passage metabolism cause the drug
to be rapidly absorbed into the brain. Methods of drug delivery through the nasal
route can transfer a wide range of therapeutic agents (small molecules and macro-
molecules) to the CNS. Several studies have shown that, when administered nasally
to the CNS, the drug offers effective therapeutic effects in lower doses (Fig. 14.1).
The transmission of therapeutic agents from nose to brain is described below.
Fig. 14.1 Schematic of the niosomal nasal delivery route for brain targeting
2.2.1 Olfactory Pathway
The olfactory area at the top of the nasal cavity is known as the possibility of drug
delivery through the nose to the brain for the treatment of various CNS diseases
[13]. Drugs can pass through the olfactory epithelial space through the tight intersti-
tial space with passive diffusion, or through transmission through cell membranes
with endocytosis, or neuron transfer [5, 14]. Most drugs that deposit in the olfactory
area are extracellularly transported between cells. Various studies on drug delivery
through the nose have suggested the role of P-glycoprotein in this pathway. In addi-
tion, a study was performed to test the penetration and transfer of the drug in the
three-dimensional culture of these cells (3D MucilAir) as a model of nasal structure
[15]. The olfactory neurons play an important role in targeting drugs to the brain
through the nasal path [16]. The path of the drugs for transmission is from the intra-
cellular axon to the olfactory bulb and then to the brain [17]. The diameter of the
olfactory axon in humans is about 0.1–0.7 micrometers, indicating that molecules
that have a diameter in this range can easily deliver their pharmaceutical agents
through this route. Since nanosystems used in drug delivery are usually nano sized,
they seem to be suitable for transmission through this pathway [7].
Drug delivery through the epithelium is faster than axonal transport. Drug
delivery from the olfactory pathways occurs through extracellular and intracellular
mechanisms. Most lipophilic drugs are transported through passive diffusion, while
most hydrophilic drugs are transported through the paracellular pathway. The
hydrophilic and molecular weight of drugs has a significant effect on drug absorp-
tion. Drugs with high lipophilicity are usually absorbed through the transcellular
pathway [5].
2.2.2 Trigeminal Pathway
The trigeminal pathway for drug delivery from the nose to the brain has been less
studied. The main function of the trigeminal nerve is to transmit chemical and ther-
mal information to the nose, mouth, and ocular mucosa [18, 19]. The trigeminal
nerve pathway can be an important site for drug delivery to the brain through the
nasal route. For example, insulin-like growth factor 1 was transmitted to the brain
through the trigeminal and olfactory pathways [20].
2.2.3 Lymphatic Pathway
Drugs can be transferred through several extracellular pathways such as perineural,

perivascular, and lymphatic channels in the olfactory region. These extracellular
pathways are connected to the olfactory bulb of the brain by olfactory nerves [7,
21]. Therefore, the lymphatic pathway also plays an important role in drug delivery
from the nose to the brain.
2.2.4 Systemic Pathway
The systemic pathway is an indirect transmission from the nose to the brain and can
be a promising approach for low molecular weight lipophilic drugs [22, 23]. Drugs
are absorbed by the vascular regions of the epithelial membrane of the nasal mucosa
and lymphatic system and then are transported to the systemic circulation to avoid
the first-pass metabolism of the drug [22, 24].
2.3 Advantages and Disadvantages of the Nasal Drug

Delivery Route
Targeted drug delivery through the nasal route to the brain reliably, effectively, non-
invasively, and directly transmits drug agents to the CNS via neural connections
between the nose and the brain [25]. The nasal cavity has high blood vessels that are
also highly permeable and therefore one of the best places to prescribe drugs [10,
26]. The following are some of the unique benefits of nasal transmission:
1. Has a large surface area for drug absorption.
2. Facilities and patients are comfortable.
3. In this method, the level of drugs in the bloodstream rises rapidly.
4. Drugs penetrate this pathway well, especially low molecular weight
lipophilic drugs.
5. Circumvent difficult conditions of absorption through the intestine.
6. Circumvent hepatic metabolism, which is the first-line metabolism for drugs
that are absorbed from the intestine.
7. It is possible to transfer the drug directly to the brain through the olfactory nerves.
8. This pathway is adjacent to the lymphatic tissue; the vaccine is administered
through the nasal route directly to the lymphatic tissue.
9. This route is suitable for people who are undergoing long-term drug treatment.
10. Suitable for prescribing drugs that have low fluid stability.
In the nasal uptake pathway, drug agents reach the olfactory bulb and brainstem
after passing through the surface of the nasal epithelium, and through the pulsating
current, that spaces around the cerebral blood vessels and contributes to drug
absorption, drug agents spread to other areas of the CNS. In some cases, nasal trans-
mission is almost equivalent to intravenous injection because there is a unique con-
nection between the nasal cavity and brain [27–29].
Nasal administration is the only way to administer directly to the brain without
non-invasive methods [22]. Because proteins, peptides, nucleic acids, and even stem
cells can be transported through the nasal passage, this route has received consider-
able attention for drug delivery. In addition, via the nasal passage drugs can be
administered both locally and systematically. Drugs in the form of suspensions,
solutions, gels, surfactants bases, and emulsions can be administered through the
nose, and administration through this route increases the efficiency of targeted
delivery and decreases the side effects of systematic administration [30–33]. Some
factors that affect nasal absorption are as follows:
1. Some physicochemical properties of drugs: including drug or nanocarriers

containing drug size, molecular weight, hydrophilic or lipophilic, and resistance
to enzymatic degradation
2. Nasal condition: rate of mucociliary clearance, nasal pH condition, and

endothelial cell permeability
3. Including drug formulation, drug solubility, and viscosity [34–37]
For example, a drug with low molecular weight, lipophilic, and resistance to
enzymatic degradation with high endothelial permeability and low clearance of the
nasal cavity is well absorbed from the nasal route. The anatomy of the nasal cavity
and the condition of the nasal mucosa can affect the process of drug absorption such
as enzymatic degradation, mucociliary clearance, nasal cavity blood flow, nasal
health conditions [38–45].
2.4 Mechanism of Drug Absorption from the Nasal Route
The main step in absorbing drugs from the nasal cavity is to cross the mucus. Large
particles find it relatively hard to pass through the mucus layer, but small particles
pass easily [46]. The nasal mucosa contains mucin, a protein that can bind to sol-
utes, and the presence of this mucin affects the absorption process. Environmental
or physiological changes cause structural changes in the mucosal layer, and this
influences the rate of absorption through the nose [47]. After the drug passes through
the mucosa, there are several mechanisms for absorption through the mucosa. These
include simple diffusion from the membrane, transcellular transcytosis by vesicular
carriers across the cell, and paracellular transmission between cells. Among the
several mechanisms mentioned, paracellular and transcellular pathways are pre-
dominant [48]. Transmission from the paracellular pathway is slow and passive.
The lower the molecular weight of the drug or nanocarriers containing the drug, the
faster it is absorbed through nasal passages. In contrast, low bioavailability has been
reported for drugs with molecular weight above 1000 Daltons [46]. Lipid drugs are
often transported through a lipoid pathway, also known as the intercellular process;
the transfer of this pathway depends on the lipophilicity of the drug. Other drug
routes include passing through the cell membrane through active transport facili-
tated by the carrier and passing through the opening of tight junctions [48]. Barriers
to drug absorption are potential metabolism before reaching systemic circulation
and improper length of stay in the nasal cavity [49]. Many water-soluble drugs are
poorly absorbed through the nose and therefore do not have sufficient bioavailabil-
ity. Penetration enhancers are often used to increase the absorption and bioavail-
ability of such drugs [50]. The mechanism of action of penetration enhancers is that
they increase the rate of drug absorption by making reversible changes in the struc-
ture of the nasal epithelial barrier [49]. Researchers have been drawn to investigate
the intranasal drug delivery method based on the findings so far. Nonetheless, it is
critical to comprehend medication uptake across the nasal mucosa. The nose is a
complex organ from a kinetic standpoint since three separate processes, such as
drug disposal, clearance, and absorption, occur simultaneously inside the nasal cav-
ity. Understanding the nasal anatomy and related physiological aspects is critical for
optimal drug absorption across the nasal mucosa.
2.5 Nasal Anatomy and Physiology of the Nose
The human nasal cavity is separated into two nasal cavities by the septum and has a
total volume of 16 to 19 mL and a total surface area of 180 cm2 [51]. Each cavity
has a volume of around 7.5 mL and a surface area of about 75 cm2 [52]. A solute can
be deposited in one or more of three anatomically distinct locations following medi-
cation administration into the nasal cavity: the vestibular, respiratory, or olfactory
regions. The vestibular area is responsible for filtering airborne particles and is
located at the entry of the nasal passages [53, 54]. When it comes to medication
absorption, it is thought to be the least essential of the three zones [55]. The respira-
tory system is the largest and most vascularized, and it is primarily responsible for
drug absorption. The olfactory region has a surface area of around 10 cm2 and is
important for drug delivery to the brain and CSF. In the nasal cavity, there are three
main anatomical zones. A mucus layer covers the epithelium of the nose canal, trap-
ping particles. Cilia clean the mucus layer from the nasal cavity, which is replen-
ished every 10 to 15 min [56]. Mucosal secretions have a pH of 5.5 to 6.5 in adults
and 5.0 to 6.7 in children [57], which retains particles and allows cilia to remove
them from the nasal cavity. The mucus moves through the nose at an approximate
rate of 5 to 6 mm/min resulting in particle clearance within the nose every 15 to
20 minutes [58, 59]. Numerous enzymes [51], for instance, cytochrome P450,
enzyme isoforms [60] (CYP1A, CYP2A, and CYP2E), carboxylesterases, and glu-
tathione S-transferases are found in the nasal cavity [61, 62].
2.6 Brain Targeting Through the Nasal Route
Because of the BBB’s poor distribution into the CNS, the development of numerous
potentially interesting CNS therapeutic candidates has been hampered for some
time. The intranasal route can deliver therapeutic drugs to the brain without passing
through the BBB because of the unique connection between the nose and the
CNS [63].
A unique characteristic and superior choice is the capacity to transfer therapeutic
drugs to the brain via drug absorption across the olfactory region of the nose [58].
When drugs were administered nasally to rats, some drugs produced significantly
higher CSF and olfactory bulb drug levels than when administered intravenously
[25]. Many scientists have identified evidence of nose-to-brain transfer [64]. Many
previously abandoned potent CNS medication candidates with intranasal delivery
have the potential to become successful CNS therapeutic drugs. Thus, several nasal
intranasal injection formulations were developed for the treatment of disorders such
as epilepsy, migraine, MS, depression, and erectile dysfunction [65].
2.7 Drugs for Glioblastoma Treatment

Administered Intranasally
Several studies have been carried out to discover the optimal intranasal treatment
for glioblastoma (GBM) utilizing monotherapy or in combination with other drugs,
including natural and/or synthetic agents. The research that was done to develop a
viable treatment for this aggressive brain tumor is summarized here.
Intranasal delivery of curcumin, as a natural compound, combined with a
glioblastoma-specific antibody was suggested by Mukherjee et al. The targeted
curcumin-CD68 Ab conjugate was intranasally administered to mice in which gli-

oma GL261 cells were xenografted in the brain. Adult male C57BL/6 mice were
given a curcumin-CD68 Ab solution in PBS intranasally every 72 h ten days after
GL261 cells were xenografted, whereas another set of animals got an intraperito-
neal injection of a commercially available lipid-complexed form of curcumin, that
is, Curcumin Phytosome. Curcumin-CD68 Ab conjugate intranasal delivery and
Curcumin Phytosome intraperitoneal injection both caused GL261 brain tumor
remission in 50% of mice, confirming that CD68 Ab could be delivered to the brain
via the intranasal route and that CD68 Ab had a targeted therapeutic effect after
intranasal delivery. Furthermore, on day 90, 70% of the animals given curcumin-
CD68 Ab intranasally and 60% of those given Curcumin Phytosome intraperitoneal
were still alive, whereas all the control group animals, that is, vehicle-treated mice,
were already dead. As the obtained results, intranasally delivered curcumin-targeted
conjugates can directly kill GBM cells and also lead to repolarizing tumor-associated
microglial cells (TAMs) to a tumoricidal state [66].
Rhein (4, 5- dihydroxyanthraquinone-2-carboxylic acid) is a natural compound
with anti-inflammatory, antioxidant, anti-fibrosis, neuroprotective, and anti-tumor
properties [67]. The CD38 enzymatic activity is inhibited by rhein, which leads to
attenuating glioma progression. To demonstrate this, Blacher et al. conducted a
study while using a syngeneic mouse glioma progression model (CD38-deficient
C57BL/6J (CD38-/-) mice) [68]. Glioma cells (GL261) were intracranially
implanted into the mice’s brains after 24 h, and vehicle or rhein was administered
three times each week for 22 days. Rhein can suppress CD38 enzymatic activity,
which leads to reduced microglia activation that is supportive of tumor progression.
The intranasal administration of rhein suppressed the glioma progression signifi-
cantly in WT mice, showing that CD38 is a therapeutic target in the tumor microen-
vironment and that small-molecule inhibitors of CD38 could be a potential treatment
for glioma [68]. Furthermore, Shingaki et al. evaluated the direct brain uptake of
5-fluorouracil (5-FU) from the nasal cavity, as well as whether the inhibition of CSF
secretion by choroid plexus could lead to increased brain concentration of the free
drug [69]. In this study, male Wistar animals were administered 5-FU intravenously
or nasally in the presence or absence of intravenous infusion of acetazolamide (AZA).
AZA (25 mg/kg) was injected for 15 min before initiating the nasal perfusion of
5-FU in the n groups of co-treatment. CSF secretion by choroid plexus epithelial
cells is inhibited by AZA. The active transport of Na+ ions is connected to CSF
secretion in these cells, and AZA significantly reduces the activity of the Na/K
ATPase [70]. The results found that intravenous administration of AZA increased
the CSF content of nasally given 5-FU by 200–300% when compared to 5-FU nasal
perfusion without pre-treatment with AZA. By reducing CSF secretion from the
choroid plexus and so maintaining the concentration of the nasally administered
drug in the CSF, AZA was able to improve nose-to-brain drug transport [69]. It was
concluded that co-administration of therapeutic agents to treat neurological diseases
with drugs that reduce CSF secretion from the choroid plexus could be an interest-
ing alternative to treating diseases of the brain, such as GBM, because the concen-
trations of therapeutic agents in the brain are improved.
In another report, the same group found a similar effect in male Wistar rats after
methotrexate (MTX) administration by nasal injection [71]. MTX is a folic acid
antagonist that inhibits the enzyme dihydrofolate reductase and has been used in
treating a variety of cancers [72]. Because MTX has a poor penetration across the
BBB, therapeutic options for GBM via oral administration are limited [73]. In the
study, MTX was administered nasally with sodium carboxymethyl cellulose (CMC)
added to improve the nasal residence time of the formulation, and AZA was given
orally 30 min later. The amount of MTX measured in the CSF was higher than that
measured in plasma 15 minutes after intranasal injection, indicating that MTX was
transported directly from the nasal cavity to the CSF. When compared to the con-
centrations found in the CSF following intraperitoneal administration, plasma had a
greater concentration. Simultaneously, the effect of oral AZA 30 min before nasal
MTX administration was investigated, and it was shown that the co-treatment
enhanced the concentration of MTX in CSF by 195% [71].
In another research, MTX-loaded chitosan microspheres were prepared by spray-
drying technique. In this way, different molecular weights of chitosan were used to
fabricate chitosan microspheres owing to promote the nose-to-brain delivery of the
MTX. The animals were given MTX solution, and MTX-loaded chitosan micro-
spheres were intranasally administered. According to the obtained results, a higher
concentration of MTX in rat brain tissues was shown after intranasal administration
of the MTX-loaded chitosan microspheres when compared to the MTX solution,
which was attributed to the presence of chitosan. In fact, chitosan is known to be a
safe mucoadhesive polymer that could effectively improve the brain hydrophilic
drug delivery, like MTX, via intranasal administration [74].
Another study [75] suggested that temozolomide (TMZ) be delivered by the
nose. After oral administration, TMZ is efficiently absorbed and is available in cap-
sule form. TMZ has also shown good penetration via the BBB and an acceptable
toxicity profile [76]. However, a significant increase in overall survival was observed
in multimodal treatment with TMZ and radiotherapy group as compared to the
radiotherapy alone group. This study suggested that 60–75% of patients with GBM
present no clinical benefit from treatment with TMZ [77]. Based on these findings,
a rat model with orthotopic C6 glioma xenografts was employed to investigate the
therapeutic efficacy of intranasal administration of TMZ to take benefit of the drug’s
brain-targeting capabilities. In fact, it was proposed that TMZ be administered intra-
nasally to restrict systemic exposure to the drug and therefore reduce toxic effects
on healthy organs. During the 40-day experiment, the rats were given saline solution
or TMZ via three distinct delivery routes: intravenous, oral, or intranasal, and tumor
size, rat survival time, and pathological changes were evaluated. When compared to
all other groups, including controls, magnetic resonance imaging revealed a signifi-
cant reduction in the volume of glioma xenografts in the intranasal TMZ group
(p < 0.05). Immunohistochemistry and a terminal deoxynucleotidyl transferase
dUTP nick end labeling (TUNEL) assay were also used to examine proliferating
cell nuclear antigen (PCNA) and tumor cell apoptosis. High tumor cell apoptosis
rate as well as a reduction in protein expression of PCNA were observed in treat-
ment by the intranasal route.
When comparing the three groups of C6 glioma-bearing rats, the intranasal TMZ
group had a significantly higher median survival time. Control animals given saline
solution survived 20 days, while animals treated with TMZ orally, TMZ intrave-
nously, and intranasally survived 21.5, 19, and 31 days, respectively [75]. The find-
ings of this study suggest that intranasal TMZ delivery can inhibit the growth of C6
glioma in vivo and that it could be an effective glioma treatment strategy.
Pineda et al. evaluated a solution of TMZ in dimethyl sulfoxide (DMSO) in nude
mice xenografted models bearing human glioblastoma tumors derived from the
human glioma stem cell lines TG16, TG1N, and TG20 [78]. TG16, TG1N, and
TG20 human glioma cell lines were injected intrastriatal into ten-week-old female
Swiss nu/nu mice. The anesthetized mice received 10 L of TMZ or vehicle intrana-
sally 1 month after the graft, and this treatment was repeated three times a week for
2 weeks. Intranasally administered TMZ slowed tumor growth and significantly
increased the lifetime of mice engrafted with TG16 and TG1N cells, without any
effect on tumors produced by TG20 cells, which are resistant to TMZ in vitro. The
findings showed that the intranasal route for TMZ delivery into the brain for the
treatment of intrastriatal brain tumors should be investigated further [78].
The studies that are reviewed above collectively demonstrate that the intranasal
administration of anticancer drugs can induce benefits in the treatment of GBM and
the intranasal route of administration might allow for direct access of the drugs to
the brain, serving as an effective strategy for glioblastoma treatment. However, the
use of nanotechnology to design a nanosystem as an intranasal drug delivery system
could be a promising strategy for clinical employment of nose-to-brain administra-
tion over more traditional methods.
3 Nanotechnology-Based Drug Delivery
The notion of medicine administration in traditional dose forms is shifting because

of nanotechnology. Nanoparticles are a type of particulate medication delivery tech-
nology in which the particle size is in the nanometer range (1–1000 nm).
Nanoparticles are being studied in great detail to develop medication delivery meth-
ods that can penetrate physiological barriers [62]. There has been a lot of interest in
developing nanotechnology by employing nanoparticles as carriers for tiny and
large molecules throughout the last few decades. Nanoparticles have been created
using a variety of polymers. The term “nano” comes from a Latin word that means
“dwarf.” A nanometer is one thousand millionth of a meter (i.e., 1n = Nanosize, size
refers to one thousand millionth of a given unit). For several decades, the word
“nanotechnology” has been most widely used in fields of science such as electron-
ics, physics, and engineering. Biomedical and pharmacological disciplines, on the
other hand, have yet to be investigated [79].
The nanomaterial has several advantages including increased surface, enhanced
solubility, increased rate of dissolution and bioavailability, rapid onset of action, and
less amount dose required in the field of pharmacy [80]. These materials and tech-
nologies can be developed to interact with a high degree of functional specificity for
applications in medicine and physiology, offering a level of interaction between

technology and biological systems not previously possible [81]. In this chapter, we
focused on niosomes as key nanocarriers in nasal drug delivery.
3.1 Structure of Niosomes
The structure of niosomes is spherical and consists of one or more microscopic

layers. This structure formed by non-ionic surfactants that cholesterol and charge
inducers can also be used in this structure [82]. Different types of surfactants used
to form niosomes differ in the number of combinations and the molar ratio [83].
Examples of surfactants used to synthesize niosomes include polyoxyethylene fatty
acid esters, sorbitan fatty acid esters, alkyl glyceryl ethers, and alkyl ethers [84].
The addition of cholesterol to the bilayer of surfactant in niosomes maintains the
strength of the bilayer and reduces its leakage. Also, charge inducers provide charge
to vesicles, and the size of these vesicles increases, thus increasing the encapsula-
tion efficiency of the drug in niosomal structures. Negative charge inducers include
lipoamino acids, dihexadecyl phosphate, and dicetyl phosphate and positive induc-
ers include cetylpyridinium chloride and stearyl amines; these compounds increase
the stability of niosomes structures [85, 86]. In an aqueous solution, non-ionic sur-
factants orient the hydrophilic end of the amphipathic surfactant molecules outward
(i.e., toward the aqueous phase), while the hydrophobic ends of the two surfactant
molecules orient each other (i.e., toward the lipophilic environment) and form a
bilayer structure resembling a cell membrane [82].
In the structure of closed niosomes, the inner and outer bilayer surfactant is an
aqueous phase and a lipophilic space is in the middle of these two phases [86].
Energy is required to form the closed bilayer structure of the niosomes, which is
supplied by thermal energy or physical stimulation. Van der Waals forces and repul-
sive forces between surfactant molecules are the most important stabilizing forces
of vesicles in the structure of niosomes. Variables such as changes in vesicle com-
ponents (type, concentration, and composition), surface charge, size, and volume
will alter the properties of the niosomes [87]. As shown in Fig. 14.2, niosomes can
be classified into three groups based on their vesicle size: small unilamellar vesicles
(SUV; 10–100 nm), large unilamellar vesicles (LUV; 100–500 nm), and multilamel-
lar vesicles (MLV > 500 nm) [88].
3.2 Advantages and Disadvantages of Niosomes-Based Drug

Delivery Systems
Niosomes have several advantages over other nano-carriers:

1. Surfactants used for synthesis niosomes are non-immunogenic, biocompatible,
and biodegradable.
2. The method used to produce niosomes does not involve very toxic solvents.
Fig. 14.2 Schematic typical vesicle size of niosomes
3. The chemical stability of niosomes components is high, so their storage and

transportation do not require special conditions.
4. By changing the structural composition and production method of niosomes,
their physicochemical properties such as size, shape, and fluidity can be easily
changed.
5. Niosomes can carry large amounts of drugs.
6. Niosomes can be used to deliver unstable and sensitive drugs because they
protect drug ingredients against heterogeneous conditions inside and outside
the body.
7. Niosomes improve the therapeutic function of drug molecules because they
have high circulation time and limit the effects of the drug on the target cell.
8. Niosomes can be administered in a variety of ways, such as topical, oral, and
injectable.
9. Niosomes have different drug formulations: semi-solid, powder, and suspension.
10. Niosomes increase the bioavailability of insoluble drugs when used orally.
11. Niosomes increase the penetration of drugs when used for skin delivery.
12. Niosomes-based drug delivery formulation leads to better patient compliance
compared to free oil forms.
13. To regulate the rate of drug release from the structure of the niosomes, becoming
an aqueous phase, it can be emulsified in the non-aqueous phase.
In contrast, niosomes have disadvantages including aggregation, physical and
chemical instability, decomposition of vesicles, and leakage encapsulated drugs.
Also, the methods required to prepare multilayer vesicles are time-consuming and
require specialized equipment [89, 90].
3.3 Formulation Components of Niosomes
3.3.1 Non-ionic Surfactants
Non-ionic surfactants are a group of surfactants that have no charge in the structure
of their head group. Non-ionic surfactants have high stability and biocompatibility
compared to positive, negative, and amphoteric surfactants. Non-ionic surfactants
have an amphiphilic structure, that is, they have a distinct hydrophilic part and a
distinct hydrophobic part [91].
Non-ionic surfactants play a major role in the structure of niosomes and are the
most abundant factor in the structure of niosomes. Non-ionic surfactants used in
niosomes synthesis have an amphipathic structure and these include polysorbates
[92], terpenoids [93], spans [94], alkyl oxyethylene [95], and so squalene belongs to
the group of terpenes, a natural lipid, used in the synthesis of niosomes.
The advantage of squalene in niosomes synthesis is that it stabilizes the structure
of niosomes and is also slightly toxic in vivo and in vitro [93]. Polysorbate is another
group of non-ionic surfactants used in the structure of niosomes; for example, nio-
somes synthesized with 80 Polysorbates are an excellent vector for gene delivery
because they have a polyethylene glycol (PEG) group in their structure. Niosomes
with a structure of Span 60/Tween 60/cholesterol are a carrier with a high percent-
age of drug encapsulation because the interaction is established between the acyl
group of 60 Span and the drug [94].
3.3.2 Cholesterol
Steroids are an important part of cell membranes, and their presence affects
membrane fluidity and permeability. Cholesterol is the most important steroid that
is often used to synthesize niosomes. Cholesterol binds to non-ionic surfactants
using a hydrogen bond.
Although cholesterol may not play a role in lipid formation, it plays a role in
stabilizing and controlling the properties of niosomes nanostructures. The addition
of cholesterol to niosomes formulation affects the properties of the nanosystem,
such as lipid layer permeability, rigidity, increased drug encapsulation efficiency,
easier hydration of frozen niosomes, and increased biocompatibility of nanocarri-
ers. Cholesterol reduces leaky niosomal nanocarriers by inhibiting phase shifts from
gel to liquid [96].
3.3.3 Charge Inducer Molecules
In the synthesis of niosomes, charge inducer molecules may also be added. Charge
inducer molecules, by inducing positive or negative electrostatic charges on the
surface of niosomal vesicles, maintain the suspension state of nanocarriers, prevent
aggregation, and ultimately increase stability. Negative inducers of electric charge
include diacetyl phosphate (DCP) and phosphatidic acid. Stearylamine (STR) and
stearyl pyridinium chloride are negatively charged inducer molecules. For niosomes
synthesis, a percentage of charge-inducing material of 2.5–5 M is acceptable; add-
ing more than this amount prevents synthesis [97].
3.4 Types of Niosomes
3.4.1 Proniosomes
Proniosomes is a new vesicle system for delivering medication to the skin and
ocular. Proniosomes overcome a variety of disadvantages of previous structures,
such as physical stability, aggregation, and leaking. Proniosomes are suitable for
drug delivery because they have no first-pass hepatic metabolism, no adverse effects
on oral delivery, and no gastrointestinal tract (GIT) [98]. Proniosomes are composed
of non-ionic surfactants whose outer part is hydrophobic and the inner part is
hydrophilic. Because peroxisomes increase the permeability of the skin layers, they
are very suitable for transporting drugs through the skin. Proniosomes are dehydrated
niosomes that become niosomes with the absorption of water. Proniosomes are
more stable than other carrier vesicles [98]. Proniosomes are inactive and must be
transformed to the active form, niosomes, to function. The proniosomes become
niosomes by passing through the skin layer or adding an aqueous solution. When
proniosomes are administered to the skin, they hydrate in the skin and form one-side
concentration on the outer surface of the skin, which increase the permeability of
the skin. When niosomes lysis into the endosomes of subcutaneous tissue, encapsu-
late drugs are released [99, 100]. Non-ionic surfactants used in the synthesis of
niosomes are also used to synthesize proniosomes. These non-ionic surfactants are
used in combination with cholesterol and lecithin, a structure-stabilizing phospho-
lipid. To hydrate the lipid layer, hot water, phosphate buffer with pH = 7.4, and 1%
glycerol are used to synthesize proniosomes [100]. Proniosomes have been success-
fully used as a carrier for better delivery of various drugs, including Roxithromicin,
Tazarotene, α-Mangostin, Tolterodine tartrate, and so on [98].
3.4.2 Ethosomes
Ethosomes were first introduced by Touitou et al. in 1997 [101]. Ethosomes arose
from the modification of liposomes and were composed of phospholipids, high con-
centrations of ethanol, and water [102]. Ethosomes nanocarriers compared to lipo-
somes have (1) reduced particle size, (2) negative zeta potential, (3) higher drug
encapsulation percentage, and (4) are more stable [103]. However, to develop a
more efficient delivery system, a new generation of ethosomes, that is, binary etho-
somes, and transethosomes developed. Zhou et al reported different form of etho-
somes such as binary ethosmes by adding alcohol to the conventional form [104]. In
the formulation of binary ethosomes, in addition to ethanol, another alcohol is usu-
ally propylene glycol (PG), and isopropyl alcohol is also present [103, 105, 106].
PG increases permeability, low toxicity, low skin irritation, high viscosity, as well as
greater stability than ethanol [103, 107]. This specificity of PG causes the drug to
increase its affinity to the skin and also the drug to accumulate into deeper levels of
the skin. Adjusting the ratio of PG and ethanol is very important to achieve proper
penetration of the drug into the skin [105, 108]. Sung et al. introduced a new genera-
tion of ethosomes in 2012. The advantages of this new generation autosome were
the same as those of previous generation ethosomes and liposomes [109].
Transethosomes are identical to ethosomes in composition, but they also contain a
penetration enhancer (surfactant) [109]. Evidence suggests that transethosomes are
smaller in size, more elastic, and have more permeability to the skin – the “more
permeability to skin” probably due to the synergistic effect between ethanol and
surfactant [110].
3.4.3 Bola-Surfactant Niosomes
Bola surfactant is used to synthesize bola niosomes. Surfactants of this type were
initially discovered in the membrane of Archaebacteria in 1980. These surfactants
have two hydrophilic heads that are connected by one or two lipophilic bonds. In
2010, Zakharova et al. showed that bola surfactants have low critical micelle con-
centrations, high surface tension, high self-assembly, and high tolerance in vitro and
in vivo compared to conventional surfactants [111, 112].
3.4.4 Aspasomes
Vesicles synthesized by supramolecular amphiphiles that have antioxidant

properties, such as aspartic acid and its derivatives, are used therapeutically for
diseases in which active oxygen species are produced. Ascorbyl palmitate (ASP)
combination with cholesterol and a negative charge inducer can be used to synthesize
bilayer lipid of niosomes. Aspasomes are prepared by film hydration method and
then hydration with aqueous solution along with sonication is synthesized.
Gopinath et al. introduced the nanoparticle formulation of aspasomes (ASC-P)
[113]. Submicron-sized aspasomes are synthesized by thin layer hydration. A lipid
film is synthesized with ascorbyl palmitate and cholesterol (27.63 to 72.18) and
dicetyl phosphate at 10% mol of total lipid and hydrated with phosphate saline buf-
fer (PBS, pH 7.4). For example, for hydration of zidovudine (AZT) hydrophilic
drug, it is first dissolved in PBS and then the solution is hydrated on a thin layer.
Then the prepared suspension is sonicated in an ultrasonicator to obtain AZT-
encapsulated aspasomes. In a study for zidovudine (AZT) encapsulation in aspa-
somes, adding cholesterol to the lipid layer showed no change in size, zeta potential,
and zidovudine (AZT) encapsulation percentage. But release rate of zidovudine
(AZT) varied with the presence of cholesterol. The antioxidant property of ascorbyl
was maintained even after ascorbyl palmitate was converted to aspasomes.
Aspasomes also showed increased skin penetration and AZT preservation proper-
ties [114].
3.5 Methods of Preparation
The qualities of niosomes can vary widely depending on how they are synthesized.
In this chapter, different approaches were reviewed:
3.5.1 Thin-Film Hydration (TFH)/Handshaking Method (HSM)
These two methods are mostly put into one category for the similarities they have
(although some articles have separated them to be two different methods) [115].
Based on the research and the lab practices done, they seem to be the most common
technique to prepare niosomes: need a round bottom flask, a volatile organic solvent
like diethyl ether or chloroform, etc. (for dissolving surfactant), cholesterol, and
charge inducers (rotary evaporator). Using TFH/HSM, the solvent will be evapo-
rated at room temperature which creates a thin dry film of dissolved components,
and then the dried film must be hydrated, so an aqueous phase will be added with
gentle agitation [116]. As can be seen in Fig. 14.3, depending on the structure of the
drug (hydrophilic or hydrophobic) decide where the aqueous phase must be added:
( A) Aqueous phase if it is hydrophilic
(B) Organic solvent if it is hydrophobic
3.5.2 The “Bubble” Method
An organic solvent won’t be used for this method, but a three-neck flask will be
needed (fabric must be glass) to be in a water bath for maintaining the temperature.
The first neck flask must be able to place the thermometer, the second one is used to
pass the nitrogen, and the third (last) one is attached for the water-cooled reflux. So,
using this method first cholesterol, then surfactant, and finally, phosphate buffer is
mixed together and then these particles are dispersed at 70 °C. Afterward, a high-
shear homogenizer will be used for 15 s and afterward, nitrogen gas will be imme-
diately supplied to the mixture (bubbling of the nitrogen gas must be at 70 °C). The
vesicles produced this way are large and unilamellar [89].
3.5.3 Ether Injection Method (EIM)
In this method, an aqueous solution is used so that the solution of cholesterol and
surfactant dissolved in diethyl ether (volatile organic solvent) will be injected with
the help of a 14 gauge needle, and then they must be put into preheated warm water
(maintained at 60 °C). Finally, niosomes are formed by vaporization of diethyl ether
(volatile organic solvent) using a rotary evaporator. These single-layered niosomes
can have a diameter varying from 50 to 1000 μm [115–117].
Fig. 14.3 Schematic non-ionic surfactant vesicles (niosomes) formation by lipid layer

