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Pharmaceutical Nanotechnology, 2019, 7, 1-16 1

REVIEW ARTICLE

Nanotechnology in Insulin Delivery for Management of Diabetes

Fatemah Bahman, Khaled Greish* and Sebastien Taurin*

Department of Molecular Medicine, Princess Al-Jawhara Centre for Molecular Medicine, School of
Medicine and Medical sciences, Arabian Gulf University, Manama, Kingdom of Bahrain

Abstract: Diabetes is a group of diseases characterized by hyperglycemia and originating

ARTICLEHISTORY
Received: January 26, 2019
Revised: February 22, 2019
Accepted: March 18, 2019
DOI:
10.2174/2211738507666190321110721
from the deficiency of or resistance to insulin, or both. Ultimately, the most effective
treatment for patients with diabetes involves subcutaneous injections of insulin. However,
this route of administration is often painful and inconvenient, as most patients will have to
self-administer it at least twice a day for the rest of their lives. Also, infection, insulin pre-
cipitation, and either lipoatrophy or lipohypertrophy are frequently observed at the site of
injection. To date, several alternative routes of insulin administration have been explored,
including nasal, pulmonary and oral. Although oral delivery of insulin is an ideal route for
diabetic patients, there are several limitations that have to be overcome such as the rapid
loss of insulin in the gastric fluid and low oral bioavailability. Numerous strategies have
been carried out to improve these limited parameters such as the use of enzyme inhibitors,
absorption enhancers, mucoadhesive polymers and chemical modification for receptor-
mediated absorption. Also, insulin-loaded nanocarriers bypass several physiological barri-
ers. This current review focuses on the various barriers existing in the delivery of insulin
through the oral route and the strategies undertaken so far to overcome those obstacles us-
ing nanocarriers as a potential vehicle of insulin.

Keywords: Diabetes mellitus, insulin analog, insulin, nanoparticles, nasal delivery, oral delivery, pulmo-
nary delivery.

1. INTRODUCTION producing β-cells in the pancreas [1, 3]. T2DM is a

Diabetes mellitus (DM) is a metabolic syn-
drome regrouping a plethora of chronic patholo-
gies characterized by hyperglycemia and denoting
the insufficient secretion and or action of insulin
[1]. DM is mainly classified into type I (T1DM)
and II (T2DM). Other types of DM, including ges-
tational diabetes and secondary forms of diabetes,
are less common [2]. T1DM is an autoimmune
condition mediated by the destruction of insulin-
*Address correspondence to these authors at the College of
Medicine and Medical Sciences, Department of Molecular
Medicine, and Nanomedicine Unit, Princess Al-Jawhara
Center for Molecular Medicine, Arabian Gulf University,
Manama, Kingdom of Bahrain; Tel: +973 17 237 393; and
+973 17 237; E-mails: khaledfg@agu.edu.bh;
sebastient@agu.edu.bh
non-insulin dependent DM and has been associated
to a combination of genetic, physiologic and envi-
ronmental factors such as the age, obesity, and the
lack of physical activities [4, 5]. TD2M arises
from the peripheral resistance to insulin, leading to
insulin overproduction by β-cells and or a relative
insulin deficiency due to impaired β-cell function
[6]. The global prevalence in adults has been in-
creasing over the last decades as 8.8% of the adult
population worldwide has developed DM [7].
T1DM represents 7-12% of the DM cases world-
wide [8], while T2DM is more common and has
increased by nearly 5-folds over the last 40 years
and is estimated to affect more than 500 million
people worldwide in 2018 [9, 10]. T2DM is ex-
pected to further spread over the next few years
among the middle and low-income counties as

