Nurul Syakirah Nor Azman 201680724

Rabiatul Adawiyah Ali 2016807248

Ruzanna Aini Zulkifli 2016807172
Safwanah Rahman 2016648398
Siti Zuhairah Zainuddin 2016807378
Advantages of Oral
Liraglutide

Active Ingredient

• Pharmacological property
• Physicochemical property

Strategies to develop Oral

Liraglutide
• Excipients & formulation
• Manufacturing process
• Quality Control
Formulate Liraglutide that is suitable for oral
consumption
 Injection Oral

pain at the site of injection no pain

inconvenience of multiple injections no contamination

insulin lipodystrophy cost-effectiveness

hyperinsulinemia better mimics insulin pathway

• Ansari MJ. Oral delivery of insulin for treatment of diabetes: classical challenges and current opportunities. Journal of Medical Sciences. 2015 Jul 1;15(5):209-20.
• Fonte P, Araújo F, Reis S, Sarmento B. Oral insulin delivery: how far are we?. Journal of diabetes science and technology. 2013 Mar;7(2):520-31.
• Gowthamarajan K, Kulkarni GT. Oral Insulin—Fact or Fiction?. Resonance. 2003 May 1;8(5):38-46.
 Limitation of Oral Insulin Delivery
poor
prone to slow onset of
absorption in
degradation action
GIT

imprecise
inaccurate
glycemic
dosing
control
• Ansari MJ. Oral delivery of insulin for treatment of diabetes: classical challenges and current opportunities. Journal of Medical Sciences. 2015 Jul 1;15(5):209-20.
• Fonte P, Araújo F, Reis S, Sarmento B. Oral insulin delivery: how far are we?. Journal of diabetes science and technology. 2013 Mar;7(2):520-31.
• Gowthamarajan K, Kulkarni GT. Oral Insulin—Fact or Fiction?. Resonance. 2003 May 1;8(5):38-46.
Active Ingredient:
GLP-1 analogues
MOA: GLP-1 receptor agonist
Marketed: Liraglutide (injection)
Class : Incretin Mimetics
Molecular Formula : C172H265N43O51

Prepared by: Safwanah Rahman

Why want to make Liraglutide (GLP-1
analogues) in oral form?
Why not other class of antidiabetic drugs?
Adherence issue to insulin pen: cost
Doesn’t cause hypoglycemia
How to synthesize oral
analogues of GLP-1:

SARs
Our analogue
of oral GLP-1

X
 Pharmacokinetics
Endogenous GLP-1 Liraglutide Oral analogue GLP-1
plasma half-life = 1.5-2 min ~ 13 hours 165-184 hours
(1 week)
Metabolised by DPP-4 enzymes (at different rates)
 BUT!

“A phase 2 clinical study showed that the

bioavailability was low following oral
administration (~1%)

https://www.frontiersin.org/articles/10.3389/fen
do.2019.00155/full
 Physical
Insulin cannot diffuse across the GI tract due to its high
molecular weight and hydrophilic properties.

BARRIER TO ORAL
DELIVERY OF
INSULIN
Enzymatic Chemical
Proteolysis of oral insulin
The stomach lumen is
formulation begins in the
highly acidic (pH 1-3.7),
stomach, and is followed
whereas that in the
by intracellular peptidase
intestine varies between
degradation in the
neutral and slightly alkaline
enterocytes.
range (pH 6-8)
 Deliver biological active and intact insulin to
the blood circulation after oral absorption
through the epithelium that can mimic
basal and post-prandial endogenous insulin
release
Enhance GI tract permeability and
insulin absorption by reversibly
loosening the tight junction, or targeting
GI receptors for insulin transcellular
absorption