hydration method
3.5.4 Sonication Method
It is one of the conventional methods; sonication is used to prepare niosomes. In this

method, solution of the drug in the buffer must be prepared so that afterward surfac-
tant and cholesterol can add up [118]. The next necessary step to produce multila-
mellar vesicles is probe sonication (they require high levels of energy) at 60 °C for
3 min. It’s possible to produce unilamellar vesicles if it would be a further ultrasoni-
cator. So, the ability to control niosomes particle sizes can be achieved by sonica-
tion of the mixture at a particular frequency, temperature, and time [116].
3.5.5 Reverse Phase Evaporation Method (REV)
In this method, an organic solvent will be used like ether and chloroform, and then
surfactant and cholesterol (as the aqueous drug solution) will be mixed in (taken in
the ratio 1:1 ratio) [116] and then the mixture will be added to aqueous phase con-
taining the drug afterward resulting in a two-phase system, then they must be
homogenized so that organic phase is evaporated under negative pressure to form
niosomes. Now large unilamellar vesicles are formed [91]. To form a semi-solid gel
of large vesicles this emulsion must be dried in a rotary evaporator at 40 C. To form
small stable uniform vesicles small quantities of the buffer will be added and the
semi-solid form is sonicated at 4–5 C [118]. REV can be the ideal method for creat-
ing niosomes of hydroxychloroquine, isoniazid, ellagic acid, and bovine serum
albumin due to: high % EE, large particles size with a small variation to encapsulate
large hydrophilic macromolecules with relatively higher EE than other methods
[97]. Keep in mind that if the structure of the used drug is deformed (for being in
temperatures greater than 50 °C or in organic solvents), direct entrapment method
cannot be used [115].
3.5.6 Micro-Fluidization Method
This method is based on the submerged jet principle. Surfactants and the drug
solution are pumped through an interaction chamber under the pressure of 100 ml/
min, and a cooling loop is required to remove the heat produced from before the
micro-fluidization. Using this method, it is possible to create different forms of
niosomes with greater uniformity, small size, unilamellar vesicles, and better
reproducibility of niosomes [91].
3.5.7 Trans-Membrane pH Gradient (Inside Acidic)
In this method, multilamellar vesicles are produced; so to create niosomal

suspension,a round bottom flask is being used. Firstly, the surfactant and cholesterol
must be mixed so they can dissolve together in chloroform, and then the mixture
must be put under pressure to evaporate chloroform. The mixture should be vor-
tex with 300 mM and citric acid (pH 4.0) to hydrate film. But still, the job is not
done; an aqueous solution containing 10 mg/ml of the drug must be added to the
solution and vortex. Set the pH of the final solution to 7.0–7.2 by adding 1 M diso-
dium phosphate. Finally heat the mixture at 60 °C for 10 min [116].
3.5.8 Single-Pass Technique
This is a patented technique for creating niosomes within the range of 50–500 nm.
It has also been mentioned as a multiple membrane extrusion. In this method, a
lipid-containing drug suspension must be passed through a porous device and then
through a nozzle. Finally, the uniform-sized niosomes are prepared [89].
3.5.9 Heating Method (HM)
The heating method was introduced by Mozafari et al. [119, 120] in 2005. In this
method, surfactants and cholesterol were hydrated in PBS (pH = 7.4) separately at
room temperature for one hour under a nitrogen atmosphere. The solution then is
stirred and heated up to 120 °C to dissolve cholesterol. At the next level, the tem-
perature must reach 60 °C. Afterward, surfactants and other additives should be
added to the buffer while stirring (meant for cholesterol) continues for another
15 min and after all niosomes nanocarriers were designed. At the end stage, created
niosomes must be kept at room temperature for 30 min, and then for future needs,
they will be stored at 4–5 °C in a nitrogen atmosphere [115].
3.5.10 Freeze and Thaw Method (FAT)
This method enables us to create frozen and thawed multilamellar vesicles (FAT-
MLVs). First, niosomes are prepared with the TFH method (thin-film hydration),
then niosomal suspensions are frozen in liquid nitrogen for 1 min and are thawed in
a water bath at 60 °C for 1 min [121].
3.5.11 Microfluidic Hydrodynamic Focusing
This method provides better-sized niosomes for distribution compared to a

conventional method. Lo et al. created niosomes out of two miscible liquids via
diffusive mixing based on microfluidic hydrodynamics [122, 123]. Hence, a rapid
and controlled manner is required to mix the miscible liquids in microchannels.
The following are the factors that can affect the assembly of niosomes:
(a) Microfluidic mixing conditions.
(b) Chemical structure of the surfactant.
(c) Material: the micro-channels fabrication, for example; large-sized niosomes
will be produced If we use aider micro-channels for it and increase the diffusive
mixing time & hence.
(d) Low-rate ratio: This factor can affect the size of the produced niosomes, for
example, if the rate increases it will be decreasing diffusive mixing time. So, the
manufactured niosomes will be small-sized [89].
3.5.12 Dehydration-Rehydration Method
The initiator of this method was Kirby and Gregoriadis in 1984 [124]. In this
method, vesicles must first be prepared by the thin-film hydration method. Next,
liquid nitrogen should be used to frizz vesicles and then it should be freeze-dried
overnight; this will form powder niosomes, and then phosphate buffer saline
(pH 7.4, at 60 °C) should be used for hydration.
3.5.13 Supercritical Carbon Dioxide Fluid Method (scCO2)
The advantages are:

(a) One-step production
(b) Easy scale-up
Manosroi et al. designed this method for creating niosomes [125, 126]. To sum
up, Tween 61, cholesterol, glucose, PBS, and ethanol must be added into the view
cell and the CO2 gas should be introduced into the view cell, next equilibrium must
be reached through magnetic stirring, and after this level, the pressure should be
released and finally, niosomal dispersions can be found (Niosomes created by this
method will be in the range of 100–440 nm) [91]. So, keep in mind that this method
requires solvents that are non-inflammable, non-toxic, and volatile.
3.5.14 The Handjani-Vila Method
In this method, the aqueous solution of the drug must be mixed with cholesterol and
surfactant. Then, ultracentrifugation or agitation should be used to homogenize the
mixture at a controlled temperature [127].
3.6 Characterization of Niosomes
The parameters that characterize niosomes are as follows:
3.6.1 Size, Morphology, and Size Distribution of Niosomes
Light microscopy, coulter counter, photon correlation spectroscopy, electron

microscopic analysis, SEM (scanning electron microscope), TEM (transmission
electron microscope), freeze-fracture replicator, light scattering, and zetasizer can
be used to determine the size and morphology of niosomes. Because the two
methods use different measurement concepts, the particle size determined by the
transmission electron microscope is smaller than the dynamic light scattering (DLS)
[115, 128–130]. Rinaldi et al. [131] investigated the size, shape, and size distribution
of the niosomes sample using atomic force microscopy.
3.6.2 Entrapment Efficiency
It can be computed by subtracting the total amount of drug added from the amount
of unloaded drug [84]. Exhaustive dialysis, filtration, gel chromatography, and cen-
trifugation can all be used to determine the unloaded drug [131]. By dissolving
niosomes in 50% n-propranolol or 0.1% Triton X–100, the concentration of loaded
medicines can be measured [132]. The percent entrapment efficiency can be calcu-
lated using the calculation below [115].
Quantity of drug loaded in the niosomes

%Entrapment Efficiency 100
The total quantity of drugs in the suspension

3.6.3 Charge on Niosomes and Zeta Potential
Because of the charge on them, niosomes repel one other. By inhibiting aggregation
and fusion, electrostatic repulsion keeps them stable [133]. The zeta potential is
used to determine the charge on niosomes. The zeta potential is determined using a
zeta potential analyzer, mastersizer, microelectrophoresis, pH-sensitive fluoro-
phores, high-performance capillary electrophoresis, and a DLS apparatus [134].
Henry’s equation is the formula for calculating zeta potential [135, 136].
E
£

where £ = zeta potential, μE = electrophoretic mobility, η = viscosity of medium,

and Σ = dielectric constant.
Because of electrostatic repulsion between particles, Bayindir and Yuksel [122]
employed dicetyl phosphate (DCP) to give the surface charge on niosomes and
found that a negative zeta potential in the range of 41.7 to 58.4 mV is enough to
keep the system stable. Manosroi et al. [137] used two different charges to manufac-
ture gallidermin niosomes (anionic and cationic). They noticed differences in nio-
somes size because, in anionic vesicles, the charge was neutralized by the positive
charge of gallidermin, resulting in small niosomes, whereas in cationic vesicles, the
charge was neutralized by repulsion between the cationic charges, resulting in big
niosomes.
3.6.4 Number of Lamellae
The number of lamellae can be determined using a variety of techniques such as

AFM, NMR, small-angle X-ray spectroscopy, and electron microscopy [138, 139].
Small-angle X-ray scattering combined with in situ energy-dispersive X-ray diffrac-
tion can be utilized to characterize the thickness of bilayers [140, 141].
3.6.5 Membrane Rigidity
The mobility of a fluorescent probe as a function of temperature can be utilized to

evaluate membrane stiffness [142]. Fluorescence polarization can be used to deter-
mine the micro-viscosity of the niosomal membrane to better understand its packing
structure [125]. The membrane characterization of pentamidine niosomes was done
by Rinaldi et al. DPH and pyrene were utilized because DPH indicates lipid order
and pyrene indicates lateral diffusion inside the bilayer [143]. The fluorescent mea-
surements (ʎ = 350–425 nm) were made with a luminescence spectrometer, and the
fluorescence anisotropy (r) was calculated using the equation below:
IVV GIVH
Fluorescence Anisotropy r

IVV 2GIVH
3.6.6 In Vitro Release
The dialysis membrane method is used to investigate in vitro release. Niosomes are
placed in a dialysis bag, which is then placed in a container with dissolving media,
usually buffer, in this procedure. This entire assembly is kept at a constant tempera-
ture of 37 °C on a magnetic stirrer. A sample is obtained from the receptor compart-
ment at specific time intervals, and drug concentration is measured using any
method described in the literature [136, 137, 144].
The dialysis approach was used to release temozolomide niosomes [145],
benazepril hydrochloride niosomes [146], paclitaxel, curcumin cationic PEGylated
niosomes [147], and diltiazem niosomes [148]. Aboul Einien [129] studied the
release of ascorbic acid derivative from aspasomes using a cellophane membrane
(mol. Wt. cut off = 500–1000) soaked in glycerin: water (1:3) for 15 min; 0.5 g of
aspasomes were packed in this membrane, firmly knotted, and placed in a USP
dissolution apparatus I. The experiment was carried out in 250 mL of phosphate
buffer (pH 7.4) at 32 °C ± 0.5 °C temperature and 50 rpm speed. At a predefined
time interval, the samples were spectrophotometrically examined. To investigate the
diffusion of morusin from niosomes, Agarwal et al. [149] utilized a different
approach. They dispersed 15 mg of preparation in 15 ml of phosphate buffer between
pH 4.5 and 7.4. This sample was taken in 15 Eppendorf tubes. These tubes were
revolved at a constant speed of 130 rpm and a temperature of 37 °C for 9 days. The
tube is removed at a predetermined time interval and centrifuged at 15000 rpm for
30 min. The drug concentration of the resulting supernatant was determined using
spectrophotometry.
3.6.7 Tissue Distribution/In Vivo Study
The method of delivery, drug concentration, effect, and present time of the drug in
tissues such as the liver, lung, spleen, and bone marrow all influence in vivo inves-
tigations for niosomes [115, 133]. Animal models can be used to investigate a drug’s
tissue distribution. Animals must be sacrificed, and various tissues such as the liver,
kidney, heart, lungs, and spleen must be taken, washed with buffer, homogenized,
and centrifuged to investigate the distribution pattern. The drug content of the super-
natant is determined [136]. Onochie et al. [150] studied the bioavailability of benzyl
penicillin niosomes in albino rats in vivo.
The intubation tube was used to administer each formulation (0.1 ml) orally.
Blood samples were taken at predefined intervals for 24 h using the retro-orbital
puncture method, and the supernatant was utilized to measure serum drug
concentration.
3.6.8 Stability Studies
On storage, the drug may leak from the niosomes, because of aggregation and fusion
[115]. Kopermsub et al. [151] performed the stability studies of niosomes by expos-
ing the preparation to different conditions of temperature (4°, room temperature,
and 45°) for 2 months. Niosomes are also exposed to various humidity and light
(UV) conditions. During stability studies, parameters like size, shape, and entrap-
ment efficiency are evaluated periodically. In the same manner stability of green tea
extract niosomes [152], lornoxicam niosomes [153], cefdinir niosomes [154], and
Ginkgo biloba [154] niosomes have been performed. Bayindir and Yuskel [122]
studied the effect of gastrointestinal enzymes on the stability of niosomes. This
study was performed by exposing the drug and drug-loaded niosomes in different
gastrointestinal enzymes like pepsin, trypsin, and chymotrypsin and found that nio-
somes protect the drug from degradation by gastrointestinal enzymes.
3.7 Routes of Administration
Drug-loaded niosomes can be supplied via a variety of routes, depending on the

condition, drug characteristics, and the site of administration. These administration
paths are briefly described below.
3.7.1 Intravenous
Intravascular delivery of niosomes is possible. The advantage of injecting the

medicine is that it enters the systemic circulation immediately; also, the niosomes
improve the drug’s stability and prolong its time in the blood. With minimal changes,
the drug can also be administered to a specific location. Many medications’ nio-
somes are delivered using the intravenous method [87, 155]. Niosomes of morin
hydrate were produced by A. Y. Waddad et al. [156] for intravascular injection. To
increase the stability and bioavailability of phenol, He et al. [157] developed
PEGylated niosomes. PEGylated niosomes can block uptake from the mononuclear
phagocytic system, allowing to improve circulation time.
3.7.2 Intramuscular
Niosomes can also be given via the intramuscular method. Jitender Singh Wilkhu
[158] fabricated niosomes for the oral and intramuscular administration of subunit
influenza antigen.
3.7.3 Dermal and Transdermal
In the event of skin problems, the dermal route is employed to deliver drugs locally.
It is solely employed for local activity. This approach has the advantage of prevent-
ing the medicine from entering the systemic circulation, resulting in fewer side
effects. The medicine enters the systemic circulation via transdermal distribution;
however, drugs confront a barrier in the form of the skin. The vesicular system is
extremely useful in enhancing medication delivery via both dermal and transdermal
routes [139]. Niosomes operate as a drug reservoir, allowing the drug to penetrate
deeper into the body. To avoid gastrointestinal problems, NSAIDs are delivered by
a transdermal administration method [84].
Clomipramine is provided encapsulated in niosomes to reduce first-pass
metabolism and increase bioavailability [159]. Manosroi et al. [137] produced
gallidermin niosomes for transdermal administration. They demonstrated improved
transdermal medication delivery with increased drug accumulation in the skin and
no systemic adverse effects. Patel et al. [160] improved lopinavir transdermal
administration from niosomal gel. They compared the niosomal gel to the ethosomal
gel of the same medication and discovered that the ethosomal gel deposition was
better using ex vivo permeation experiments. Niosomes were found to be safer than
ethosomes in histopathological investigations and their in vivo bioavailability was
substantially higher than the oral suspension of lopinavir. A papain-loaded elastic
niosomal gel with a molecular mass of 23.5 kDa was effectively developed for scar
therapy by transdermal application [161]. Sandeep et al. [162] produced a
fluconazole proniosomal gel for topical use. Ex vivo skin penetration and permeation
experiments revealed that a large amount of drug has collected in the skin, improving
local drug delivery for a longer period. Abdelbary et al. [163] developed methotrexate
niosomes for topical administration of methotrexate to patients with psoriasis. This
preparation has the highest proportion of drug deposition in the skin (22.45%). To
achieve continuous medication delivery, Narayana Charyulu R et al. [164] combined
penetration enhancers with methotrexate. Junyaprasert et al. [165] used different
surfactants (Span 60 and Tween 60) and solubilizers to make ellagic acid niosomes
(propylene glycol 400, propylene glycol, and methanol).
The niosomes size, entrapment efficiency, and drug permeability were all
modified by the formulation. Junyaprasert et al. [166] investigated the effects of
chemical penetration enhancers on ellagic acid skin permeability. The penetration
enhancer has altered the permeation of ellagic acid from niosomes at 24 hours,
according to in vitro skin permeation tests in the human epidermis. The DMSO
niosomes have the highest drug concentration in the epidermis, while N-methyl-2-
pyrrolidone niosomes have the highest concentration in the acceptor compartment.
This research shows that DMSO niosomes are effective for epidermal distribution
of ellagic acid, while N-methyl 2-pyrrolidone (NMP) niosomes are effective for
dermal delivery. Niosomes of the following drugs are also made and evaluated:
ascorbic acid derivative (topical delivery) [129], green tea extract (transdermal)
[152], diacerein (topical) [89], etodolac (topical) [167], celecoxib (transdermal)
[168], baclofen (topical) [169], and resveratrol (topical) [170]. For transdermal drug
delivery, phenol ethosomes [130] and pentazocine proniosomes [171] are also
created.
3.7.4 Oral
As the oral route is the preferred approach for drug administration, niosomes are
also given this way. The acidic environment and digestive enzymes, which may
degrade the medication, are a difficulty in the oral distribution of the medicine
[122]. However, niosomes have been demonstrated to successfully carry the medi-
cation to the gastric mucosa [172]. To improve oral bioavailability, niosomes con-
taining tenofovir disoproxil fumarate [173], cefdinir [154], paclitaxel [122], and
Ginkgo biloba extract are produced. To improve the oral activity, microbiological
activity, and duration of action, Onochie et al. [150] developed benzylpenicillin
niosomes. Lornoxicam niosomes were developed to prolong the drug’s action when
taken orally [153]. Samyukta Rani et al. [174] made orlistat niosomes from pronio-
somes to improve solubility, regulate release, and length of action. To improve insu-
lin penetration through the intestinal membrane, Moghassemi et al. [175] produced
trimethyl chitosan (TMC)-coated niosomes of insulin.
3.7.5 Ocular
When a medicine must be given in the anterior location of the eye, topical ocular
administration is usually preferred [115]. Drugs delivered in conventional forms
have a bioavailability of just 1–3%, and they are subject to precorneal loss due to
tear production and insufficient residence time in the conjunctival sac [176, 177]. To
distribute naltrexone via the ocular pathway, Abdelkader et al. [178] produced
controlled release niosomes and discomes. They discovered that anionic niosomes
outperform neutral niosomes in improving naltrexone penetration across the cornea.

By covering tacrolimus niosomes with mucoadhesive hyaluronic acid, Zeng et al.
[179] created tacrolimus niosomes. Because of its high lipophilicity and molecular
weight, tacrolimus has a poor corneal penetration (822.5 D). The hyaluronic acid–
coated niosomes improve corneal permeability and ocular contact time. Abdelkader
et al. [180] also created unique nano-sized elastic niosomes for ocular delivery of
prednisolone acetate and sodium phosphate. They tested for ocular irritation, bio-
availability, and anti-inflammatory properties, as well as compared the results to
those of traditional eye drops (both suspension and solution). Using a modified
Draize test, researchers discovered that both forms of prednisolone have good ocu-
lar tolerability and bioavailability. A side effect elevation in intraocular pressure
created by prednisolone was greatly reduced by niosomes preparations.
3.7.6 Pulmonary
The pulmonary route of administration of niosomal drugs has various advantages,

including enhanced mucus permeability, sustained drug delivery, targeting, and
superior therapeutic outcomes. The interaction of niosomes for pulmonary gluco-
corticoid administration with human lung fibroblasts is created and tested. At all
incubation durations, these niosomes showed no appreciable toxicity in the concen-
tration range of 0.01 to 1 M. Vesicular carriers have been discovered to be located
in the cytoplasm using confocal laser scanning (site for glucocorticoid receptors).
These vesicles were shown to significantly increase drug absorption by human lung
fibroblasts as well as drug activity [181].
3.7.7 Nasal Administration
Nasal delivery is an excellent option for medicines with a high first-pass metabolism.
Diltiazem is rapidly absorbed from the mouth, although its bioavailability is only
30–60% due to substantial hepatic first-pass metabolism by cytochrome P450
enzymes. Nasal administration has some drawbacks, such as a short residence time
in the nasal cavity due to mucociliary clearance, airflow restriction, and nasal
mucosa sensitivity, all of which impair drug penetration and systemic bioavailabil-
ity. Nasal niosomal diltiazem has been demonstrated to have higher absorption and
less elimination [148].
3.8 Applications of Niosomes
Niosomes can be employed as a delivery device for a range of pharmaceutical

reasons. Table 14.1 provides a summary of some of the prior studies on the
application of niosomes in a tabular format.
Table 14.1 Recent studies in drug delivery using niosomes and applications
Route
Application Surfactant Method Therapeutic agent administration Reference
Protein delivery Tween 60 Lipid layer Glutathione In vitro [182]
hydration
Span 60 Lipid layer Insulin In vitro [175]
hydration
Brij 92 Lipid layer Insulin Oral [183]
hydration
Span 60 Lipid layer Insulin Oral [184]
hydration
Span 40 Lipid layer N-acetyl Topical [185]
hydration glucosamine
Span 60 Lipid layer Bovine serum Oral [186]
hydration albumin
Anticancer Span 60 Lipid layer Cisplatin [187]
drugs delivery hydration
Span 60 Lipid layer 5-Flourouracil Topical [188]
hydration
Span 80 Sonication Curcumin [188]
Bola Lipid layer 5-Fluorouracil Intravenous [111]
surfactant hydration
Span 60 Lipid layer 5-Fluorouracil Topical [189]
hydration
Span 60 Lipid layer Flutamide Oral [190]
hydration
Pluronic Lipid layer Doxorubicin In vitro [191]
P123 hydration
Tween 80 Lipid layer Curcumin Vein injection [192]
hydration
Tween-100/ Lipid layer Curcumin In vitro [193]
span 80 hydration
Span 60 Ethanol Gambogenic acid In vitro [194]
injection
Tween 80 Lipid layer Paclitaxel and In vitro [147]
hydration curcumin
Tween 80 Lipid layer Doxorubicin and In vitro [195]
hydration curcumin
Carrier for Span 60 Lipid layer Hemoglobin Intravenous [196]
hemoglobin hydration
Treatment of Span 60 Lipid layer Lamivudine [197]
HIV-AIDS hydration
Span 60 Ether injection Stavudine [198]
Span 60 Lipid layer Stavudine [199]
hydration
Span 80 Ether injection Zidovudine [200]
(continued)
Route
Application Surfactant Method Therapeutic agent administration Reference
Vaccine and Span 60 Lipid layer Tetanus toxoid [201]
antigen delivery hydration
Span 20 Lipid layer Newcastle Parenteral [202]
hydration disease vaccine
Span 60 Lipid layer Ovalbumin [203]
hydration
Span 60/ Reversed Bovine serum Topical vaccine [204]
Span 85 phase albumin
evaporation
Management of Span 60 Lipid layer Methotrexate Topical [163]
psoriasis hydration
Free Lipid layer Methotrexate Topical [205]
surfactant hydration
Span 60 Lipid layer Acitretin Topical [206]
hydration
Treatment of Span 40/ Lipid layer Selenium and In vitro [207]
leishmaniasis Tween 40 hydration glucantime
Span 40/ Lipid layer Amphotericin B intramuscularly [208]
Tween 40 hydration and glucantime
3.8.1 Delivery of Proteins and Peptides
Protein and peptide medications have long been challenging to deliver orally due to
their degradation by the acidic environment and enzymes of the gastrointestinal
tract. Niosomes, on the other hand, shield these drugs from protolithic enzymes [84,
132]. Moghassemi et al. [153] developed bovine serum albumin niosomes (BSA).
The formulation was tuned for loading and release as a function of cholesterol to
span 60 M ratios, and an inverted light microscope was utilized to monitor the posi-
tion of protein in the vesicle. To improve insulin penetration, niosomes of trimethyl
chitosan-coated insulin are also produced for oral delivery [175].
3.8.2 Delivery of Anticancer Drugs
Niosomes can deliver anticancer drugs to a specific organ. This targeting could be
passive [209] (deposition of niosomes within the tumor due to unique properties of
tumor cells not found in normal cells) [210], physical (delivery based on specific
environmental conditions such as pH or magnetic fields) [118], or active [209]
(delivery based on specific environmental conditions such as pH or magnetic fields)
(active uptake of niosomes by the tumor cell). Active targeting can be accomplished
by altering the surface’s structural features or by binding the ligand to the niosomes.
Curcumin, which is hydrophobic, and doxorubicin hydrochloride, which is hydro-
philic, were encapsulated in niosomes for anticancer treatment in this study. They
observed two distinct release phases: doxorubicin release during the first 2 days,
followed by curcumin release for 7 days. Against HeLa cell lines, the cytotoxic
impact was amplified (synergistic). For the co-administration of curcumin and
paclitaxel, Alemi et al. [147] developed cationic PEGylated niosomes. The improved
synergistic anticancer effects of these niosomes were reported. Agarwal et al. [149]
created the morusin niosomes for anticancer therapy potentiation. He noticed that
the drug was released in a dependent manner. The release of morusin from niosomes
was lower at pH 7.4 than it was at pH 4.5. In acidic settings (pH 4.5), drug release
was 58.1% after 120 hours, but it was only 43.3% at physiological pH 7.4. It sug-
gests that in the acidic environments of cancer cells, significant drug release can be
achieved.
3.8.3 Delivery of Vaccine and Antigen
Wilkhu et al. [211] developed bilosomes for vaccine administration orally. Bile salt
is incorporated into the bilayer of vesicles to manufacture bilosomes. The antigens
are protected by these bilosomes from being degraded by enzymes found in the
gastrointestinal system (GIT).
3.8.4 Carrier for Hemoglobin
Because of their strong oxygen absorptive capabilities, niosomes can also be used
as a hemoglobin carrier in the blood [212].
3.8.5 Treatment of HIV-AIDS
Niosomes can be used to deliver drugs for sustained delivery in AIDS patients. The
low efficacy and toxicity of these drugs pose an issue in their delivery, which could
be solved by constructing a niosomal system. Due to dose-dependent hematological
toxicity, significant first-pass metabolism, short biological half-life, and poor
absorption, zidovudine is an anti-HIV drug with limited therapeutic efficiency [146,
213]. Niosomes have been reported to solve zidovudine issues [199]. Lopinavir is
an HIV protease inhibitor that is reversible. Because of its low aqueous solubility,
high log P value, cytochrome P450 3A4 sensitivity, and susceptibility to
P-glycoprotein efflux transporters, its systemic bioavailability via the oral route is
limited. Transdermal niosomes were created and compared to the ethosomal gel to
address these concerns. Ex vivo skin permeation experiments revealed that the etho-
somal gel deposition of a drug into the skin was higher than the niosomal gel, but
niosomes permeated deeper through the skin and had a better drug release profile
[160]. Kamboj et al. developed niosomes to improve the oral bioavailability of teno-
fovir disoproxil fumarate [173]. They discovered a twofold increase in bioavailabil-
ity and a considerable improvement in the drug’s mean residence time, indicating a
longer drug release time. Stavudine niosomes were manufactured by Shreedevi

et al. [214] for targeting and controlled release.
3.8.6 Management of Psoriasis
Psoriasis is an inflammatory skin condition that lasts for a long time. It has been
reported to affect joints and is recurring [180, 215]. Topical treatment is often used
for mild to moderate psoriasis [173]. When more than 20% of the patient’s body is
affected, systemic therapy is recommended. Emollients, keratolytic agents, coal tar,
anthralin, calcipotriene, and corticosteroids are some of the topical treatments for
psoriasis. Phototherapy may be combined with systemic therapy. Systemic therapy
for psoriasis includes methotrexate, cyclosporine, corticosteroids, and etretinate
[152]. Nausea, diarrhea, dizziness, and mouth ulcers are all common side effects of
systemic methotrexate treatment [133, 216]. Hematological and liver damage are
potentially possible side effects [217]. Topical methotrexate may help prevent these
issues. For better psoriasis care, Abdelbary and AbouGhaly [163] created and opti-
mized niosomes containing methotrexate for topical application. In comparison to
the oral solution, the niosomes were optimized using the Box-Behnken design and
reported to have a much higher area under curve and skin deposition amount of
drug. The safety of niosomes was proven by histopathological examinations.
Hashim et al. [206] developed an acitretin nano-vesicular gel for topical use to com-
bat the drug’s low solubility, stability issues, skin irritation, and substantial systemic
adverse effects. Moghaddam et al. [89] used topical application to prepare the dia-
cerein niosomes for targeted distribution.
3.8.7 Treatment of Leishmaniasis
Because niosomes are taken up by the reticuloendothelial system and accumulate

there, they can be used to treat disorders like leishmaniasis [87, 218]. Niosomes
have also been utilized to treat malignancies that have spread to the liver and spleen
[132]. The Leishmania parasite primarily infects the liver and spleen. Antimonial
(drugs used to treat leishmaniasis) might affect the liver, kidneys, and other organs
[219]. The niosomal formulation can increase the drug’s absorption in the liver,
reducing the drug’s negative effects on other organs [132]. Positively charged nio-
somes entrapped with autoclaved Leishmania major against cutaneous leishmania-
sis had a moderate effect and successfully delayed the formation of lesions in
BALB/c mice, according to Pardakhty et al. [220]. For Leishmania tropica,
Mostafavi et al. [207] produced selenium niosomes with glucantime. In vitro testing
revealed that selenium niosomes combined with glucantime have effective anti-
leishmanial action and improved potent lethal activity. For Leishmania tropica,
Parizi et al. [207] investigated the immune-modulatory and antileishmanial action
of benzoxonium chloride niosomes. They discovered that as the concentration of the
drug was increased, the expression of interleukin IL-10 decreased while that of
interleukin-12 increased.
3.8.8 Diagnostic Imaging
Niosomes have the potential to be employed as a carrier for radiopharmaceuticals,

making them valuable in diagnostic imaging of organs such as the liver and spleen.
For imaging, 99mTc labeled DTPA is utilized [136, 221]. Iobitridol (diagnostic agent)
is utilized with niosomes for x-ray imaging [222]. Gadobenate dimeglcemine in a
conjugated niosomal formulation with [N-palmitoylglucosamine (NPG)], PEG
4400, and both PEG and NPG have been found to increase tumor targeting of an
encapsulated paramagnetic drug as measured by MR imaging [138, 223]. By adding
contrast agents or dyes (near-infrared) in the inner aqueous or non-aqueous com-
partment or conjugating onto the surface of niosomes, A. Massotti [224] created
unique biconjugate niosomes for imaging. Gd (EDTA) 2- may be utilized as a con-
trast agent for incorporation [225]. Optical imaging combined with magnetic reso-
nance imaging is also a useful method for tumor diagnosis [226–228]. In vivo
imaging can be achieved by combining polyethylene amino groups with near-
infrared probes [229].
3.8.9 Enhancement of Bioavailability
Drug bioavailability can be improved with niosomes. To improve oral bioavailability,