2211-7385/19 $58.00+.00 © 2019 Bentham Science Publishers

2 Pharmaceutical Nanotechnology, 2019, Vol. 7, No. 2 Bahman et al.

more people adopt a sedentary lifestyle (Table 1)
[11, 12]. In 2016, the World Health Organization
(WHO) estimated that DM was the 7th cause of
death worldwide, 1.6 million deaths were directly
related to DM [13]. Furthermore, the total health
care budget associated with DM increased from
the US $246 in 2007 to $673 billion in 2015 [14]
and is estimated to rise to $2.2 trillion by 2030
[15].
Table 1. Prevalence increased % of DM worldwide
in 2045 according to IDF 2018.
Prevalence
Area
Increased %
Asia
South East Asia
Middle East countries and North Africa
Africa
North America & Caribbean
South and Central America
Europe
2. INSULIN
In 1921, F.G. Banting, C.H. Best, and J.J.R.
Macleod were the first to discover insulin. In
1922, a 14-year-old patient with T1DM received
the first injection of a pancreas extract, transform-
ing a deadly condition into a manageable disease
[16]. The insulin hormone is a 51 amino acids pro-
tein [17], forming an “A” (21 amino acids) and
“B” (30 amino acids) chain connected with a di-
sulfide bond. Insulin is synthesized into a single
chain precursor called preproinsulin; the removal
of the signal peptide during insertion into the en-
doplasmic reticulum creates proinsulin [18]. The
proinsulin contains three domains, an amino-
terminal B chain, a carboxy-terminal A chain and
a connecting peptide called the C peptide. Within
the endoplasmic reticulum (ER), the C peptide is
cleaved by endopeptidases, thereby producing the
mature form of insulin. Then, both insulin and free
C peptide are packaged in the Golgi as a micro-
crystalline zinc insulin hexamer into secretory
granules which accumulate in the cytoplasm [19].
The stimulation of β-cell leads to the exocytosis of
insulin and diffusion into the islet blood capillaries
[18]. Physiologically, the secretion of insulin is
activated by the blood glucose level [19]. Some
amino acids such as glutamine and leucine can in-
directly influence β-cell insulin secretion. Leucine
can stimulate glutamate dehydrogenase, which
converts glutamate into α-ketoglutarate. Gluta-
mine, after being transformed into glutamate by
glutaminase and then α-ketoglutarate can enter the
citric acid cycle resulting in ATP production and
enhancement of insulin secretion [20]. Also, free
amino acids including alanine and glutamine, are
released into the blood and served as signals to
glucagon secretion. This results in the elevation of
blood glucose levels (BGLs), which then triggers
insulin release into the circulation [21, 22].
15%
3. DIFFERENT TYPE OF RECOMBINANT
84% INSULIN
110%
The insulin used for the treatment of DM was
156% provided initially from porcine or cow pancreas
35% extracts. However, the growing number of diabetic
patients and the concern over the development of
62%
allergic reaction led to the development of recom-
16% binant human insulin. In 1978, Genentech and City
of Hope National Medical Center produced the
human insulin by inserting the genes into Esche-
richia coli [23]. The first recombinant human insu-
lin, Humulin, was approved by the FDA in 1982
[24]. Recently, genetic engineering allowed the
development of insulin analogs to provide the
better control of blood glucose. There are various
human insulin analogs in the market as shown in
Table 2. Modifications made on the insulin back-
bone to produce the different types of insulin ana-
logs are shown in Fig. (1). Rapid-acting insulins,
such as Lispro [25], Aspart [26], and glulisine [27]
have a rapid onset of action of approximately 15-
30 min and a duration of action of about 4 to 5 h.
Short-acting insulins such as Actrapid, Humulin,
Hypurin and Neutral [28] have an onset of action
of around 30 min to 1 h and a duration of action of
approximately 6-8 h, with a peak at 2-4 h. Inter-
mediate-acting insulins such as Neutral Protamine
Hagedorn (NPH) [29] and Lente [30] have an on-
set of action of 1-2 h, a peak action of 6-10 h and
approximately 10-16 h duration of action. Long-
acting insulins such as an Insulin Detemir (desB30
insulin) [26, 31], Glargine and Ultralente [32] have
an onset of activity of 2 h, a peak of action of
about 6-20 h and a duration of around 36 h.
Nanotechnology in Insulin Delivery for Management of Diabetes Pharmaceutical Nanotechnology, 2019, Vol. 7, No. 2 3

Fig. (1). Amino acid sequence of human insulin and its analogs. a: Insulin lispro, the sequence changes from proline
B28 and lysine B29 to Lysine B28 and proline B29. b: Insulin glargine, sequence addition of arginine B31 and ar-
ginine B32, plus substitution alanine A21 to glycine. c: insulin aspart, substitution of proline B28 to aspartate B28
d: Insulin glulisine, substitutions of asparagine B3 to lysine B3 and lysine B29 to glutamate B29. e: Insulin detemir.
Acylation of lysine B29 with C14 fatty acid chain, and removal of threonine B30. Adapted from [33].

Table 2. Human insulin analogs on the market.