Prolong intestinal Biocompatible

residence time of and safe
insulin

IDEAL
Cost-effective with
Resist acidic gastric NANOPARTICLE high insulin loading
and intestinal pH, and CARRIERS FOR ORAL and loading efficiency
GI enzymatic
degradation INSULIN DELIVERY
 PH-RESPONSIVE NANOPARTICLE SYSTEM
EXCIPIENTS FUNCTION
Chitosan Bio-polymer, mucoadhesive¹ ²
Poly (g-glutamic acid) Bio-polymer, cryoprotectant³
EGTA (ethylene glycol Chelating agent
tetraacetic acid) Enzyme inhibitory (against protease), Enhance
permeation (high bioavailability) ²
Magnesium sulphate and Increase insulin load⁴
sodium tri-polyphosphate (TPP)
Trehalose Cryoprotectant¹
Eudragit Coating to dissolve at pH 7 (jejunum, ileum) & 6.6
(duodenum)²
¹Sonaje, K., Chen, Y.-J., Chen, H.-L., Wey, S.-P., Juang, J.-H., Nguyen, H.-N., Hsu, W.-C., Lin, J.-K., Sung, H.-W. (2010). Enteric-coated capsules filled with freeze-dried chitosan/poly(γ-glutamic acid) nanoparticles for oral insulin delivery.
Biomaterials, 31(12), 3384–3394. doi:10.1016/j.biomaterials.2010.01.042
²Wong, C. Y., Al-Salami, H., & Dass, C. R. (2017). Potential of insulin nanoparticle formulations for oral delivery and diabetes treatment. Journal of Controlled Release, 264, 247–275. doi:10.1016/j.jconrel.2017.09.003
³Najar IN and Das S: Poly-Glutamic Acid (PGA) - Structure, Synthesis, Genomic Organization and Its Application: A Review. Int J Pharm Sci Res 2015; 6(6): 2258-80.doi: 10.13040/IJPSR.0975-8232.6(6).2258-80.
⁴Akagi, Takami & Matsusaki, Michiya & Akashi, Mitsuru. (2010). Pharmaceutical and Medical Applications of Poly-Gamma-Glutamic Acid. 10.1007/978-3-642-12453-2_7.
SAR
CHITOSAN EGTA

Chuang, E.-Y., Lin, K.-J., Su, F.-Y., Chen, H.-L., Maiti, B., Ho, Y.-C., … Sung, H.-W.
(2013). Calcium depletion-mediated protease inhibition and apical-junctional-
complex disassembly via an EGTA-conjugated carrier for oral insulin delivery.
Journal of Controlled Release, 169(3), 296–305. doi:10.1016/j.jconrel.2012.11.011
Figure 2. Primary approaches for the peptides/proteins
loaded nanoparticles traversing the epithelium.⁵

⁵Han, Y., Gao, Z., Chen, L., Kang, L., Huang, W., Jin, M., … Bae, Y. H. (2019). Multifunctional oral
delivery systems for enhanced bioavailability of therapeutic peptides/proteins. Acta Pharmaceutica
Sinica B. doi:10.1016/j.apsb.2019.01.004
IDEAL PACKAGING
• Brown blister pack: protect from light
Ingredients: magnesium sulphate, EGTA
• Temperature: 2-8°C (refrigerate) @
15-30°C (room temp. For 30days only)
 Manufacturing Process
Enteric-coated capsules filled with freeze-dried chitosan/poly(g-glutamic
acid) nanoparticles
 Protein (GLP-1) is
premixed with aqueous γ-
PART 1: PGA

Preparation Sodium Triphosphate (TPP),

of Insulin MgSO₄ & EGTA are added

Loaded NP
*Using ionic Stirred for 30mins & added
to CH solution to form NP
gelation
method¹ Collected at 8000rpm for
50mins using centrifugation
& stored at 4⁰C
¹Sonaje, K., Chen, Y. J., Chen, H. L., Wey, S. P., Juang, J. H., Nguyen, H. N. Sung, H. W. (2010). Enteric-coated capsules filled with freeze-dried chitosan/poly(γ-glutamic acid) nanoparticles for oral insulin delivery.
Biomaterials, 31(12), 3384–3394. https://doi.org/10.1016/j.biomaterials.2010.01.042
 The schematic illustration of internal structures of NP