niosomes of paclitaxel [122], cefdinir [154], benzylpenicillin [150], and tenofovir
disoproxil fumarate [173] are produced. Diltiazem niosomes were developed for
nasal delivery to improve bioavailability [148].
3.9 Targeted Drug Delivery
Tavano et al. [209] and A. Massotti [224] prepared niosomes for targeted delivery of
drugs to tumor cells. Tavano et al. prepared to transfer conjugated pluronic nio-
somes of doxorubicin for delivery to tumor cells. A. Massotti prepared pH-sensitive
niosomes for delivery of a drug to hepatoblastoma. Targeting was done using sur-
face modification and no pH-sensitive molecule was used. These niosomes undergo
protonation of amino groups present on their surface after penetration into the cell
and release their cargo by “sponge effect.”
3.10 Brain Targeting
As illustrated above, niosomes can entrap lipophilic or hydrophilic drugs and deliver
the drug molecules to the target site in a sustained and/or controlled way [75, 115].
Drug organ distribution and metabolic stability have been reported to be affected by
niosomes. Surface modification of niosomes has been shown to improve target
selectivity for cancer drug delivery systems [118]. De et al. reported that modifica-
tion of temozolomide-loaded niosomes with chlorotoxin, a target-specific peptide,
significantly improved the temozolomide glioma targeting efficacy [145]. In com-
parison to the intranasal solution of the drug, surface-modified niosomes containing
olanzapine (an atypical antipsychotic medicine) demonstrated a three-fold increase
in olanzapine concentration in the brain [230]. Pentamidine is an antiprotozoal
drug, also having an anti-inflammatory and neuroprotective effect in Alzheimer’s
disease [231–233]. Its clinical efficacy is limited due to poor permeability across
blood-brain-barrier and high hepatotoxicity. To overcome these issues chitosan-
glutamate coated pentamidine niosomes were prepared for intranasal drug delivery
to reach the brain. Approach to the brain via intranasal delivery bypasses the first-
pass hepatic metabolism and blood-brain barrier [11, 17, 22].
4 Summary
Despite the great advances in drug discovery, still, neurologic diseases are the
second cause of death around the world [234]. Conventional drug delivery methods
such as peripheral routes including oral and parenteral administrations are one of
the most common routes for drug delivery whenever systemic effects are intended
[235]. Besides the fact that the parenteral route is usually painful and also requires
technical assistance, conventional methods also showed other major drawbacks in
the efficient delivery of therapeutic agents to the brain. First of all, due to the pres-
ence of the blood-brain barrier (BBB), the drug administered via conventional
routes results in dramatically lower drug concentration in the brain [5]. Secondly,
systemic clearance as well as first-pass metabolism and enzymatic degradation hin-
der the efficacy of the drug and significantly reduce the drug bioavailability [236].
There are two barriers between blood and brain extracellular fluids: BBB and the
blood-cerebrospinal fluid barrier (BCSFB) which mediate communication between
the central nervous system (CNS) and the periphery [237, 238]. The BBB consists
of a tight layer of endothelial capillary cell junctions which are surrounded by astro-
cyte foot processes. The BBB plays a key role in regulating CNS homeostasis and
function by protecting the CNS from pathogens, toxins, inflammation, and injury. It
is a highly regulated barrier that allows highly selective transport of essential mol-
ecules to the brain [239]. BBB loss or dysfunction by various diseases such as brain
traumas, stroke, multiple sclerosis (MS), and neurodegenerative disorders could
result in neuronal dysfunction and degeneration. Although the BBB is a critical
component of CNS, it is a significant barrier for drug transport from the blood to the
brain, and just the drugs with molecular weight less than 400 Da, high polarity, and
not multicyclic can across the BBB successfully [234, 240]. All of these factors trig-
ger the hunt for an alternative delivery system that directly reaches the brain.
Different strategies that are mostly invasive such as intraventricular, intraparenchy-
mal, and intrathecal delivery (disruption of the BBB) have been investigated for this
purpose [240]. Over the past decades, several studies have described the
nose-to-brain route as a promising approach that could offer an opportunity to serve

as a noninvasive direct route to the CNS. The concept of the nose-to-brain drug
delivery, for the first time, was introduced by W. H. Frey II in 1989 (William H, Frey
I. Inventor Neurologic Agents for nasal administration to the brain. US1991
1990-12-04). This method previously was often used for brain targeting of insulin
or insulin-like growth factor and later was developed for delivery of larger molecu-
lar weight substances like proteins, peptides, and bioactive [236].
As previously mentioned, nose-to-brain drug delivery is an invasive alternative
and patient-friendly route over the traditional and invasive drug administration
routes which could provide faster onset of action, high blood flow, and porous endo-
thelial membrane to absorb drugs efficiently while circumventing the hepatic first-
pass metabolism, BBB, and potentially lowering the systemic exposure, which
enables the easy and self-administration possibilities. Furthermore, this route is
optimal for drugs that are susceptible to enzymatic degradation and gastrointestinal
tract acidic environment [239]. Generally, there are two main pathways for drugs to
reach the brain from the nasal cavity: (1) neuronal pathway as the major route and
(2) crossing the BBB through systemic circulation as the minor route.
Desired drugs could be located in the deeper region of the nasal cavity which is
firstly absorbed by olfactory and trigeminal neurons, and then through cellular
transport can reach the olfactory bulbs [235, 236]. The nasal cavity includes three
main regions: vestibular, respiratory, and olfactory regions. After a drug enters the
nasal cavity, it encounters mucociliary clearance and then moves forward to reach
respiratory and olfactory regions. From these regions, depending on the formula-
tion, physiological condition, and the administration device, the drug can be trans-
ported to the brain by several mechanisms such as trigeminal nerve pathway,
lymphatic and vascular pathway, olfactory nerve pathway, and cerebrospinal fluid.
When the drug reaches the brain, it is distributed throughout the CNS by perivascu-
lar transport [235]. Although the nasal to brain route is a promising method for fast,
easy, efficient, and targeted drug delivery, some limitations must be acknowledged
when developing new therapeutics to be administered via this route. First of all, this
route can be used just for potent drugs with a dose volume of 100–250 ml for liquids
and 20–50 mg for powders [235]. Secondly, there is a possibility of poor drug per-
meation through the nasal mucosa due to the low drug retention caused by mucocili-
ary clearance and enzymatic degradation. For this reason, drugs to be delivered by
this route should be protected from degradation [236]. Thirdly, due to high vascu-
larization, there is a possibility of peripheral side effects through systemic absorp-
tion which also reduces the drug concentration in the brain [236]. Finally, there is a
need to use the proper nasal delivery device to install the right amount of drug cor-
rectly in the nasal cavity [234]. However, different approaches such as permeation
enhancers, protective drug capsulation and colloidal carriers, suitable mucoadhe-
sive system, controlled delivery system, and other novel approaches have been
employed to improve the drug delivery through the nasal to brain route [236].
Niosomes are non-ionic surfactant vesicles fabricated by hydrating synthetic non-
ionic surfactants, either with or without cholesterol or lipids. They’re vesicular
structures that appear similar to liposomes and can transport both amphiphilic and
lipophilic medicines. Niosomes are a viable vehicle for drug administration because
of their non-ionic nature and are biodegradable, biocompatible, non-immunogenic,
and structurally flexible. For the treatment of cancer, viral infections, and other
microbial diseases, niosomes have been extensively studied for controlled release
and targeted administration. Niosomes can entrap both hydrophilic and lipophilic
drugs, allowing them to circulate in the body for a prolonged period. Encapsulation
of a drug in the vesicular system is expected to prolong its presence in the systemic
circulation and improve penetration into a target tissue, perhaps reducing toxicity if
selective absorption is possible. This chapter focuses on the nasal drug delivery
route, advantages and disadvantages of niosomes, types of niosomes, methods of
preparation, characterization, routes of administration, and applications of niosomes.
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Chapter 15
Nanosuspension – A Novel Drug Delivery
System via Nose-to-Brain Drug Delivery
Hemant K. S. Yadav and Raghad Zain Alabdin
Abstract Nanosuspension administration via the nasal route of administration

appears to be an effective pathway of nose-to-brain drug delivery. The nasal cavity
is highly vascularized and innervated by nerves, making it an attractive route of
administration that bypasses many oral or parenteral limitations and challenges,
thus having more advantages and being an effective treatment choice, especially for
neuronal diseases. Nanosuspension was found to be an enhancer of drug absorption
though the nasal route of administration due to its small particle size and its capabil-
ity of optimizing the hydrophilic drugs within it to increase its solubility. Intranasal
nanosuspensions are currently being investigated to improve nose-to-brain drug
delivery. Nose-to-brain drug delivery and nanosuspension application (insight for
nose-to-brain drug delivery) are discussed in this chapter.
Keywords Nanosuspension · Drug absorption · Brain and intranasal
H. K. S. Yadav (*)
School of Pharmacy, Suresh Gyan Vihar University, Jagatpura, Jaipur, Rajasthan, India
R. Z. Alabdin
Department of Business Development, Gulf Pharmaceutical Industries (Julphar),
Ras Al khaimah, UAE
Department of Pharmaceutics, College of Pharmacy, RAK Medical and Health Sciences
University, Ras Al khaimah, UAE
https://doi.org/10.1007/978-3-031-23112-4_15
326 H. K. S. Yadav and R. Z. Alabdin
1 Introduction
Brain is the most important organ that is considered as a complex organ, and the
deaths due to neuronal diseases are considered the seventh leading cause of death
globally [1]. There are several limitations to the therapeutic effects of neuronal
medications, such as the presence of blood-brain barrier. The blood-brain barrier
blocks more than 95% of the molecules used for the treatment of neuronal diseases
[1]. In this regard, many research works were conducted to find solutions to enhance
the bioavailability at the central nervous system. Nanosized molecules and the nasal
route of administration are the most attractive solutions to pass the blood brain bar-
rier and achieve the best therapeutic effect of the neuronal medications due to its
many favorable characteristics which include better patient compliance, non-inva-
siveness, and ease of administration. Intranasal delivery has also been used for sys-
temic delivery for other diseases treatments or preventions such as flu vaccines and
other areas. Formulations such as nanosuspensions not only solve many issues
related to the solubility and bioavailability of molecules, but they also enhance pen-
etration through the blood-brain barrier and the blood-cerebrospinal fluid barrier.
This chapter focuses on intranasal nanosuspensions intended for nose-to-brain
delivery.
2 Nanosuspension
Nanosuspension is a colloidal suspension where the particle size of molecules is

less than 1 micrometer. Nanosuspension has many advantages such as solubility and
bioavailability enhancement, suitable for both hydrophilic and hydrophobic mole-
cules, better drug loading than other nanoparticles, the possibility of dose reduction,
physical and chemical stability enhancement, and the possibility of passive drug
targeting [2]. There are two methods of nanosuspension preparation, bottom-up and
top-down technology. Bottom-up technology involves the processes of microemul-
sion, precipitation, and melt-emulsification. In the bottom-up technology, the drug
is first dissolved in an organic solvent and then it is precipitated while the antisol-
vent is added in the presence of a stabilizer. This technique includes various adapta-
tions, which are solvent-anti-solvent method, supercritical fluid processes, spray
drying, and emulsion-solvent evaporation. Top-down technique is majorly depen-
dent on the reduction of the particle size of larger drug particles into smaller drug
particles and this process is accomplished by using various milling techniques such
as media milling, microfluidization and high pressure or shear homogenization [3].
Nanosuspension has promising efficacy and several therapeutic applications due
to its properties and several merits and due to the fact that it can be administered
through different routes. Based on that, many drugs in the form of nanosuspensions
are intended for different routes of administrations for different therapeutic areas
and some of them are presented in Table 15.1 [2, 4].
15 Nanosuspension – A Novel Drug Delivery System via Nose-to-Brain Drug Delivery 327
Table 15.1 Nanosuspensions intended for different routes of administrations for different

therapeutic areas
Route Drugs Company/Author
Oral route Triglide® (fenofibrate) Sciele Pharma/Skyepharma
Intravenous Abraxane® (paclitaxel) Abraxis Bioscience/Astrazeneca
Ophthalmic Hydrocortisone M. A. Kassem
Pulmonary Budesonide Jerry Z. Yang
Intrathecal Busulfan Skyepharma
Topical Silver Nucryst
3 Nose-to-Brain Drug Delivery
To understand the concept of nose-to-brain drug delivery, it is crucial to know the

anatomy of the nasal cavity and the different pathways of drug absorption through
it. It is well known that the nose has two functions, olfaction and respiration. The
nose has two parts, the external and the internal nose. Bones and cartilages are the
components of the external nose and are located in the center of the face. As for the
nasal cavities, they are composed of three regions, the vestibular, the olfactory, and
the respiratory regions. The outermost part of the nasal cavity is the smallest region,
which is called the vestibular region. This region is composed of nasal hair and
sebaceous glands. Due to the structural features of this region, this region is not suit-
able for drug absorption. However, the nasal septum contains a small opening to the
vomeronasal organ where the drug can be absorbed directly into the brain via the
terminalis nerve [5]. The respiratory region forms 80% of the nasal cavity and it is
composed of three turbinates. This region is very attractive to be targeted for drug
absorption because of its high surface area. It is highly vascularized and it is sup-
plied by the branch of the maxillary artery, which is innervated by the trigeminal
nerve, which contributes to the nose-to-brain drug delivery [6]. Lastly, the superior
part of the nasal cavity is the olfactory region, that is surrounded by the olfactory
neural cells, which are bipolar neurons providing direct portal between the nose and
the central nervous system. Specifically, their unmyelinated axons are covered by
the olfactory cells and nerve fibroblasts, which are in continuity with meninges and
subarachnoid space [7]. Crowe et al. classified the mechanisms of nose-to-brain
drug delivery into two pathways – intracellular and extracellular. Intracellular
mechanism is preferred when the drug is internalized by an olfactory neuron and
traffics the endocytic vesicle within the cell to the projection site of the neuron by
exocytosis. As for the extracellular mechanism, it is where the drug crossing the
nasal epithelium to the lamina propiato, the neurons and then gets transported exter-
nally along the length of the axon via bulk flow process which finally lead into the
central nervous system where the drug is distributed via fluid movement. It is crucial
that the drug’s ability to cross the blood-brain barrier and the blood-CSF barrier is
determining the drug penetration. Thus, the drug properties, characteristics and for-
mulation parameters may determine the drug absorption pathway from the nasal
cavity [8]. The different pathways of drug absorption through the nasal route of
administration are demonstrated in Fig. 15.1.
Mucociliary
Clearance
Nose
Respiratory Olfactory Region

Region
Trigeminal Olfactory Transcellular Paracellular

nerve nerve transport transport
Blood Axonal transport Subarachnoid space
Brain Olfactory
Stem Bulb CSF
BBB
Brain Clearance
Fig. 15.1 Different pathways of drug absorption through the nasal route of administration
There are many advantages of the intranasal route of administration such as non-
invasiveness, avoidance of BBB, avoidance of first pass and gastrointestinal degra-
dation, high bioavailability, and the direct connection to the central nervous system.
However, the small surface area, mucociliary clearance, enzymatic degradation, the
possibility of nasal toxicity, and the poor permeation are the limitations of this route
of drug administration.
4 Permeation Enhancer Techniques Through the Nasal

Route of Drug Administration
Different techniques were studied to enhance drug penetration through the nasal
route of administration to achieve best solutions for different disease treatments.
Some of these techniques use the drug in the form of solutions as small lipophilic
molecules can easily penetrate the tissues and blood-brain barrier. Nanoparticle for-
mulations include nanosuspensions in which their small particle size makes them an
optimum choice to enhance the drug penetration. Also, Mucoadhesive agents can be
used in the formulations to increase the residence time and reduce the nasal clear-
ance. Different polymers can be used as mucoadhesive agents in the formulations.
Some of these polymers are discussed below.
5 Types of Polymers Used in the Formulation

of Nanosuspensions
5.1 Chitosan
It is a biodegradable and non-toxic polymer which is obtained from hydrolysis reac-

tion of chitin, a natural polysaccharide that is a major component of crustacean
exoskeleton. Unmodified chitosan is soluble in acidic medium and has great muco-
adhesive properties. The size of chitosan nanoparticles depends on its concentra-
tion, molecular weight, and surface charge [9, 10].
5.2 Gelatin
It is formed by either alkaline or acidic hydrolysis reaction of collagen. It has a tri-

ple helical structure and high content of proline, glycine, and hydroxyproline resi-
dues. Gelatin formed by the alkaline treatment of collagen has a lower isoelectric
point and more carboxyl groups than that formed by the acidic hydrolysis reac-
tion. [9, 10]
5.3 Sodium Alginate
Sodium alginates are unbranched and linear polymers which consist of ß-(1-4)
linked mannuronic acid and residues of α-(1-4) linked guluronic acid which are
arranged in blocks, also called G blocks or M blocks, and they can alternate with
each other. The nature of this polymer is hydrophilic, anionic, and has different
molecular weights. This polymer is unable to reswell in an acidic environment,
which helps in the incorporation of acidic drugs into the bead, which protects it
from gastric juice [9, 10].
5.4 Albumin
Human serum albumin is the most prominent plasma protein which consists of
nearly 585 amino acids and has an α-helical tertiary structure. Some of albumin’s
properties are that it is positively charged and multifunctional as it is involved in
transportation, enzymatic activities, and ligand binding [9, 10].
5.5 Tamarind
Tamarind, scientifically called “Tamarindus indica,” and known as “Imli,” is a

member of the dicotyledonous family Leguminosae. Tamarind seeds consist of
20–30% of the outer coat (testa) and 70–80% of the kernel that also called “endo-
sperm.” It consists of crude fibers which have higher carbohydrate percentage in the
form of sugars. Tamarind seeds are used as raw materials in the manufacture of dif-
ferent materials such as tamarind seed kernel powder (TKP), polysaccharide, adhe-
sive, and tannin. Tamarind seeds and the gum extracted from them are also used for
other purposes and the gum is currently gaining importance in pharmaceutical man-
ufacturing such as in the preparation of mucoadhesive formulations [11, 12].
6 Characterization and Evaluation of Nanosuspensions

for Nose-to-Brain Drug Delivery
Identification of nanoparticle properties is required to design the nanoparticles to be

suitable for different applications. In general, determination of particle size is the
most important characteristic as it not only determines the efficacy of the products
but it can also lead to irritation if the particle size is large. There are many available
tools to measure particles smaller than 1 μm. For example, light scattering (LS),
also called photon correlation spectroscopy, is a rapid method for determining the
particle size, size distribution, and polydispersity index.
This technique is well adapted for routine measures. Electron microscopes use
electromagnetic radiations of shorter wavelengths and provide magnification that
can disclose details with a resolution of up to approximately 0.1 nm. Scanning elec-
tron microscopy (SEM) allows for a resolution between 3 and 5 nm and even 1 nm
in some advanced microscopes. Images of the sample are taken; hence, it is possible
to study the external morphology of the particles along with the determination of
their size. Nanoparticles prepared using different methods can produce particles of
various morphologies. The internal morphology of substances as well as their par-
ticle size can be measured using Transmission Electron Microscopy (TEM). TEM
and high-resolution TEM are more powerful than SEM in providing details at the
atomic level and can yield information regarding the crystal structure, quality, and
grain size. Potentiometric techniques are used to characterize the phenomenon of
counter condensation, the nature of interactions with oppositely-charged surfactant

molecules, and the stoichiometry of polyelectrolyte complexes. Zeta potential mea-
surements are useful to detect the absorption of polyelectrolytes on the particle
surface.
6.1 Fourier-Transform Infrared Spectroscopy (FT-IR)

and Differential Scanning Calorimetry (DSC)
FT-IR spectra is the study where compatibility is investigated by using IR spectro-

photometer. Firstly, the solid pellet is prepared using KBr at room temperature and
then analyzed. FTIR study can be performed on pure polymers, pure drugs, and
formulations.
It is also important to study the dynamic DSC studies to determine the resulted
molecule’s nature and confirm its compatibility [13].
6.2 Particle Size Analysis, Zeta Potential,

and Polydispersity Index
The average particle size distribution, zeta potential, and polydispersity index of the
resulting nanoparticles can be determined using zeta potential analyzer. Prior to
testing the samples, the nanoparticle dispersion should suitably diluted [13].
6.3 Scanning Electron Microscope (SEM)
It is also critical to study the size and morphology of nanoparticles, which can be
done by scanning electron microscope (SEM) or transmission electron microscopy
(TEM). Nanoparticles should be suitably diluted with a solvent prior to placing the
sample [13].
6.4 Encapsulation Efficiency
The amount of encapsulated drug in nanoparticles can be determined by different

methods. One of these methods is to dissolve a known amount of the prepared
nanoparticles in a few milliliter of methanol and diluted using distilled water.
Methanol action is to lyse the nanoparticles and release the drug into the solution.
The released amount of the drug can be determined by UV spectrophotometer. The
amount of the encapsulated drug can be determined by dividing the amount
encapsulated by the amount of the drug added and then multiplying by 100 to get
the percentage encapsulated [14].
Encapsulation efficiency was calculated as:
Encapsulation efficiency%

Amount of encapsulated drug / Amount of added drug 100.
6.5 Stability Testing
The nanosuspension should be studied for physical and chemical stability under
several conditions for a known period to determine the proper storage conditions
and shelf life of the product. Also, it is very critical to study the stability of the prod-
uct to determine the suitable packaging, instructions of use, and other factors [15].
6.6 Testing Direct Nose-to-Brain Delivery
Different models to test nose-to-brain drug delivery as described by Franciska Erdo

are summarized below:
6.6.1 In Vivo Models
Mouse and rat models are the most useful models for the purposes of the preliminary
studies of drug absorption. However, it does not always correlate with humans because
of the physiological and anatomical differences. The study is based on administering
the drug into the nostril by using a micropipette and keeping the animal in supine
position to allow the drug to reach the olfactory region. The olfactory region in mice
or rats covers 50% of the nasal the cavity, but in humans, it is only 10%, which is simi-
lar in monkeys. The structure of the conchae also is different, where human and mon-
keys only have a single scroll conchae but the nasal epithelium of monkeys has a
smaller surface area than the nasal epithelium of humans. Thus, animal studies cannot
be that accurate to determine the intranasal drug administration [8, 16].
6.6.2 In Vitro Models
The drug transport can be studied more clearly and controlled through the in vitro
studies. The major studies are described below:
A. RPMI 2650 – a cell culture model of nasal barrier.
RPMI 2650 is extracted from the human nasal epithelial tissue, specifically,
a human nasal squamous cell carcinoma of the nasal septum. It is suitable for
testing nasal metabolism and toxicity assays. RPMI 2650 in ALI cultures can be
used as a screening tool for cytotoxicity and permeability in pre-clinical stud-
ies [17].
B. CaCo-2 cell line.
CaCo-2 cell line is extracted from the human colon carcinoma and separated
into different monolayers. It is used to study drug absorption through the nasal
route of administration as it is the most suitable model for this purpose [18, 19].
6.6.3 Ex Vivo Models
This model is used to study a drug’s permeability through the nasal cavity using
Ussing chamber. It is also useful for quantifying the active transport, passive diffu-
sion efflux transport, and identifying the carrier-mediated transports. It is a useful
model to compare drug transport through respiratory and olfactory pathways [20].
Different preclinical and clinical studies are discussed below in the coming sections
of this chapter.
7 Different Drugs Used and Current Research
There is a criterion for the selection of the drug in the formulation of nanosuspen-
sions, that is, when formulating the nanosuspension, the API should have one of the
following characteristics:
• The drug should be water insoluble but which are soluble in oil (having high Log
P), API which are insoluble in both water and oil, drugs in crystalline form with
reduced tendency to dissolve, regardless of the solvent.
• Potent APIs.
There are many drugs which have been studied for nose-to brain delivery wherein
researchers conducted several studies on these molecules viz. Valproic acid,
Duloxetine, Artemether,Temozolomide, Curcumin, Asenapine and Embedin. These
are used in treatment of different central nervous system diseases as shown in
Table 15.2 [21].
As noticed, most researchers target the smallest possible particle size of the for-
mulation to enhance drug penetration through the nasal cavity. The different drugs
and diseases which were targeted by different scientists through the nasal route of
administration are explained below.
Valproic acid nanostructured lipid carrier was developed by Eskandari et al. by
the solvent diffusion and ultrasonication method and it was characterized for differ-
ent parameters. Eskandari observed that the ratio of the plasma concentration was
higher when the drug was administered intranasally to rats compared to when it was
administered intra-peritoneally. Thus, it proved to be a better seizure therapy as a
preliminary evaluation [22].
Table 15.2 Different drugs used for nose-to-brain delivery

Particle size of the
Selected drug Targeted disease formulation (nm)
Valproic acid Epilepsy 154
Duloxetine Depression 100
Artemether Cerebral malaria 124.3
Temozolomide Brain tumour 100
Asenapine Schizophrenia 167.3
Cytokine intereferon- Multiple sclerosis
beta-1 b
Rosuvastatin Pentylenetetrazole (PTZ) induced 219.15
seizures
Duloxetine nanostructured lipid carrier formulated by Alam et al. using homog-

enization and ultrasonication method was evaluated for the in vivo nasal infusion
study. It was observed that the permeability was only 2.5 times higher in case of
intranasal administration as compared to the drug solution [23].
Artemether nanostructured carriers were developed by Jain et al. using micro-
emulsion method. It was studied for understanding the brain-to-blood ratio for dif-
ferent routes. It was observed that the intranasal route of administration has a higher
targeting efficiency and drug penetrating ability to the brain [24].
Temozolomide nanostructured lipid carriers were developed and studied by
Khan et al. It was observed that the particle size was less than 100 nm and the
entrapment efficiency reached up to 81.64%. The in vivo studies and the scintigra-
phy images showed high concentrations of the drug in the mouse brain and higher
brain-to-blood ratio [25].
Asenapine nanostructured lipid carriers were developed and studied by Singh
et al. It was observed that the particle size was 167.3 nm and the entrapment effi-
ciency reached up to 83.5%. The biodistribution study showed a higher drug con-
centration peak. As for the behavioural study, it was observed that the intranasal
treatment promised a better choice for schizophrenia treatment where the extra-
pyramidal side effects were reduced and the anti-psychotic effects were
increased [26].
Cytokine intereferon-beta-1 b was investigated by Ross et al. for the nose-to-
brain delivery. Intranasal administration of cytokine intereferon-beta-1 b showed
higher brain concentrations when compared to the intravenous administration [27].
Intranasal rosuvastatin liquid crystalline nanoparticle was developed by
Mohammad Zubair Ahmed for the treatment of pentylenetetrazole-induced sei-
zures, increasing current electroshock (ICES) induced seizures, and PTZ-induced
status epilepticus. It was observed that intranasal rosuvastatin at lower dose was
more effective than the oral and intraperitoneal administration of rosuvastatin. The
nanoparticles were developed by hydrotrope method using glyceryl monooleate as
the lipid phase. The Transmission electron microscopy revealed that the formed
nanoparticles were cubic in shape and multivesicular with a mean size of
219.15 ± 8.14 nm. The entrapment efficiency of 70.30 ± 1.84% was achieved.
Table 15.3 Different clinical trials investigating the nose-to-brain drug delivery for the treatment
of different central nervous system diseases
Pharmaceutical
Drug form Medical condition Clinical trial status
Oxytocin Nasal spray, Benzodiazepine withdrawal Ongoing, 2020
suspension and craving
Clonazepam Nasal spray Recurrent acute repetitive Ongoing, 2007
seizures (ARS)
Insulin Nasal spray, Phelan-McDermid syndrome Ongoing, 2012
solution
INSULIN Nasal spray Early Alzheimer’s dementia Prematurely Ended,
ASPART (eAD) 2015
Oxytocin Nasal spray Psychotic disorders Completed &
Favorable, 2014
Intranasal administration of rosuvastatin nanoparticles showed that there was a sig-

nificant increase in latency to PTZ-induced seizures and ICES seizure threshold
compared to control and intranasal rosuvastatin solution [28].
8 Clinical Trials Investigating the Nose-to-Brain

Drug Delivery
A lot of clinical trials are investigating the treatments of central nervous system
diseases by using the nose-to-brain drug delivery. Some of these clinical trials are
shown in Table 15.3 according to the EU Clinical Trials Register [29].
9 Recent Clinical Trials of Nanosuspensions
Use of ivermectin mucoadhesive nanosuspension as nasal spray was investigated

for the management of early covid-19. The clinical trial was completed on February
2021 and as per EMA revision, it was concluded that the available data did not sup-
port its use for COVID-19 outside well-designed clinical trials due to the much
higher ivermectin concentrations than those achieved with the currently authorized
doses [29, 30].
10 Conclusion
Intranasal route of administration is a promising route of administration for nose-to-

brain drug delivery. It has many advantages like non-invasiveness, avoidance of
BBB, avoidance of first pass and gastrointestinal degradation, high bioavailability,
and the direct connection to central nervous system. However, the small surface
area, mucociliary clearance, enzymatic degradation, the possibility of nasal toxicity
and the poor permeation are the limitations of this route of drug administration. One
of the effective solutions to overcome these challenges and limitations of the nasal
route of administration is administering nanosuspensions. Nanosuspension can be
considered the most suitable formulation choice for hydrophobic drugs which are
restricted by molecular weight, high log P, and melting point. Also, nanosuspension
method of formulation and preparation is simple and has scope for scaling up.
Intranasal nanosuspension is a very attractive choice for the administration of drugs
targeting the central nervous system by passing the blood-brain barrier, which
makes it an attractive area for investigation by several pharmaceutical companies
and researchers.
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jneuroim.2004.02.011.
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Chapter 16
Nasal Delivery of Micro and Nano
Encapsulated Drugs
Muhammad Sarfraz, Sara Mousa, Ranim Al Saoud, and Raimar Löbenberg
Abstract Drug encapsulation in nanoparticles and microparticles has been shown

to be a promising application for nasal drug delivery. Drug encapsulation and sur-
face modification of nano and microparticles can enhance drug bioavailability by
controlling drug release, decreasing mucociliary clearance, and enzymatic degrada-
tion. The drug particle size, the nature of the polymer, and the surface decoration on
the polymer play key roles in overcoming the physiological challenges of nasal drug
administration. This chapter cites several polymers used to formulate nanoparticles
and microparticles for nasal drug delivery. It describes techniques used to modify
the surface of polymers to improve physiochemical properties such as swelling,
gelling, wetting, mucoadhesiveness, and permeation. Polymer surface modifica-
tions can achieve safer drug delivery. Unlike other means of drug delivery, nasal
drug delivery can be effective at low concentrations. Several candidates that have
demonstrated effective intranasal drug delivery are described.
Keywords Drug encapsulation · Polymers · Surface modification · Drug delivery
1 Introduction
The nasal drug delivery approach has conventionally been confined to local or
topical acting therapeutic agents that address minor nasal problems. However, in
recent years, the nasal route of drug administration has emerged as an alternative to
oral and parenteral drug delivery. The therapeutic effects of drugs applied through
the nose have some physiological challenges [1]. Enzymatic drug degradation in the
nasal mucosa, low drug retention times, mucociliary clearance, poor permeability of
nasal mucosa, and nasomucosal toxicity are the factors that limit the application of
intranasal drug delivery [1–3]. In contrast, intranasal drug delivery provides
M. Sarfraz (*) · S. Mousa · R. Al Saoud