Onset (min-hrs) Duration (hrs) Chemical Structure

Rapid-acting insulin
Insulin Lispro 28(B)-L-lysine-29(B)-L-proline-human insulin
Insulin Aspart 15 - 30 min 4h-5h AspB28
Insulin glulisine 3(B)-Lys, 29(B)-Glu-human insulin
Short-acting insulin
Actrapid
Humulin
0.5 - 1 h 6h-8h
Hypurin
Neutral
(Table 2) contd…
4 Pharmaceutical Nanotechnology, 2019, Vol. 7, No. 2
Onset (min-hrs)
Intermediate-acting insulin
Neutral Protamine Hagedorn (NPH)
1 -2 h
Lente
Long-acting insulin
Insulin detemir
Insulin glargine 2h
Insulin Ultralente
4. DIFFERENT STRATEGIES FOR INSULIN
DELIVERY
Subcutaneous insulin delivery is currently the
preferred method of insulin delivery [34, 35]. Nu-
merous alternative strategies have been investigat-
ed for insulin delivery than subcutaneous route
including oral, rectal, ocular, buccal, intravaginal,
transdermal, pulmonary and nasal [36]. The intra-
muscular route (IM) of administration is also not a
reliable route as the IM delivery route cannot
mimic the regular secretion pattern of insulin and
leads to change the pharmacokinetic and pharma-
codynamic properties of insulin [37]. The IM insu-
lin injections could lead to hypoglycemia immedi-
ately after injection and hyperglycemia before the
next dose [38].
4.1. Subcutaneous Insulin Injections
Subcutaneous injections called preprandial in-
jections of rapid-acting insulin and predormital
injections of long-acting insulin are the regimens
for patients with T1DM to maintain BGL when the
preprandial blood glucose is 3.9 to 7.2 mmol/L (70
to 130 mg/dl) and the 2 h postprandial blood glu-
cose less than 10 mmol/L (<180 mg/dl) [1]. The
insulin is administered directly to the fatty layer
which is less vascularized and therefore slows the
absorption rate. However, non-adherence and poor
compliance are associated with insulin subcutane-
ous injections [39].
Insulin pumps usually work by delivering insu-
lin continuously through the subcutaneous route.
The pump mimics the body basal insulin release in
24 h cycle. Bolus insulin can be administered to
correct the high glycemic levels after meals. The
Bahman et al.
Duration (hrs) Chemical Structure
Proline at position 29 in the B-chain is replaced
10 h - 16 h by Lysine.
LysB29(Nε-tetradecanoyl) des(B30)
36 h Gly A21, Arg B31, Arg B32
advantages of this method include that the precise
insulin dosages can be administered and calculat-
ed. However, the disadvantages include mechani-
cal failure and inflammation at the injection site
[40]. Usage of the insulin pump is generally for
adults suffering from T1DM [41].
4.2. Nasal Insulin
Intranasal insulin delivery has advantages be-
cause the nasal cavity is the largest absorptive sur-
face area in the body accounting for approximately
150 cm2. Also, the mucosa is highly vascularized
and may promote the direct protein transport into
the systemic circulation bypassing the first-pass
metabolism by the liver and intestine [42]. How-
ever, several factors may delay the intranasal ab-
sorption such as nasal cavity clearance, infiltration
across the mucus coating and enzymatic degrada-
tion similar to oral insulin delivery [43]. Intranasal
delivery of insulin has been reported by using li-
pid-based particles including liposomes, micro-
emulsions and solid lipid nanoparticles (SLNs).
Furthermore, insulin-loaded liposomes showed an
effective absorption enhancer for the nasal insulin
in rabbits and reduced BGL for 8 h with an insulin
bioavailability of more than 13% [44, 45].
4.3. Pulmonary Insulin
Insulin is usually delivered to the lungs in pow-
der form and is delivered to the lungs with the help
of an inhaler. The mode of delivery is comparable
but not superior to the traditional subcutaneous
injectable route [34]. It is documented that inhaled
insulin can also achieve a normal BGL of 70 to
130 mg/dl [46]. The lungs are well perfused hence
increasing the surface area for absorption. Some of
Nanotechnology in Insulin Delivery for Management of Diabetes
the disadvantages are associated with the inhaled
insulin that can precipitate acute episodes of bron-
chospasm in asthmatic patients and may also cause
hypoglycemic episodes and throat irritation [47].
Liposomes loaded with insulin enhance the pul-
monary uptake of insulin and improve hypogly-
cemic effect when compared with the free insulin
[48]. Insulin-encapsulated in PLGA microcapsules
demonstrated a constant insulin release with a
longer period of 4 to 5 fold longer than respirable
spray dried insulin powder, as well as a stable gly-
cemic level up to 24 h after administration [49].
The Exubera® was the first inhaled product ap-
proved by the FDA in 2006. It is a dry powder
product available in 1 mg and 3 mg doses that can
be taken with the help of an Inhance™ inhaler de-
vice. The pharmacokinetic and pharmacodynamic
(PK/PD) properties were similar to the insulin As-
part with a 10-15 min onset of action [50]. In clin-
ical trials with uncontrolled T1DM and T2DM pa-
tients, Exubera showed a significant reduction in
postprandial BGL and Hb A1c [51]. Exubera was
discontinued by Pfizer in 2007 due to its high
price, huge size and concerns over its effect on the
lungs [52]. Afrezza is another dry powdered for-
mulation of inhaled insulin developed. Afrezza has
an onset of action of 15 min, duration of 2-3 h, and
was ideal for controlling postprandial BGL. There-
fore, it completed phase III clinical trials for pa-
tients with T1DM and T2DM and was approved
by the FDA in 2014 [53-55].
4.4. Oral Insulin
Oral insulin administration is the most conven-
ient and ideal route for DM. However, the oral bi-
oavailability of insulin is usually less than 10%,
and it is reported that less than 0.5% of orally ad-
ministered insulin enters the systemic circulation
[56]. Also, there are many challenges associated
the oral method delivery. Some of these challenges
include the activity of the proteolytic enzymes in
the GIT [57] and the poor bioavailability of oral
insulin due to the low permeability of the intestine
[58].
5. OVERCOMING PHYSIOLOGICAL BAR-
RIERS AND ENZYMATIC BARRIERS FOR
ORAL DELIVERY
New studies geared towards changing the struc-
ture of the insulin analogs to be more stable in
Pharmaceutical Nanotechnology, 2019, Vol. 7, No. 2 5
acidic condition. The gastric environment is char-
acterized by a harsh environment caused by
peptidase activities and the acidic pH [59, 60].
One study showed that HP55 modification, one
kind of hydroxypropyl methylcellulose phthalate,
an enteric coating, protected insulin against acidic
