Overcoming oral insulin delivery barriers: application of cell penetrating peptide and silica-based nanoporous composites

Abstract

Oral insulin delivery has received the most attention in insulin formulations due to its high patient compliance and, more importantly, to its potential to mimic the physiologic insulin secretion seen in non-diabetic individuals. However, oral insulin delivery has two major limitations: the enzymatic barrier that leads to rapid insulin degradation, and the mucosal barrier that limits insulin’s bioavailability. Several approaches have been actively pursued to circumvent the enzyme barrier, with some of them receiving promising results. Yet, thus far there has been no major success in overcoming the mucosal barrier, which is the main cause in undercutting insulin’s oral bioavailability. In this review of our group’s research, an innovative silica-based, mucoadhesive oral insulin formulation with encapsulated-insulin/cell penetrating peptide (CPP) to overcome both enzyme and mucosal barriers is discussed, and the preliminary and convincing results to confirm the plausibility of this oral insulin delivery system are reviewed. In vitro studies demonstrated that the CPP-insulin conjugates could facilitate cellular uptake of insulin while keeping insulin’s biologic functions intact. It was also confirmed that low molecular weight protamine (LMWP) behaves like a CPP peptide, with a cell translocation potency equivalent to that of the widely studied TAT. The mucoadhesive properties of the produced silica-chitosan composites could be controlled by varying both the pH and composition; the composite consisting of chitosan (25 wt-%) and silica (75 wt-%) exhibited the greatest mucoadhesion at gastric pH. Furthermore, drug release from the composite network could also be regulated by altering the chitosan content. Overall, the universal applicability of those technologies could lead to development of a generic platform for oral delivery of many other bioactive compounds, especially for peptide or protein drugs which inevitably encounter the poor bioavailability issues.

Key words： insulin; cell penetrating peptide; mucoadhesive composites; oral delivery

Acknowledgments

This work was sponsored by Grant R31-2008-000-10103-01 from the World Class University (WCU) project of South Korea. Victor C. Yang is currently a participating faculty member in the Department of Molecular Medicine and Biopharmaceutical Sciences, Seoul National University, Korea. It was also partially supported by the National Key Basic Research Program of China (2013CB932502), School of Pharmacy, Fudan University and The Open Project Program of Key Laboratory of Smart Drug Delivery, Fudan University, MOE & PLA, China (SDD2011-02).

References

1 Capaldi B. Treatments and devices for future diabetes management. Nursing Times, 2005, 101(18): 30–32

2 Cobble M E. Initiating and intensifying insulin therapy for type 2 diabetes: why, when, and how. American Journal of Therapeutics, 2009, 16(1): 56–64

3 Prevention Cf DCa. National diabetes fact sheet general information and national estimates on diabetes in the United States. Centers for Disease Control and Prevention, 2003

4 Heinemann L. New ways of insulin delivery. International Journal of Clinical Practice. Supplement, 2011, 65(170): 31–46

5 Gordon Still J. Development of oral insulin: progress and current status. Diabetes/Metabolism Research and Reviews, 2002, 18(S1): S29–S37

6 Heinemann L. New ways of insulin delivery. International Journal of Clinical Practice. Supplement, 2010, 64: 29–40

7 Reis C P, Damge C. Nanotechnology as a promising strategy for alternative routes of insulin delivery. Methods in Enzymology, 2012, 508: 271–294

8 Fonte P, Andrade F, Araujo F, Andrade C, Neves J, Sarmento B. Chitosan-coated solid lipid nanoparticles for insulin delivery. Methods in Enzymology, 2012, 508: 295–314

9 Card J W, Magnuson B A. A review of the efficacy and safety of nanoparticle-based oral insulin delivery systems. American Journal of Physiology. Gastrointestinal and Liver Physiology, 2011, 301(6): G956–G967

10 He P, Tang Z, Lin L, Deng M, Pang X, Zhuang X, Chen X. Novel biodegradable and pH-sensitive poly(ester amide) microspheres for oral insulin delivery. Macromolecular Bioscience, 2012, 12(4): 547–556

11 Cefalu W T. Concept, strategies, and feasibility of noninvasive insulin delivery. Diabetes Care, 2004, 27(1): 239–246

12 Krishnankutty R K, Mathew A, Sedimbi S K, Suryanarayan S, Sanjeevi C B. Alternative routes of insulin delivery. Zhong Nan Da Xue Xue Bao. Yi Xue Ban, 2009, 34(10): 933–948

13 Bellary S, Barnett A H. Inhaled insulin: new technology, new possibilities. International Journal of Clinical Practice, 2006, 60(6): 728–734

14 Cefalu W T. Evolving strategies for insulin delivery and therapy. Drugs, 2004, 64(11): 1149–1161

15 Sajeesh S, Bouchemal K, Marsaud V, Vauthier C, Sharma C P. Cyclodextrin complexed insulin encapsulated hydrogel microparticles: An oral delivery system for insulin. Journal of Controlled Release, 2010, 147(3): 377–384