College of Pharmacy, Al Ain University, Al Ain Campus, Al Ain, United Arab Emirates
e-mail: muhammad.sarfraz@aau.ac.ae
R. Löbenberg
Faculty of Pharmacy and Pharmaceutical Sciences, Katz Centre for Pharmacy & Health
Research, University of Alberta, Edmonton, AB, Canada
https://doi.org/10.1007/978-3-031-23112-4_16
340 M. Sarfraz et al.
enhanced patient compliance and offers a greater potential for targeted drug deliv-
ery, a faster onset of drug action, and accessibility to the central nervous system
(circumventing the first pass effect). These advantages make the nose an attractive
route for drug administration [1, 3].
The nose is a complex organ designed to provide filtered respiration, a sense of
smell, and protection against environmental threats [4]. Drug delivery formulations
must meet the challenging task of avoiding the disruption of these complex func-
tions while providing the necessary medication. The response of ciliated cells
exposed to a nasal drug formulation can provide a test for functional disruption. A
dose at which the cilia continue to beat, i.e., ciliary beat frequency (CBF), at a con-
stant rate and the ciliated cells are not affected by the drug or formulation is consid-
ered to be a compatible dose. Disruptions of ciliary function by altering the beat
frequency or by increasing exfoliation have a negative impact on the body’s immune
system and can lead to mucosal irritation [5]. The status of cilia can be determined
by the in vitro cilia beat frequency or it can be measured with a confocal laser scan-
ning microscope or an electron microscope [6]. The presence of mucosal irritation
is another indicator of bio-incompatibility. In large species, such as a beagle dog,
mucosal irritation can be observed visually [7]. In smaller species, the visualization
of mucosal irritation is difficult, so histopathological methods must be utilized [8].
To overcome some of these physiological challenges, researchers use absorption
enhancers in nasal formulations and study its influence on mucosal irritation, enzy-
matic activity, and to clear nasal passages [9]. Absorption enhancers facilitate the
transport of the drug through the nasal epithelium and lower the effective drug expo-
sure. As absorption enhancement can change the optimal drug exposure, judicious
adjustments are required [9].
Other researchers have tried to use the appropriate mucoadhesive systems—such
as hydrogel [10], viscous formulations [11], mucoadhesive polymers [12], in situ
gelling [13]—that can improve the drug retention time and decrease mucociliary
clearance. Drug encapsulation into a nanoparticle system is a precaution that must
be taken to prevent enzymatic degradation [14, 15]. However, the clinical success of
intranasal therapy is limited by the dose provided and the frequency of dosing.
The characteristics of a drug and its excipient (an inactive substance that serves
as a vehicle or medium for a drug) need to be taken into account. Direct delivery to
the delicate nasal epithelium of non-biocompatible, unprotected drugs, and certain
excipients like preservatives can upset the nasal homeostasis and result in nasal
irritation. Drug formulations must be compatible with nasal physiology if the nasal
route of administration is to be used successfully. In contrast to other routes of drug
administration, the nasal cavity permits a limited amount (25–200 μL) [15] of drug
in a single dose per nostril. Therefore, only the use of potent drugs is favored for
intranasal drug delivery. Drug excipients should be biocompatible and odor and
taste-free to enhance patient compliance [16]. As the drug is absorbed primarily in
the nasal mucosa, pH, viscosity, and tonicity must also be considered in intranasal
formulations [10].
If the challenges described above can be overcome, the intranasal route can
provide remarkable advantages over oral and parenteral routes of drug administration.
16 Nasal Delivery of Micro and Nano Encapsulated Drugs 341
This chapter briefly describes different procedures used to develop effective and
successful intranasal drug delivery with the help of micro and nanoparticulate sys-
tems. At present, several drugs are being investigated and clinically developed for
intranasal delivery. Intranasal drug delivery is a promising approach to treat central
nervous system (CNS) diseases because of its potential to deliver drugs across the
blood-brain barrier (BBB).
2 Polymers Used to Encapsulate Drugs
Nasal drug formulations can be encapsulated in micro or nano particles. The size of
the drug particles plays an important role, as it influences the interaction and trans-
port of particles across mucosal surfaces. Studies have shown that a particle size of
10 μm deposited in front of the nose, moreover the particle size of 1 μm can effi-
ciently cross intra-mucosal barriers [17]. To target the blood-brain barrier (BBB)
using the nasal drug delivery system, the particle size should be less than 100 nm [18].
The surface of drug particles—the surface charge and the hydrophobic or
hydrophilic nature of the particle surface—affects their interactions with the
biological environment. By encapsulating the drug with a polymer, we can control
the drug release from the polymer and achieve a low concentration of the drug in the
nasal epithelium, which eventually leads to a biocompatible formulation. The slow
release of the drug and the use of absorption enhancers prevent the exposure of
mucosal epithelial cells in the nasal cavity to a high concentration of the drug, as the
drug is absorbed quickly as soon as it is released from the formulation. Biodegradable
and biocompatible polymers that have been used extensively in nano and micro
encapsulations of drugs for nasal drug delivery include chitosan and poly(lactic-co-
glycolic acid) (PLGA).
3 Chitosan
Widder and his colleagues first reported chitosan micro-particles in 1983 [19].
Chitosan is a non-toxic natural polymer made of alkaline polysaccharides; it is the
second most abundant natural polysaccharide (cellulose is the most abundant natu-
ral polysaccharide) [20]. Chitosan’s properties of mucoadhesion, biodegradability,
and biocompatibility make it a good candidate for nano and micro encapsulations of
a wide range of nasal formulations of different drugs (Table 16.1) [21].
The positive charge of an amino acid group on chitosan interacts with a negative
charge on a sialic acid in the mucous membrane in the nose, and their coupling
enables a mucoadhesion for a prolonged time in the nasal cavity. This coupling can
also interfere with mucociliary clearance and increase the residence time of drug
[22–24]. Extensive toxicity studies have concluded that chitosan is safe in nano-
formulations using various routes of administration [25, 26]. Attempts to modify the
Table 16.1 Drugs encapsulated in chitosan nano and microparticles with or without surface
modification for intranasal delivery
Chitosan polymer modification
Drugs Particle size strategies References
Carbamazepine 216–221 nm Carboxymethyl chitosan [29]
Rasagiline 153 nm Chitosan glutamate [30]
Desvenlafaxine 151.5 nm PLGA/Chitosan [59]
Simvastatin 200 nm Lecithin/chitosan [54]
Estradiol 237.7– Thiolated-chitosan [60]
300.9 nm
Leuprolide 170–334 nm Thiolated-chitosan [48]
Buspirone 224.18– Thiolated Chitosan [33]
229.22 nm
Duloxetine 212.84– Proniosomes of thiolated [34]
234.98 nm chitosan
Donepezil 410.4–467 nm Liposomes dispersed into [35]
thiolated chitosan hydrogel.
Terbutaline sulphate 345.5 nm Lipid-chitosan hybrid [53]
Cetirizine 120 nm Deoxycholate-chitosan- [31]
hydroxybutyl nanoparticles
pH/thermo-responsive
Chitosan-coated liposome
Fexofenadine 716 nm [51]
Risperidone 86 nm and [36, 37]
132.7 nm
Quetiapine fumarate 131.08 nm [39]
Selegiline 341–502 nm [40]
Rotigotine 75.37 nm [41, 42]
Methotrexate 189 nm [43]
Bromocriptine 149.8– [44]
164.1 nm
Pramipexole dihydrochloride 292.5 nm [45]
Midazolam 144.99– [49]
149.41 nm
Catechin hydrate 93.46 nm [50]
Eugenol 224.5 nm [57]
Alpha-cyano-4-hydroxycinnamic 213–875 nm [36]
acid and cetuximab
Tapentadol hydrochloride 199.7– [46]
202.7 nm
chitosan polymer to improve its physiochemical properties, control drug release,

inhibit its enzymatic degradation, and prevent its toxicological effect are outlined in
the following sections.
4 Chitosan Surface Modifications
4.1 Carboxymethyl Chitosan (CMC)
Chitosan is soluble only in acidic medium, below pH 6.5 [27]. Chemical
modifications such as quaternization or the introduction of hydrophilic or
carboxyalkyl groups can enhance the solubility of chitosan. Carboxymethyl chitosan
(CMC) is one of the most studied chemically modified derivatives, and it is easy to
synthesize [28]. In an intranasal formulation, carbamazepine, an antiepileptic drug,
can be loaded in CMC nanoparticles to enhance its bioavailability and enable easy
and quick administration in emergencies; than conventional formulation [29].
4.2 Glutamate Chitosan
Chitosan nanoparticles administered intranasally can absorb water from the mucus
present in the nasal mucosa, drying the nasal passages and causing discomfort. In an
attempt to improve the in vitro dissolution rate of chitosan, chitosan glutamate
nanoparticles were synthesized and loaded with rasagiline to enhance the wetting
effect. The results showed that glutamate nanoparticles have a better wetting effect,
decreased particle size, and enhanced bioavailability [30].
4.3 Deoxycholate-Chitosan-Hydroxybutyl Nanoparticles
Chemical modifications of chitosan can promote stimuli-responsive derivatives.

Deoxycholate-chitosan-hydroxybutyl nanoparticles were tested for thermo and pH-
responsive behaviors. This system promotes quick action with a prolonged release
time upon hydrolysis of the chitosan backbone [31].
4.4 Thiolated Chitosan
Thiolated chitosan offers further enhanced bioadhesive properties via the formation
of a covalent bond—disulfide bond—between the thiol on chitosan and the mucus,
which is rich in cysteine, in the nose. Thiolated chitosan has gelling features in situ
that can strengthen the stability and cohesion of the carrier. In addition to their abil-
ity to open tight junctions, thiolated polymers inhibit enzymatic activity by chelat-
ing metals essential for their activity. Thiolated polymers do not disturb the ciliary
beat frequency (CBF), thus, they have no toxicological effect [32]. When buspirone
hydrochloride was added to thiolated chitosan, a biphasic drug release was observed,
starting with a burst release followed by a sustained release for 24 h [33].
Thiolated chitosan was also incorporated with other nanoformulations to enhance
their properties in nasal delivery. Proniosomes of duloxetine were formulated in
thiolated chitosan in a gel form that promoted the sustained release of the drug in
the nose, enhancing the mucoadhesion and permeation of the drug by over 1.86
times compared to proniosomes only, and providing an optimal particle size and a
high drug loading efficiency [34]. Dispersing donepezil liposomes in thiolated chi-
tosan hydrogel increased the content of the drug in the brain up to 107% compared
to oral delivery [35].
4.5 Deacetylated Chitosan
The positively charged surface of chitosan enables its compartmentalization and

conjugation with different molecules, conferring a targeting effect. Chitosan oligo-
saccharide is a shorter and deacetylated chain of chitosan. This derivative offers
higher solubility and lower viscosity than chitosan. The higher conjugation capacity
of chitosan oligosaccharide was proposed to act as a targeting ligand for encapsu-
lated drugs. Alpha-cyano-4-hydroxycinnamic acid was encapsulated in chitosan
oligosaccharide nanoparticles when the chitosan oligosaccharide surface contained
the covalently conjugated monoclonal antibody cetuximab as targeting ligand. This
delivery system provided a nose-to-brain delivery that has significantly reduced the
cytotoxicity and antiangiogenic effects of the drug in patients with glioblas-
toma [36].
5 Drug Encapsulation with Chitosan Derivatives
The nasal delivery system has been widely used to enhance the bioavailability of a
wide range of drugs. Chitosan is an attractive candidate for nasal delivery as com-
pared to conventional formulations. The conventional formulations of chitosan lack
the ability to deliver the required amount of drug to the brain due to multiple chal-
lenges. Table 16.1 shows drugs that are encapsulated in chitosan to overcome differ-
ent challenges.
5.1 Risperidone
The extensive first pass metabolism and the lipophilicity of risperidone make it an
attractive drug candidate for researchers to encapsulate risperidone using chitosan.
Risperidone-loaded chitosan nanoparticles were delivered with a higher yield than
risperidone alone [37]. Risperidone encapsulated in chitosan nanoparticles had a

particle size of 86 nm, when formulated by ionic gelation method using poloxamer
188 as a stabilizer. Risperidone-loaded chitosan nanoparticles had better pharmaco-
kinetic and pharmacodynamic properties than risperidone alone [37]. Moreover,
risperidone loaded in lipid-based chitosan nanoparticle escapes mucociliary clear-
ance due to its bioadhesive properties [38].
5.2 Quetiapine Fumarate
The Box-Behnken experimental design was developed to encapsulate quetiapine

fumarate in chitosan nanoparticles; this increased its bioavailability around two-
fold and conferred a prolonged retention time [39]. Pharmacokinetic studies showed
a high brain-targeting efficiency. These advantages resulted from chitosan’s ability
to pass through tight junctions, which increased its brain-targeting delivery via the
intranasal route [39].
5.3 Selegiline
Selegiline, a medication used for Parkinson’s disease, showed optimized

bioavailability with in vitro and ex-vivo studies. In addition, when formulated in
chitosan nanoparticles, selegiline offered more than 90% drug loading [40].
Compared to oral administration, selegiline concentrations increased around 20-fold
in the brain and 12-fold in the plasma after the intranasal administration of selegiline
in chitosan nanoparticles.
5.4 Rotigotine
The 1% bioavailability of rotigotine limits its physiological effectiveness. Intranasal

approaches using rotigotine formulated in chitosan nanoparticles showed promising
in vivo results [41], which were further confirmed by the results on cell lines and in
animal models. Pharmacokinetic studies suggest that the intranasal route is the best
route for direct transport of rotigotine to the brain [42].
5.5 Methotrexate
Encapsulations of drugs in chitosan-based formulations enhance their bioavailability

through their high drug loading efficiencies. Methotrexate-loaded chitosan-based
hydrogel nanoparticles resulted in a drug-loading efficiency of 72.03 ± 0.85% and
produced a significantly higher brain concentration of methotrexate compared with
simple solution [43].
5.6 Bromocriptine
Encapsulation of bromocriptine in chitosan-based formulations to treat Parkinson’s

disease achieved 84.2% ± 3.5% drug loading efficiency [44]. Moreover, pramipex-
ole dihydrochloride-loaded chitosan nanoparticles showed a drug-loading efficiency
up to 91 ± 0.95%, with adequate particle size and better in vivo activity compared
to the conventional formulation [45].
5.7 Tapentadol Hydrochloride
Nasal delivery is a promising approach for centrally-acting analgesics that are

quickly metabolized. Tapentadol hydrochloride acts centrally with a half-life of 4 h;
it requires frequent administration because it is extensively metabolized into inac-
tive moieties that are rapidly eliminated, and its hydrophilic nature decreases its
concentration in the central nervous system (CNS). Nasal delivery of tapentadol
hydrochloride encapsulated in chitosan nanoparticles provided a quick onset of
drug action, increased the time of tapentadol bioavailability compared to the intra-
venous route, and avoided the bitter taste associated with oral formulations [46].
5.8 Estradiol
A significant increase in hormonal drug bioavailability was achieved when the

drugs were loaded onto chitosan nanoparticles and administered nasally. Estradiol
loaded into chitosan nanoparticles and delivered nasally achieved higher plasma
and tissue concentrations than corresponding oral formulations. Leuprolide loaded
onto thiolated chitosan demonstrated a —two to five fold enhanced bioavailability
and a prolonged half-life [47, 48].
5.9 Midazolam
Midazolam loaded onto chitosan nanoparticles and administered intranasally was

superior in brain-targeting efficiency to a midazolam solution (IV) formulation,
offering a non-invasive approach to treat life-threatening seizures in status epilepti-
cus [49].
5.10 Catechin Hydrate
Catechin hydrate was formulated in chitosan nanoparticles to test its anticonvulsant

effect. This formulation delivered promising in vivo results for the treatment of
epilepsy as a result of its enhanced nose-to-brain delivery and a sustained release of
over 24 h [50].
5.11 Fexofenadine
Allergic rhinitis was treated with fexofenadine in liposomes coated with chitosan
and administered nasally. There were few systemic side effects, and the formulation
offered a prolonged release of the drug that can reduce the dosing frequency [51].
5.12 Astragalus Polysaccharides
Encapsulation of astragalus polysaccharides (APS) as microspheres in chitosan

provided a local extended release in the nasal delivery of the anti-inflammatory drug
for severe asthma, and alleviated allergic symptoms over an extended period [52].
6 Drug Encapsulation with a Lipid Polymer Hybrid
6.1 Terbutaline Sulphate
Terbutaline sulphate, a drug used to relieve bronchial asthma, was hybridized with
chitosan, a linear polymer saccharide, to enhance the mucoadhesive properties of
the drug’s formulation. The hybrid was formulated as a phospholipid coated with
chitosan/pectin (CS/PC) nanoparticles which offer a prolonged residence time [53].
6.2 Simvastatin
Simvastatin-loaded lecithin/chitosan nanoparticles were prepared in an attempt to

develop a new formulation for statins that would promote their use in neuro-
degradative diseases. Conventional formulations of statins do not guarantee the
delivery of the drug to the brain [54].
6.3 Lorazepam
A benzodiazepine derivative used for status epileptics, lorazepam was encapsulated

with a chitosan polymer to offer safer, more effective, and more convenient delivery,
especially for children. Microspherical hydrogels made of chitosan and pluronic
moieties and loaded with lorazepam offered thermo-sensitive properties. They
become viscous at 37 °C, allowing the microspheres to disperse before instillation.
This approach showed a sustained release over 24 h. Further in vivo studies are
needed to establish the efficacy of this formulation and its superiority over conven-
tional formulations [55].
7 Vaccine Encapsulation with Chitosan Derivatives
Recently, the nasal delivery of vaccines has attracted the attention of many research
groups. Polymeric nanoparticles have been investigated as carriers for different anti-
gens. Besides the advantages of chitosan and its water-soluble derivative, trimethyl
chitosan (TMC), as carriers, they could act as adjuvants, making this class a polymer
of choice. In an attempt to study nasal polymeric vaccines, inactivated PR8 influenza
virus was loaded into trimethyl chitosan (TMC) nanoparticles and coated chitosan
(CHT) nanoparticles respectively. Both TMC and CHT nanoparticles were coated
with sodium alginate (ALG) to study immune-stimulation potential after intranasal
administration. PR8-TMC-ALG formulation showed significantly higher immuno-
stimulatory response, compared with PR8-CHT-ALG and PR8 virus alone [56].
8 Drug Encapsulation with Co-polymers
8.1 Eugenol
Many reports indicate that eugenol is effective in the treatment of cerebral ischemia.
However, eugenol is volatile and susceptible to oxidation, which affects its stability
and contributes to its low bioavailability. To address these issues, eugenol was
encapsulated in nanoparticles and administered intranasally to improve its
bioavailability. Chitosan-coated poly(ε-caprolactone) nanoparticles loaded with

eugenol were prepared. The encapsulation of eugenol in chitosan increased the drug
concentration in the brain through chitosan mucoadhesive properties [57].
8.2 Ropinirole
Ropinirole is used to treat individuals with Parkinson’s disease. In an attempt to

improve the permeation of ropinirole hydrochloride through the nasal route of
administration, trimethyl chitosan was co-formulated with dipalmitoylphosphati-
dylcholine and poly(lactic-co-glycolic acid) (PLGA) as a penetration enhancer.
This mixture has been shown to enhance ex vivo nasal drug permeation by 2.35 fold
compared to the 50% bioavailability of the oral form [58].
8.3 Desvenlafaxine Succinate
Desvenlafaxine succinate was an antidepressant drug and attempts were made to

encapsulate it in copolymer and determine its intranasal potential. Desvenlafaxine
(DVF) was loaded in PLGA/chitosan nanoparticles and evaluated for in vitro and
in vivo studies. In vitro results showed a biphasic release pattern with 30% (pH 7.4)
and 34% (pH 6.0) drug release within 1 h which may be due to the DVF attached to
the surface of the nanoparticle, followed by characteristic sustained release for more
than 24 h that may be due to swelling and hydration PLGA/Chitosan nanoparticles
from the core matrix. In vivo results in rodents’ models demonstrate that DVF
loaded PLGA/Chitosan nanoparticles administered intranasally had higher drug
concentration in brain (954.56 ± 126.63 ng/ml) in comparison with the IV adminis-
tration (396.91 ± 64.34 ng/ml) [59].
9 Poly (Lactic-Co-Glycolic Acid): PLGA
The first report concerning poly lactic acid (PLA) biocompatibility and
biodegradability was published in 1966. Afterward, polyglycolic acid (PGA) was
discovered. PLA was formulated in long-acting microparticle formulations due to
its slow rate of degradation. Unlike PLA, PGA was not formulated in microparticle
formulations because of its fast degradation rate and its high hydrophilicity. As a
result, poly lactic co-glycolic acid (PLGA) was synthesized from PLA and PGA to
develop a customizable polymer. PLGA can be customized based on the PLA:PGA
ratio used to synthesize it. The PLA:PGA ratio will control whether the rate of
degradation is fast or slow, and it will control the lipophilicity/hydrophilicity ratio,
because PLA is lipophilic whereas PGA is hydrophilic. Moreover, the crystallinity
of PLGA depends on the lactic acid isomers that are used. For instance, the
enantiomers of lactic acid are semi-crystalline but the racemic mixture is amorphous.
Since the amorphous polymers provide a homogenous dispersion of the active
ingredient used, the D,L-PLGA is preferred [61]. PLGA is synthesized through
ring-opening polymerization to produce a high molecular weight copolymer. It is a
useful copolymer due to its biodegradability and biocompatibility. Variations in the
ratio of lactic acid to glycolic acid affect the melting point, the solubility, and the
glass transition temperature [62]. Mainly, PLGA is degraded in vivo by hydrolysis,
but enzymatic degradation could take place due to the presence of ester bonds.
Hydrolytic degradation can occur through surface degradation and bulk degrada-
tion, with or without autocatalysis. Polymer degradation can be detected by measur-
ing changes in the molecular weight of the polymer or by evaluating changes in the
physical properties of the polymer [63]. The degradation of PLGA is influenced by
factors such as the PLA:PGA ratio, the hydration rate of the polymer, the crystallin-
ity of the polymer, and the pH of the environment [64, 65].
Due to the properties of PLGA, it is widely used to form nanoparticles (NPs)
[66]. PLGA NPs can be prepared using nanoprecipitation, double-emulsion solvent
evaporation, and phase separation technique [67–69]. Because PLGA NPs can be
used in passive or active transport to cross the blood-brain barrier, they are of great
interest in the intranasal delivery of drugs targeting the brain. However, low brain
uptake was observed with the passive transport of PLGA NPs. Active transport of
PLGA NPs is mediated through carrier transport, adsorption, or receptor mediated
transcytosis. The mode of active transport depends on the type of modification per-
formed on the PLGA NPs. When PLGA NPs are modified with membrane perme-
able molecules, these molecules are transported with the PLGA NPs across the
blood-brain barrier (BBB). When the surface of the PLGA NPs is modified with
positively charged molecules, the positively charged molecules are adsorbed on the
negatively charged surface of the neurons, resulting in cell endocytosis. Modifications
of PLGA NPs using ligands specific to blood-brain barrier cell surface receptors
enable the transportation of PLGA NPs into neurons through receptor-mediated
transcytosis [70]. The use of PLGA NPs in intranasal formulations enhances the
drug targeting efficiency (%DTE). For instance, an intranasal formulation of PLGA
NPs was found to have a drug targeting efficiency of 129.81% [68]. PLGA NPs
administered intranasally provide a prolonged duration of drug release in compari-
son to intravenous administration [67, 68, 71].
10 PLGA Surface Modifications
10.1 Pegylation
To increase the penetrating ability and stability of NPs in mucus, their surface is
coated with polyethylene glycol (PEG). The PEG coating prevents the charges on
the surface of the NPs from forming hydrophobic or electrostatic interactions;
consequently, mucoadhesion is decreased [72]. Specific physicochemical properties,

such as low molecular weight PEG, must be met to promote the formation of a
dense PEG coating [73]. Another study demonstrated that a PEG coating improved
the retentivity of an intranasal formulation [74].
10.2 Lactoferrin
The modification of PLGA nanoparticles with lactoferrin is useful due to the

overexpression of lactoferrin receptors on the surface of respiratory epithelial cells,
neurons, and capillaries in neurodegenerative diseases [74, 75]. To provide targeted
brain delivery through intranasal administration, a research group successfully for-
mulated PEG-PLGA nanoparticles modified with lactoferrin. The formulation
exhibited small particle size, narrow size distribution, low cytotoxicity, and enhanced
cellular accumulation due to lactoferrin [74]. PLGA nanoparticles co-modified with
lactoferrin and N-trimethyl chitosan (TMC) were formulated. Chitosan was modi-
fied to TMC by reductive methylation to enhance its solubility and adhesion. The
modification resulted in better cellular uptake when compared with PLGA nanopar-
ticles. Also, a higher brain distribution was observed over a longer period of time
[75]. In another study, PLGA NPs modified with lactoferrin and folic acid increased
the blood-brain barrier permeability coefficient by two-fold [76].
10.3 Peptides
10.3.1 RVG29
RVG29, an amino acid derived from the rabies virus, was reported to enhance the
accumulation of PEG-PLGA nanoparticles in the brain. RVG29 is attached to the
surface of PLGA NPs through avidin-biotin interactions [75]. The RVG29 modified
PEG-PLGA NPs exhibited higher brain distribution than unmodified PEG-PLGA
[76]. In another study, RVG29 modified PEG-PLGA NPs targeted specific tissues
near the trigeminal nerves [75].
10.3.2 Octa-Arginine
Octa-arginine, a cell-penetrating peptide, can increase CaCO2 cellular uptake and

cell permeability when conjugated on the surface of PLGA NPs. Also, conjugation
of a 12 amino acid peptide denoted by (Pep TGN i.e., bacteriophage clone for brain-
targeted delivery) with PEG-PLGA NPs results in a two to three-fold increase in
brain accumulation. PLGA NPs modified with octa-arginine can be prepared using
the solvent evaporation technique. Intranasal administration of the formulation
leads to a high cellular uptake and a high rate and extent of drug delivery. It is
important to mention that the zeta potential increases due to the neutralizing interac-
tion between the positive charge of octa-arginine and negative charges of carboxylic
acid groups on the NP surfaces. Due to the hydrophilicity of octa-arginine, the
enhanced wetting results in a higher burst release of the drug from the formulation.
Octa-arginine modified PLGA NPs are thought to be transported through receptor-
mediated mechanisms [77].
10.3.3 RGD Tripeptide
The RGD (Arg-Gly-Asp) tripeptide can be used as a surface modification to target

cancer cells. PLGA NPs modified with the RGD (Arg-Gly-Asp) tripeptide decrease
enzymatic degradation and provide higher drug targeting ability. Administering the
formulation intranasally controls cancer growth and provides a localized effect, thus,
it is a promising formulation to target brain diseases [78]. PLGA NPs modified with
RGD were well distributed in a tumor location; their presence was accompanied by an
increased inhibition of tumor cells and a higher reduction in tumor volume [69].
10.3.4 Lectins
Odorranalectin (OL), the smallest lectin discovered, has low toxicity and low
immunogenicity. OL can help nanoparticles to avoid cilia clearance and enzymatic
degradation, and can improve membrane permeability through binding to highly
selective glycosylated receptors expressed on the nasal mucosa. OL is attached to
PEG-PLGA NPs through an interaction between the thiol group in OL and the
maleimide group in PEG-PLGA. OL PEG-PLGA NPs have some toxic effects on
the cilia, but no cytotoxicity was observed in intranasal administrations [79]. This
modification can enhance cellular uptake, prolong the mean residence time of the
drug, and increase the bioavailability of drug-loaded nanoparticles [79].
10.4 Monoclonal Antibodies
Monoclonal antibodies can be used to modify the surface of nanoparticles (NPs). The
anti-EPHA3 (Ephrin Type A Receptor 3) monoclonal antibody was utilized as a sur-
face ligand in an intranasal formulation to enhance the targeting of glioblastoma mul-
tiforme. Anti-EPHA3 was attached to the nanoparticle surface by thiolation to interact
with the maleimide in trimethyl chitosan (TMC). The modification is safe and pro-
vides high cellular uptake and efficient targeting. A comparison between intranasal
and intravenous administrations revealed that intranasal administration delivered
more drug to the brain than intravenous administration. In vivo evaluation of the anti-
glioma activity indicated a significant increase in apoptotic glioma cells [80].
11 Drug Encapsulation with PLGA Derivatives
11.1 Lamotrigine
To provide a targeted and prolonged drug delivery method for lamotrigine, PLGA
nanoparticles encapsulating lamotrigine were prepared through a modified nanopre-
cipitation method. The formulation increased the accumulation of the drug in the
brain at levels higher than the levels observed in intravenous administration. The
formulation followed the Korsmeyer Peppas model, in which the drug release from
the formulation is due to the initial swelling of the polymer followed by a gradual
drug release from the matrix [68].
11.2 Haloperidol
Intranasal administration of haloperidol enhanced the efficacy of this antipsychotic

drug which is used to treat schizophrenia. Haloperidol was encapsulated in PEG-
PLGA nanoparticles coated with Solanum tuberosum lectin (STL). The emulsion/
solvent evaporation technique was successful at producing Haloperidol-STL-PEG-
PLGA nanoparticles with a high entrapment efficiency (EE) of 73–85%. The drug
reached the brain at higher concentrations when administered intranasally in com-
parison to other routes of administration. Haloperidol loaded in STL-PEG-PLGA
nanoparticles showed a positive response at a lower dose than unencapsulated halo-
peridol. The observed result is attributed to the selective binding of STL, which
improved nasal cellular uptake [81].
11.3 Oxcarbazepine
The first line treatment for focal seizures is the lipophilic drug oxcarbazepine. The
free drug is effective when administered intranasally, but it requires frequent dosing.
To decrease the number of doses, increase brain targeting, and prolong the drug’s
effect, the drug was loaded in PLGA nanoparticles for intranasal administration.
The PLGA nanoparticles provided a neuroprotective effect for a period of 16 days
and more than 24-h decrease in dosing frequency. The formulation provided epilep-
tic patients with prolonged protection against seizures [82].
Other drugs formulated for intranasal delivery include rotigotine (an
enantioselective dopamine agonist), huperzine A, lamotrigine, lorazepam, curcumin
(a hydrophobic drug with low oral bioavailability), doxorubicin, paclitaxel, and
temozolomide. Table 16.2 contains a list of drugs encapsulated with PLGA deriva-
tives to overcome uptake and therapeutic barriers.
Table 16.2 Drugs encapsulated in PLGA-based nano and microparticles with surface modification
for intranasal delivery
Drug Particle size PLGA polymer modification strategies References
Rotigotine 122 nm Lactoferrin [83]
Huperzine A 153 nm Lactoferrin [84]
N-Trimethyl chitosan (TMC)
miR-124 204 nm RVG29 [76]
Loperamide 328 nm Octa-arginine (R8) [77]
Curcumin 97.1 nm Odorranalectin [79]
Doxorubicin 180–200 nm RGD tripeptide [78]
Temozolomide 145.9 nm Anti-EPHA3 monoclonal antibody [80]
Trimethylated Chitosan (TMC)
Haloperidol 121 nm PEG and lectin [81]
Oxcarbazepine 256.15 nm [82]
12 Other Polymers Used for Intranasal Delivery
12.1 Polycaprolactone Polymers
To extend its half-life and to protect it from oxidation, melatonin was formulated in
polycaprolactone-based nanoparticles for intranasal delivery to treat glioblastoma.
This formulation enabled a direct transport into the brain. Interestingly, the solubil-
ity of melatonin was increased ≈35-fold and its antitumor effect was demonstrated
at very low doses of the formulation. These advantages were attributed to a con-
trolled drug release over time and a high cellular uptake [85].
Carboplatin was encapsulated in polycaprolactone-based nanoparticles to treat
glioblastoma. In situ nasal perfusion studies showed a better absorption of carboplatin
and in vitro studies revealed a promising carboplatin antitumor effect [17]. In another
study, to stabilize and enhance the mucoadhesion properties of polycaprolactone, an
amphiphilic methacrylic copolymer composed of methyl methacrylate (MMA) and
2-(dimethylamino)ethyl methacrylate (DMAEMA) was synthesized, and an emulsi-
fier was added to olanzapine polycaprolactone-based nanoparticles. This combina-
tion produced a pH-sensitive cationic coating that offered controlled release and an
increased olanzapine concentration in the brain. In vivo studies strongly suggest that
this formulation offers a safe and effective nose-to-brain delivery [86].
12.2 Cellulose Derivatives
Cellulose derivatives are biocompatible in general. They are easily formulated and
provide a cost-effective choice for nasal delivery. Several studies noted that hydroxy-
propyl methylcellulose (HPMC)-based microspheres delivered nasally offered a
rapid onset of action, mainly due to the low viscosity of HPMC. The encapsulation
of tramadol in HMPC-based microspheres provided a quick onset of action; in vitro
drug release was 94% after 90 min [87].
Table 16.3 Drugs encapsulated in polymer as nano and microparticles for intranasal delivery
Drug Particle size Polymer References
Olanzapine 327–469 nm Polycaprolactone [86]
Melatonin 100–400 nm Polycaprolactone [85]
Carboplatin 306.9–316.3 nm Polycaprolactone [17]
Tramadol 12–18 μm HPMC [87]
Valsartan 10–22 μm HPMC [88]
Ropinirole HCL 2.35 μm Sodium Alginate [89]
Valsartan formulated as HPMC-based microspheres provided a quick onset of