gastrointestinal conditions and then dissolved in
the small intestine environment. This modification
reduced the cumulative release rate of insulin in
simulated gastric fluid from 33.24 to 15.87%,
while cumulative release rates of insulin in simu-
lated intestinal fluid increased from 15 to 58.06%
[61, 62]. Also, different studies have confirmed
that the use of digestive enzyme inhibitors can
overcome this challenge such as sodium glycocho-
late, bacitracin, soybean trypsin inhibitors, apro-
tinin, and camostat mesilate [63]. One approach
uses different types of protease inhibitors such
aprotinin, soybean trypsin inhibitors, and camostat
mesilate may result in the deficiency of these en-
zymes in humans if they administrated for long
period [64, 65]. One study found that tryp-
sin/chymotrypsin inhibitor aprotinin was used to
shield insulin and protect it from degradation [66].
Also, a new class of enzyme inhibitors called
chicken and duck ovomucoids has been docu-
mented, these formulations offered complete pro-
tection against the action of both enzymes trypsin
and chymotrypsin [67]. Moreover, polymer inhibi-
tor conjugates such as carboxymethyl cellulose-
elastin (CMC-Ela) have demonstrated protection
against enzymes trypsin and elastase in vitro. After
4 h of incubation, about 33% of the therapeutic
protein was found to be active against the elastase
[68]. Another approach to protect proteins is using
Serpin (Serine protease inhibitor) that forms cova-
lent complexes with the protease [69].
6. DIFFERENT FORMULATIONS FOR
ORAL INSULIN NANOPARTICLES
Nanoparticles provide potent tools for the oral
delivery of insulin such as 1) insulin encapsulation
within nanoparticle carriers protect it from acidic
pH and enzymatic degradation in the gastrointesti-
nal tract (GIT); 2) the mucoadhesive coating of the
nanoparticles increases the efficient drug delivery
near the site of absorption in the intestine; 3)
mucus-penetrating particles or coated insulin na-
noparticle may improve transportation of insulin
6 Pharmaceutical Nanotechnology, 2019, Vol. 7, No. 2
and 4) stabilization of the poorly soluble insulin by
encapsulation into nanoparticle such as PLGA [57,
70].
6.1. Nano-Medicine and Nano-Particles (Nano-
Carriers) that used for Insulin
Recently, nanoparticles were developed to
promote the oral delivery of insulin. The nanopar-
ticles were either nanospheres where the insulin is
uniformly distributed within the polymeric matrix
or nanocapsules where the insulin is within the
membrane polymers (polymeric film) [71-73]. Ta-
ble 3 presents some of these nanoparticles used for
the oral delivery of insulin.
6.1.1. Natural Polymers
Polysaccharides (chitosan, dextran, and algi-
nate) and proteins (casein and gelatin) are the most
known naturally occurring polymers used in the
oral delivery of insulin. They are hydrophilic, non-
toxic, biocompatible and biodegradable polymers.
6.1.1.1. Chitosan-Insulin Nanoparticles
Chitosan is a type of a linear cationic non-toxic
polysaccharide, that adheres to the mucosal sur-
face, hence causing the opening up of the tight epi-
thelial junctions [74]. Chitosan has a strong elec-
trostatic affinity towards mucin, is the only natural
polysaccharide with good biodegradability, bio-
compatibility, and has an excellent adhesion prop-
erty [75]. The encapsulation of insulin in chi-
tosan/heparin nanoparticles demonstrated a normal
glycemic control [76]. This study used a dose of
60 IU/kg that lowered the mean BGL by 30% for 2
h after insulin administration and lasted for 4 h;
also it provided 50% lower BGL at 3 h after ad-
ministration that lasted for 5 h [76]. Another study
showed that chitosan nanoparticles encapsulating
insulin (CS-NPs) had a size between 265.3 ± 34 to
387.4 ± 35.6 nm, and administration of 21 IU/kg
resulted in the hypoglycemic effect that prolonged
up to 15 h and an average pharmacological bioa-
vailability relative to subcutaneous injection of
insulin up to 14.9% [77].
Lin et al. developed a nanoparticle of chitosan
within Poly (γ-glutamic acid) sized between 110 to
150 nm that resulted in a hypoglycemic effect after
oral administration of doses of 15 and 30 IU/kg,
which showed a significant decrease of BGL that
Bahman et al.
persisted up to 10 h in diabetic rats [78]. Cui et al,
formulated an insulin pH sensitive carboxylated
chitosan grafted poly(methyl methacrylate) nano-
particles that lowered BGL after oral administra-
tion at 50 and 25 IU/kg and the pharmacological
bioavailability was 9.7% at 25 IU/kg, 4.4% and
3.2% at 50 and 100 IU/kg respectively, in diabetic
rats [79].
Elsayed et al., prepared insulin-chitosan com-
plexed with oleic acid, plurol oleique as cosurfac-
tant and labrasol as a surfactant and showed a sig-
nificant hypoglycemic effect in diabetic rats after
oral insulin dose of 50 IU/kg throughout 24 h [80].
6.1.1.2. Alginate
Alginate is a nontoxic and biodegradable nano-
carrier obtained from the marine brown algae. It is
composed of 1,4-linked-β-D-mannuronic acid and
α-L-guluronic acid residues that form polyelectro-
lyte complexes with polycations such as chitosan.
Insulin-loaded nanoparticles were prepared by
the ionotropic pre-gelation of alginate with calci-
um chloride followed by complexation between
alginate and chitosan and led to the formation of
nanoparticles with a size of 748 nm [81]. The
pharmacological effect of these nanoparticles was
evaluated in diabetic rats at 50 and 100 IU/kg dos-
es and showed a significant reduction in BGL by
more than 40% with 50 IU/kg and 100 IU/kg dos-
es, while the hypoglycemic effect was sustained
over 18 h [82].
The alginate/chitosan nanoparticles that form
complexes with cationic β-cyclodextrin polymers
will be able to protect insulin against degradation
in simulated gastric fluid [83]. Also, insulin-
loaded dextran sulfate/chitosan nanoparticles with
a size of 527 nm, when given orally at a dose of 50
and 100 IU/kg resulted in low BGL of 35%. The
rats experienced the hypoglycemic effects for over
18 h [82].
Ma et al. prepared a chitosan insulin nanoparti-
cle composed of chitosan, and TPP (pentasodium
tripolyphosphate) sized 269 to 688 nm, respective-
ly. These nanoparticles were effective in lowering
the BGL of diabetic rats when taken orally at insu-
lin doses of 50 and 100 IU/kg. In particular, the
oral insulin dose of 100 IU/kg was able to main-
tain the BGL for at least 11 h [84].
Nanotechnology in Insulin Delivery for Management of Diabetes Pharmaceutical Nanotechnology, 2019, Vol. 7, No. 2 7