16 Yadav N, Morris G, Harding S E, Ang S, Adams G G. Various non-injectable delivery systems for the treatment of diabetes mellitus. Endocrine, Metabolic & Immune Disorders Drug Targets, 2009, 9(1): 1–13

17 Banting F G, Best C H, Collip J B, Campbell W R, Fletcher A A. Pancreatic extracts in the treatment of diabetes mellitus. Canadian Medical Association Journal, 1922, 7: 6

18 Del Curto M D, Maroni A, Palugan L, Zema L, Gazzaniga A, Sangalli M E. Oral delivery system for two-pulse colonic release of protein drugs and protease inhibitor/absorption enhancer compounds. Journal of Pharmaceutical Sciences, 2011, 100(8): 3251–3259

19 Jelvehgari M, Milani P Z, Siahi-Shadbad M R, Monajjemzadeh F, Nokhodchi A, Azari Z, Valizadeh H. In vitro and in vivo evaluation of insulin microspheres containing protease inhibitor. Arzneimittel-Forschung, 2011, 61(1): 14–22

20 Su F Y, Lin K J, Sonaje K, Wey S P, Yen T C, Ho Y C, Panda N, Chuang E Y, Maiti B, Sung H W. Protease inhibition and absorption enhancement by functional nanoparticles for effective oral insulin delivery. Biomaterials, 2012, 33(9): 2801–2811

21 Marschutz M K, Bernkop-Schnurch A. Oral peptide drug delivery: polymer-inhibitor conjugates protecting insulin from enzymatic degradation in vitro. Biomaterials, 2000, 21(14): 1499–1507

22 Saudek C D. Novel forms of insulin delivery. Endocrinology and Metabolism Clinics of North America, 1997, 26(3): 599–610

23 Avadi M R, Sadeghi A M, Mohammadpour N, Abedin S, Atyabi F, Dinarvand R, Rafiee-Tehrani M. Preparation and characterization of insulin nanoparticles using chitosan and Arabic gum with ionic gelation method. Nanomedicine; Nanotechnology, Biology, and Medicine, 2010, 6(1): 58–63

24 Cui F, He C, He M, Tang C, Yin L, Qian F, Yin C. Preparation and evaluation of chitosan-ethylenediaminetetraacetic acid hydrogel films for the mucoadhesive transbuccal delivery of insulin. Journal of Biomedical Materials Research. Part A, 2009, 89A(4): 1063–1071

25 Cui F, Qian F, Zhao Z, Yin L, Tang C, Yin C. Preparation, characterization, and oral delivery of insulin loaded carboxylated chitosan grafted poly(methyl methacrylate) nanoparticles. Biomacromolecules, 2009, 10(5): 1253–1258

26 Schilling R, Mitra A. Degradation of insulin by trypsin and alpha-chymotrypsin. Pharmaceutical Research, 1991, 8(6): 721–727

27 Nishihata T, Rytting J H, Kamada A, Higuchi T. Enhanced intestinal absorption of insulin in rats in the presence of sodium 5-methoxysalicylate. Diabetes, 1981, 30(12): 1065–1067

28 Cui C Y, Lu W L, Xiao L, Zhang S Q, Huang Y B, Li S L, Zhang R J, Wang G L, Zhang X, Zhang Q. Sublingual delivery of insulin: effects of enhancers on the mucosal lipid fluidity and protein conformation, transport, and in vivo hypoglycemic activity. Biological & Pharmaceutical Bulletin, 2005, 28(12): 2279–2288

29 Muranishi S. Delivery system design for improvement of intestinal absorption of peptide drugs. Yakugaku Zasshi, 1997, 117(7): 394–414

30 Chung S W, Hil-lal T A, Byun Y. Strategies for non-invasive delivery of biologics. Journal of Drug Targeting, 2012, 20(6): 481–501

31 Schwarze S R, Ho A, Vocero-Akbani A, Dowdy S F. In vivo protein transduction: delivery of a biologically active protein into the mouse. Science, 1999, 285(5433): 1569–1572

32 Schwarze S R, Dowdy S F. In vivo protein transduction: intracellular delivery of biologically active proteins, compounds and DNA. Trends in Pharmacological Sciences, 2000, 21(2): 45–48

33 Cooper I, Sasson K, Teichberg V I, Schnaider-Beeri M, Fridkin M, Shechter Y. Peptide derived from HIV-1 TAT protein, destabilizes a monolayer of endothelial cells in an in vitro model of the blood-brain barrier, and allows permeation of high molecular weight proteins. Journal of Biological Chemistry, 2012, •••:

34 Yu R, Zeng Z, Guo X, Zhang H, Liu X, Ding Y, Chen J. The TAT peptide endows PACAP with an enhanced ability to traverse bio-barriers. Neuroscience Letters, 2012, 527(1): 1–5

35 Elliott G, O'Hare P. Intercellular trafficking and protein delivery by a herpesvirus structural protein. Cell, 1997, 88(2): 223–233

36 Min S H, Kim D M, Kim M N, Ge J, Lee D C, Park I Y, Park K C, Hwang J S, Cho C W, Yeom Y I. Gene delivery using a derivative of the protein transduction domain peptide, K-Antp. Biomaterials, 2010, 31(7): 1858–1864