action. Animal models showed that nasal delivery enhanced activity compared to
oral formulations. Further studies are needed to establish whether hypertension can
be managed by drugs delivered through the intranasal route [88].
12.3 Alginate Derivatives
Ropinirole hydrochloride, an anti-Parkinson drug, was encapsulated in sodium

alginate microparticles by spray drying for nasal delivery. The formulation had no
toxic effect on the nasal mucosa and provided a rapid drug release. It is important to
mention that the drug:polymer ratio affects the time and quantity of drug release
from the formulation [89]. Table 16.3 lists a number of drugs that are encapsulated
in a polymer to overcome different nasal delivery challenges.
13 Conclusion
Intranasal drug administration is a non-invasive technique to deliver small drug

molecules and biologicals to the central nervous system. Successful intranasal for-
mulations require appropriate adjuvants to enhance drug absorption and decrease
mucociliary clearance. To augment drug bioavailability, a drug can be encapsulated
in nanoparticles. Polymeric encapsulation with chitosan, PLGA, and their deriva-
tives helps to control drug release; encapsulation decreases enzymatic drug degra-
dation and increases cellular uptake. This might enable low drug concentrations to
be more effective compared to other routes of administration.
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Chapter 17
Different Strategies for Nose-to-Brain
Delivery of Small Molecules
Smita P. Borkar and Abhay Raizaday
Abstract The intranasal (IN) route of drug administration has emerged as an

alternative route over the systemic (oral and parenteral) drug delivery to the brain.
The intranasal route of drug administration exhibits as a non-invasive technique
to bypass the BBB for the delivery of drugs inside the brain and the CNS. This
method is helpful for those drugs that are unable to invade the BBB to show its
action in the CNS and thus erase the demand of systemic delivery and shrink sys-
temic side effects. Drug delivery from the nose to the brain/CNS takes very less
time through both olfactory and trigeminal nerves. Intranasal delivery does not
require the involvement of any receptor as it occurs through an extracellular route.
The delivery of the drug via an IN route offers various advantages over a systemic
drug delivery system as it directly delivers the drug into the brain via the olfactory
route. The presence of drugs in the olfactory bulb, in turn, increases the drug bio-
availability in the brain and reduces degradation as well as wastage of the drug
through systemic clearance. However, there are some limitations associated with IN
like poor drug permeation through the nasal mucosa and mucociliary clearance.
There are many novel drug delivery strategies (nano-drug carrier system, colloidal
carriers, mucoadhesive devices, controlled delivery system, pro-drug, etc.) are
adapted to overcome the above-stated limitations. Nose-to-brain delivery also
involves nasal-associated lymphatic tissues (NALT) and deep cervical lymph nodes.
This review focuses on different strategies for nose-to-brain delivery of small
molecules.
Keywords Intranasal delivery · Systemic delivery through nasal passage · Nano

drug delivery for quick absorption · Mucoadhesive drug delivery for nasal passage
S. P. Borkar
College of Pharmacy, JSS Academy of Technical Education, Noida, Uttar Pradesh, India
Arvind Gavali College of Pharmacy, Jaitapur, Satara, Maharashtra, India
A. Raizaday (*)
College of Pharmacy, JSS Academy of Technical Education, Noida, Uttar Pradesh, India
https://doi.org/10.1007/978-3-031-23112-4_17
362 S. P. Borkar and A. Raizaday
1 Introduction
Intranasal drug delivery is emerging as a reliable and promising pathway to deliver

a wide range of therapeutic agents, including small and large molecules, peptides
and proteins, and genes to the central nervous system for the treatment of brain
diseases such as Alzheimer’s disease, Parkinson’s disease, depression, migraine,
schizophrenia, and glioma. This presents non-invasive entry into the brain via direct
nose-to-brain and/or indirect nose-to-blood-to-brain routes. The nasal mucosa was
seen as a potential route of administration to achieve a faster and higher level of
drug absorption as it is permeable to more compounds than the gastrointestinal
tract. In recent years, many drugs have been shown to achieve better systemic bio-
availability via the nasal route than when administered orally.
The unique relationship between nasal cavity and cranial cavity tissues in
anatomy and physiology makes intranasal delivery to the brain feasible. Intranasal
delivery provides some drugs with short channels to bypass the blood-brain barrier
(BBB), especially for those with fairly low brain concentrations after a routine
delivery, thus greatly enhancing the therapeutic effect on brain diseases. In the past
two decades, a good number of encouraging outcomes have been reported in the
treatment of diseases of the brain or the central nervous system (CNS) through nasal
administration. In spite of the significant merit of bypassing the BBB, direct nose-
to-brain delivery still bears the problems of low efficiency and volume for capacity
due to the limited volume of the nasal cavity, the small area ratio of olfactory mucosa
to nasal mucosa and the limitations of low dose and short retention time of drug
absorption. It is crucial that selective distribution and retention time of drugs or
preparations on olfactory mucosa should be enhanced so as to increase the direct
delivery efficiency.
Several nanocarrier-based strategies have been developed to transport therapeutic
agents to the brain, including nanoparticles, liposomes, and exosomes following
intranasal delivery. However, the multiple barriers in nose-to-brain route - including
rapid mucociliary clearance in the nasal cavity, enzyme degradation, and the blood-
brain barrier (BBB) - pose serious challenges to brain-targeted drug delivery. The
potential for treatment possibilities with intranasal transfer of drugs will increase
with the development of more effective formulations and delivery devices.
2 Mechanism of Nasal Absorption
Several mechanisms have been proposed for absorption of drugs through the
nasal route,
1. The first mechanism involves an aqueous route of transport, which is also known
as the paracellular route. This route is slow and passive. There is an inverse log–
log correlation between intranasal absorption and the molecular weight of water
soluble compounds. Poor bioavailability was observed for drugs with a molecu-
lar weight greater than 1000 Daltons.
17 Different Strategies for Nose-to-Brain Delivery of Small Molecules 363
2. The second mechanism, which involves transport through a lipoidal route, is also
known as the transcellular process and is responsible for the transport of lipo-
philic drugs that show a rate dependency on their lipophilicity.
3. Drugs also cross the cell membranes by an active transport route via carrier-
mediated means or transport through the openings of tight junctions.
3 Strategies for Improving Nasal Drug Delivery
Various strategies used to improve availability of the drug in the nasal mucosa
include:
1 . To improve the nasal residence time
2. To enhance nasal absorption
3. To modify drug structure to change physicochemical properties
3.1 Improve the Nasal Residential Time
Mucociliary clearance works to remove foreign bodies and substances from the
nasal mucosa as quickly as possible. One way to delay clearance is to apply the drug
to the front of the nasal cavity, an effect that is largely determined by the type of
dosage form used. The preparation could also be formulated with polymers such as
methyl cellulose, hydroxypropyl methylcellulose or polyacrylic acid, in which the
incorporation of the polymer increases the viscosity of the formulation and also acts
as a bioadhesive with the mucus. Increasing residence time does not necessarily
lead to increased absorption; this concept can be illustrated by considering an insu-
lin solution with a similar viscosity containing carbopol and carboxy methylcellu-
lose (CMC). Carbopol improves absorption where CMC solution does not improve
insulin absorption. The increase in viscosity retards drug diffusion from the matrix
and causes retention in absorption with CMC. In case of carbopol, it causes an
increase in absorption due to the opening of intracellular junctions.
Another lucrative way to increase nasal resistance time is to use biodegradable
microspheres as carriers for drug delivery. Biodegradable microspheres swell in the
presence of water, thereby increasing viscosity. This phenomenon leads to an
increase in the nasal residence time.
3.2 Enhancing the Nasal Absorption
The term “absorption enhancer” generally refers to an agent whose function is to

increase absorption by improving membrane permeation rather than by increasing
solubility. Thus, these agents are sometimes referred to more specifically as
permeability enhancers, wherein the drug passes through the nasal mucosa, altering
in some way the structure of the epithelial cells (permeation enhancers). This must
be accomplished without causing damage or permanent changes to the nasal mucosa.
In general, absorption enhancers can act through one of the following mechanisms:
(a) Open tight junctions
(b) Decrease mucociliary clearance
(c) Inhibit enzyme Activity
(d) Reduce mucus viscosity or elasticity
The mechanism of action of the absorption enhancer increases the rate at which the
drug passes through the nasal mucosa. Many activators work by altering the struc-
ture of epithelial cells in one way or another, but they should do so without causing
damage or permanent changes to the nasal lining. The general requirements of an
ideal penetration enhancer are as follows:
(a) This should lead to an effective increase in the absorption of the drug.
(b) It should not cause permanent damage or tissue damage.
(c) It should be effective in small amounts.
(d) It must be non-irritant and non-toxic.
(e) The enhancing effect should occur when absorption is required.
(f) The effect must be temporary and reversible. It must be compatible with other
excipients.
Types of penetration enhancers:
(a) Solvents
(b) Alkyl methyl sulphoxides
(c) Pyrrolidones
(d) Dodecyl azacycloheptan-2-one
(e) Surfactants
Mechanisms of penetration enhancers:
(a) Increasing cell membrane permeability by opening tight junctions and formation
of intracellular aqueous channels
(b) Increasing lipophilicity of the charged drug by forming ion pairs
(c) Inhibiting proteolytic activity
3.3 To Modify the Structure of the Drug to Change

the Physicochemical Properties
Modifying the structure of the drug without altering the pharmacological activity is
a lucrative method of improving nasal absorption. Here, the modification of physi-
cochemical properties such as molecular size, molecular weight, PKa, and solubil-
ity are favorable for nasal absorption of the drug.
4 Nose-to-Brain Delivery
Nose-to-brain delivery poses a big challenge. In fact, a large number of neurological

diseases require therapies in which the drug must reach the brain, avoiding the dif-
ficulties due to the blood–brain barrier (BBB) and the problems connected with
systemic administration, such as drug bioavailability and side-effects. For these rea-
sons, the development of nasal formulations able to deliver the drug directly into the
brain is of increasing importance.
The blood–brain barrier (BBB) separates the central nervous system (CNS) from
the systemic circulation. The barrier characteristics of BBB depend on the proper-
ties of the brain endothelial cells that constitute the walls of the blood vessels. There
are many neurological diseases such as neurological infections, Parkinson’s disease,
Alzheimer’s disease, multiple sclerosis, age-related neurodegenerative diseases,
and cerebral ischemia that require a therapy in which the drug must reach the brain.
Furthermore, many of these diseases need chronic therapies. Drug targeting to
the brain poses a big challenge because many of these drugs cannot cross the
BBB. Therefore, many efforts must be made to design strategies to solve this prob-
lem. The use of the nose-to-brain delivery route is an important and non-invasive
method of drug delivery to bypass the BBB. In fact, it is well-known that there is an
intranasal direct anatomical connection between the nasal cavity and the CNS,
which suggests the development of nasal formulations for brain targeting of drugs.
Different strategies have been developed for nose-to-brain drug delivery and involve
nanomedicine with different kinds of nanocarriers: polymeric nanoparticles, nano-
emulsions, dendrimers, and nano-micelles. The development of new nasal systems
poses a great challenge in the field of controlled drug targeting and delivery.
5 Factors Relating to the Rate and Capacity of Drug

Transport from Nose to Brain
5.1 Physicochemical Properties of the Drug
The rate and capacity of drug transport from the nasal mucosa to the brain depends
primarily on the drug’s physicochemical properties, especially its molecular weight,
lipophilicity, and degree of dissociation.
5.1.1 Relative Molecular Weight
Most small molecular weight (<400 Dalton) drugs can be freely transported into the
brain through the nasal mucosa epithelium, specifically for odorant molecules.
Generally, drugs with a molecular weight above 1000 Dalton show poor capability
in penetrating the physiological barrier, and the rate of mucosa permeation is highly
sensitive to molecular size [36, 37].
Sakane et al. examined the influence of molecular weight (4400 Dalton
fluorescent-labeled dextrans [FD4], 9400 [FD10], 18,900 [FD20] and 40,500
[FD40]) on the nose-to-brain transport of FD by determining their concentrations in
cerebrospinal fluid (CSF) following i.v. and intranasal administration. As a result,
no FD could be detected in CSF after i.v. administration. FD4, FD10 and FD20
were detected in CSF upon intranasal administration, and their concentration
decreased with increasing molecular weight. Meanwhile, their concentrations in
plasma were much lower than those after i.v. administration. Nevertheless, it is
imperative to understand the uptake of higher molecular weight materials like some
peptides and viruses to the brain through their special pathways [38].
Born et al. and Fehm et al. administered three types of peptides intranasally:
melanocortin (4 – 10) (MSH/ACTH (4 – 10)), vasopressin and insulin, and found
that intranasal administration could significantly increase drug concentration in the
brain as compared to i.v. administration, indicating that the nasal mucosa pathway
was more advantageous for hydrophilic high molecular weight drugs. Many large
water-soluble drugs also reach the brain following intranasal administration by trav-
eling along the olfactory and trigeminal neural pathways. Most viruses, when
absorbed by nasal mucosa, could be taken up by neuron axon endings and trans-
ported to olfactory bulbs through axoplasma flow in olfactory nerve cells to further
reach the olfactory brain. However, the rate and capacity of large molecular drug
transport into the brain also had a relationship with drug transport pathways and
whether there were specific receptors. For example, with the addition of wheat germ
agglutinin (WGA) to horseradish peroxidase (HRP), which could not cross the
NBB, WGA-HRP could be transported into the brain by conjugating with the WGA
receptors on the mucosa [56, 57].
5.1.2 Lipophilicity
Sakane et al. investigated the influence of lipophilicity on drug transport from nose
to brain using sulfonamides as model drugs and found that there existed a direct
transport pathway for sulfonamides from nose to brain, and within a certain range,
drug concentrations in CSF increased with their elevating lipophilicity. Our group
studied the absorption of diltiazem and acetaminophen under a series of pH values
by the method of classical rat in situ nasal recirculation, and found that like most
other biological membranes, the nasal mucosa presented a “lipid sieve” feature,
making itself easy to penetrate by highly lipophilic drugs, with a well-defined linear
correlation between the drug’s oil-water distribution coefficient and its absorption
rate constant [58, 59].
5.1.3 Degree of Dissociation
Sakane et al. studied the relationship between the degree of dissociation of the
model drug sulphisomidine (p K a = 7.5) and its direct transport from the nasal cav-
ity to CSF by measuring its concentrations in plasma and CSF after rat nasal perfu-
sion with a series of buffers (pH 5.5, 6.5, 7.4, 8.7 and 9.4). The nasal clearance
increased with the elevation in the un-ionized fraction of the drug and the ratio of
the drug concentration in CSF to that in the nasal perfusion fluid changed in accor-
dance with the un-ionized fraction of the drug, showing that both the nasal absorp-
tion and the drug transport conformed to the pH partition theory. Namely, the degree
of ionization of an intranasally administered drug could affect both the absorption
across the nasal epithelium and its transport into the CSF. The drug concentration in
the CNS inversely correlated with its dissociation [60].
5.2 Drug Concentration, Dosage, and Dosing Volume
Drug concentration, dosage, and dosing volume are three major factors impacting
drug nasal absorption in correlation to one another. The nasal absorption of most
drugs increases with the increase in concentration, especially those with an absorp-
tive mechanism of passive diffusion. In animal experiments, drugs were ordinarily
administered by immersing the olfactory region. Within a certain dose range, the
drug absorption and therapeutic effects would rise with increasing dosage. However,
the volume of the nasal cavity is limited and the dosage for nasal administration is
relatively low at 25–200 μl, thus, constricting to a certain degree the amount of drug
transport from nose to brain [47].
5.3 Nasal Mucous Membrane Cilia Clearance

and Dosage Form
Following intranasal administration, the precipitation, clearance, and absorption of

a drug or its preparation are all completed in the nasal passage. The epithelium of
the nasal passage is covered by a mucus blanket, which entraps particles and bacte-
rium to be cleared from the nasal cavity and renewed by cilia. The nasal mucous
cilia play a defensive role in maintaining the normal physiological environment of
the nasal cavity, and in the meantime, they are capable of clearing the drug or prepa-
ration particles together. Except for the nasal vestibule, olfactory membrane, and a
small anterior part of the conchae, the remainder is ciliated epithelium with numer-
ous surface microvilli in the nasal cavity, including the epithelium of the respiratory
system. Mucociliary clearance limits the time available for drug or drug preparation
absorption, although in contrast, the numerous microvilli on the ciliated nasal
epithelium providing a huge membrane area greatly enhance drug absorption

compared with non-ciliated epithelium.
6 Nanocarrier-Based Strategies Promoting Nose

to Brain Delivery
The development of nanocarrier-based delivery systems has created exciting

opportunities for the management of brain diseases [1–8]. At present drugs can be
delivered to the brain by loading them into a nanocarrier-based system, which would
interact well with:
1 . Nasal mucosa to increase drug absorption time
2. The olfactory nerve fibers to promote direct nose-to-brain delivery
3. The endothelial microvessel cells at the BBB; this would produce higher drug
concentrations in brain parenchyma through indirect nose-to-blood-to-brain
pathway.
Nanocarriers could be further modified with targeting moieties to preferentially bind
to specific receptors of transporters presented at respiratory epithelial cells, neurons,
and the BBB for enhanced brain selectivity and affinity. Hence, they can be exploited
for efficient drug trafficking across the barrier structure through membrane trans-
cytosis processes. Overall, more favorable drug pharmacokinetics, improved effi-
cacy, and safety have been achieved using nanocarrier-based delivery systems. In the
following section, we will review and discuss how and why nanocarriers (mainly
nanoparticles, liposomes, and exosomes) do facilitate nose-to-brain delivery.
6.1 Nanoparticles
Nanoparticles are colloidal systems with compact structure where the therapeutic
agent is either entrapped within the matrix or bound on the particle surface by con-
jugation or adsorption. Nanoparticles are mostly made of polymers, lipids, or a
combination of both to produce sustained and controlled drug release. In the next
section, we will review different nanoparticle-related approaches to convey thera-
peutics to the brain following intranasal administration and highlight the surface
modifications (Fig. 17.1).
6.1.1 Residence Time Increasing Nanoparticles
Mucociliary clearance in the nasal cavity is a significant barrier that restricts

therapeutics delivery to the brain through intranasal administration. Mucoadhesive
and viscosity increasing agents have been used in hopes of increasing drug residence
time in the nasal cavity to allow for better absorption.
Fig. 17.1 Four types of nanoparticles promote nose-to-brain delivery: lectin-modified

nanoparticles, lactoferrin-modified nanoparticles, cell-penetrating peptide-modified nanoparticles,
and residence time increasing nanoparticles
Mucoadhesive polymers like chitosan, alginate, gelatin, cellulose, and

polyacrylate were often used to fabricate nanoparticles, producing prolonged
retention time at the absorption site and therefore increasing brain delivery efficiency
[9, 27–29].
Didanosine, piperine, and quetiapine fumarate have been loaded into chitosan
nanoparticles to achieve brain delivery through the nasal route. Haque et al. reported
that venlafaxine-loaded nanoparticles made by alginate and chitosan were able to
effectively deliver the drug to the brain in a significant quantity to treat depression,
following nasal administration. This could be the summation of the attribution of a
number of factors such as the increase in absorption time due to reduction in nasal
mucociliary clearance, increase in permeation across nasal mucosa, modulation of
P-gp efflux transporters present on the BBB that prevent the entry of drugs into the
brain, and contribution of paracellular transport through tight junction modulation
between the cells. Not only small molecules, but peptides were also delivered by
this approach. Lu et al. described gelatin nanoparticles as carriers of substance P, a
peptide able to increase the dopamine content in the brain. The developed system
increased functional improvement and decreased levels of apoptosis on rats with
hemiparkinsonism, indicating the potential application for Parkinson’s disease. In
addition, most mucoadhesive nanoparticles were hydrophilic and prepared in mild
process, making them adapted for gene delivery. For the first time, chitosan nanopar-
ticles were able to complex siRNA targeting galectin-1, which is overexpressed in
glioblastoma, and protect them from RNAse degradation. Moreover, highly concen-
trated chitosan nanoparticles effectively and rapidly conveyed siRNA through nasal
mucosa into the olfactory bulbus and the hindbrain for glioblastoma treatment [10–
15, 33].
6.1.2 Lectin-Modified Nanoparticles
Lectins are sugar-binding proteins that specifically recognize sugar molecules and
therefore, are capable of binding glycosylated membrane components. A novel
polymeric nanoparticle developed by Gao et al. was composed of wheat germ
agglutinin conjugated to poly (ethylene glycol)-poly (lactic acid) (PEG-PLA) in an
effort to increase absorption of nanoparticles to the brain. They used the nanoparti-
cle carrier to encapsulate coumarin and found a two-fold increase in brain concen-
trations after intranasal administrations when compared to intranasal administration
of unmodified PEG-PLA nanoparticles. In a later study, they determined if the
nanoparticle carrier would be applicable to transport peptides to the brain. They
incorporated a vasoactive intestinal peptide into the wheat germ agglutinin conju-
gated PEG-PLA nanoparticles. When given intranasally, the authors reported 5.6- to
7.7-fold higher brain levels from the surface-modified nanoparticles when com-
pared to vasoactive intestinal peptide given intranasally as a solution or delivered in
unmodified nanoparticles. The novel carrier was assessed for toxicity issues during
intranasal use by analyzing concentrations of surrogate markers, such as tumor
necrosis factor alpha and wheat germ agglutinin-specific antibodies; it was con-
cluded that the nanoparticles were a safe agent for use in intranasal therapy target-
ing the brain [16–18].
However, one main challenge for various lectin members is their serious
immunogenicity in vivo caused by the relative large molecular weight, which
remarkably limits their potential application in drug delivery systems. Odorranalectin
(OL), identified from skin secretions of Odorrana grahami, has so far been proven
to be the smallest peptide with lectin-like activity and presented much less
immunogenicity than wheat germ agglutinin. OL is able to specifically recognize
L-fucose, which is widely located on the olfactory epithelium of nasal mucosa,
suggesting it as a promising ligand to promote intranasal drug delivery [19–24].
Wen et al. established a novel (OL)conjugated poly(ethyleneglycol)–poly(lactic-
co-glycolic acid) (PEG–PLGA) nanoparticle to load urocortin peptide, which is
able to provide a long-term restoration of nigrostriatal function. The OL modifica-
tion on the surface increased the brain uptake of PEG–PLGA nanoparticles, and
thus, enhanced the neuroprotective effects of urocortin following intranasal applica-
tion. In further research, OL-modified cubosomes presented good transportation
ability of peptides into the brain ameliorating learning impairment in rats with
Alzheimer’s disease through nasal delivery [25, 26, 46].
6.1.3 Lactoferrin-Modified Nanoparticles
Lactoferrin is a natural iron binding cationic glycoprotein generally present in

mammalian brain cells, having the ability to bind to the lactoferrin receptor at BBB
firstly and thereafter getting converted to a positively-charged group which binds to
negatively-charged BBB at physiological condition. Especially, lactoferrin receptor
has been demonstrated to be overexpressed in age-related neurodegenerative
diseases, including Alzheimer’s disease, Parkinson’s disease, Huntington disease,

and amyotrophic lateral sclerosis. In addition, high lactoferrin receptor expression
was observed in the nasal and bronchial epithelial cells, making it reasonable to
modify nanoparticles with lactoferrin as a ligand to promote membrane penetration
[27–31].
Liu et al. developed poly(ethyleneglycol)-poly(ε-caprolactone) (PEG-PCL)
nanoparticles modified with lactoferrin as the carriers of neuroprotective peptides
for the treatment of Alzheimer’s disease. Cellular accumulation was enhanced by
lactoferrin-modified nanoparticles, contrary to the non-modified ones via clathrin/
caveolae-mediated endocytosis and direct translocation. A desirable brain biodistri-
bution profile with significantly increased coumarin (a marker) delivery in the rat
olfactory bulb and olfactory tract was also demonstrated. Moreover, in the pharma-
codynamic experiment, neuroprotective peptide-loaded lactoferrin-modified
nanoparticle treatment showed significant improvement in both behavioral studies
and histology studies. The accumulative evidences indicated that the lactoferrin-
modified nanoparticles were adapted to deliver peptides for brain disease treatment
following intranasal administration [32].
6.1.4 Cell Penetrating Peptides-Modified Nanoparticles
Cell penetrating peptides (CPPs), composed of short, polycationic or amphipathic

peptides have significantly improved the delivery of a variety of molecules, includ-
ing siRNA, plasmid DNA, antisense ONs, PNA, proteins, and peptides [33–39].
CPPs have been designed to overcome both extracellular and intracellular barriers,
by promoting the movement of cargo across cell membranes, and in some cases,
enabling the release of molecules trapped inside endosomes into the cytoplasm. Due
to the partly hydrophobic and cationic nature of some CPPs, they are able to pene-
trate the negatively charged cell membrane effectively without inflicting mem-
brane damage.
CPP-modification on nanoparticle surface might offer a safe and effective
approach to enhance nose-to-brain drug delivery in potential diagnostic and thera-
peutic applications of brain disease treatment.
6.2 Liposomes
Liposomes have received widespread attention as another type of nanocarrier for

therapeutically-active compounds due to their unique characteristics such as the
capability to incorporate hydrophilic and hydrophobic drugs, good compatibility,
low toxicity, lack of immune system activation, and targeted delivery of bioactive
compounds to the site of action [65]. Liposomes are spherical vesicles that resemble
cells in that they contain an inner hydrophilic core and a relatively impermeable
outer lipophilic phospholipid bilayer. Their unique structure makes it possible to
entrap drugs both in an aqueous and a lipid phase. Lipophilic drugs are generally
Fig. 17.2 Three types of liposomes promote nose to brain delivery: (a) stealth liposomes, (b) cell-
penetrating peptide-modified liposomes, and (c) flexible liposomes
entrapped almost completely in the lipid bilayers of liposomes, and since they are
poorly water soluble, problems like loss of an entrapped drug during storage are
rarely encountered. Hydrophilic drugs may either be entrapped inside the aqueous
cores of liposomes or be located in the external water phase.
The nasal clearance half-life of liposome was fourfold longer than normal
clearance half-life of the human nose, thus suggesting the mucoadhesion ability of
liposomal systems and the potentiality of nasal applications. Furthermore, liposomes
have an additional advantage since they may be modified to efficiently target a par-
ticular site of interest like BBB. Hence, targeted liposomes were mostly studied for
the diagnosis and treatment of brain diseases.
The possible mechanism for the transportation of drug across the BBB is due to
the phospholipid bilayer of liposome facilitating the permeation of drug across vari-
ous biological membranes. Hence, various surface modifications have been made
with a view to enable the transfer of liposomal carrier via the BBB [25].
There are a number of receptors present on the surface of the BBB, particularly
for different proteins, peptides, and antibodies. Such molecules are used as surface-
active ligands and assist the translocation via receptor-mediated transcytosis. At the
same time, the cationic liposomes may cross the BBB via absorption-mediated tran-
scytosis. Another strategy is carrier-mediated transcytosis that utilizes some nutri-
ents like glucose and glutathione, which are able to bind to the surface of liposome
and facilitate its translocation. Once the liposome enters into the brain, it releases
the entrapped drug to the target site, initially through passive diffusion, where the
drug release is triggered by general passive efflux. This does not control the release
rate; hence, some more progressive approaches have been developed that respond to
the changes in the physiological environment and release the drug in a controlled
manner. In such a system, the drug release from the liposomal vesicle is triggered
by a change of pH, enzymatic stimulus, or change in the level of some redox agents
like glutathione. In the next section, we emphasize such modifications on the sur-
face of liposomes (Fig. 17.2) for improving bioavailability of therapeutics in the
brain through the nasal route [40, 41].
6.2.1 Stealth Liposomes
Stealth liposome technology is one of the most frequently used liposome-based

systems for delivery of active molecules. This strategy was achieved by modifying
the surface of the liposome membrane with hydrophilic polymer conjugates. The
employed hydrophilic polymers were natural or synthetic polymers such as PEG,
chitosan, and polyvinyl alcohol (PVA). PEG remains the most widely used polymer
conjugate with a high biocompatibility, non-toxicity, low immunogenicity, and anti-
genicity. The presence of those hydrophilic polymers prevents the aggregation of
liposomes in suspension and decreases liposomal uptake by the reticuloendothelial
system. Specifically, in the nasal route, PEG is also beneficial to increase the muco-
sal retention time, thereby boosting the liposomal brain targeting efficiency. Zheng
and co-workers have prepared a stealth liposomal system for the intranasal delivery
of peptides for the treatment of Alzheimer’s disease [42].
6.2.2 Flexible Liposomes
One more adapted approach in the case of intranasal delivery of the therapeutic
agents to the brain is using flexible liposomes. Flexible liposomes are referred to as
a liposomal system with high elasticity to facilitate the drug absorption over skin
and mucous membrane. The method adopted for the preparation of flexible lipo-
somes was thin film homogenization by using PEG as an edge activator, the agent
added to enhance the elasticity and flexibility as well as the hydrophilicity of lipoi-
dal membrane.
Li et al. reported the preparation of flexible liposomes to deliver galantamine
hydrobromide, which was very potent for the treatment of Alzheimer’s disease. In
fact, this molecule has the ability to specifically and reversibly inhibit acetylcholin-
esterase to reduce the formation of Aβ aggregates. Still, when administered orally or
intravenously, the drug produced systemic/gastrointestinal (GI) side effects as well
as some degradation by the GIT enzymes. Hence, the drug is delivered through the
intranasal route that offers the advantage of bypassing the GI effects, thereby reduc-
ing the systemic side effects by directly delivering the drug to the brain (bypassing
the BBB) via the olfactory route. At the same time, the hydrophilic drug encounters
some problem in absorption through nasal mucosa. To enhance the nasomucosal
absorption, the drug was loaded into flexible liposomes [43–46].
6.2.3 Cell Penetrating Peptides Modified Liposomes
The same scheme has been followed by another author to enhance the pharmacological
properties and drug targeting to brain while reducing the drug clearance by first pass
metabolism and systemic side effect of rivastigmine. Rivastigmine is able to inhibit
the activity of both the acetylcholinesterase and butylcholinesterase, thereby
increasing the neurotransmitter level in brain and reducing the severity of loss due
to neurodegeneration.
The drug transportability of liposomal formulations across the BBB was
investigated in vitro and it was observed that the CPP-modified liposome offered an
enhanced BBB permeation than the conventional liposome due to the high perme-
ability of CPP. The pharmacokinetic study was performed on rats, and the drug
concentration was observed in various regions of the brain as well as in peripheral
organs after intranasal administration of liposomal formulations. The result reveals
that the maximum rivastigmine concentration was observed in the entire brain
region after intranasal administration of CPP-modified liposomes, which retain for
sufficient periods of time compared to unmodified liposomes. Hence, CPP-modified
liposomes, when administered through the intranasal route, offer a promising
approach to enhance the brain targeting of rivastigmine while removing all the asso-
ciated side effects [47–51].
6.3 Exosome
6.3.1 Natural Bioactivities of Exosomes
Exosomes are naturally formed membranous nanosized vesicles 50–150 nm in

diameter. They are derived from the endosomes of mos T-cell types, especially cells
of the immune system like dendritic cells, macrophages, B-cells, and T-cells; as
well as mesenchymal stem cells, cancer cells, endothelial cells, and epithelial cells.
An exosome is a “lipid nanosphere” with a bilayered membrane containing various
types of proteins and lipids originated from the parent cell, thus, it presents specific
cell tropism. Exosomes can carry cell-type-specific proteins found in the parent
T-cell membrane, such as myelin proteins derived from oligodendrocytes, with spe-
cific homing selectivity [7, 47–50].
The unique properties of exosomes can be attributed to their biogenesis. In
general, the formation of exosomes consists of three different processes: the first
step is the formation of endocytic vesicles from plasma membrane; the second step
is the inward budding of the endosomal vesicle membrane resulting in multivesicular
bodies (MVBs), which consist of intraluminal vesicles; and the last step is the fusion
of these MVBs with the plasma membrane, releasing the vesicular contents that are
known as exosomes. Exosomes play a significant and diverse role in intercellular
communication regulating the development and function of multicellular organ-
isms. These extracellular vesicles are specialized in long-distance intercellular com-
munications facilitating protein transference and functional mRNAs and microRNAs
for subsequent protein expression in large T-cells. In fact, this is a highly efficient,
smart, and economic manner of exchanging information between cells through
secretion of exosomes. Thus, exosomes alone present unique biological activities
that may be used for therapeutic purposes, especially for immune regulation and
regenerative effects.
6.3.2 Methodologies of Loading Drugs into Exosomes
Three distinct strategies were mainly reported for the loading of exosomes with
therapeutic agents (loading naïve exosomes isolated from parental cells ex vitro; (B)
loading parental cells with a drug, which is then released in exosomes; and finally,
(C) transfecting/infecting parental cells with DNA encoding therapeutically active
compounds which are then released in exosomes. Each strategy has its advantages
and limitations, and may be dictated by the type of therapeutic agent, site, and the
type of disease, and conditions suitable for a specific type of exosome-encapsulated
agent [53, 54].
6.3.3 Therapeutic Effects of Intranasally Administrated Exosomes
Exosomes present as carriers capable of traveling from one cell to another, easily
passing their content across the cell membrane due to their unique characteristics,
delivering their cargo-preserving bioactivities. It is noteworthy that exosomes pos-
sess an intrinsic ability to cross biological barriers, including the BBB, and offer the
potentialities for brain targeting. In one of the first reports, exosomes loaded with an
anti-inflammatory small molecule compound, curcumin, were shown to protect
mice from brain inflammation. The application of exosomes for curcumin-loading
improved its solubility, increased circulation time, preserved drug therapeutic activ-
ity, and improved brain delivery [55].
7 Nose to Brain Formulations
Sr. No. Drug Formulation Treatment for the disease