Table 3. Different types of oral insulin nanocarrier.

Route of Dose (I.U./kg)

Nano-Carrier Components Size (nm)
Administration (min/max)

Carboxylated chitosan + methyl

251 to 319 Oral 15 (min)- 100 (max)
methacrylate
Polysaccharide chitosan nanoparticules 265.3 ± 34 to
Oral 21 (max)
(CS-NPs) 387.4 ± 35.6
Chitosan + Oleic acid + Plurol oleique +
108 Oral 50
Labrasol
25 (min)
Chitosan + Alginate 748 Oral
100 (max)
50 (min)
Chitosan + Dextran sulfate 527 Oral
100 (max)

Chitosan-insulin Chitosan + TPP (pentasodium 7 (min)

250 to 400 Oral
tripolyphosphate) + Poloxamer 188 21 (max)
50 (min)
Chitosan + TPP(pentasodium
269 to 688 Oral
tripolyphosphate) 100 (max)

15 (min)
Chitosan + Poly(ç-glutamic acid) 110 to 150 Oral
30 (max)
Chitosan + Alginate + Calcium chloride + 25 (min)
Labrafac CC + Phospholipid + Span 80 + 488 Oral
Cremorphor EL 50 (max)

Chitosan + γ-PGA 185.1 to 198.4 Oral 30

PLGA + Phospholipid + PVA 102 to 428 Oral 20
PLGA + Hp55 169 Oral 20
PLGA + Chitosan + Pluronic 188 134.4 Oral 15
PLGA-Insulin
PLGA + Pluronic 188 121.3 Oral 15
PLGA + Sodium oleate + PVA 161 Oral 20
PLGA + PEG + Folate ∼260 Oral 50
25 (min)
PCL-Insulin PCL and Eudragit® RS 331 Oral
100(max)
Dextran + Epichlorohydrin + vitamin
160 to 250 Oral 20
B(12)
Dextran + Alginate + Poloxamer + Chito-
Dextran- Insulin 396 Oral 50
san + BSA

Dextran + Alginate + Chitosan + PEG + >1842 (90 %) 25 (min)

Oral
BSA >812 (50 %) 100 (max)
(Table 3) contd…
8 Pharmaceutical Nanotechnology, 2019, Vol. 7, No. 2 Bahman et al.

Route of Dose (I.U./kg)

Nano-Carrier Components Size (nm)
Administration (min/max)

Isopropyl myristate + Labrasol + Plurol

200 to 400 Oral 100
Oleique + butyl (2) cyanoacrylate

Polyalkylcy- Polybutylcyanoacrylate + Tween 20 78 Oral 50

anoacrylate-insulin Polybutylcyanoacrylate + Tween 20 +
67 Oral 50
Soyabean oil + vitamin E
Isobutyl cyanoacrylate (IBCA)+ insulin 145 Oral 100
Lecithin + stearic acid + ploxamer + wheat
germ agglutinin-N-glutamyl-phosphatidyl- 75.3 Oral 50
ethanolamine
Stearic acid/octadecyl alcohol/cetyl palmi-
Solid lipid-insulin tate/glyceryl monostearate/glyceryl palmi- 213 to 444.8 Oral 50
tostearate/glyceryl tripalmitate/glyceryl
behenate
Witepsol 85E 243 ± 10 Oral 25
Witepsol 85E + chitosan 470 ± 32 Oral 25
N-trimethyl chitosan chloride +
Targeted insulin 342 Oral 50
CSKSSDYQC peptide +
nanoparticle
PLA-PEG + human polyclonal IgG Fc 63 Oral 1.1