Sonaje, K., Chen, Y. J., Chen, H. L., Wey, S. P., Juang, J. H., Nguyen, H. N. Sung, H. W. (2010). Enteric-coated capsules filled with freeze-dried chitosan/poly(γ-glutamic acid) nanoparticles for oral insulin delivery.
Biomaterials, 31(12), 3384–3394. https://doi.org/10.1016/j.biomaterials.2010.01.042
 Aqueous NP added with Trehalose
& mixed for 30 mins

PART 2:
Preparation of The mixture was frozen at -20⁰C
freeze-dried overnight
NPs¹
The frozen NP is lyophilized using
freeze dryer to obtain in powder
form
¹Sonaje, K., Chen, Y. J., Chen, H. L., Wey, S. P., Juang, J. H., Nguyen, H. N. Sung, H. W. (2010). Enteric-coated capsules filled with freeze-dried chitosan/poly(γ-glutamic acid) nanoparticles for oral insulin delivery.
Biomaterials, 31(12), 3384–3394. https://doi.org/10.1016/j.biomaterials.2010.01.042
 The capsules are
The powder is filled immersed in
inside caps size 9 methanol (Eudragit
PART 3: S100)

Preparation
of Enteric-
Coated Caps¹ Drying at room
temperature

¹Sonaje, K., Chen, Y. J., Chen, H. L., Wey, S. P., Juang, J. H., Nguyen, H. N. Sung, H. W. (2010). Enteric-coated capsules filled with freeze-dried chitosan/poly(γ-glutamic acid) nanoparticles for oral insulin delivery.
Biomaterials, 31(12), 3384–3394. https://doi.org/10.1016/j.biomaterials.2010.01.042
Liraglutide Analytical
Method
By Siti Zuhairah
 • Disintegration test
• Dissolution test

RP-HPLC
Quality
Zetasizer
Purity control
Particle Test
size
analysis
 1) Particle Size Analysis

• Size of the nanoparticles was determined by a Zetasizer Nano ZS instrument1,2.

• Measure particle & molecular size – Dynamic light scattering
• Zeta potential & electrophoretic mobility – Electrophoretic light scattering
• Molecular weight – Static light scattering.2

1Huang W, Tsui CP, Tang CY, Gu L. Effects of Compositional Tailoring on Drug Delivery Behaviours of Silica Xerogel/Polymer Core-shell Composite Nanoparticles. Sci Rep
[Internet]. Springer US; 2018;8(1):1–13. Available from: http://dx.doi.org/10.1038/s41598-018-31070-9
2Sonaje K, Chen YJ, Chen HL, Wey SP, Juang JH, Nguyen HN, et al. Enteric-coated capsules filled with freeze-dried chitosan/poly(γ-glutamic acid) nanoparticles for oral
insulin delivery. Biomaterials [Internet]. Elsevier Ltd; 2010;31(12):3384–94. Available from: http://dx.doi.org/10.1016/j.biomaterials.2010.01.042
 2) Reverse Phase HPLC

Chromatographic conditions as
optimized above in the table. These
optimized conditions were followed for
the determination of Liraglutide in bulk
samples and its combined formulations1

1 Satyanarayana PV V, Madhavi AS. VALIDATED RP - HPLC METHOD FOR THE ESTIMATION OF

LIRAGLUTIDE IN TABLET DOSAGE Instrumentation and Analytical Conditions : Int J Sci Invent
Today. 2012;1(1):17–26.
 Fig 2: Typical chromatogram of Liraglutide standard1
1 Satyanarayana PV V, Madhavi AS. VALIDATED RP - HPLC METHOD FOR THE ESTIMATION OF LIRAGLUTIDE IN TABLET DOSAGE Instrumentation and
Analytical Conditions : Int J Sci Invent Today. 2012;1(1):17–26.
 3) Final Product Quality Control
i) Disintegration test.
• Using bracket rack assembly.
• Place 1 capsule in each tubes.
• Using simulated gastric fluid > after 1 hour, observe the capsule1.
Acceptance criteria
No evidence of disintegration, cracking, or softening.