1 Valproic acid Lipid nanoparticles Epilepsy
2 Pramipexole Chitosan nanoparticles Parkinson’s disease
3 Donepezil Nanoemulsion Alzheimer’s disease
4 Venlafaxine Nanogel Antidepressant
5 Risperidone Nanoemulsion Neuroprotective & antitumor
8 Future Prospective of Nose-to-Brain Delivery
Treatment of neurological diseases remains one of the most significant challenges,

and advances in nanotechnology have provided promising solutions to this chal-
lenge. Based on the past few years’ research, we can conclude that nanotechnology
has gained considerable focus. Multiple nanocarriers such as solid lipid nanoparticles,
liposomes, polymeric nanoparticles, dendrimers, nanogels, micelles, nanoemulsions,

and nanosuspensions have been studied for the delivery of brain therapeutics.
It is expected that in the near future, more drugs in the form of nasal formulations
intended for brain disorders will be commercially available. However, this func-
tional drug delivery mechanism to the brain is a potential area of research because
there are still certain unresolved challenges during intranasal delivery. These include
handling large molecular weight polar drugs such as peptides and proteins, low
membrane permeability, mucociliary clearance, and the possibility of enzymatic
degradation of the molecule in the lumen of the nasal cavity. These problems can be
solved by focusing on bioadhesive excipients and absorption enhancers in the for-
mulation. The current nanoparticle-based drug delivery technology should be
improved further, so that it can be target-oriented, safe, effective, and cost-effective.
Additionally, development of CNS nanoformulations needs to focus on improving
their BBB permeability, reducing neurotoxicity, and increasing their drug-tracking
performance and specificity for brain tissue using novel targeting moieties.
Furthermore, adequate clinical and preclinical trials to improve the intranasal deliv-
ery system are required. It is also not entirely clear how drugs are delivered directly
to the brain; thus, further research is required to better understand the exact mecha-
nism of drug passage through the intranasal route to specific brain areas. It is also
important to pay attention to formulation strategies, drug delivery devices, develop-
ment of new excipients, and mucoadhesive characteristics of polymers, all of which
could potentially improve bioavailability, prolong retention, and maximize the
effects of the drugs. Additionally, toxicodynamic studies of drug excipients and
nanotoxicity of nanocarriers should also be extensively investigated.
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Chapter 18
Is There a Global Market
and Opportunities for Nasal Drug
Delivery? Recent Trends in Global Nasal
Delivery Market
Abdullah Abdelkawi, Jean Pierre Perez Martinez, and Yashwant V. Pathak
Abstract Many civilizations around the world have used nasal drug delivery
systems within their medicinal practices throughout history, such as Indian and
Persian civilizations. This medicinal technology is prevalent throughout the industry
and continues to affect today’s medicine as a result of continuous innovation. With
the growing amount of research conducted throughout the years within this field,
there has been a multitude of uses discovered for nasal drug deliveries. There is also
a better understanding of how nasal drug deliveries affect the body and the benefits
that it has for treating various diseases. Because of this, more efficient and safe
technologies have been developed for nasal drug deliveries. Nasal drug deliveries
have been recently implemented in many different fields of medicine as new tech-
nologies for the administration of traditional medicine treatments. One of them is a
technology that focuses on nasal-CNS delivery by utilizing the nasal cavity to help
transport the active components of the medication and bypass the blood-brain bar-
rier. Similarly, a new technology that allows insulin to be administered through the
nasal cavity is being explored. Diseases affecting the brain and respiratory system
are especially susceptible to treatment by nasal drug delivery systems. Brain tumors,
Alzheimer’s, and Parkinson’s disease are just some of the possible types of diseases
that can be treated effectively with nasal drug deliveries. Nanotechnology can be
applied to nasal drug delivery due to its ability to better bypass the membranes that
would normally block the absorption of most drugs. Because of this, nanotechnol-
ogy can be used within the nasal cavity to better target brain receptors and poten-
tially bypass some layers, such as the blood-brain barrier, due to the increased
A. Abdelkawi · J. P. Perez Martinez
Y. V. Pathak (*)
Faculty of Pharmacy, Airlangga University, Surabaya, Indonesia
e-mail: ypathak1@usf.edu
https://doi.org/10.1007/978-3-031-23112-4_18
382 A. Abdelkawi et al.
permeability that these technologies can provide. Because of many technological

advances within nasal drugs delivery systems in recent years, there are many market
forecasts showing the potential growth for this market in the future. There are many
possibilities and possible applications for nasal drug delivery systems, which allows
for a greater degree of versatility for the distribution and development of these
systems.
Keywords Nasal drug delivery systems · Nanotechnology · Blood-brain barrier ·

Global market trends
1 Introduction
Nasal drug delivery systems have been present throughout history, even though it
has had most of its growth within recent years. Multiple cultures and civilizations
across the globe have incorporated nasal drug deliveries into their medicinal prac-
tices. Due to their efficiency and versatility, nasal drug delivery systems have been
an effective remedy for multiple diseases across history. Two prominent cultures
throughout history that have used nasal drug delivery systems have been Persian
and Indian cultures. Persian cultures mainly utilized nasal drug delivery systems to
administer some of their herbal medicines. Persians utilized all three forms of nasal
drug deliveries: powders, liquids, and gaseous forms to treat a plethora of diseases.
Many of their pharmaceutical manuscripts detailed the different applications for
these various forms of nasal drug delivery. It also explained their effects on various
diseases and their efficiency. The fumigation approach was used to treat facial palsy,
headaches, common colds, and rhinitis. Similarly, powders were utilized to treat
headaches, epistaxis, and epilepsy. Lastly, liquid forms of nasal drug delivery were
applied to treat insomnia, epilepsy, headaches, and vertigo. Persians administered
these forms of drugs by utilizing sacred and ancient plants or fruits. Specifically,
seeds, exudates, leaves, barks, and vinegar are common materials acquired from
these medicinal herbs and plants that were used to manufacture the early nasal drug
delivery systems [50]. Persians saw the value of nasal drug delivery systems for
treating nasal infections, diseases, and even diseases affecting the CNS. Today,
many manufacturers and researchers are looking for ways to effectively implement
nasal drug delivery systems to treat CNS diseases, which makes it even more
astounding that Persians were able to identify the efficacy of nasal drug delivery
systems in treating the CNS [6].
Additionally, the use of nasal drug deliveries has been greatly documented in
Indian medicine. Indian medicine has frequently referred to nasal drug delivery
systems as Nasya Karma within many historical manuscripts, and nasal drug deliv-
ery systems were especially prevalent in the Ayurvedic systems of Indian medicine
[36]. Nasya Karma is derived from the words Nasa (nose) and Shodhana Karma
18 Is There a Global Market and Opportunities for Nasal Drug Delivery… 383
(purification procedures) and the combination of these words roughly means to

purify the nose by administering drugs through the nasal cavity. Nasya Karma was
specifically invented to treat diseases above the nasal clavicle. Indian manuscripts
also detail different times of the day and seasons in which different forms of Nasya
Karma are the most effective for treating specific diseases [37]. Similar to Persian
cultures, Nasya Karma has different forms of drug delivery systems, such as medi-
cated oils, ghee, decoction, powders, and smoke. These different techniques of
administering Nasya Karma treat those conditions and diseases indirectly by
enhancing the function of the endocrine glands and the nervous system. Nasya
Karma was recorded to aid the central nervous system and multiple applications
were explored in how Nasya Karma could aid the CNS, similar to how Persian
manuscripts also explored this possibility. As seen throughout Indian and Persian
history, the effectiveness and efficiency of nasal drug deliveries has been greatly
detailed in many cultures of the world. Because of this, modern medicine has taken
great interest in the development of nasal drug deliveries and in effectively distribut-
ing them across a global scale. This chapter will explore the current marketed prod-
ucts in nasal drug delivery systems, the growth of these products, new technologies
recently introduced in the market, specific diseases targeted by nasal drug delivery
systems, nanotechnology applications within nasal drug delivery systems, and the
forecasted market scenario for nasal drug delivery systems [15].
2 Current Products in Market
The market for pharmaceuticals has taken great interest in nasal drug delivery
systems over the years, with many effective products being readily available in the
current market. The versatility and opportunities for nasal drug delivery systems
have allowed for the creation of many different methods of administering nasal drug
delivery systems [28]. There are many different kinds of nasal drug delivery prod-
ucts currently available in the market, such as drops, mechanical spray pumps,
medicinal atomizers, mechanical powder sprayers, etc [27]. Many of these products
fall into three main categories: liquid delivery, powdered delivery, and vapor deliv-
ery. Medicinal atomizers/nebulizers allow liquid drugs to be converted into droplets
to allow for easier inhalation of the medicine. Atomizers/nebulizers can be
electrically-driven or gas-driven, depending on the mechanism used. Atomizers/
nebulizers typically use topical steroids or drugs as their main way of administra-
tion, with some of them specifically using insulin [46]. Atomizers/nebulizers are
used to treat sinusitis, Alzheimer’s, nasal polyps, etc. Moreover, mechanical spray
pumps are currently infiltrating the market. Mechanical spray pumps can include
squeeze bottles, multi-dose metered-dose spray pumps, and single/duo-dose spray
pumps. These products utilize substances such as oxytocin, topical steroids, and
triptans to treat headaches, migraines, allergic and perennial rhinitis, induction of
lactation and labor as well as a multitude of other uses [12]. Using medicinal drops
is another common form for administering nasal drug delivery systems. Drops are
usually administered within pipettes, catheters, or droppers. They are mainly com-
posed of substances such as decongestants, desmopressin, and topical steroids [33].
They are mainly used to treat illnesses such as rhinitis, diabetes, nasal polyps, and
the common cold. Lastly, there are vapor products that are incorporated for nasal
drug delivery. These products utilize vapor inhalers to administer the dosage. Methol
is the most common substance found within these products. They are often used in
multiple doses and can tackle illnesses such as rhinitis or the common cold. Many
of these products overlap on what illnesses they treat, so it allows for a greater
viability in the administration of these drugs.
3 How the Growth of These Products Has Happened
Due to the viability and efficacy of nasal drug delivery systems, there has been a
great deal of interest and growth in this market over the last two decades. More dos-
age forms have become readily available and new methods of administering nasal
drug delivery systems are constantly being researched. Nasal drug delivery systems
continue to grow because they are more advantageous than many other forms of
drug delivery systems [24]. Achievement of improved drug transport, increased
drug absorption and bioavailability, and enhancement of the medication’s physico-
chemical qualities, such as water solubility and masking of the drug’s odors, are
reasons that have contributed to the rapid growth of nasal drug delivery products on
the market [4]. Additionally, the nasal cavity is a convenient drug administration site
for patients who want to self-administer their own medication. Patients are much
more likely to comply with taking their medication through nasal drug delivery
systems and they face fewer risks encountered with other forms of administering
medications, such as reduced risk of overdose, mitigation of pain, non-invasive
delivery, and mitigation of needle infections [49]. On the other hand, the expensive
cost of producing most of the nasal drug delivery devices has stagnated the wide-
spread production of these products. Because of this, cost effective alternatives are
constantly explored in order to increase the availability and accessibility of these
drugs [17].
4 Any New Technologies Introduced in Last Two Decades
New technologies and possibilities for nasal drug delivery systems are constantly
being researched, especially due to their projected growth and viability. One of
these new technologies that were recently introduced is the possibility of insulin
administration through nasal drug delivery systems. Most insulin delivery sys-
tems currently being used have issues with patient compliance due to the complex
method of administering the drug. Some insulin delivery systems rely on needles
to administer the insulin into the body, which may cause issues for patients who
have a fear of needles. Aditionally, drug administration through needles has an

increased risk of infection. Due to this, research has been conducted into alterna-
tive ways of administering insulin, with one of the most successful methods so far
being inhaled insulin [22]. Inhaled insulin as a nasal drug delivery system has
seen some success, especially for mitigating some of the negative effects of using
needles for insulin intake. However, inhaled insulin has been found to cause some
negative side effects for people without adequate lung health, such as smokers,
because it could worsen the symptoms of some lung diseases and cancers.
Furthermore, high levels of exercise while taking inhaled insulin could lead to
hypoglycemia. Containers of many insulin inhalers have not been optimized in
terms of their shape, with many of the current designs being bulky or clunky [43].
Despite all of this, there is a future for inhaled insulin delivery systems due to the
potential benefits and research that is constantly conducted to streamline inhaled
insulin delivery systems as much as possible in order to improve their efficacy and
reduce any negative side effects [40]. Moreover, nanotechnology has become a
field of interest within many disciplines in the last two decades due to its possible
application toward nasal drug delivery systems. With nanotechnology, there can
be many more applications for nasal drug delivery systems, and it has the poten-
tial to greatly increase the viability and efficiency of these systems.
Moreover, researchers have tested the efficacy of two forms of modified micelles
for being a viable method for nose-to-brain drug administration. Recently intro-
duced within the nasal drug delivery system space is the use of poly(ethylene
glycol)-polycaprolactone block polymeric micelles modified by a cell-penetrating
peptide: Tat (PEG-PCL-Tat). This new technology has allowed for better delivery of
drugs directly to the brain [29]. The utilization of the nasal cavity within this system
delivers anticancer drugs such as camptothecin with high efficacy. Additionally, this
specific modified cell-penetrating peptide provided improved therapeutic efficacy
within a brain tumor model [19]. Furthermore, in order to utilize and optimize this
technology, a different modified peptide in (Bom/PEG-PCL-Tat) was designed and
tested. Using bombesin, an optimization of GRPR selectivity, cellular uptake, and
cytotoxicity in C6 glioma cells as well as the intracerebral drug distribution and
therapeutic efficacy in a nose-to-brain delivery system was achieved. The results
were obtained through tests on rats with brain tumors and it showed that when rats
were given camptothecin after a brain tumor graft, the medication administered by
Bom/PEG-PCL-Tat mixed micelles allowed the rats to live longer rather than ones
administered by PEG-PCL-Tat micelles [19].
Lastly, the use of chitosan has been newly incorporated within the nasal drug
delivery field. This unique natural cationic polysaccharide has brought many medic-
inal benefits within this field. Chitosan can be obtained by the deacetylation of chi-
tin and by introducing new physical and chemical features. Chitosan provides
mucoadhesive characteristics, permeation-enhancing capabilities, and controlled
drug release qualities. Additionally, it shares non-toxic, biocompatible, and biode-
gradable qualities [45]. Chitosan has been used across various cultures to treat high
blood pressure, high cholesterol, obesity, wounds, and other health issues. Chitosan
application through the nasal route has been very promising and continues to break
ground within the nasal drug delivery market [14].
5 Some Specific Diseases Where Nasal Drug Delivery is

Most Applicable
Nasal drug delivery systems have a wide range of medical uses, but nasal drug
delivery is most applicable for diseases such as rhinitis, sinusitis, and other nasal
diseases/infections [31]. Nasal drug delivery systems can also be used for treating
health complications related to smoking, such as nicotine addiction and lung can-
cers [9]. Furthermore, extensive research has been conducted on the possibility of
using nasal drug delivery systems to target CNS diseases, especially as a possible
method of treating Alzheimer’s. Due to the limited barriers between the nasal cavity
and the CNS, nasal drug delivery systems have been predicted to be one of the best
ways of targeting CNS diseases. Many possible drugs that are currently being
researched to treat Alzheimer’s have been tested as nasal drug delivery systems due
to these advantages of the nasal cavity pathway [10]. A major advantage of using the
nasal cavity for treatment of Alzheimer’s and other diseases affecting the brain is
the ability of nasal drug delivery systems to potentially circumvent the blood-brain
barrier via the olfactory bulb, which increases the bioavailability of the drug com-
pared to oral administration [1].
6 Nanotechnology Applications in Nasal Drug Delivery
One of the fields in which nasal drug delivery can benefit the most is the field of
nanotechnology. Nanotechnology lends itself to be very useful within nasal drug
delivery systems due to the small amount of barriers that nasal drugs have to go
through to reach the target sites within the body [34]. Intranasal drug delivery has
been especially useful for allowing better drug absorption by circumventing the
blood-brain barrier [20]. The blood-brain barrier limits the administration of many
types of drugs towards the brain, which decreases the efficacy and absorption of
these drugs [26]. By using nasal drug delivery systems, the blood-brain barrier
could be avoided and better absorption of the drug through the nasal cavity can be
achieved [44]. The nasal cavity allows for direct transportation into the olfactory
bulb, in which the drug can be absorbed and transported towards the brain [30].
However, the nasal cavity still hosts its own set of biological barriers and complex
structures, especially when there are infections or diseases present within the nasal
cavity. This is one of the scenarios in which nanotechnology can greatly aid the
development of nasal drug delivery systems by allowing more ways to circumvent
the biological barriers present within the nasal cavity, thus increasing the efficacy of
these drug delivery systems [21]. Solid lipid nanoparticles (SLNS) have been on the
forefront of lipid nanotechnology [2]. This novel finding is composed of a solid
lipid core matrix that can solubilize lipophilic molecules. Its efficacy has already
been shown in the Pfizer and Moderna iterations of the SARS COVID-19 vaccine.
This novel and highly efficient technology promises to push nasal drug delivery
systems forward in an attempt to globalize this market and give better alternatives
to drug delivery. Solid lipid nanoparticles have shown to be beneficial in treating
CNS diseases [38]. In particular, it is efficient with neurodegenerative diseases [7].
Because the intranasal route provides direct nose-to-brain drug delivery, SLNs show
promise to combat other diseases using intranasal medication delivery for systemic
absorption like cardiovascular diseases, infections, severe pain, and menopausal
syndrome [5]. Nonetheless, further research is being conducted to test key factors
such as drug absorption at subtherapeutic levels and rapid mucociliary clearance in
order to fully incorporate it within the intranasal drug delivery field [41]. Intranasal
drug delivery provides key advantages with this novel nanotechnology because it
allows it to avoid first-pass metabolism, gastrointestinal degradation, and the blood-
brain barrier [11]. Nanoemulsion is another novel nanotechnology that is providing
key breakthroughs within nasal drug delivery systems. Nanoemulsions are nano-
sized emulsions that aim to improve the delivery of active pharmaceutical ingredi-
ents [8]. It is aided in doing so due to its thermodynamically stable isotropic system
in which two immiscible liquids are mixed to form a single phase by means of an
emulsifying agent [18]. The mechanism of nanoemulsions has already proven its
efficacy in distributing active drug components through the nasal cavity into the
brain in multiple tests conducted on rats. Due to these results, nanoemulsions are
considered one of the more promising candidates for nanotechnology application to
nasal drug delivery treatment of Alzheimer’s and Parkinson’s disease [42]. Polymeric
nanoparticles are nanocarriers that can encapsulate an active compound within their
polymeric cores, with the nanoparticle itself ranging from 1 to 100 nm [51]. The
compounds can also be surface absorbed by the polymeric core, which allows for a
greater degree of versatility with the administration of this type of drug nanocarrier
within nasal drug delivery systems [23]. For example, peptide delivery through the
use of polymeric nanoparticles in nasal drug delivery systems has been recently
researched as a possible way to treat epilepsy due to the projected efficiency of this
method of treatment [39]. Polymeric micelles are nanocarriers that are created
through amphiphilic block copolymers. This particular type of nanocarrier has a
hydrophobic core–hydrophilic shell structure that allows for hydrophobic drugs to
be encapsulated within the core [3]. Polymeric micelles have seen effectiveness in
nasal drug delivery systems by showing their effectiveness in bypassing the blood-
brain barrier [48]. Polymeric micelles allow for more effective distribution of active
drugs across different biological barriers present around the CNS, which shows
their viability as an effective nanocarrier of nasal drug administration [35].
Nanotechnology proves to be at the forefront of innovation and leads the future of
nasal drug delivery systems. All these different kinds of nanotechnological advance-
ments allow for a greater degree of viability and a variety of methods that can be
used for enhancing nasal drug delivery systems [16]. As a result, many possibilities
are likely to be discovered as more research is conducted to treat diseases that were
otherwise blocked by physiological barriers.
7 Forecasted Market Scenario for Nasal Drug Delivery
Nasal drug delivery systems have a very fortuitous future due to their viability and
efficiency. Even though the field is seeing rapid growth in North America, many
other parts of the world are also experiencing growing interest and availability for
nasal drug delivery systems. This shows the increasing potential for a global market
for nasal drug delivery systems and the possibilities for having a global distribution
network for nasal drug delivery systems. Traditionally, pharmaceutical products
take about 10–15 years to create a new product from the ground up and effectively
introduce it into the market. However, with the current advancements in this field,
development time could be shortened. Extensive research is also being conducted in
reducing the cost and maximizing the efficiency of these drugs. One of these poten-
tially cost-effective and efficient nasal drug delivery methods that could be refined
in the future is exhalation delivery systems (EDS) technologies [13]. Nasal drug
delivery systems are also expected to overtake the injectable method of drug deliv-
ery for many current medications due to better patient compliance and drug bio-
availability [47]. Nasal drug delivery systems are currently being developed as
alternative forms of treatment for many chronic diseases, such as diabetes, osteopo-
rosis, and endometriosis [36]. To illustrate the increasing demand of nasal drug
delivery systems, Mordor Intelligence conducted a market study over the predicted
growth of nasal drug delivery systems from 2021 through 2026. The scope of the
report included a collection of data on multiple regions across the globe and on
multiple dosage methods for nasal drug delivery systems. Their research indicated
that drops and liquids are going to be the fastest growing dosage methods of nasal
drug delivery systems, and North America is most likely going to dominate the
global market for nasal drug delivery systems. Although results are also showing
that the Asian Pacific region is forecasted to experience significant growth as well
because nasal drug delivery systems are already established as a prominent form of
treatment in their traditional medicine. Nonetheless, the research highlights that the
North American region is shown to have a monopoly over the current market in
terms of growth, earnings, and forecasted implementation of new products.
However, the monopoly is expected to be broken and the research projects that there
is going to be intense competition in the market between multiple international
companies, which will help to propel even more innovation within nasal drug deliv-
ery systems [25]. Collaboration between these prominent international companies,
such as through joint ventures, is also going to foster more growth within the nasal
drug delivery systems market and drive innovation to the overall medicinal market
over the coming years [32]. However, it is important that this growth is regulated
and maintained correctly in order to push effective, health changing, and adaptable
medicine that can help millions of individuals without primarily focusing on mon-
etary gains.
8 Conclusions
The prospects for a global market for nasal drug delivery systems are very promising.
Due to the extensive amount of research and rich history surrounding nasal drug
delivery systems, significant advancements have been made within the field of nasal
drug delivery systems [25]. These advancements have improved the viability and
efficiency of these medical products, which has allowed for a greater degree of
marketability within the pharmaceutical market. The nasal drug delivery systems
are also much more user friendly than many other traditional treatments, such as
injections, which improves patient compliance with administering the drug. These
pharmaceutical products can also be self-administered, which removes the hin-
drance of having to go to a medical professional to receive and utilize this kind of
treatment option. Nasal drug delivery systems have also shown to be suitable
replacements for the traditional treatment of many chronic conditions, such as dia-
betes. Nasal drug delivery systems have also shown promise as possible treatment
for Alzheimer’s disease due to the reduced biological barriers encountered in the
nose-to-brain route.
Nanotechnology has also shown great promise towards being applicable and
beneficial towards drug delivery systems because of the possibility to refine the
benefits of nasal drug delivery systems even more. Nanotechnology also provides
an increased amount of variability for different forms of administration of nasal
drug delivery systems, which increases the range of diseases and conditions that can
be treated by nasal drug delivery systems. Overall, nasal drug delivery systems have
an auspicious future ahead because of their immense versatility. Many places across
the globe already have an extensive history with nasal drug delivery systems, lead-
ing to greater ease in marketing these products globally. There is extensive research
being carried out globally due to the high demand for nasal drug delivery systems,
therefore improving the market prospects of these products worldwide. Many mar-
ket trends show nasal drug delivery systems to be one of the fastest growing sectors
within the pharmaceutical industry. Based on the sources collected throughout this
chapter, the authors believe that there is a global market for nasal drug delivery
systems.
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Chapter 19
Nasal Drug Delivery System: Regulatory
Perspective
Sudhir Sawarkar and Julie Suman
Abstract Drugs administered intranasally are a viable alternative to parenteral

delivery. The nasal route could be a viable option for medicines with low oral bio-
availability due to GI tract breakdown or substantial mucosal or first-pass metabo-
lism. Enzymatic deactivation is a significant issue for polypeptide medicines. As a
result, the nasal route, which has low enzymatic activity and many vascular tissues
with strong blood flow, may be a viable alternative to injections. The nasal mem-
brane virtually absorbs small drug molecules totally. Increased drug delivery,
reduced mucociliary clearance, and/or coadministration with non-toxic permeation
enhancers can all help to increase the absorption of bigger molecules. The nasal
route appears to be highly attractive for the delivery of peptides and medications
that must demonstrate quick therapeutic efficacy due to its accessibility and excel-
lent patient acceptance as well as its familiarity with the general population (Zia
et al. Clin Res Regul Aff 10:99–135, 1993). The global regulatory viewpoint for
nasal drug delivery systems will be discussed in this chapter. We can clearly see how
authorities like the European Commission (EC), the US Food and Drug
Administration (USFDA), and the MHLW/PWDA (Japan) set the bar for laws that
are adopted by many countries. The International Council for Harmonization of
Technical Requirements for Pharmaceuticals for Human Use (ICH) is unique in that
it brings regulatory agencies and the pharmaceutical industry together to debate
scientific and technical aspects of pharmaceuticals and establish ICH standards
(https://www.dgra.de/media/pdf/studium/masterthesis/master_da-silva_lais_2018.
pdf). The state of regulatory control over digital or smart medical devices used to
deliver medicines to patients is a topic that is discussed extensively in this chapter.
Rules of most countries have failed to keep up with scientific and technological
advancements as medical devices become more sophisticated and inventive. This
S. Sawarkar (*)
QRServes Global LLC, Sharjah, United Arab Emirates
e-mail: sudhir.sawrakar@qrserves.com
J. Suman
Next Breath, Halethorpe, MD, USA
https://doi.org/10.1007/978-3-031-23112-4_19
394 S. Sawarkar and J. Suman
predicament has been made worse by the growing use of digital technologies. As a
result, concerns about medical device safety and data security have prompted
authorities to enact or plan to enact legislation and regulations to help with the
management and implementation of medical device safety, accountability, cyberse-
curity, and interoperability.
Keywords Nasal · Regulatory · ICH · Digital technology
1 Introduction
In terms of burden on the healthcare system, the years 2019 to 2021 were the most
difficult for humanity. The greatest pandemic due to COVID-19 (SARS-CoV-2
virus) was fought by humanity. A face mask, vaccines, and a few medications were
part of the mitigation plan. Almost all vaccines have been cleared for emergency
use. Nasal drug delivery was employed for a few of the vaccines under develop-
ment, and the results were promising. Intranasal vaccination is just one example that
highlights the importance of this route of administration. Nasal drug delivery is a
typical route of administration for a variety of medical conditions ranging from
allergic rhinitis to cancer pain. The nose is the entrance point for systemic distribu-
tion of a wide range of medications and therapies for a wide range of disorders.
Fentanyl (breakthrough cancer pain), Triptans (migraine pain management), nico-
tine (smoking cessation), Calcitonin (Osteoporosis), Metoclopramide, and
Esketamine (depression) are some of the medications that utilize the nose for sys-
temic delivery. Medications addressing different therapeutic domains and diseases,
such as pharmaceuticals that work on the CNS to treat disorders like Alzheimer’s
disease and obesity [2], may join the growing ranks of marketed products for sys-
temic distribution via the nasal route. The advantage of delivering therapies straight
from the olfactory region into the brain, allowing the drug to bypass the blood-brain
barrier, is the fundamental rationale for this trend in attaining systemic distribution
via the nasal route. Furthermore, nasal vaccination is a popular alternative to injec-
tion because it causes patients little discomfort. Mucosal vaccinations promote not
only effective local immune protection but also a systemic immunological response
similar to injections. Intranasal vaccines also have the potential to prevent viral
shedding. At the moment, a lot of effort is being put into building smart devices. As
the movement toward data-connected health gets traction and the benefits for
patients, physicians, payers, and manufacturers become obvious, one thing becomes
clear: integrating electronics into a device is not an easy task, but it is very necessary
that regulatory coverage be expanded to include intelligent controls and data man-
agement. This expansion is already happening in countries like the United States,
the European Union, Australia, Japan, and Brazil.
19 Nasal Drug Delivery System: Regulatory Perspective 395
2 The Global Regulatory Perspective Over Nasal Drug

Delivery System
Currently, there are no international regulations specific to orally inhaled and nasal
medication products. The regulatory framework for nasal spray medications in the
Western world is well-established. As the usage of nasal sprays, particularly gener-
ics, increases in the BRIC nations (Brazil, Russia, India, and China), regulatory
organizations such as ANVISA in Brazil and the CFDA in China are contemplating
adopting FDA-like control strategies. Below are included the regulatory agencies
for each location as well as the suggested safety, quality, and efficacy requirements.
While the aforementioned nations have particular rules for nasal medication deliv-
ery devices, their recommendations are frequently based on those of the International
Conference on Harmonization. Following are references to the fundamental prin-
ciples of product development and quality assurance.
2.1 ICH
The International Conference on Harmonization of Technical Requirements for