Li, X et al. prepared a nanoparticle by prepar-
ing an insulin solution composed of chitosan, algi-
nate and calcium chloride supplemented to the oily
phase which was composed of 68.5% Labrafac
CC, 25% Span™ 80, and 6.5% phospholipid; Then
this emulsion was kept into an aqueous solution
containing 3% Cremophor EL (castor oil deriva-
tive) and low-viscosity sodium alginate and when
administered orally at the insulin doses of 12.5
IU/kg, 25 IU/ kg, and 50 IU/kg, it resulted in lower
BGL and pharmacological bioavailability of 25
and 50 IU/kg was 8.19% ± 0.58% and 7.84% ±
0.29%, respectively [85]. Lin Y et al,. prepared a
nanoparticle composed of chitosan and poly (ç-
glutamic acid) sized 185.1 to 198.4 nm that
showed a reduction in BGL at a dose of 30 IU/kg
and hypoglycemic effects were present for up to
12 h [78].
6.1.1.3. Dextran-Insulin Nanoparticles
The dextran sulfate is an exocellular bacterial
polysaccharide that is a nontoxic, highly water-
soluble, biodegradable and biocompatible poly-
mer. Dextran is chemically coupled to insulin and
acts by protecting the insulin against the GIT pro-
teases [86]. Dextran is normally conjugated to vit-
amin B-12, then when this complex reaches the
intestines, it escapes the obstacles in the digestion
system [72, 87].
Chalasani et al. developed a VB12 nanoparticle
system that improves the uptake capacity of both
nanoparticles and VB12 transport to deliver insu-
lin orally. The results of this study concluded that
20 IU/kg of insulin conjugated with VB12-
Dextran NPs showed a significant BGL reduction
up to 24 h [87].
Reis et al. created an oral insulin nanoparticle
based on alginate‐dextran sulfate core, capsulated
with a chitosan‐polyethylene glycol albumin shell.
Albumin was supplemented to avoid protease ac-
tion on the destruction of insulin and chitosan be-
cause of mucoadhesive properties, while PEG
helped the stabilizer to improve the insulin half-
life. Chitosan-PEG-albumin nanoparticle mini-
mized the BGL up to 70%. Therefore, the insulin‐
loaded chitosan-PEG-albumin nanoparticle with
25, 50, 100 IU/kg administered orally to diabetic
rats caused a reduction in BGL, lasted for 24 h
with a maximal effect after 14 h [88].
Nanotechnology in Insulin Delivery for Management of Diabetes
Woitiski et al. produced a multilayer nanoparti-
cle sized 396 nm that consisted of alginate and
dextran sulfate nucleating around calcium and
binding to poloxamer, stabilized by chitosan, and
coated with albumin. This nanoparticle adminis-
tered orally to diabetic rats (50 IU/kg) reduced
BGL to 40% and continued hypoglycemic effect
for over 24 h [89].
6.1.2. Synthetic Polymers
Poly ɛ-caprolactone (PCL), poly (lactic-co-
glycolic acid) (PLGA) and polylactides (PLA) are
synthetic polymers and were used in oral insulin
delivery due to their good biocompatibility and
biodegradability.
6.1.2.1. Poly (Lactic-co-Glycolic Acid) (PLGA)
Poly (lactic-co-glycolic acid) (PLGA) is an ali-
phatic polyester synthetic biodegradable biopoly-
mer. Yang et al. prepared PLGA nanoparticles
loaded with insulin. The administration of insulin-
loaded PLGA nanoparticles induced a rapid drop
in BGL for up to 24 h [90]. PLGA has the ad-
vantage of passing through the GIT and be ab-
sorbed by the Peyer’s patches by the M cells in the
intestine. PLGA nanoparticles have been exten-
sively used because of their encapsulating proper-
ties and their ability to release a drug in a con-
trolled way [49]. Although there is a prolonged
release with the use of PLGA nanoparticles, it has
been challenging how to monitor the amount of
drug released per day and compromise the glyce-
mic control [91]. PLGA has shown good bioavail-
ability when administered with chitosan [92].
PLGA contributes to the stability of the colloidal
system and its transport across the mucosal surfac-
es [93].
Cui F et al. prepared a novel nanoparticle com-
posed of dichloromethane (DCM). Ethyl acetate
(EAc) containing 2% polymer (w/v) was added to
the insulin -phospholipid (Ins-SPC) complex, and
poured into an aqueous solution containing 2%
Poly (vinyl alcohol), in which soybean phosphati-
dylcholine (SPC) was used to improve the lipo-
solubility of insulin. This formulation showed that
oral administration of 20 IU/kg (Ins-SPC) nano-
particles reduced the fasting BGL to 57.4% within
the first 8 h and this continued for 12 h [94].
Pharmaceutical Nanotechnology, 2019, Vol. 7, No. 2 9
Also, the same group prepared PLGA nanopar-
ticles (PNP) and PLGA-Hp55 (PHNP) nanoparti-
cles sized 150 ± 17 and 169 ± 16 nm, respectively,
and resulted in an effective reduction in BGL after
oral administration [95]. Zhang et al. developed a
mucoadhesive PLGA nanoparticle coated with chi-
tosan or Eudragit® RS to enhance the oral bioa-
vailability. These nanoparticles were prepared by
water-in oil-in-water solvent evaporation tech-
nique; insulin was dissolved in an aqueous solu-
tion of Pluronic 188, PLGA and an aqueous solu-
tion of Pluronic 188, to form a double emulsion
(w/o/w). This preparation was tested orally on dia-
betic animals which resulted in hypoglycemia, and
pharmacological availability of both nanoparticles,
PLGA-NP and CS-PLGA-NP, relative to the sub-
cutaneous injection was calculated to be 7.6% and
10.5%, respectively, at the dose of 15 IU/kg [96].
Wu Z et al. concluded that the pharmacological
availability of the 50 IU/kg Eudragit® RS (RS)
coated PLGA nanoparticle was 9.2% [97].
Sun et al. prepared an insulin-S.O (Ins-S.O) co-
mplex by a hydrophobic ion pairing (HIP) method
of size 161 nm. These nanoparticles reduced BGL
to 23.85% for up to 12 h post-oral administration
and the effect was prolonged for 24 h [98].
Jain S et al. showed that the oral administration
of (50 IU/kg) of insulin encapsulated in folate-
(FA) coupled with polyethylene glycol (PEG)
ylated polylactide-co-glycolide (PLGA) nanoparti-
cles (NPs; FA-PEG-PLGA NPs) exhibited a hypo-
glycemic effect for 24 h [99].
6.1.2.2. Polylactide (PLA)
PLA has similar characteristics to PLGA but
they are hydrophobic, and they degrade more
slowly because of the crystallinity (100). As men-
tioned before, Cui et al. reported enhanced insulin
efficiency (up to 90%) in PLA and PLGA nano-
particles, where insulin was encapsulated with
phosphatidylcholine (SPC) to improve the lipo-
solubility and oral bioavailability of 7.7% [94].
6.1.2.3. Poly-ɛ caprolactone (PCL)
It is a biodegradable polyester polymer with a
semi‐crystalline feature. It has superior viscoelastic
properties and the advantage that it generates a
less acidic environment during degradation as
10 Pharmaceutical Nanotechnology, 2019, Vol. 7, No. 2
compared with PLGA [100]. Damgé et al. group
developed a nanoparticle from a mixture of poly-
ester poly (ε-caprolactone), and a polycationic
acrylic polymer (Eudragit® RS) sized 331nm. The-
se carriers used for delivering insulin orally proved
a significant prolonged hypoglycemic effect in
both diabetic and normal rats [101].
6.1.3. Polyalkylcyanoacrylate-Insulin Nanoparti-
cles
Initially, it was used in tissue “glue” during
surgery. However, many studies have demonstrat-
ed its importance in the delivery of insulin orally,
because of its stability and its biodegradable char-
acter [86]. It is a polymeric colloidal drug carrier,
with isobutyl cyanoacrylate nanocapsules with less
than 300 nm in diameter. These nanoparticles can
pass the intestinal epithelium cells and protect in-
sulin from proteolytic degradation [102]. Also, it
was designed by interfacial emulsion polymeriza-
tion of a cyanoacrylic monomer where the oil
phase (Miglyol 812) containing insulin was mixed
to an aqueous medium in the presence of polox-
amer 188 (surfactant agent). Damgé et al. group
prepared insulin nanospheres composed of a pol-
ymeric carrier formed by polymerization of isobu-
tyl cyanoacrylate (IBCA) in an acidic pH of size