• After 1 hour, operate in stimulated intestinal fluid > after 1 hour, observe the tablet.
Acceptance criteria:
All of the capsule disintegrate completely. If > 2 capsule fail to disintegrate completely,
repeat the test on 12 additional capsule. Not fewer than 16 of the total of 18 capsule
tested disintegrate completely1, 2

1Al-Gousous, J., & Langguth, P. (2015). Oral Solid Dosage Form Disintegration Testing — The Forgotten Test. Journal of Pharmaceutical Sciences,
104(9), 2664–2675. doi:10.1002/jps.24303
2Silva DA, Webster GK, Bou-Chacra N, Löbenberg R. The significance of disintegration testing in pharmaceutical development. Dissolution Technol.

2018;25(3):30–8.
 ii) Dissolution test.
Acid Stage Buffer Stage
• Hydrochloric acid in the vessel, and assemble the • Use Tribasic Sodium phosphate (Temp=
apparatus (Tempt: 37 ± 0.5) 37 ± 0.5) . Continue to operate the
• Place 1 capsule and operate the apparatus at the apparatus (individual monograph).
specified rate given in the monograph 1 . • At the end of the time period, withdraw
• After 2 hours, withdraw an aliquot of the fluid, an aliquot of the fluid
• Perform an analysis of the aliquot using a suitable • Perform the analysis using a suitable
assay method in individual monograph 1,2 assay method.

1Miller, D. A., Gamba, M., Sauer, D., Purvis, T. P., Clemens, N. T., & Williams, R. O. (2007). Evaluation of the USP dissolution test method A for enteric-
coated articles by planar laser-induced fluorescence. International Journal of Pharmaceutics, 330(1-2), 61–72. doi:10.1016/j.ijpharm.2006.08.047
2Lo L, Lu X, Lloyd D. Dissolution testing of a controlled release capsule formulation: Challenges and solutions using a semi-automated dissolution system. Dissolution

Technol. 2013;20(2):6–13.
REFERENCES
• Sonaje, K., Chen, Y. J., Chen, H. L., Wey, S. P., Juang, J. H., Nguyen, H.
N. Sung, H. W. (2010). Enteric-coated capsules filled with freeze-dried
chitosan/poly(γ-glutamic acid) nanoparticles for oral insulin delivery.
Biomaterials, 31(12), 3384–3394.
https://doi.org/10.1016/j.biomaterials.2010.01.042
REFERENCES
• Ahrén, B. (2011) ‘GLP-1 for type 2 diabetes’, Experimental Cell Research,
317(9), pp. 1239–1245. doi: 10.1016/j.yexcr.2011.01.010.
• Araújo, F. et al. (2012) ‘Oral delivery of glucagon-like peptide-1 and
analogs: alternatives for diabetes control?’, Journal of diabetes science and
technology. Diabetes Technology Society, 6(6), pp. 1486–97. doi:
10.1177/193229681200600630.
• Evaluation, D. (no date) ‘Chemistry review(s)’.
• Knudsen, L. B. and Lau, J. (2019) ‘The Discovery and Development of
Liraglutide and Semaglutide.’, Frontiers in endocrinology. Frontiers Media
SA, 10, p. 155. doi: 10.3389/fendo.2019.00155.
• PRATLEY, R. E. et al. (2019) ‘55-OR: Oral Semaglutide vs. Liraglutide and
Placebo in T2D: PIONEER 4’, Diabetes. American Diabetes Association,
68(Supplement 1), pp. 55-OR. doi: 10.2337/db19-55-OR.