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Polyglutamic Acid Functionalization of Chitosan Nanoparticles Enhances the Therapeutic Efficacy of Insulin Following Oral Administration

AAPS PharmSciTech volume 20, Article number: 131 (2019) Cite this article

Abstract

In the present study, stable chitosan nanoparticles (Ch-NPs) were developed using the ionotropic gelation method, where poly(sodium 4-styrenesulfonate) (PSS) was used as a cross-linking agent while polyglutamic acid (PGA) for functionalization to improve the oral uptake through calcium-sensing receptors and amino acid transporters present in intestinal epithelium. Formulation was optimized by the design of experiments (DoE) approach using a three-level central composite design and characterized for in vitro parameters such as morphology, particle size, polydispersity index (PDI), entrapment efficiency and zeta potential. Morphological analysis demonstrated the formation of spherical NPs with particle size, zeta potential, and entrapment efficiency in the range of 210 nm ± 2.8 nm, 18.1 mV ± 0.14 mV, and 85.9% ± 0.28%, respectively. The developed NPs exhibited sustained release at different pH conditions and almost threefold higher uptake in comparison with non-functionalized NPs in Caco-2 cell uptake studies. In vivo studies in diabetic animals demonstrated low levels of plasma glucose for almost 24 h. Pharmacological availability (PA) of insulin administered through Ch-PSS-PGA NPs (17.28 ± 0.9) was significantly higher as compared to that of insulin administered through control NPs, i.e., Ch-PGA NPs (10.9 ± 1.5) and Ch-PSS NPs (12.9 ± 1.8). Data on hand suggest the ability of the developed NPs in overcoming the poor stability and, thus, poor therapeutic efficacy following oral administration.

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References

  1. Mukhopadhyay P, Mishra R, Rana D, Kundu PP. Strategies for effective oral insulin delivery with modified chitosan nanoparticles: a review. Prog Polym Sci. 2012;37(11):1457–75.

    CAS  Article  Google Scholar 

  2. Sung H-W, Sonaje K, Liao Z-X, Hsu L-W, Chuang E-Y. pH-responsive nanoparticles shelled with chitosan for oral delivery of insulin: from mechanism to therapeutic applications. Acc Chem Res. 2012;45(4):619–29.

    CAS  PubMed  Article  Google Scholar 

  3. Khafagy E-S, Morishita M, Onuki Y, Takayama K. Current challenges in non-invasive insulin delivery systems: a comparative review. Adv Drug Deliv Rev. 2007;59(15):1521–46.

    CAS  Article  Google Scholar 

  4. TenHoor C, Dressman J. Oral absorption of peptides and proteins. STP Pharm Sci. 1992;2(4):301–12.

    CAS  Google Scholar 

  5. Goldberg M, Gomez-Orellana I. Challenges for the oral delivery of macromolecules. Nat Rev Drug Discov. 2003;2(4):289–95.

    CAS  Article  Google Scholar 

  6. Woodley JF. Enzymatic barriers for GI peptide and protein delivery. Crit Rev Ther Drug Carrier Syst. 1994;11(2–3):61–95.

    CAS  PubMed  Google Scholar 

  7. Aoki Y, Morishita M, Takayama K. Role of the mucous/glycocalyx layers in insulin permeation across the rat ileal membrane. Int J Pharm. 2005;297(1):98–109.

    CAS  PubMed  Article  Google Scholar 

  8. Yin L, Ding J, He C, Cui L, Tang C, Yin C. Drug permeability and mucoadhesion properties of thiolated trimethyl chitosan nanoparticles in oral insulin delivery. Biomaterials. 2009;30(29):5691–700.

    CAS  PubMed  Article  Google Scholar 

  9. Fonte P, Nogueira T, Gehm C, Ferreira D, Sarmento B. Chitosan-coated solid lipid nanoparticles enhance the oral absorption of insulin. Drug Deliv Transl Res. 2011;1(4):299–308.