145 nm. These nanocapsules showed a hypogly-
cemic effect after 2 h of nanosphere administration
of 100 IU/kg [102].
Hou et al. group prepared two formulations of
insulin-polybutylcyanoacrylate nanoparticle (IPN)
in soybean oil containing 0.5% (v/v) Tween-20
and 5% (v/v) Vitamin E (size 67 nm) and poly-
butylcyanoacrylate containing 0.5% (v/v) Tween-
20 (size 78 nm), which showed the hypoglycemic
effect following oral administration of (50 IU/kg)
to diabetic rats [103].
Graf et al. group prepared a nanoparticle of in-
sulin in poly(alkyl cyanoacrylate) nanoparticles
composed of isopropyl myristate, caprylocaproyl
macrogolglycerides, polyglyceryl oleate and insu-
lin of size 200-400nm that showed a hypoglycemic
effect for up to 36 h after oral administration of
100 IU/Kg [104].
6.1.4. Solid Lipid-Insulin Nanoparticles (SLN)
In 1990, colloidal nanocarriers prepared from
solid lipids have been used as nanoparticles [105].
Bahman et al.
The main advantage of SLNs as nanocarrier sys-
tems is their physiological lipid components that
reduce the risk of toxicity and protect the proteins
from degradation by GIT enzymes [106].
Zhang et al. prepared lectin-modified SLN to
entrap insulin. These SLNs were further modified
with wheat germ agglutinin-N-glutaryl-phospha-
tidylethanolamine (WGA). Both formulations pro-
tected insulin from degradation by GIT enzymes.
Furthermore, a reduction in BGL following oral
gavage was observed in rats with both formula-
tions. The pharmacological bioavailability follow-
ing oral administration of insulin-SLNs and WGA-
modified insulin-SLNs was 4.46% and 6.08%, re-
spectively [107].
Fonte et al. developed chitosans covered with
Witepsol 85E solid lipid nanoparticles (SLN) to
encapsulate insulin. The diameters of SLN and
chitosan-SLN were 243 ± 10 nm and 470 ± 32 nm,
respectively. The oral administration of chitosan-
coated insulin SLN to diabetic rats decreased the
BGL up to 24 h [108].
Yang et al. group created a solid lipid nanopar-
ticle composed of stearic acid, octadecyl alcohol,
cetyl palmitate, glyceryl monostearate, glyceryl
palmitostearate, glyceryl tripalmitate and glyceryl
behenate of size about 213 to 444.8 nm. This for-
mulation showed a hypoglycemic effect that lasted
for 24 h after oral administration of 50 IU/kg
[109].
More recently, Xu et al. developed an insulin-
loaded SLN with endosomal escape peptide. The
oral administration of this insulin-loaded SLN (50
IU/kg) transiently decreased BGL in diabetic by
nearly 35% after 3 h which was still low by almost
20% after 12 h. In comparison, insulin (5 IU/kg)
given subcutaneously decreased BGL by 70% af-
ter 2 h but returned to its original level after 12 h
[110].
6.1.5. Targeted Insulin Nanoparticles
Jin et al. developed a goblet cell-targeting na-
noparticle that consisted of N-trimethyl chitosan
chloride (TMC) modified with a CSKSSDYQC
peptide (CSK) targeting insulin. The results of in
vivo studies on diabetic animals showed better hy-
poglycemic effect 1.5 fold compared to the un-
modified formulation [111].
Nanotechnology in Insulin Delivery for Management of Diabetes
Pridgen et al. prepared PLA-PEG, and human
polyclonal IgG Fc targeted to the neonatal Fc re-
ceptor (FcRn) of size 63 nm. This nanoparticle
showed a significant hypoglycemic effect in dia-
betic mice after oral administration [112].
7. ORAL INSULIN NANOFORMULATIONS
IN CLINICAL TRIALS
Over the last few years, laboratories have de-
veloped various nanoformulations to improve the
oral delivery of insulin. However, the biocompati-
bility of these nanocarriers, low oral bioavailabil-
ity and large interindividual variations in insulin
absorption have limited their transition to the clin-
ic. Few oral insulin formulations developed by
pharmaceutical companies are now being assessed
in clinical trials (Table 4) [113, 114]. Oral-lyn is
an insulin spray, developed by Generex Biotech-
nology, that has been approved in Ecuador and
Lebanon but still in phase III clinical trials in Eu-
rope, Canada, and the USA. Oral-Lyn was granted
approval by the FDA under the Investigational
New Drug (IND) program for the treatment of
T1DM and T2DM patients with severe or life-
threatening conditions [115]. The Oral-lyn was
developed for delivery of insulin via the buccal
mucosa to bypass the liver and GI tract. The for-
mulation consists of insulin-loaded micelle with a
size above 7 µm and a surfactant to enhance ab-
sorption [115, 116]. The other formulations devel-
oped for oral delivery of insulin are still in early
phases I or II. IN-105 developed by Biocon is a
modified insulin conjugated through a spacer to a
small polyethylene glycol molecule which im-
proves its stability. The drug is currently in phase
II clinical trial [117]. Diasome Pharmaceutical has
developed an insulin-loaded liposome harboring
on its surface a biotin-phosphatidylethanolamine
to target the liver [115]. The insulin-loaded
hepatocyte-directed vesicle (HDV-I) has a diame-
ter below 150 nm and is stable in the blood and the
upper GI. The low amount of insulin in each cap-
sule (5U) and the targeting to the hepatocytes are
designed to maximize efficiency and decrease the
risk of insulin overdosing [116]. HDV-I was as-
sessed in several clinical trials in phase II and one
in phase III for T1DM and T2DM (Table 4). NOD
pharmaceuticals developed Nodlin which consists
of insulin nanoparticles embedded in bio-adhesive
Pharmaceutical Nanotechnology, 2019, Vol. 7, No. 2 11
calcium phosphate enteric-coated capsules. The
formulation was tested in phase I clinical trial
[118]. Macrulin is a lecithin-based microemulsion
developed to enhance the bioavailability of insulin
[119]. Macrulin is currently in phase II clinical
trial in Europe for the treatment of T1DM and
T2DM. Jordanian Pharmaceutical Manufacturing
Co has developed an oral liquid insulin delivery
system in which insulin-loaded chitosan nanopar-
ticles are dispersed in an oily vehicle [120]. The
preparation was assessed in phase I clinical trial
and demonstrated promising bioavailability [121].
Novo Nordisk A/S developed several insulin for-
mulations utilizing Merrion’s GIPET® Technology
consisting of a lipid mixture or lipid microemul-
sion with a surfactant in an enteric-coated gel cap-
sule [122]. The various formulations were tested in
phase I clinical trials for patients with T2DM (Ta-
ble 4). The assessment of NN1952 ceased due to
the interaction with the food intake [123]. Oshadi
Drug Administration Ltd. has developed Icp-
insulin, proinsulin, and C-peptide in Oshadi carrier
for the oral delivery of insulin [115]. The Oshadi
carrier is composed of pharmacologically inert sil-
ica nanoparticles with a hydrophobic surface and
branched polysaccharide [115]. The formulation was
assessed in phase I and II clinical trials (Table 4).
In addition to the new era of nanomedicine to
treat diabetes, few alternative insulin delivery sys-
tems are being developed such as a glucose-
responsive closed-loop insulin delivery system via
a painless microneedle array patch that contains
insulin encapsulated vesicles sensitive to both hy-
poxia and hydrogen peroxide [124]. Another sys-
tem conjugated glucosamine and insulin (Glc- in-
sulin) to promote binding on red blood cell mem-
branes. The binding is reversible and releases insu-
lin in the setting of hyperglycemia [125].
CONCLUSION
Oral insulin administration constitutes a tre-
mendous challenge to the management of DM and
the improvement in patient compliance. Nanomed-
icine-based drug delivery systems are designed to
overcome barriers including the enzymatic degra-
dation, low intestinal permeability and bioavaila-
bility, which may contribute to overcome these
challenges. This review has presented several
nanocarriers and strategies tested in vivo for the
12 Pharmaceutical Nanotechnology, 2019, Vol. 7, No. 2 Bahman et al.