37 Derossi D, Joliot A H, Chassaing G, Prochiantz A. The third helix of the antennapedia homeodomain translocates through biological membranes. Journal of Biological Chemistry, 1994, 269(14): 10444–10450

38 Jin G S, Zhu G D, Zhao Z G, Liu F S. VP22 enhances the expression of glucocerebrosidase in human Gaucher II fibroblast cells mediated by lentiviral vectors. Clinical and Experimental Medicine, 2012, 12(3): 135–143

39 Tanaka M, Kato A, Satoh Y, Ide T, Sagou K, Kimura K, Hasegawa H, Kawaguchi Y. Herpes simplex virus 1 VP22 regulates translocation of multiple viral and cellular proteins and promotes neurovirulence. Journal of Virology, 2012, 86(9): 5264–5277

40 Chang L C, Lee H F, Yang Z, Yang V. Low molecular weight protamine (LMWP) as nontoxic heparin/low molecular weight heparin antidote (I): Preparation and characterization. AAPS PharmSci, 2001, 3(3): 7–14

41 Chang L C, Liang J, Lee H F, Lee L, Yang V. Low molecular weight protamine (LMWP) as nontoxic heparin/low molecular weight heparin antidote (II): In vitro evaluation of efficacy and toxicity. AAPS PharmSci, 2001, 3(3): 15–23

42 Chang L C, Wrobleski S, Wakefield T, Lee L, Yang V. Low molecular weight protamine as nontoxic heparin/low molecular weight heparin antidote (III): Preliminary in vivo evaluation of efficacy and toxicity using a canine model. AAPS PharmSci, 2001, 3(3): 24–31

43 Park Y J, Chang L C, Liang J F, Moon C, Chung C P, Yang V C. Nontoxic membrane translocation peptide from protamine, low molecular weight protamine (LMWP), for enhanced intracellular protein delivery: in vitro and in vivo study. FASEB Journal, 2005, 19(11): 1555–1557

44 Xia H, Gao X, Gu G, Liu Z, Zeng N, Hu Q, Song Q, Yao L, Pang Z, Jiang X, Chen J, Chen H. Low molecular weight protamine-functionalized nanoparticles for drug delivery to the brain after intranasal administration. Biomaterials, 2011, 32(36): 9888–9898

45 Ramadas M W P, Dileep K J, Ramadas M, Anitha Y, Sharma C P, 0. Lipoinsulin encapsulated alginate-chitosan capsules: intestinal delivery in diabetic rats. Journal of Microencapsulation, 2000, 17(4): 405–411

46 Kimura T, Sato K, Sugimoto K, Tao R, Murakami T, Kurosaki Y, Nakayama T. Oral administration of insulin as poly(vinyl alcohol)-gel spheres in diabetic rats. Biological & Pharmaceutical Bulletin, 1996, 19(6): 897–900

47 Mitchell D J, Steinman L, Kim D T, Fathman C G, Rothbard J B. Polyarginine enters cells more efficiently than other polycationic homopolymers. Journal of Peptide Research, 2000, 56(5): 318–325

48 Futaki S, Nakase I, Suzuki T, Zhang, Sugiura Y. Translocation of branched-chain arginine peptides through cell membranes: flexibility in the spatial disposition of positive charges in membrane-permeable peptides. Biochemistry, 2002, 41(25): 7925–7930

49 Wong T W. Chitosan and its use in design of insulin delivery system. Recent Patents on Drug Delivery & Formulation, 2009, 3(1): 8–25

50 Damge C, Maincent P, Ubrich N. Oral delivery of insulin associated to polymeric nanoparticles in diabetic rats. Journal of Controlled Release, 2007, 117(2): 163–170

51 Sarmento B, Ribeiro A, Veiga F, Sampaio P, Neufeld R, Ferreira D. Alginate/chitosan nanoparticles are effective for oral insulin delivery. Pharmaceutical Research, 2007, 24(12): 2198–2206

52 Liang J F, Zhen L, Chang L C, Yang V C. A less toxic heparin antagonist—low molecular weight protamine. Biochemistry. Biokhimiia, 2003, 68(1): 116–120

53 Tsui B, Singh V K, Liang J F, Yang V C. Reduced reactivity towards anti-protamine antibodies of a low molecular weight protamine analogue. Thrombosis Research, 2001, 101(5): 417–420

54 Carlsson J, Drevin H, Axén R. Protein thiolation and reversible protein-protein conjugation.N-Succinimidyl 3-(2-pyridyldithio)propionate, a new heterobifunctional reagent. Biochemical Journal, 1978, 173(3): 723–737

55 Chickering D E, Mathiowitz E. Bioadhesive microspheres I. A novel electrobalance-based method to study adhesive interactions between individual microspheres and intestinal mucosa. Journal of Controlled Release, 1995, 34(3): 251–262

56 Sudhakar Y, Kuotsu K, Bandyopadhyay A K. Buccal bioadhesive drug delivery—a promising option for orally less efficient drugs. Journal of Controlled Release, 2006, 114(1): 15–40