Registration of Pharmaceuticals for Human Use (ICH) is a unique organization that
convenes regulatory agency officials from the European Union (EMA), Japan
(MHLW), and the United States (FDA) in a single platform. As an organization,
ICH has expanded to include 20 Members and 35 Observers.
ICH has developed the following quality-related recommendations which can be
applied to nasal drug delivery products.
(a) ICH Q8 (R2) Pharmaceutical Development [3]
This Guideline’s purpose is to provide information on the contents of Section
3.2.P.2 (Pharmaceutical Development) for drug items that come within Module 3 of
the Common Technical Document (ICH topic M4) [4]. This guideline is not appli-
cable to the contents of drug product submissions made during the clinical research
phase of drug development. During these phases, however, it is essential to bear in
mind the concepts indicated in this guidance. This guidance may also apply to other
product categories. In November 2008, the appendix to the Harmonized ICH text
was finished and included into the core guideline, which was referred to as Q8 at the
time (R1). The addendum elucidates some of the major concepts mentioned in the
main guideline. In addition, this appendix describes the concepts of excellence
through design (QbD). The appendix is not intended to establish new standards;
instead, it explains how the applicant might apply concepts and methodologies
(such as design space) from the original Q8 paper to all dosage forms. When a com-
pany uses quality by design and quality risk management (Q9: Quality Risk
Management) in conjunction with a suitable pharmaceutical quality system, there
are opportunities to enhance science- and risk-based regulatory procedures (Q10:
Pharmaceutical Quality System). In August 2009, minor modifications were made

to the Q8(R1) Guideline.
(b) ICH Q9 Quality Risk Management [5]
Under Step 4, the ICH Harmonized Guideline was completed in November 2005.
This guideline covers ideas and approaches for quality risk management that
may be applied to a variety of pharmaceutical quality elements. Throughout the
lifetime of drug substances, drug (medicinal) products, biological and biotechno-
logical goods, these components include development, manufacture, distribution,
and inspection and submission/review procedures.
(c) ICH Q10 Pharmaceutical Quality System [6]
In June 2008, the ICH Harmonized Guideline was finalized as part of Step 4.
Throughout the product lifecycle, this guideline applies to the systems that enable
the development and production of pharmaceutical drug substances and drug prod-
ucts, including biotechnology and biological products, throughout the product life-
cycle. Aspects of Q10 should be applied proportionally and appropriately to each
step of the product lifecycle, taking into consideration the different objectives of
each stage. The parts of Q10 should be implemented in a manner that is suitable and
proportional to each step of the product lifecycle, taking into account the distinc-
tions between the phases and their respective objectives.
(d) ICH Q11 Development and Manufacture of Drug Substances [7]
This document describes how to construct and comprehend the drug substance’s
manufacturing process as well as what information should be included in Module 3
of the Common Technical Document (CTD) Sections 3.2.S.2.2–3.2.S.2.6 (ICH
M4Q) [4]. It addresses aspects of the creation and production of pharmacological
substances, such as the availability of steps to eradicate impurities. This Guideline
pertains to drug substances as stated in the scope sections of ICH Guidelines Q6A
and Q6B, but following consultation with regulatory authorities, it may also be
applied to other types of products.
(e) ICH Q12 Pharmaceutical Product Lifecycle Management [8]
The purpose of this guideline is to offer a framework for more predictable and
efficient management of post-approval Chemistry, Manufacturing, and Controls
(CMC) modifications throughout the product lifecycle. Adoption of this new ICH
Guideline will encourage innovation and continual improvement in the biopharma-
ceutical business as well as enhance quality assurance, consistent product delivery,
and proactive supply chain planning. It helps regulators (assessors and inspectors)
to have a better knowledge of the Pharmaceutical Quality Systems (PQSs) utilized
by companies for post-approval CMC alterations management. The purpose of this
new guideline, which consists of a core guideline and annexes, is to augment the
current ICH Q8 through Q11 Guidelines.
All Above ICH guidelines are well adapted by developed countries for nasal drug
delivery product registration, monitoring, and control in respective countries.
2.2 USFDA: United States Food and Drug Administration
The US Food and Drug Administration is one of the most powerful regulatory
agencies in the world. Its rules and regulations serve as a standard for many nations
throughout the world. The FDA is made up of the Office of the Commissioner and
four directorates that handle the agency’s major functions: Medical Products and
Tobacco, Foods and Veterinary Medicine, Global Regulatory Operations and Policy,
and Operations. The FDA has a wide range of regulatory authorities. There is a
strong relationship between the FDA’s responsibilities and those of several other
government bodies. The FDA’s headquarters are in the unincorporated area of White
Oak, Maryland. In addition, the agency maintains 223 field offices and 13 labs
spread across the 50 states, the United States Virgin Islands, and Puerto Rico.
USFDA monitors the utilization of unapproved medicinal compounds and formula-
tions [12–16]. This results in the United States adopting far tighter regulations.
The Center for Drug Evaluation and Research (CDER) and the Center for
Devices and Radiologic Health (CDRH) evaluate medicines and medical devices in
the United States for the FDA. The following regulatory guidance for the industry
further defines the drug product performance parameters.
These referred guidelines represent the FDA’s current position on orally inhaled
and nasal medicinal products.
(a) CDER 2002 Nasal Spray and Inhalation Solution, Suspension and Spray Drug
Products, Chemistry, Manufacturing and Controls [11]
This document offers industry guidance on the chemistry, manufacturing, and
controls (CMC) documentation that should be included in new drug applications
(NDAs) and abbreviated new drug applications (ANDAs) for nasal spray and inha-
lation solution, suspension, and spray drug products with local and/or systemic
effects. This guidance covers, for each of these categories, the CMC information
that should be included in the application for the drug product components, manu-
facturing process, and related controls, but not for the manufacture of drug sub-
stances. The rules also include requirements for labeling. This guideline does not
apply to inhalation powders (commonly known as dry powder inhalers, DPIs) or
nasal powders. Intranasal delivery of nasal powders such as glucagon (BAQSIMI®,
Eli Lilly) is an emerging field. The 2002 CMC Guidance emphasizes more on aque-
ous systems. Nonetheless, there is a standard for inhalation powders. Certain char-
acteristics, such as aerodynamic particle size (APSD) measurements, are necessary
for nasal powders, as outlined in FDA-2018-D-1098 Metered Dose Inhaler (MDI)
& Dry Powder Inhaler (DPI) Products – Quality Considerations Guidance for
Industry, Pharmaceutical Quality/CMC Documentation – Draft [9].
(b) CDER 2003 Bioavailability and bioequivalence studies for nasal aerosols and
nasal sprays for local action, Biopharmaceutics – Draft [10]
Bioavailability and Bioequivalence Studies for Nasal Aerosols and Nasal Sprays
for Local Action is the title of a proposed industry guideline issued by the Food and
Drug Administration (FDA). This draft document provides guidance to applicants

contemplating product quality studies to demonstrate bioavailability (BA) or bio-
equivalence (BE) in support of new drug applications (NDAs) or abbreviated new
drug applications (ANDAs) for locally acting drugs in nasal aerosols (metered-dose
inhalers) and nasal sprays (metered-dose spray pumps). The guideline is being reis-
sued as a level 1 draft guidance for input since it has been modified in significant
respects.
(c) CDER 2017 Combination Product guideline [17]
This guideline describes and explains the final rule on CGMP requirements for
combination goods (final rule as codified in 21 CFR part 4) that the FDA released
on January 22, 2013 (final rule as codified in 21 CFR part 4). Prior to the issuance
of the final rule, although CGMP regulations existed to establish requirements for
drugs, devices, biological products, and Human Cells, Tissues, and Cellular and
Tissue-Based Products (HCT/Ps), there were no regulations to clarify and explain
how these CGMP requirements applied to combination products. The purpose of
the final rule was to provide this clarification and detail how to verify compliance
with applicable CGMP standards.
(d) CDER 2017 Draft Guidance for Industry – Comparative Analyses and Related
Comparative [18]
This guideline is designed to assist potential applicants who want to submit an
abbreviated new drug application (ANDA) for the approval of a planned combina-
tion product that comprises both a drug constituent part and a delivery device con-
stituent part. The majority of the suggestions in this guideline are centered on a
comparison of the proposed user interface for the generic drug-device combination
product (generic combination product) to the user interface for the reference listed
drug (RLD). User interface refers to all components of the combined product with
which a user interacts for the purposes of this advice. This comprises the delivery
mechanism that is a component of the combination product, any controls and dis-
plays linked with it, as well as the product’s labeling and packaging.
(e) CDER 2016 Human Factors Studies and Related Clinical Study Considerations
in Combination Product Design and Development: Draft Guidance for Industry
and FDA Staff [19]
This guideline instructs industry and FDA staff on the fundamentals of human
factors (HF) studies in the development of combination products as specified by 21
CFR Part 3. This guidance clarifies the different types of HF studies, the recom-
mended timing and sequencing of HF studies, and how HF studies are part of the
process to maximize the likelihood that the combination product user interface is
safe and effective for use by the intended users, uses, and environments. The advice
also explains how HF studies relate to other clinical trials so that safe and effective
combination medicines can be made and reviewed quickly. The advice also talks
about how to handle HF information in experimental or marketing applications.
(f) CDER 2016 List of Highest Priority Devices for Human Factors Review: Draft
Guidance for Industry and Food and Drug Administration Staff [20]
This guideline is intended to assist potential applicants who want to submit an
abbreviated new drug application (ANDA) to seek approval for a proposed combi-
nation product containing both a drug constituent part and a delivery device con-
stituent part. This guideline focuses largely on comparing the proposed user
interface for a generic drug-device combination product (generic combination prod-
uct) to the user interface for the reference listed drug (RLD). For the purposes of this
advice, the word “user interface” refers to all product components with which a user
interacts. This includes the delivery mechanism, any associated controls and dis-
plays, and the labeling and packaging of the product.
(g) CDER 2018 Allergic Rhinitis: Developing Drug Products for Treatment

Guidance for Industry [21]
This guideline is designed to assist sponsors in the development of pharmacological
treatments for allergic rhinitis in children and adults. There are two forms of allergic
rhinitis: seasonal allergic rhinitis (SAR) and perennial allergic rhinitis (PAR). The
recommendations encompass trial design, efficacy, and safety (PAR) for novel SAR
and PAR therapies.
(h) CDER 2018 Draft Guidance: Contents of a Complete Submission for Threshold
Analyses and Human Factors Submissions to Drug and Biologic Applications [22]
This document offers industry and FDA officials with information on the contents
of threshold analyses and human factors (HF) submissions as well as submission
methods and FDA review dates.
(i) September 2018 Non-allergic Rhinitis: Developing Drug Products for

Treatment [23]
This guidance is intended to assist applicants for new drug and biologic licenses
in developing drug products for the treatment of non-allergic rhinitis (NAR) in chil-
dren and adults. The advice discusses clinical phenotypic definition, trial design,
effectiveness, and safety for novel therapeutic medicines under development. The
recommendations address the implementation of programs for the treatment of
vasomotor rhinitis (VMR), a subtype of NAR.
(j) CDER 2016 Applying Human Factors and Usability Engineering to Medical
Devices – Guidance for Industry and Food and Drug Administration Staff [24]
The FDA developed this guideline paper to help the medical device industry in
using human factors and usability engineering techniques that will raise the likeli-
hood of new medical devices being safe and effective for their intended users, uses,
and usage conditions. This paper’s recommendations are intended to aid device
manufacturers in enhancing the design of their devices to lessen the potential of user
mistake and consequent damage. These recommendations, according to the FDA,
will enable manufacturers to examine and decrease the risks associated with medi-
cal device usage.
(k) USFDA Product Specific Guideline [25]
FDA publishes product-specific guidances describing the agency’s current
thinking and expectations on how to develop generic drug products that are
therapeutically equivalent to specific reference listed drugs in order to assist the
generic pharmaceutical industry in identifying the most suitable methodology for
developing drugs and generating evidence required to support ANDA approval.
With enhanced clarity on product-specific instructions, applicants wishing to
manufacture generic drugs will have a stronger chance of allocating resources
efficiently. The agency’s mission is to ensure that legislation and regulations as well
as scientific standards reflect the most recent scientific knowledge. The FDA’s goal
of providing patients with access to high-quality inexpensive medicines is advanced
by increasing patient access to such treatments. These guidelines can be accessed on
https://www.accessdata.fda.gov/scripts/cder/psg/index.cfm [25]
• CDER 2020 Draft Guidance for Sumatriptan, Nasal, Spray
• CDER 2019 Draft Guidance on Beclomethasone Dipropionate (Aerosol,
Metered; Inhalation)
• CDER 2019 Draft Guidance on Fluticasone Propionate (Powder; Inhalation)
• CDER 2019 Draft Guidance on Fluticasone Propionate; Salmeterol Xinafoate
(Aerosol, Metered; Inhalation)
• CDER 2019 Draft Guidance on Budesonide (Spray, Metered; Nasal)
• CDER 2019 Draft Guidance on Fluticasone Furoate (Metered, Spray; Nasal)
• CDER 2019 Draft Guidance on Azelastine Hydrochloride and Fluticasone
Propionate (Spray, Metered; Nasal)
• CDER 2019 Draft Guidance on Fluticasone Propionate (Metered, Spray; Nasal)
• CDER 2019 Draft Guidance on Mometasone Furoate Monohydrate (Metered,
Spray; Nasal)
• CDER 2019 Draft Guidance on Triamcinolone Acetonide (Metered, Spray; Nasal)
• CDER 2019 Draft Guidance on Fluticasone Propionate and Salmeterol Xinafoate
(Powder; Inhalation)
• CDER 2019 Draft Guidance for Dihydroergotamine Mesylate, Nasal,
Spray, Metered
• CDER 2018 Draft Guidance for Talc, Inhalation, Aerosol, Metered
• CDER 2018 Draft Guidance for Zolmitriptan, Nasal, Spray
• CDER 2017 Draft Guidance on Mometasone Furoate (Powder; Inhalation)
• CDER 2017 Draft Guidance on Tiotropium Bromide (Powder; Inhalation)
• CDER 2018 Draft Guidance on Beclomethasone Dipropionate (Aerosol,
Metered; Nasal)
• CDER 2017 Draft Guidance on Azelastine Hydrochloride (Metered Spray; Nasal)
• CDER 2017 Draft Guidance on Fluticasone Propionate (Powder; Inhalation)
• CDER 2017 Draft Guidance for Naloxone Hydrochloride, Nasal, Spray
• CDER 2016 Draft Guidance for Buprenorphine Hydrochloride; Naloxone

Hydrochloride, Oral, Sublingual Tablet
• CDER 2016 Draft Guidance on Albuterol Sulfate (Aerosol, Metered; Inhalation)
• December 2016 Draft Guidance on Budesonide (Powder; Inhalation)
• CDER 2016 Draft Guidance on Olopatadine Hydrochloride (Metered
Spray; Nasal)
• CDER 2016 Draft Guidance on Umeclidinium Bromide (Powder; Inhalation)
• CDER 2016 Draft Guidance on Fluticasone Furoate (Powder; Inhalation)
• CDER 2016 Draft Guidance on Fluticasone Furoate; Vilanterol Trifenatate
(Powder; Inhalation)
• CDER 2016 Draft Guidance on Indacaterol Maleate (Powder; Inhalation)
• CDER 2016 Draft Guidance on Mometasone Furoate (Aerosol; Metered;
Inhalation)
• CDER 2016 Draft Guidance on Ciclesonide (Aerosol; Metered; Inhalation)
• CDER 2016 Draft Guidance on Formoterol Fumarate; Mometasone Furoate
(Aerosol; Metered; Inhalation)
• CDER 2015 Draft Guidance on Budesonide; Formoterol Fumarate Dihydrate
(Aerosol; Metered; Inhalation)
• CDER 2013 Draft Guidance on Fluticasone Propionate; Salmeterol Xinafoate
(Powder/inhalation)
• CDER 2012 Draft Guidance on Budesonide (Suspension/Inhalation)
• CDER 2012 Draft Guidance on Ciclesonide (Aerosol, Metered/nasal)
• CDER 2015 Draft Guidance on Aclidinium Bromide (Powder, Metered;
Inhalation)
• CDER 2015 Draft Guidance on Fluticasone Propionate (OTC; Metered,
Spray; Nasal)
• CDER 2015 Draft Guidance on Formoterol Fumarate (Powder; Inhalation)
• CDER 2015 Draft Guidance on Levalbuterol Tartrate (Aerosol, Metered;
Inhalation)
• CDER 2015 Draft Guidance on Ipratropium Bromide (Aerosol, Metered;
Inhalation)
2.3 European Medicines Agency
The European Medicines Agency (EMA) communicates with the European

Directorate for the Quality of Medicines and Healthcare (EDQM), a Council of
Europe directorate, regarding medication quality and protection of public health.
Orally inhaled and nasal pharmaceuticals are governed by the following
recommendations:
(a) June 2006 Guideline on the Pharmaceutical Quality of Inhalation and Nasal
Products [26]
This article relates to human medical devices intended to deliver a medicine into
the lungs or nasal mucosa in order to have a local or systemic effect. The paper
addresses the required quality attributes of commercially available medicinal prod-
ucts. However, the general principles mentioned here should also be applicable to
products utilized in clinical studies. It is unlikely that all of the aforementioned tests
will be performed on all batches of clinical trial samples. To qualify the drug for
commercialization, however, a thorough analysis of the drug substance and drug
product batches utilized in major clinical studies is necessary. Only inhalation and
nasal product quality concerns are addressed, together with the need for safety test-
ing (e.g., for excipients and leachables). In addition to safety and efficacy, other
guidance papers such as ICH guidelines address other quality problems (e.g., con-
taminants, process validation, stability testing, and specifications). Pharmaceutical
development research approaches (e.g., priming studies) and analytical techniques
(e.g., cascade impactor analysis) applied mostly for inhalation and nasal drugs lack
specific guidelines. This material may appear in other sources (e.g., United States
Pharmacopeia, European Pharmacopeia, ISO standards). It is well known that the
extensive variety of formulation and delivery device characteristics of inhalation
and nasal products necessitates a degree of testing procedure flexibility.
(b) January 2009 Guideline on the Requirements for Clinical Documentation for
Orally Inhaled Products (OIP) Including the Requirements for Demonstration
of Therapeutic Equivalence Between Two Inhaled Products for Use in the
Treatment of Asthma and Chronic Obstructive Pulmonary Disease (COPD) in
Adults and for Use in the Treatment of Asthma in Children and Adolescents [27]
This guideline is a revision of the CPMP Points to Consider on the Requirements
for Clinical Documentation for Orally Inhaled Products (OIP) CPMP/EWP/4151/00.
It clarifies the clinical documentation requirements for abridged applications for
orally inhaled formulations and variations/extensions to a marketing authorization,
including both single active substance products and combination products, in terms
of demonstrating therapeutic equivalence between two inhaled products for the
management and treatment of asthma and chronic obstructive pulmonary disease in
adults, in addition to the management and treatment of bronchitis.
(c) June 2017 Concept Paper on Revision of the Guideline on the Requirements for
Clinical Documentation for Orally Inhaled Products (OIP) Including the
Requirements for Demonstration of Therapeutic Equivalence Between Two
Inhaled Products for Use in the Treatment of Asthma and Chronic Obstructive
Pulmonary Disease (COPD) in Adults and for the Treatment of Asthma in
Children and Adolescents [27]
This concept paper aims to update the criteria for showing therapeutic equivalence
between two inhaled drugs. The guideline is primarily concerned with hybrid
applications, but it may also be applicable to other applications such as line exten-
sions and variants that are predicated on showing therapeutic equivalence in com-
parison to a reference product. First issued in September 2000, the guideline was
amended between September 2007 and January 2009. (referred to as Revision).
2.4 European Directorate for the Quality of Medicines &

HealthCare
EDQM has published following important guidelines for Orally Inhaled and Nasal
Drug products:
• Monographs applicable to Orally Inhaled and Nasal Drug Products from The
European Pharmacopoeia* 2019, 10th Edition, published by EDQM.
• Nasal Preparations, Dosage Forms 0676, European Pharmacopoeia: 2019 10th
Edition (10.3) 01/2021:0676, Pages 4823–4825.
• Inhalation Preparations, Dosage Forms 0671 European Pharmacopoeia: 2019
10th Edition (10.5) 07/2021:0671, Pages 5706–5711.
• The European Pharmacopoeia, a legal and scientific reference for pharmaceutical
quality control, is published by EDQM. All makers of pharmaceuticals and
pharmaceutical ingredients must adhere to these rules and include the mono-
graphs in the quality portion of their EMA applications.
2.5 UK Medicines and Healthcare Products

Regulatory Agency
The Medicines and Healthcare Products Regulatory Agency, part of the United
Kingdom’s Department of Health and Social Care, is responsible for guaranteeing
the safety of medicines and medical devices.
(a) June 2018 Guidance: Medical device stand-alone Software Including Apps
(including IVDMDs) [28]
This guideline supersedes the previous MHRA guidance entitled “Medical
device standalone software, including applications.” In addition to the rise of medi-
cal device apps in hospitals and community-based healthcare management, the
importance of fitness and social care applications is rising. However, in the United
Kingdom, standalone software and apps that meet the definition of a medical device
are still required to be UKCA-marked in accordance with the Medical Device
Regulations 2002 (as amended) (UK MDR 2002) to ensure that they are regulated,
acceptably safe for use, and perform as the manufacturer/developer intends.
(b) September 2017 Human Factors and Usability Engineering – Guidance for
Medical Devices Including Drug-device Combination Products [29]
Manufacturers of all device classes, developers of medical devices and drug-
device combination products, and UK Approved Bodies responsible for guaranteeing
the quality of these devices should examine this guideline. Those involved in the
procurement and risk management of medical device-related activities may find this
guidance useful. This guideline may be valuable to physicians, the NHS, NICE, and
other stakeholders, but it does not apply to them or other professionals making treat-
ment choices.
In the United Kingdom, medical devices are controlled by the UK Medical
Devices Regulations 2002 (SI 2002 No 618 as amended), which implements three
EU Medical Device Directives:
• Directive 90/385/EEC on active implantable medical devices (AIMDD).
• Directive 93/42/EEC on medical devices (MDD).
• Directive 98/79/EC on in vitro diagnostic medical devices (IVDD).
• Part II of the UK Medical Devices Regulations (MDR) 2002, Annex I (for
general medical devices) and Part III of the UK Medical Devices Regulations
(MDR) 2002, Annex 1 (for active implantable medical devices) [as amended by
Schedule 2A to the UK Medical Devices Regulations (MDR) 2002] specify the
essential requirements for medical devices to ensure adequate safety and
performance.
2.6 Health Canada
In Canada, Health Canada evaluates the safety, effectiveness, and quality of

pharmaceutical goods.
(a) October 2001 Guidance for Industry – Pharmaceutical Quality of Inhalation and
Nasal Products [30]
The fourth version of the Guidance for Industry Pharmaceutical Quality of
Inhalation and Nasal Products was issued by Health Canada in 2006. Representatives
from Health Canada’s Therapeutic Products Directorate (TPD) and the European
Medicines Agency’s Quality Working Party authored this joint guideline paper
(QWP). This document pertains to medical devices intended to administer a phar-
macological agent to the lungs or nasal mucosa for a local or systemic impact in
humans. The paper focuses on the required quality characteristics of commercially
available drugs. These essential principles should also apply to clinical trial prod-
ucts. It is unlikely that all of the aforementioned tests will be done on all clinical
trial batches. To qualify the therapy for sale, comprehensive characterization of the
drug material and drug product batches employed in crucial clinical trials is neces-
sary. The research focuses exclusively on new marketing authorization applications
(including generic drugs) and does not discuss in detail the predicted quality char-
acteristics of existing inhalation and nasal therapies. However, while modifying
existing products, the aforementioned concepts should be addressed. This guidance
was developed for items containing synthetic or semisynthetic pharmacological
substances. However, the core ideas discussed here should also be applicable to
various inhalation and nasal solutions. This document addresses pressurized metered
dose inhalers, dry powder inhalers, nebulization products, and non-pressurized
metered dose inhalers as well as pressurized metered dose nasal sprays, nasal
powders, and nasal liquids, excluding liquid inhalation anesthetics, nasal ointments,
creams, and gels. In addition to addressing only the quality of inhalation and nasal
goods, the demand for safety testing (e.g., for excipients and leachables) is also
emphasized.
In addition to safety and efficacy, additional quality problems (e.g., contaminants,
process validation, stability testing, and specifications) are addressed in other
guidance papers, such as ICH guidelines.
(b) April 1999 Guidance to Establish Equivalence or Relative Potency of Safety
and Efficacy of a Second Entry Short-Acting Beta2-Agonist Metered Dose
Inhaler (MDI) [31]
This guideline paper for second entry applicants outlines the steps to demonstrate
the equivalence or relative potency of the safety and efficacy of short-acting beta2-
agonist bronchodilators delivered by inhalation aerosol (metered dose inhaler;
MDI). The publication combines input from a range of sources, including the
Standards Committee of the Canadian Thoracic Society, the Canadian Pharmaceutical
Manufacturers Association, the Canadian Drug Manufacturers Association, and a
number of people. Other study designs or kinds may be considered provided the
technique has been adequately verified and Therapeutic Products Program approval
has been acquired beforehand (TPP).
(c) October 2018 Guidance Document: Data Requirements for Safety and

Effectiveness of Subsequent Entry Inhaled Corticosteroid Products Used for the
Treatment of Asthma – Summary [32]
The purpose of this advice is to aid sponsors in the gathering and analysis of
comparative clinical data for Inhaled Corticosteroid (ICS) medications used in the
treatment of asthma. This advice applies to products with the same active compo-
nent and circumstances of use as the Canadian Reference Product (CRP).
2.7 The Standards Council of Canada
The Standards Council of Canada is the approval body for national standards
(a) August 2011 CAN/CSA-Z264.1-02 (R2011) Spacers and Holding Chambers
for Use with Metered-Dose Inhalers [33]
This standard outlines the specifications for spacers and/or holding chambers
(S-HCs) for use with pressurized metered-dose inhalers. It details the material and
components, packaging and labeling, and in vitro-determined aerosol properties.
Regulatory standards for in vitro testing of OINDPs offer general specifications for
measuring OINDP performance under set laboratory conditions to determine essen-
tial end-point focused critical quality characteristics (CQA) that drive product
quality.
The following are some of the most widely used tests: Single Actuation Container
(SAC) Lifetime Aerodynamic Particle Size Distribution (APSD) via Cascade
Impaction (NGI, ACI), Geometry of Spray Pattern Puff Priming/Re-priming
Delivered Dose Mass Uniformity Valve/Pump Delivery Priming/Re-priming (Shot
Weight) Size of Drug Particles via Microscopy (Suspensions) Droplet Size
Distribution (DSD) using Laser Diffraction for Through-Life Testing Product
Wasting Through-Life Inter-/Intra-container Product Testing.
2.8 TGA Australia
TGA is the apex body for Registration of Inhalation and nasal sprays. A prescription
drug registration procedure exists for the registration of all medications [34].
The Therapeutic Goods (Classifications of Therapeutic Goods) Instrument 2018
[35] (established under section 23A) specifies several therapeutic goods classes.
Prescription and other drugs listed by the instrument, including (1) Metered-dose
asthma inhalers, are one of the groups of therapeutic commodities in the instrument.
(2) Nasal corticosteroids.
This implies that applications under section 23 for these medications are
considered using the process for prescription drugs [36].
(a) New Registrations
For new registrations of over-the-counter medications classified as prescription
drugs: Utilize the prescription medicine registration process [34], including the pre-
scription medicine form and payment of prescription drug costs. Use prescription
pharmaceutical data requirements [37], including requirements for reporting infor-
mation on the source of the active ingredient [38]. Do not use prescription medica-
tion labeling requirements Use OTC labeling requirements [39], including: Product
Information, Consumer Medicine Information, carton and product label, and any
product inserts. OTC medicines that are not classed with prescription medicines use
the OTC medicines registration process [40] and apply OTC data requirements for
over-the-counter (OTC) inhalation and nasal spray medications not classified as
prescription drugs.
The information below on quality, therapeutic equivalency of locally acting
medicines, and modifying the formulation or delivery device often does not apply
to over-the-counter medications that are not classified as prescription drugs.
However, there may be instances where portions of this advice apply.
Quality guidelines
• Guidelines on the pharmaceutical quality of inhalation and nasal products
(EMEA/CHMP/QWP/49313/2005 Corr) were adopted by the TGA [26]
• Stability testing of current active chemicals and associated formulations (CPMP/
QWP/122/02, rev C) [41]
• Except for nebulization solutions, inhalation and nasal spray medications are
crucial dosage forms for batch size and stability investigations.
• Information and data should be supplied in regulatory submissions for process
validation of finished goods (EMA/CHMP/CVMP/QWP/BWP/70278/2012-
Rev1) [42]
• Certain documents include useful information on the quality control of these
products:
• BP’s general monograph for inhalation preparations [43]
• USP monograph general 5> Inhalation and nasal medication products -
informational generalities and testing of product quality [44]
• USP chapter <610> Performance Quality Tests for Inhalation and Nasal Drug
Products: Aerosols, Sprays, and Powders [44]
2.9 Middle East/GCC
(a) Central Registration

The GCC Area’s consolidated procedure outlines the possibilities of registering
pharmaceutical items within the region. The Cooperation Council for the Arab
States of the Gulf, often known as the “Gulf Cooperation Council” (abbreviated
“GCC”), was established on May 25, 1981. The original member states are the
United Arab Emirates, the Kingdom of Saudi Arabia, the Sultanate of Oman, Qatar,
Bahrain, and Kuwait. To initiate the centralized procedure, the applicant must sub-
mit an application for registration for each production location that is not GCC-DR-
accredited. In addition, the applicant must submit one application for product
marketing authorization for each intended manufacturing line together with the reg-
istration application. The applicant must submit an application for registration for
each production site that has not been accredited by the GCC-DR in order to initiate
the centralized procedure. In addition to the previously indicated application for
registration, the applicant must submit one application for product marketing autho-
rization for each anticipated manufacturing line to the appropriate regulatory body.
A product dossier, created in compliance with the GCC CTD format criteria, is
initially submitted with product samples to the executive office of the GCC-DR,
where it is reviewed for completeness before transmission to the technical review
committees of the member states. Individual member-state evaluations are con-
ducted initially, followed by a discussion of the received comments at the subse-
quent committee meeting. The authorities will contact the applicant if more
information is requested. In such a case, the Committee will repeatedly reexamine
the submitted responses until they are complete and acceptable. Beginning in
February 2019, all submissions must be done solely through the GHC Electronic
Gateway in e-CTD format. To assist applicants with the submission procedure,
GHC has released many documents, including a tutorial on “How to Register for the
GHC Web Client,” the “GHC Electronic Submission Portal-Naming Convention
Files,” and the “Web Client User Guide.” The region-specific Module 1 must be
produced in compliance with the “GCC Module 1 Specification and the Baseline
eCTD Submission Requirements” per the December 2018 directive. After receiving
a favorable decision on the product application from the Central Committee, the
applicant sends samples, procedures, and materials for testing to a Central
Committee-accredited laboratory (MAA). If the committee reaches a positive deci-
sion, the executive office contacts the applicant and issues registration certificates
for the product as well as the manufacturing firm or location. The applicant may
seek an appeal within 2 months after obtaining notice of the judgment.
(b) Registration requirements in the leading GCC nations
Despite the fact that the registration procedure for medications in Gulf
Cooperation Council (GCC) nations is centralized and relatively similar, the regula-
tory requirements of a few big countries, such as Saudi Arabia and the United Arab
Emirates, are separate.
This regulation’s framework is identical to that of the centralized product
registration system, which covers nasal medicines. In each nation, classification,
site registration, and product registration are conducted differently. These nations
have a well-established regulatory structure as well as enforcement measures: (1)
Saudi Arabia (2) Bahrain (3) Qatar (4) Kuwait (5) United Arab Emirates (6) Oman
(7) Qatar.
2.10 Indian Perspective
The Drugs and Cosmetic Act of 1940 and Drugs and Cosmetic Rules of 1945
oversee drug registrations, import, production, distribution, and sale in India. This
act also established the Central Drugs Standard Control Organization (CDSCO) and
the Drug Controller General of India’s office (DCGI). For its functions, the CDSCO
has six zonal, four sub-zonal, eleven port/airport, and six laboratories. DCGI has not
established any particular guidelines for evaluating the effectiveness and safety of
orally inhaled products. The CDSCO has developed “Guidelines for bioavailability
and bioequivalence studies” for generic medication applications. Guidelines/rules
such as Rule 122A to E of the Drugs and Cosmetics Act Schedule Y of the Drugs
and Cosmetics Act and Rules thereunder (Amended in 2005), Good Clinical Practice
(GCP) guidelines issued by CDSCO, and Ethical Guidelines for Biomedical
Research Involving Human Subjects govern all clinical trials in India. When evalu-
ating the regulatory procedure for registering a second-entry orally inhaled product
in India, the following categories are significant.
(a) The reference drug is not approved in India
If the reference product has not been authorized in India, the second-entry orally
inhaled product would be designated as a novel drug since it would be deemed the
first market entry of the drug substance. Other instances that the Substances and
Cosmetics Act classifies as novel drugs include the following:
Drugs that have been approved by the DCGI but are now intended to be marketed for other
indications; and fixed-dose combinations of two or more drugs that have been individually
approved but are now proposed to be combined in an unapproved ratio.
All new medications must undergo clinical testing to assess their safety and efficacy
for Indian patients. If India is participating in a worldwide clinical trial, no more
than 20% of the total number of participants can be recruited from Indian locations.
These tests are required for both domestically produced and imported
pharmaceuticals.
For approval of all strengths of a second-entry medicine or a novel drug with
multiple strengths, the following requirements must be met: The qualitative content
of the respective strengths is virtually identical. b. The ratio of active components to
excipients is basically the same across all strengths. c. The manufacturing process is
generally same, and all strengths are produced by the same producer. d. Where rel-
evant, a suitable research has been conducted on at least one of the formulation’s
strengths. e. It has been demonstrated that the pharmacokinetics of systemic avail-
ability are linear across the therapeutic dosage range.
(b) The reference drug is approved in India
Bioequivalence based on pharmacokinetics alone is unsuitable for orally breathed
medications [non-solution pharmaceutical products] that are intended to operate
locally in the lungs; comparative clinical trials or PD studies are essential to show
equivalence. In India, the idea of PK bioequivalence for orally inhaled medicines is
not well-established, as the correlation between systemic levels and lung deposition
is still growing. No precedent exists for the approval of a second-entry orally inhaled
medication based only on PK bioequivalence. However, since additional research
demonstrates a strong link between Cmax and AUC0-t and lung deposition, PK
bioequivalence studies are widely recognized as adequate for establishing the
equivalence of orally inhaled drugs. The pulmonary accessible dosage is indicated
by the AUC0-t measurement, whereas the regional deposition is shown by the Cmax
measurement. With sufficient rationale, then, an evaluation of “interchangeability”
utilizing “pharmacokinetic equivalence” or “PK bioequivalence” between the sec-
ond entry medicine and the reference product may serve as an alternative to clinical
trials. When the second entry medicine is an inhalation aqueous solution containing
the same active substance(s) in the same concentration and virtually the same excip-
ients in comparable concentrations as the reference product, bioequivalence is self-
evident and no further in vivo investigations are necessary. The gadget could or
might not be comparable to the standard model. To establish equivalent device per-
formance between the reference inhalation product and the second-entry medica-
tion product, more in vitro testing is necessary.
2.11 China’s Perspective
(a) Regulatory Framework for Generic Drug Registration in China