Table 4. Lists of oral insulin formulation systems in different clinical phases.

S. No. Company Name Product Name Product Technology Action Development Phase

1 Generex Oral-Lyn ™ Micelle-Penetration enhancers Short III (NCT00668850)

Biocon/
2 IN-105 PEG Conjugated insulin Short II (NCT03430856)
Bristol-Myers Squibb
II (NCT02794155)
Diasome Pharmaceuticals, HDV-insulin Hepatic-directed vesicle-insulin II-III (NCT00814294)
3 Short
Inc (HDV-1) (liposomal nanocarrier) II (NCT03156361)
II (NCT03096392)
Insulin nanoparticles with
NOD Pharmaceuticals, I (ChiCTR-TRC-
4 Nodlin bioadhesive nanoencapsulation Intermediate
Inc./ Shanghai Biolaxy, Inc 12001872)
(NOD Tech)
5 Provalis PLC Macrulin ™ Lipid-based microemulsion Short II
Jordanian Pharmaceutical Liquid delivery system with
6 JPM oral insulin I
Manufacturing Co. PLC insulin-chitosan nanoparticles
Insulin analog with an oral delivery
7 Novo Nordisk A/S NN1952 Short I (NCT01028404)
system GIPET®
I (NCT01334034)
OI338GT Insulin analog with an oral delivery
8 Novo Nordisk A/S Long I (NCT01931137)
(NN1953) system GIPET®
I (NCT01796366)
OI362GT Insulin analog with an oral delivery
9 Novo Nordisk A/S Long I (NCT01597713)
(NN1954) system GIPET®
OI287GT Insulin analog with an oral delivery
10 Novo Nordisk A/S Long I (NCT01809184)
(NN1956) system GIPET®
I (NCT01120912)
Oshadi Drug Insulin, proinsulin, and C-peptide in
11 Oshadi Icp. Short II (NCT01973920)
Administration Ltd. Oshadi carrier
I-II (NCT01772251)

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