TUGAS BIOPHARMACEUTICAL

Niosome dan liposom

AFRIYANI
1821012027

Dosen Pengampu
Dr. Muslim Suardi, M.Si, Apt

PROGRAM PASCASARJANA
FAKULTAS FARMASI
UNIVERSITAS ANDALAS
 NIOSOM

1 • A niosome is a non-ionic surfactant-based liposome. Niosomes are

formed mostly by cholesterol incorporation as an excipient.

• The sizes of niosomes are microscopic and lie in nanometric scale.

The particle size ranges from 10nm-100nm A typical niosome vesicle
would consist of a vesicle forming amphiphile i.e. a non-ionic
2
surfactant such as Span-60, which is usually stabilized by the
addition of cholesterol and a small amount of anionic surfactant such
as dicetyl phosphate, which also helps in stabilizing the vesicle.

(Priyanka, Beloshe. et.al., 2019)

ADVANTAGES OF NIOSOMES
▪ The vesicles may act as a depot,
releasing the drug in a controlled
manner.
▪ The surfactants used are
biodegradable, biocompatible and
non-immunogenic.
▪ They can be made to reach the site
of action by oral, parenteral as well
as topical routes.
▪ Due to the unique infrastructure
consisting of hydrophilic,
They are osmotically active and
stable, and also they increase the
stability of entrapped drug.
▪ They improve oral bioavailability
of poorly absorbed drugs and
enhance skin penetration of drugs.
The vesicles may act as a depot,
releasing the drug in a controlled
manner.
▪ Niosomal dispersion in an aqueous
phase can be
(Priyanka, Beloshe. et.al., 2019)
COMPONENTS OF NIOSOMES [

Non-ionic
Surfactants

Charged
Cholesterol
Molecule

(Priyanka, Beloshe. et.al., 2019)

 1. NON-IONIC SURFACTANTS

• The role surfactants play a major role in the

formation of niosomes.
• The following non-ionic surfactants are
generally used for the preparation of niosomes.
E.g. –
1. Spans (span 60, 40, 20, 85, 80)
2. Tweens ( tween 20, 40, 60, 80)
3. Brijs ( brij 30, 35, 52, 58, 72, 76).
4. Alkyl amide (e.g. galactosidesand, glucosides)

(Priyanka, Beloshe. et.al., 2019)

 • Cholesterol is used to provide rigidity and proper shape, conformation to the niosomes preparations. Cholesterol
is a steroid derivative, which is mainly used for the formulation of niosomes.
2. Cholesterol

• Some charged molecules are added to niosomes to increase stability of niosomes by electrostatic repulsion which
prevents coalescence. The negatively charged molecules used are diacetyl phosphate (DCP) and phosphatidic
acid. Similarly, stearylamine (STR) and stearyl pyridinium chloride are the well known positively charged
3. Charged Molecule molecules used in niosomal preparations.

(Priyanka, Beloshe. et.al., 2019)

 METHOD OF PREPERATION

• 1. Ether Injection Method

(Priyanka, Beloshe. et.al., 2019)

2. HAND SHAKING METHOD (THIN FILM
HYDRATION TECHNIQUE)

(Priyanka, Beloshe. et.al., 2019)

3. Sonication Method 4. Micro fluidization Method

(Priyanka, Beloshe. et.al., 2019)

5. REVERSE PHASE EVAPORATION METHO

(Priyanka, Beloshe. et.al., 2019)

 TYPES OF NIOSOMES

1. According to the nature of lamellarit 2. According to the size

▪ Multilamellar vesicles (MLV) 1 - 5 μm in ▪ Small niosomes (100 nm – 200 nm)

size.
▪ Big niosomes (2 μm – 4 μm)

▪ Large unilamellar vesicles (LUV) 0.1 - 1 μm Large niosomes (800 nm – 900 nm)
in size

Small unilamellar vesicles (SUV) 25 - 500

▪ Big niosomes (2 μm – 4 μm)
nm in size.