    CAS  PubMed  Article  Google Scholar 

  10. Sharma G, Sharma AR, Nam J-S, Doss GPC, Lee S-S, Chakraborty C. Nanoparticle based insulin delivery system: the next generation efficient therapy for type 1 diabetes. J Nanobiotechnol. 2015;13(1):74.

    Article  Google Scholar 

  11. Zhang X, Qi J, Lu Y, He W, Li X, Wu W. Biotinylated liposomes as potential carriers for the oral delivery of insulin. Nanomedicine. 2014;10(1):167–76.

    CAS  PubMed  Article  Google Scholar 

  12. Rekha M, Sharma CP. Synthesis and evaluation of lauryl succinyl chitosan particles towards oral insulin delivery and absorption. J Control Release. 2009;135(2):144–51.

    CAS  PubMed  Article  Google Scholar 

  13. Sarmento B, Ribeiro A, Veiga F, Sampaio P, Neufeld R, Ferreira D. Alginate/chitosan nanoparticles are effective for oral insulin delivery. Pharm Res. 2007;24(12):2198–206.

    CAS  PubMed  Article  Google Scholar 

  14. Wedmore I, McManus JG, Pusateri AE, Holcomb JB. A special report on the chitosan-based hemostatic dressing: experience in current combat operations. J Trauma Acute Care Surg. 2006;60(3):655–8.

    Article  Google Scholar 

  15. Pangburn S, Trescony P, Heller J. Lysozyme degradation of partially deacetylated chitin, its films and hydrogels. Biomaterials. 1982;3(2):105–8.

    CAS  PubMed  Article  Google Scholar 

  16. Dhawan S, Singla AK, Sinha VR. Evaluation of mucoadhesive properties of chitosan microspheres prepared by different methods. AAPS PharmSciTech. 2004;5(4):122–8.

    PubMed Central  Article  Google Scholar 

  17. Szczepanowicz K, Hoel H, Szyk-Warszynska L, Bielanska E, Bouzga A, Gaudernack G, et al. Formation of biocompatible nanocapsules with emulsion core and pegylated shell by polyelectrolyte multilayer adsorption. Langmuir. 2010;26(15):12592–7.

    CAS  PubMed  Article  Google Scholar 

  18. Yazdimamaghani M, Razavi M, Mozafari M, Vashaee D, Kotturi H, Tayebi L. Biomineralization and biocompatibility studies of bone conductive scaffolds containing poly (3,4-ethylenedioxythiophene):poly(4-styrene sulfonate) (PEDOT:PSS). J Mater Sci Mater Med. 2015;26(12):274.

    PubMed  Article  Google Scholar 

  19. Desai MP, Labhasetwar V, Amidon GL, Levy RJ. Gastrointestinal uptake of biodegradable microparticles: effect of particle size. Pharm Res. 1996;13(12):1838–45.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  20. Agrawal AK, Kumar K, Swarnakar NK, Kushwah V, Jain S. “Liquid crystalline nanoparticles”: rationally designed vehicle to improve stability and therapeutic efficacy of insulin following oral administration. Mol Pharm. 2017;14(6):1874–82.

    CAS  PubMed  Article  Google Scholar 

  21. Agrawal AK, Urimi D, Harde H, Kushwah V, Jain S. Folate appended chitosan nanoparticles augment the stability, bioavailability and efficacy of insulin in diabetic rats following oral administration. RSC Adv. 2015;5(127):105179–93.

    CAS  Article  Google Scholar 

  22. Agrawal AK, Aqil F, Jeyabalan J, Spencer WA, Beck J, Gachuki BW, et al. Milk-derived exosomes for oral delivery of paclitaxel. Nanomedicine. 2017;13(5):1627–36.