The China Food and Drug Administration (CFDA) regulates and approves drugs
sold in China. Formerly known as the State Food and Drug Administration (SFDA),
the CFDA reports directly to the State Council. It consists of 19 departments and
bureaus and 18 affiliates, some of which are directly involved in drug regulation and
approval processes, including the Center for Drug Evaluation (CDE), National
Institute for Food and Drug Control (NIFDC), Center for Certification of Drugs
(CCD), Chinese Pharmacopeia Commission (CPC), and Center for Medical Device
Evaluation (CMDE). The Provisions for Drug Registration (PDR) were issued by
the CFDA in 2007 to establish the regulatory framework for drug registration in
China. A revised draft of the PDR, released for public comment in March 2014,
integrates the recent reorganization of the CFDA and proposes considerable
revisions.
As detailed on the CFDA website, applications for respiratory products follow
the same registration process, regulatory criteria, and review and approval schedule
as all other chemical products. In some situations, breathing equipment must be
individually registered in accordance with the application procedure for medical
devices. The CFDA’s 2006 “Principles and Technical Guidelines for the Research of
Chemicals with Existing National Standards” must be followed for generic medi-
cine applications. CFDA/CDE also released/published a number of specific guide-
lines and articles for orally inhaled products, including the Technical Guideline for
Research on Quality Control of Inhalation Products in 2007, the Technical
Requirements for Inhaled Drug Research in 2009, and the Clinical Trial
Considerations for Inhaled Drugs for Asthma and COPD in 2009. The CFDA has
not yet set clear development standards for generic orally breathed medicines.
Therefore, businesses must work closely with the CFDA/CDE to define the strategy
and requirements for their specific applications.
(b) Bioequivalence Requirements in China
In 2008, the CFDA convened professionals and academics (both local and
international) from fields such as medicine, pharmaceutics, pharmacology, and
toxicology. CDE published two articles in 2009, providing for the first time the
detailed technical requirements for inhaled formulations (with the main emphasis
on the switch from CFC to HFA propellant) and clinical aspects of technical
requirements for the development of generic orally inhaled drugs for the treatment
of asthma and/or chronic obstructive pulmonary disease (COPD). The requirements
were derived from the then-draft EMEA guidance and the agency’s experience with
inhaled medication development and clinical trials. The regulations are applicable
to both imported and domestically manufactured generic medications. The CDE
papers discuss fundamental principles but offer little specifics about the criteria. The
CFDA/CDE bioequivalence standards involve evaluations of both pulmonary
deposition and systemic drug exposure using pharmacokinetics (PK),

pharmacodynamics (PD), and/or clinical trials (CT).
2.12 Nasal Vaccines and Regulations
In recent years, nasal vaccines have become frequent; nevertheless, while choosing
a device for nasal administration, it is essential to keep in mind that the administra-
tion volume is rather small. An efficient spraying apparatus will lower the amount
of antigens required for successful protection. It is a question of preference whether
the immunization is delivered by one or both nostrils. Patients tend to have more
confidence in the second alternative, which will boost its acceptance. The initial
packaging of the vaccine (dried powder or liquid) must be optimized for easy, auto-
mated filling (of both small and large volumes) and dependable storage and trans-
port protection. Single-dose devices will provide the maximum level of vaccine
protection, but their fillings will necessitate the use of highly sophisticated tech-
nologies. These systems are only suitable for nations with a well-developed infra-
structure because of their high cost and size. Liquid vaccines may be packaged with
multi-dose spray pumps if microbial contamination of the bottle’s contents can be
prevented during use. This need may be met by so-called “preservative-free pump
systems,” which are also extremely cost-efficient. Disposable sleeves or protective
caps can successfully limit the transfer of diseases from one patient to another.
Following are the major regulations which regulate vaccines, including nasal
vaccines.
(a) All vaccines type
(i) EMA: Note for Guidance on Preclinical Pharmacological and Toxicological
Testing of Vaccines (1997)
(ii) Worldwide: WHO Guidelines on Nonclinical Evaluation of Vaccines (2005)
(iii) China: State Food and Drug Administration, China Technical guidelines
for preclinical research on preventive vaccines. Notice No. 140 (April 2010)

(iv) Japan: Japanese Guideline for Non-clinical Studies of Vaccines for
Preventing Infectious Diseases, (PFSB/ELD Notification No. 0527-1, May
27 2010)
(v) India: Drug and Cosmetics Act, 1940 and Drug and Cosmetics Rule,
1945 (2005)
(b) Vaccines for pregnant women and WCBP
(i) FDA: Guidance for Industry. Considerations for Developmental Toxicity
Studies for Preventative and Therapeutic Vaccines for Infectious Disease
Indications (2006)
(c) Adjuvants
(i) EMA: Guideline on Adjuvants in Vaccines for Human Use (2005)
(d) DNA vaccines

(i) FDA: Guidance for Industry. Considerations for Plasmid DNA Vaccines for
Infectious Disease Indications (2007)
(ii) WHO: Guidelines for Assuring the Quality and Nonclinical Safety

Evaluation of DNA Vaccines (2005)
(e) Recombinant DNA vaccines
(i) FDA: DRAFT Points to Consider in the Production and Testing of New
Drugs and Biologicals Produced by Recombinant DNA Technology (1985)
(f) Viral vectored vaccines
(i) Guideline on Quality, Nonclinical and Clinical Aspects of Live Recombinant
Viral Vectored Vaccines (2010)
(g) Combination vaccines
(i) EMA: Note for Guidance on Pharmaceutical and Biological Aspects of
Combined Vaccines (1998)
2.13 Digital Medical Devices for Nasal Drug Delivery
The Internet of Things (IoT) – simply described as a system of internet-connected

gadgets that gather and send data over a wireless network – has altered the health-
care industry, from electronic health records and patient portals to telemedicine.
The launch of 5G wireless technology of the next generation in 2019 is advancing
the immense potential of digital health technology. Many medical equipment may
now communicate with and connect to other devices and systems. FDA-approved,
permitted, or cleared devices are gaining digital capabilities. New types of devices
with these features are being researched. Participators in digital health activities
include patients, health care practitioners, researchers, conventional medical device
sector enterprises, and firms new to FDA regulatory norms, such as mobile applica-
tion developers. The FDA’s Center for Devices and Radiological Health (CDRH) is
enthusiastic about these developments and the convergence of networking and con-
sumer technologies with medical devices [45].
The FDA has endeavored to provide clarification on the following issues within
the digital health business, employing pragmatic approaches that balance benefits
and risks:
• Software as a Medical Device (SaMD)
• Artificial Intelligence and Machine Learning (AI/ML) in Software as a
Medical Device
• Wireless Medical Devices
• Cybersecurity
• Device Software Functions, including Mobile Medical Applications
• Health IT
• Medical Device Interoperability
• Medical Device Data Systems
• Telemedicine
CDRH has established the Digital Health Center of Excellence, which aims to
enable digital health stakeholders to advance health care as another critical step in
fostering the progress of digital health technology.
(a) Benchmark Regulations for Digital Health [46]
Following is an overview of major laws and rules of three federal agencies:
• Act on the Portability and Accountability of Health Insurance (HIPAA)
• The Office for Civil Rights (OCR) of the United States Department of Health and
Human Services is responsible for enforcing the HIPAA standards, which pro-
tect the privacy and security of some health information and require some firms
to provide breach notifications (HHS).
• Act Concerning Food, Drugs, and Cosmetics (FD&C Act)
• The FDA enforces the FD&C Act, which controls the safety and efficacy of
medical devices, including some mobile medical applications. The FDA focuses
its regulatory oversight on a small subset of health applications that represent a
bigger danger if they malfunction.
• Act of the Federal Trade Commission (FTC Act)
• The FTC is responsible for enforcing the FTC Act, which prohibits deceptive or
unfair acts or practices in or affecting commerce, including those involving pri-
vacy and data security as well as false or misleading statements about the safety
or performance of mobile applications.
• The Health Breach Notification Rule of the FTC
• The FTC is responsible for implementing the FTC Act, which prohibits deceptive
or unfair acts or practices in or affecting commerce, such as those involving false
or misleading claims about an app’s safety or performance.
3 Summary
Patient safety is of utmost importance to all regulatory agencies around the globe,
with the USFDA, EMA, and Canada Health leading the way in the development of
legislation. These nations are also members of ICH, which guarantees regulatory
uniformity. These rules are adapted by developing nations in Asia and the Middle
East, either as-is or with revisions based on their needs. Regulations pertaining to
nasal drugs center on New Drug Applications, Bioequivalence, and Medical
Devices. These constitute most of the traditional drug systems. The USFDA, the
EMA, and the ICH have created ingredient- and delivery-device-specific recom-
mendations for nasal medication delivery devices. Clinical trials and equivalency
studies are also crucial components of nasal medication approval. Regulation man-
dates three-phased research on the efficacy of all medications and dosage formula-
tions. Vaccinations administered by nasal route are also strictly controlled by rules
and standards. With the emergence of IoT, AI, and augmented reality, there have
been significant advances in nasal medication delivery systems & medical devices/
equipment. Developed nations are also focused on smart gadgets, which provide
unique issues owing to a combination of software technology, data protection, and
complicated software and device component combinations. In the United States and
Europe, the regulation of medical devices is undergoing significant changes. This is
a very dynamic sector that requires quick extra regulatory attention and utilization
of data by technology businesses to help patients with better options.
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Index
A C
Absorption, 9, 17, 26, 44, 90, 102, 131, 144, Challenges, 2, 27–28, 44, 45, 53, 60–77, 95,
171, 190, 235, 255, 280, 340, 362, 384 129, 135, 136, 149, 160, 171, 223, 224,
Absorption enhancers, 20, 21, 28–33, 45, 134, 242, 260, 281, 336, 339, 340, 344, 355,
144, 176, 192, 194, 196, 199, 202, 254, 362, 365, 370, 375
262–269, 271, 340, 341, 363, 364, 376 CNS disorders, 60, 65–69, 73, 74, 83, 148
Alzheimer’s disease (AD), 60, 69, 70, 73, 74, CNS targeting, 66
118, 181, 183 Coronavirus disease-19 (COVID-19), 17, 18,
23, 44, 102, 134, 242, 335, 386, 394
B
Bioavailability, 11, 12, 20, 21, 26, 29, 33, 46, D
50, 53, 61, 66–72, 74, 91, 102, 103, Design of nasal drug delivery, 43–55
105, 109, 113, 115, 118, 119, 142, 144, Digital technology, 412, 413
146–148, 150–153, 157, 160, 170, 171, Drug absorption, 2, 10, 11, 25–34, 45–47, 63,
173, 176, 177, 179–181, 184, 190–192, 86, 87, 132–134, 144, 145, 149, 152,
194–197, 199, 201, 202, 205, 206, 174–183, 195, 218, 242, 244, 254, 260,
208–210, 214, 218–221, 223, 237, 242, 265, 283–286, 305, 327, 328, 332, 333,
244–246, 254, 255, 259–261, 264, 265, 355, 362, 367, 368, 373, 384, 386, 387
267–269, 280, 285, 289, 291, 302–305, Drug delivery, 9, 16, 26, 44, 84, 106, 128, 142,
308, 310, 311, 326, 328, 335, 343–346, 170, 191, 236, 254, 280, 339, 362,
348, 349, 352, 353, 355, 362, 365, 372, 382, 394
376, 384, 386, 388, 397, 398, 408 Drug encapsulation, 157, 292, 293, 340, 344–349
Biomedical applications, 102–121, 158 Drug formulations, 10, 11, 28, 33, 111, 149,
Blood-brain barrier (BBB), 10, 12, 16, 18, 19, 247, 284, 291, 340, 341
21, 23, 44, 47–52, 55, 60, 66, 68, 70, 72,
77, 84, 85, 90, 92, 95, 102, 105, 107–111,
115, 119, 147, 148, 153, 155, 157, 160, E
170, 171, 173–175, 180, 182, 183, 190, Enzyme inhibitors, 12, 20, 31, 33, 134, 159,
205, 214, 245, 280, 281, 286, 288, 311, 173, 254, 268
312, 328, 335, 341, 350, 351, 362, 365,
368–370, 372, 373, 375, 376, 394
Brain, 2, 16, 47, 60, 83, 102, 143, 170, 190, G
242, 261, 280, 326, 344, 362, 385, 394 Global market trends, 381–389
© The Editor(s) (if applicable) and The Author(s), under exclusive license to 417
Springer Nature Switzerland AG 2023
https://doi.org/10.1007/978-3-031-23112-4
418 Index
H N
High molecular weight drugs, 179, Nanocarriers, 50, 73, 102–121, 145–147,
253–271, 366 150–152, 154–160, 171, 180, 181, 184,
200, 220, 243, 245, 261, 284, 285, 290,
292, 293, 298, 365, 368, 371, 375,
I 376, 387
In situ gelification, 241 Nanoemulsions, 10, 11, 60–77, 110, 112,
International Conference on Harmonization of 113, 146, 152–154, 243, 245, 280,
Technical Requirements for 375, 387
Registration of Pharmaceuticals for Nanoparticles, 19–21, 50, 52, 60, 70, 73, 91,
Human Use (ICH), 54, 395–396, 402, 102, 105, 106, 111, 113–115, 118–120,
405, 413 134, 135, 146, 152, 156, 157, 159, 180,
Intranasal, 9, 21, 26, 45, 62, 95, 103, 134, 170, 182, 183, 191, 194–200, 202, 205,
205, 236, 254, 280, 326, 339, 362, 214–217, 220, 261, 262, 264, 265, 289,
386, 394 294, 326, 329–331, 334, 335, 340,
Intranasal administration, 26–28, 30, 45, 47, 342–355, 362, 365, 368–371, 375,
49, 52, 61, 64, 65, 67, 68, 70, 71, 73, 386, 387
75, 77, 90, 91, 94, 105, 106, 110, 113, Nanosuspension, 111, 241, 326–327, 329, 333,
120, 144, 148, 149, 151, 153–155, 157, 335, 336, 375
158, 180–183, 202, 207, 212, 216, 217, Nanotechnology, 11, 60, 142, 150, 184, 281,
241, 242, 244, 245, 261, 281, 287–289, 289, 375, 383, 385–387, 389
334, 335, 345, 348, 351–353, 366–368, Nasal, 2, 15, 25, 44, 60, 86, 102, 129, 142,
370, 371, 374 171, 190, 235, 254, 280, 339, 362,
Intranasal delivery, 9, 21, 64–69, 71, 106–108, 382, 394
148, 150, 156–158, 198, 205, 218, 246, Nasal absorption, 20, 30, 46, 95, 132–135,
270, 280, 286, 287, 326, 341, 342, 350, 150, 158, 175–181, 184, 191, 192,
353–355, 362, 373, 375, 376, 397 194–199, 207, 209, 214, 215, 219, 242,
Intranasal drug administration, 52, 92, 355 255, 258, 259, 265–269, 284,
362–364, 367
Nasal cavity, 2–4, 7–12, 15–18, 20–23, 26, 27,
L 31, 33, 44–47, 49–51, 53, 60, 70, 71,
Liposomes, 50, 52, 60, 73, 74, 102, 106, 107, 86–91, 95, 103, 106, 112, 115, 129,
114, 119, 134, 135, 146, 150–152, 154, 131, 132, 134, 143–145, 149, 154, 155,
180, 194, 198, 242, 243, 261, 263, 280, 172–177, 179, 190, 191, 193, 195–197,
293, 294, 312, 342, 344, 347, 199, 200, 204–206, 209, 213–218, 220,
362, 371–375 223, 235, 236, 238, 240, 241, 246,
259–261, 265, 266, 268, 269, 280–288,
305, 312, 327, 333, 340, 341, 362, 363,
M 365, 367, 368, 376, 383–387
Macromolecular drugs, 142, 224 Nasal delivery, 21–22, 53–55, 66, 72, 95, 102,
Mucoadhesives, 10, 11, 28, 32, 45, 48, 51, 52, 105, 118, 119, 142–160, 175, 191, 196,
55, 63, 66–69, 71–73, 105, 111, 119, 198, 204, 207, 209, 210, 215, 216, 220,
134, 146, 150, 152, 155, 159, 190–224, 221, 223, 245, 253, 254, 256, 260–270,
241, 246–248, 254, 260–266, 288, 305, 282, 305, 310, 312, 339–355,
312, 329, 330, 335, 340, 349, 368, 369, 370, 382–389
376, 385 Nasal drug delivery, 1–2, 15–23, 28–33, 44,
Mucociliary, 8, 10–12, 26, 27, 44, 45, 51–53, 45, 53–55, 73, 95, 113, 135, 142, 144,
66, 70, 73, 88, 90, 95, 103, 105, 112, 190, 192–195, 197, 205–208, 212–214,
115, 119, 129, 132, 135, 144, 145, 149, 217, 221, 223, 224, 246, 253, 280–290,
153, 155, 156, 171, 172, 176, 178, 179, 313, 339, 341, 363–365, 382–388, 394,
191, 209, 220, 236, 247, 248, 259, 266, 395, 412–413
284, 305, 312, 339–341, 345, 362–364, Nasal drug delivery system, 26–28, 43–55,
367–369, 387 119, 144, 159, 191, 193, 196, 206, 209,
Index 419
213, 221, 223, 224, 237, 281, 341, P

382–389, 394–414 Parkinson’s disease (PD), 47, 48, 55, 60, 66,
Nasal formulations, 28, 44, 54, 55, 113, 142, 67, 70, 73, 83, 93, 114, 115, 182, 190,
147, 178, 199, 200, 203, 206, 217, 224, 245, 255, 258, 265, 345, 346, 349, 362,
236, 238, 244, 247, 248, 267, 269, 271, 365, 369, 370, 375, 387, 409, 411
280, 340, 341, 365, 375 Peptide-protein drugs, 254, 259, 261,
Nasal passageways, 8, 15–21, 23 265, 267–269
Nasal pathways, 31, 96, 175, 176 Peptides, 11, 12, 26, 28, 31, 33, 44, 45, 48–50,
Nasal route, 1, 2, 23, 25–27, 30, 32–34, 43–45, 60, 74, 95, 103, 107, 109, 111, 112,
53, 74, 102, 103, 114–117, 120, 121, 117, 133, 135, 142, 144, 145, 147, 148,
127–136, 142–147, 151, 156, 160, 150, 151, 158, 160, 170–184, 190, 191,
170–173, 178, 190, 200, 209, 213, 219, 194, 195, 197, 198, 201, 214, 236, 246,
237, 242, 246, 269, 281, 283, 284, 327, 254–257, 259, 263, 265–269, 284, 307,
328, 333, 336, 339, 340, 349, 362, 369, 311, 312, 351–352, 362, 366, 369–374,
372, 373, 385, 394, 414 376, 385, 387
Nasal toxicity, 71, 72, 179, 216, 328, 336 Physicochemical, 44, 46, 53, 54, 84, 95, 112,
Nasal transport, 148, 149 133, 134, 145, 154, 155, 158, 159, 171,
Natural, 5, 10, 29, 53, 71, 105, 113, 135, 154, 177, 193, 217, 259, 284, 291, 351,
156, 157, 177, 183, 190, 192–195, 363–367, 384
200–202, 213, 218, 223, 224, 239, 241, Polymers, 28, 31–34, 48, 51, 52, 66, 67, 109,
247, 255, 286, 287, 292, 329, 341, 370, 113, 115, 117, 119, 134, 135, 146, 150,
372, 374, 385 151, 155, 156, 158, 159, 181, 190–224,
Niosomes, 242–244, 280, 290–313 238–248, 254, 260–267, 288, 289, 329,
Nose-to-brain, 2, 9–11, 13, 26, 44, 47–53, 55, 331, 340–343, 347–350, 353–355, 363,
60, 61, 63–70, 72–75, 77, 86–88, 91, 368, 369, 372, 373, 376
95, 96, 108–110, 115, 118, 121, Proteins, 26, 28, 29, 31, 33, 44, 45, 48–50, 60,
147–149, 151, 153, 156, 157, 160, 62, 72, 84, 88, 95, 102, 103, 111, 114,
170, 171, 180, 181, 183, 184, 190, 118, 120, 142–145, 147, 148, 151, 160,
191, 194, 195, 197–199, 205, 208, 170–184, 190–195, 207, 215, 216, 236,
212, 214, 216, 218, 219, 223, 242, 246, 256–258, 261, 267, 281, 284, 285,
261, 281, 286–289, 326, 334, 335, 288, 306, 307, 312, 330, 362, 369, 371,
344, 347, 354, 362, 365–369, 371, 372, 374, 376
372, 375–376
Nose-to-brain delivery, 47–51, 86, 87,
116–117, 147–149, 153, 156, 157, 160, R
170–184, 190, 191, 194, 198, 199, 212, Regulatory, 258, 394–414
216, 218, 219, 261, 332–333, Residence time, 50, 51, 53, 103, 106, 110,
362–376, 385 113, 149, 154, 157, 192, 195, 197, 200,
Nose-to-brain drug delivery, 44, 47–49, 52, 53, 204, 206, 209, 215, 217, 218, 236, 238,
55, 88, 96, 171, 184, 191, 208, 212, 241, 242, 248, 260, 269, 288, 304, 305,
223, 312, 326–336, 387 308, 329, 341, 347, 352, 363, 368–369
O S
Olfactory, 2–6, 8–10, 16, 19, 44, 47, 49–52, Safety, 28, 29, 53, 66, 72, 77, 102, 110, 121,
63, 69–71, 86–92, 94, 95, 103–106, 145, 159, 203, 205, 217, 223, 224,
109–113, 118, 129, 130, 143, 147–149, 246–249, 254, 261, 309, 368, 395, 399,
153, 170, 172–176, 180, 181, 190, 191, 402–405, 408, 409, 412, 413
206, 213, 214, 216–218, 223, 241, 242, Spreadability, 155, 191
280–286, 312, 327, 332, 333, 362, Surface modification, 157, 310, 342–344,
366–371, 373, 386, 394 350–352, 354, 368, 372
420 Index
Surfactants, 20, 27–30, 33, 34, 45, 46, 48, 50, 336, 344, 345, 350, 352, 353, 365,
60, 62, 63, 65, 71, 73, 75, 95, 102, 108, 368–370, 373–376, 386
152, 155, 156, 159, 181, 239, 248, 266, Tumors, 2, 21, 83, 114, 115, 148, 190, 238,
269, 280, 284, 290, 292–299, 304, 306, 245, 257, 286–289, 307, 310, 334, 352,
307, 312, 331, 364 370, 385
T V
Targeting, 60, 64, 66, 68, 69, 73, 74, 77, 86, Vaccines, 26, 44, 95, 102, 119–120, 127–136,
87, 95, 96, 106–109, 111, 113, 119, 142, 145, 146, 154–156, 158, 190, 192,
121, 147–149, 153, 154, 170, 171, 196, 201, 236, 237, 241, 261, 281, 284,
180–183, 190, 214, 216, 217, 280, 307, 308, 326, 348, 386, 394, 411–412
282, 305, 307, 309–312, 333, 334,

Yashwant V. Pathak, Hemant K. S. Yadav - Nasal Drug Delivery - Formulations, Developments, Challenges, and Solutions-Springer (2023)

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Yashwant

Nasal Drug Delivery

ISBN 978-3-031-23111-7 ISBN 978-3-031-23112-4 (eBook)

formulation and challenges are covered comprehensively in their respective

Tampa, FL, USA Yashwant V. Pathak

1 An Overview of the Anatomy and Physiology of Nasal

Abdullah Abdelkawi Taneja College of Pharmacy, University of South Florida,

Yahya E. Choonara, PhD Wits Advanced Drug Delivery Platform Research Unit,

Luana Perioli, PhD Department of Pharmaceutical Sciences, University of

Hemant K. S. Yadav, Allyson Lim-Dy, and Yashwant V. Pathak

Keywords Nasal · Drug Delivery · Intranasal · Targeting and physicochemical

1.1 Nasal Drug Delivery Systems [1]

© The Author(s), under exclusive license to Springer Nature Switzerland AG 2023 1

metabolism avoidance, noninvasiveness and ease of access, no sterilization require-

1.1.1 Nasal Anatomy and Physiology

4 Abnormal Nasal Physiology

drug-related factors. Increases in blood flow or parasympathetic tone, as well as

5 Tests of Nasal Physiology

Computational fluid dynamic studies of nasal airflow and physiology have

6 Factors Affecting Intranasal Delivery

Intranasal delivery includes numerous benefits like non-invasiveness, convenient

7 Nasal Passage Targeting the CNS

8 Barriers to Drug Transport from Nose to Brain

8.2 Physico-Chemical Properties of the Drugs

9 The Sensitivity of the Nasal Mucosa as a Limiting Factor

10 Impact of Delivery Instructions, Patient Compliance,

ensure a patient receives optimal treatment. Non-invasive administrations display

Keywords Nasal passageway · Drug delivery · Blood-brain barrier · COVID-19 ·

1 What Is the Nasal Passageway?

M. Mathur · Y. V. Pathak (*)

© The Author(s), under exclusive license to Springer Nature Switzerland AG 2023 15

2 What Is the Blood-Brain Barrier?

3 Properties of Nasal Passages

4 Current Relevance with COVID-19

5 Benefits of Utilizing the Nasal Passageway

There are multiple options by which medications can be delivered to individuals,

Fig. 2.2 Flow of medication via nasal spray delivery [9]

6 Possible Barriers to the Utilization

Another obstacle that a medication must overcome is enzyme degradation. The

7 Properties of an Effective Nasal Delivery Drug

Fig. 2.3 Characteristics of a successful nasal delivery drug [15]

Keywords Absorption · Challenges · Enzyme inhibitors · Nasal route · Surfactants

S. K. Amponsah (*) · I. Adams

© The Author(s), under exclusive license to Springer Nature Switzerland AG 2023 25

2 Challenges Associated with Absorption of Nasal Drug

2.1 Barriers Against Mucosal Drug Absorption

Absorption of the active pharmaceutical ingredient (API) of a drug would require

2.2 Toxicity Associated with Intranasal Applications

When developing a pharmacological formulation for intranasal administration,

3 Drug Absorption Enhancers as Opportunity for Improving

Surfactants are amphiphilic molecules with lipophilic and hydrophilic residues

Natural pulmonary surfactant could be a mixture of about 90% phospholipids and

3.1.2 Bile Salts and Their Derivatives

Non-ionic surfactants are relatively non-toxic in nature [83]. Alkylglycosides (AGs)

Biosurfactants are surface-active chemicals produced by living organisms like bac-

mucoadhesive polymers because of their biocompatibility, biodegradability, and

3.5 Tight Junction Modulators

Table 3.1 Absorption enhancers and their potential mechanisms of action

diabetes, cardiovascular, and CNS diseases. Despite its merits, administration of

Jéssica Bassi da Silva, Maria Vitoria Gouveia Botan,

Keywords Design of nasal drug delivery · Blood-brain barrier · Nasal cavity ·

J. B. da Silva · M. V. G. Botan · M. L. Bruschi (*)

© The Author(s), under exclusive license to Springer Nature Switzerland AG 2023 43

Tampa, FL, USA Yashwant V. Pathak

1 An Overview of the Anatomy and Physiology of Nasal

1.1 Nasal Drug Delivery Systems [1]

1.1.1 Nasal Anatomy and Physiology

4 Abnormal Nasal Physiology

5 Tests of Nasal Physiology

6 Factors Affecting Intranasal Delivery

7 Nasal Passage Targeting the CNS

8 Barriers to Drug Transport from Nose to Brain

8.2 Physico-Chemical Properties of the Drugs

9 The Sensitivity of the Nasal Mucosa as a Limiting Factor

10 Impact of Delivery Instructions, Patient Compliance,

1 What Is the Nasal Passageway?

2 What Is the Blood-Brain Barrier?

3 Properties of Nasal Passages

4 Current Relevance with COVID-19

5 Benefits of Utilizing the Nasal Passageway

6 Possible Barriers to the Utilization

7 Properties of an Effective Nasal Delivery Drug

2 Challenges Associated with Absorption of Nasal Drug

2.1 Barriers Against Mucosal Drug Absorption

2.2 Toxicity Associated with Intranasal Applications

3 Drug Absorption Enhancers as Opportunity for Improving

3.1.2 Bile Salts and Their Derivatives

3.5 Tight Junction Modulators

2 Factors Related to Nasal Anatomy and Physiology

3 Factors Related to the Biologically Active Agent

4 Factors Related to the Formulation

4.1.2 Protein Inhibitors (Enzyme and Glycoprotein)

4.1.4 Mucoadhesive and Mucus-Penetrating Formulations

2 Composition of Nanoemulsion for Nasal Administration

3 Factors that Influence Nanoemulsion Transport from Nose

4 Methods of Preparation of Nanoemulsion

5 Intranasal Delivery of Nanoemulsion for CNS Disorders

5.1 Nanoemulsion in the Treatment of Alzheimer’s Disease

5.2 Nanoemulsion in the Treatment of Parkinson’s Disease

5.3 Nanoemulsion in the Treatment of Migraine

5.4 Nanoemulsion in the Treatment of Psychosis

5.5 Nanoemulsion in the Treatment of Epilepsy

6 Recent Patents on Nose-to-Brain Delivery

7 Current Challenges and Future Prospective

8 Conclusion and Future Prospective

mini-pump-assisted intracranial delivery; electromagnetic force-field techniques;

2 Drug Delivery Pathways

2.1 The Nasal Cavity

2.2 The Respiratory Region and Epithelium

2.3 The Olfactory Region and Epithelium

2.3.1 Olfactory Sensory Neurons

3 Drug Delivery Pathways/Brain Targeting Sites via

3.1 The Olfactory Pathway

3.2 The Trigeminal Pathway

3.3 The Lymphatic Pathway

3.4 The Systemic Pathway

4 Factors Affecting Nasal Absorption

Keywords Nasal drug delivery · Biomedical application · Nanocarriers · Nose-to-