(Priyanka, Beloshe. et.al., 2019)

(Priyanka, Beloshe. et.al., 2019)
 CHARACTERISATION OF
Niosomes

size

Bilayer Formation

Number of Lamellae

Membrane Rigidity

Entrapment Efficiency

(Priyanka, Beloshe. et.al., 2019)

 IN VITRO RELEASE STUDY
• With the help of dialysis tubing in vitro release rate study can be done. A dialysis sac was washed and soaked in
distilled water. The suspension of vesicle was pipetted into a bag made up of the tubing and then sealed and placed
in 200 ml buffer solution in a 250 ml beaker with constant shaking at 25°C or 37°C. The buffer was analysed at
various time intervals, for the drug content
Dialysis • by an appropriate assay method

• In this technique, niosomes are placed in a number of small dialysis tubes containing 1 mL of dissolution
medium and the niosomes are then displaced from the dissolution medium
Reverse
dialysis:

• In a Franz diffusion cell, the cellophane membrane is used as the dialysis membrane. The niosomes are
dialyzed through a cellophane membrane against suitable dissolution medium at room temperature
Franz
diffusion cell

(Priyanka, Beloshe. et.al., 2019)

 IN VIVO RELEASE STUDY

For In vivo study niosomal suspension was injected intravenously (through tail vein) to the
albino rats using appropriate disposal syringe. These rats were subdivided into groups

(Priyanka, Beloshe. et.al., 2019)

 ROUTES OF DRUG ADMINISTRATION AND
EXAMPLES OF DRUGS
1. Intravenous route: Doxorubicin,
Methotrexate, Sodium Stibogluconate, 3. Transdermal route:
Iopromide, Vincristine, Diclofenac Sodium, 2. Peroral route: DNA Flurbiprofen, Piroxicam,
Flurbiprofen, Centchroman, Indomethacin,
Colchicine, Rifampicin, Tretinoin, Transferrin
vaccines,Proteins,Peptides,Erg Estradiol Levonorgestrol,
and Glucose ligands, Zidovudine, Insulin, ot,Alkaloids,Ciprofloxacin, Nimesulide, Dithranol,
Cisplatin, Amarogentin, Amphotericin B, 5- Norfloxacin, Insulin Ketoconazole, Enoxacin,
Fluorouracil, Camptothecin, Adriamycin, Ketorolac
Cytarabine Hydrochloride

6. Inhalation: All trans 5. Nasal route: Sumatriptan, 4. Ocular route: Timolol

retinoic acid Influenza Viral Vaccine Maleate, Cyclopentolate

(Priyanka, Beloshe. et.al., 2019)

 LIPOSOM

• Liposomes were first described by Bangham in

1965 while studying cell membranes. He found that
lipsomes are vesicular structures consisting of
hydrated bilalyers which form spontaneously when
phospholipids are dispersed in water. Since this, further
studies into liposomes and their application in various
fields such as medicine and research have been
explored. Liposomes are defined as structure
consisting of one or more concen spheres of lipid
bilayers separated by water or aqueous buffer
compartments.

Varun, Thakur.et.al., 2005.

 ADVANTAGES OF LIPOSOMES
 Provide controlled drug delivery
 Biodegradable, biocompatible, flexible
 Non ionic
 Can carry both water and lipid soluble drugs
 Drugs can be stabilized from oxidation
 Improve protein stabilization
 Controlled hydration
 Provide sustained release
 Targeted drug delivery or site specific drug delivery
 Stabilization of entrapped drug from hostile
environment
 Alter pharmacokinetics and pharmacodynamics of
drugs
 Can be administered through various routes
 Can incorporate micro and macro molecules
 Act as reservoir of drugs
 Therapeutic index of drugs is increased
 Site avoidance therapy
 Can modulate the distribution of drug
Varun, Thakur.et.al., 2005.
CLASSIFICATION LIPOSOMES

I. Based on composition and

mode of
drug delivery

II. Based on Size and Number of

Lamellae

Varun, Thakur.et.al., 2005.

 I. BASED ON COMPOSITION AND MODE OF
DRUG DELIVERY

• 1. Conventional liposomes
• Composed of neutral or negatively charged phospholipids and cholesterol.
1
• 2. pH sensitive liposomes : Composed of phospholipids such as phosphatidyl ethanolamine, dioleoyl phosphatidyl
ethanolamine. Subjected to coated pit endocytosis at low pH, fuse with cell or endosomes membrane and release their
2 contents in cytoplasm; suitable for intra cellular delivery of weak base and macromolecules

• 3. Cationic Liposomes: Composed of cationic lipids. Fuse with cell or endosome membranes; suitable for delivery of
negatively charged macromolecules (DNA, RNA); ease of formation, structurally unstable; toxic at high dose, mainly
3 restricted to local administration

Varun, Thakur.et.al., 2005.