    CAS  PubMed  Article  Google Scholar 

  23. Jung T, Kamm W, Breitenbach A, Kaiserling E, Xiao J, Kissel T. Biodegradable nanoparticles for oral delivery of peptides: is there a role for polymers to affect mucosal uptake? Eur J Pharm Biopharm. 2000;50(1):147–60.

    CAS  PubMed  Article  Google Scholar 

  24. Bernkop-Schnurch A, Walker G. Multifunctional matrices for oral peptide delivery. Crit Rev Ther Drug Carrier Syst. 2001;18(5):43.

    Article  Google Scholar 

  25. Makhlof A, Tozuka Y, Takeuchi H. Design and evaluation of novel pH-sensitive chitosan nanoparticles for oral insulin delivery. Eur J Pharm Sci. 2011;42(5):445–51.

    CAS  PubMed  Article  Google Scholar 

  26. Lin Y-H, Mi F-L, Chen C-T, Chang W-C, Peng S-F, Liang H-F, et al. Preparation and characterization of nanoparticles shelled with chitosan for oral insulin delivery. Biomacromolecules. 2007;8(1):146–52.

    CAS  PubMed  Article  Google Scholar 

  27. Chen M-C, Mi F-L, Liao Z-X, Hsiao C-W, Sonaje K, Chung M-F, et al. Recent advances in chitosan-based nanoparticles for oral delivery of macromolecules. Adv Drug Deliv Rev. 2013;65(6):865–79.

    CAS  PubMed  Article  Google Scholar 

  28. Conigrave AD, Brown EM. Taste receptors in the gastrointestinal tract II. L-amino acid sensing by calcium-sensing receptors: implications for GI physiology. Am J Physiol Gastrointest Liver Physiol. 2006;291(5):G753–61.

    CAS  PubMed  Article  Google Scholar 

  29. Conigrave AD, Quinn SJ, Brown EM. L-amino acid sensing by the extracellular Ca2+-sensing receptor. Proc Natl Acad Sci. 2000;97(9):4814–9.

    CAS  PubMed  Article  Google Scholar 

  30. Agrawal AK, Harde H, Thanki K, Jain S. Improved stability and antidiabetic potential of insulin containing folic acid functionalized polymer stabilized multilayered liposomes following oral administration. Biomacromolecules. 2013;15(1):350–60.

    PubMed  Article  Google Scholar 

  31. Greenfield NJ. Using circular dichroism spectra to estimate protein secondary structure. Nat Protoc. 2006;1(6):2876.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  32. D’Souza S. A review of in vitro drug release test methods for nano-sized dosage forms. Adv Pharm. 2014.

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Acknowledgments

The authors are thankful to the Director of the National Institute of Pharmaceutical Education and Research (NIPER) for providing the infrastructure facilities and to Mr. Rahul Mahajan for the technical assistance in the SEM analysis.

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  1. Ashish Kumar Agrawal

    Present address: Department of Pharmaceutical Engineering and Technology, Indian Institutes of Technology, Banaras Hindu University, Varanasi, UP, 221005, India

Affiliations

  1. Centre for Pharmaceutical Nanotechnology, Department of Pharmaceutics, National Institute of Pharmaceutical Education and Research, S.A.S. Nagar, Nagar, Punjab, 160062, India

    Dileep Urimi, Ashish Kumar Agrawal, Varun Kushwah & Sanyog Jain

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  1. Dileep Urimi
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Correspondence to Ashish Kumar Agrawal or Sanyog Jain.

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Urimi, D., Agrawal, A.K., Kushwah, V. et al. Polyglutamic Acid Functionalization of Chitosan Nanoparticles Enhances the Therapeutic Efficacy of Insulin Following Oral Administration. AAPS PharmSciTech 20, 131 (2019). https://doi.org/10.1208/s12249-019-1330-2

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  • DOI: https://doi.org/10.1208/s12249-019-1330-2

KEY WORDS

  • insulin
  • oral delivery
  • chitosan nanoparticles
  • design of experiment
  • Caco-2 cell uptake