 • 4. Long circulating or stealth liposomes: Composed of neutral high transition temperature lipid, cholesterol and 5-10% of PEG-DSPE.
Hydrophilic surface coating, low opsonisation and thus low rate of uptake by RES long circulating half life (40 hrs); Dose independent
4 Pharmacokinetics

• 5. Immuno liposomes: Conventional or stealth liposomes with attached Antibody or Recognition Sequence. Subject to receptor
mediated endocytosis, cell specific binding (targeting); can release contents extra cellularly near the target tissue and drugs diffuse
5 through plasma membrane to produce their effects.

• 6. Magnetic Liposomes: Composed of P.C, cholesterol and small amount of a linear chain aldehyde and colloidal particles of
magnetic Iron oxide. These are liposomes that indigenously contain binding sites for attaching other molecules like antibodies on their

6 exterior surface

• Temperature (or) heat sensitive liposomes: Composed of Dipalmitoyl P.C. These are vesicles showed maximum release at 41º the
phase transition temperature of Dipalmitoyl P.C. Liposomes release the entrapped content at the target cell surface upon a brief
heating to the
7 • phase transition temperature of the liposome membrane

Varun, Thakur.et.al., 2005.

 II BASED ON SIZE AND NUMBER OF
LAMELLAE
1. Multi lamellar vesicles (M.L.V): ( Size
0.1 - 0.3 micro meter)

2. Large Unilamellar Vesicles (L.U.V):

(Size 0.1 - 10 micro meter)

3. Small Unilamellar Vesicles (S.U.V):

(Size 0.1 micro mete

Varun, Thakur.et.al., 2005.

 PREPARATION OF LIPOSOMES

• An important parameter to consider when addressing the formation process of liposomes is the
rigidity of the bilayer. Hydrated-single component phospholipid bilayers can be in a liquid- crystalline
(‘fluid’) state or in a gel state. By increasing the temperature, the gel state bilayer melts and is
converted into the liquid state. This occurs at a temperature known as the transition temperature
(Tc). The Tc of a bilayer depends on:

• 1. Acyl chain length.

• 2. Degree of saturation.
• 3. Polar head group

Varun, Thakur.et.al., 2005.

 CHARACTERISATION OF LIPOSOM

1. Particle Size and Surface Charge

2. Transmission Electron Microscop

Entrapment Efficiency (%EE) and

Loading Efficiency

4. Stability Evaluation of Liposomes

Varun, Thakur.et.al., 2005.

 APPLICATIONS OF LIPOSOMES

The following are

some properties 5. Molecules with
which make wide range of
3. Enhanced drug
liposomes 2. Localized drug solubility and 5. Cancer
uptake
applicable in effect molecular weight chemotherapy
various fields. can be
1. Cell -liposome accommodated
interaction
 INTRODUCTION

Severe vision loss from posterior segment diseases such as age-

related macular degeneration and diabetic macular edema
accounts for most cases of irreversible blindness worldwide.
Recent studies have demonstrated the usefulness of nonsteroidal
antiinflammatory drugs (NSAIDs) for the treatment of these
diseases(Warren et al., 2010)

The NSAID diclofenac is a potent agent for the treatment of

macular edema of various etiologies (Soheilian et al.,

Fujisawa, Takuya. et.al. 2012.

 A liposomal eye drop formulation for retinal delivery needs to fulfill
several key requirements

1) high efficiency of drug entrapment ;

(2) particle size should be controlled to around 100 nm;

(3) the drug leakage rate from the liposomes should be slow in the tear fluid ;

(4) the liposomal formulation should be chemically and physically stable during storage. In the present study, we
demonstrate the feasibility of surface modification of liposomes in liposomal eye drop formulations. A weakly acidic
form of diclofenac was incorporated into liposomes by the calcium acetate gradient method.To improve physical
stability, the liposomal surface was modified with hydrophilic polymers. The delivery efficiency of diclofenacloaded
liposomes to the retina was compared with its molecular form (Diclod ophthalmic solution) after eye drop
administration to
rabbits

Fujisawa, Takuya. et.al. 2012.

 PREPARATION OF DICLOFENAC-LOADED LIPOSOMES

• Diclofenac-loaded liposomes were prepared by using the calcium acetate gradient method described previously (Hironaka et al.,2011).

• Thin lipid film composed of DSPC and cholesterol (8:1 molar ratio) was hydrated with calcium acetate solution (120 mM).The resulting
multilamellar vesicles were freeze–thawed and downsized using an extruder (LipoFastTM-Pneumatic; AvestinInc., Ottawa, Canada) with a
polycarbonate membrane (poresize: 0.1 m)

• The external liposome medium was replaced with HBSS–Mes buffer (pH 6.0) in two dialysis steps. For surface modification of
liposomes, PVA 205 or PVA-R was dissolved in HBSS–Mes buffer and mixed with an equal volume of liposome suspension. Diclofenac
was subsequently added into the liposome suspension and incubated at 37◦C for 10 min

• Conventional liposomes were prepared as a reference by a hydration method. The lipid film was hydrated with diclofenac (0.1%) dissolved
in HBSS–Hepes buffer (pH 7.4)

Fujisawa, Takuya. et.al. 2012.

CHARACTERIZATION OF LIPOSOMES

• The particle size and zeta potential of liposomes were measured by Zetasizer Nano ZS (Malvern, UK). The entrapment
efficiency was determined by the method described in the previous report (Hironaka et al., 2011). Release of diclofenac
from the liposomes was determined in phosphate buffered saline (PBS) at pH 7.4. A 0.5 ml aliquot of liposomes containing
diclofenac was transferred to a dialysis bag. The bag was then immersed in PBS at 37◦C (49.5 ml). At a predetermined
time, 0.2 ml of the PBS was removed and replaced with fresh PBS of equal volume. The released diclofenac was
quantified by HPLC.
• To evaluate the stability of diclofenac-loaded liposomes, the samples were stored at 4◦C or 25◦C for periods of 30 or 60
days. Aliquots were extracted at regular intervals and particle size, zeta potential, and entrapment efficiency were
determined.

Fujisawa, Takuya. et.al. 2012.

IN VIVO TISSUE DISTRIBUTION STUDY
Japanese albino rabbits (Oriental Yeast Co. Ltd., Tokyo, Japan) weighing 2.0–2.5 kg were used. This experiment was carried
out in adherence to the Guidelines for Animal Experiments of Wakamoto Pharmaceutical Co. Ltd. A single dose containing 50
l diclofenac (0.1%) was dropped into the rabbit’s eyes.

After 30 min, the rabbits were sacrificed by intravenous injection of 50 mg/ml sodium pentobarbital. Aqueous humor
samples were collected using the paracentesis technique

The eyes were enucleated and immediately frozen in liquid nitrogen. The posterior segment of the enucleated eye was
dissected to obtain samples of retina–choroid

Drug levels were determined in retina–choroid and aqueous humor by LC–MS/MS (Waters Corporation, Milford, MA, USA)

Fujisawa, Takuya. et.al. 2012.

 RESULTS AND DISCUSSION

• Particle size and entrapment efficiency of diclofenac-loaded liposomes before and after the
drug loading process are listed in Table 1.

Fujisawa, Takuya. et.al. 2012.

• The release profiles of diclofenac from the liposomes were investigated in PBS at 37◦C (Fig. 1). During the
first hour more than 80% of the diclofenac was released from the formulation prepared by the hydration
method

Fujisawa, Takuya. et.al. 2012.

Fujisawa, Takuya. et.al. 2012.
Fig. 2. Differences in tissue distribution of diclofenac after eye drop administration of different formulations in rabbits. The concentration of
diclofenac in (A) posterior retina–choroid and (B) aqueous humor 30 min after eye drop administration. Data are shown as mean ± S.E.M. (n =
4). Phospholipid, polymer, and diclofenac concentration in the resultant liposome suspension was 20.4 mol/ml, 1.0%, and 0.1%, respectively

Fujisawa, Takuya. et.al. 2012.

 KESIMPULAN

• the calcium acetate gradient method provides a higher entrapment efficiency of diclofenac
compared to the conventional hydration method. Modification of the liposome with PVA
or PVA-R improved the physical stability of diclofenac-loaded liposomes. PVA-R-
liposomes showed effective retinal delivery of diclofenac by promoting non-corneal drug
penetration after eye drop administration
 DAFTAR PUSTAKA

• Priyanka, Beloshe. et.al., 2019. Niosomes: As novel drug delivery system. International Journal of Research
i Volume 4; Issue 2; March 2019; Page No. 73-78
• Varun, Thakur.et.al., 2005. Niosomes and Liposomes - Vesicular Approach Towards Transdermal Drug
Delivery. International journal of pharmaceutical and chemical sciences issn: 22775005
• Fujisawa, Takuya. et.al. 2012. Liposomal diclofenac eye drop formulations targeting the retina:
Formulation stability improvement using surface modification of liposomes. International Journal of
Pharmaceutics 436 (2012) 564– 567