Abstract
The objective of this study was to prepare a stable self-nanoemulsifying formulation of exendin-4, which is an antidiabetic peptide. As exendin-4 is commercially available only in subcutaneous form, several attempts have been made to discover an effective oral formulation. Self-nanoemulsifying drug delivery systems are known to be suitable carriers for the oral administration of peptide drugs. Various ratios of oil, surfactant, and co-surfactant mixtures were used to determine the area in the pseudoternary phase diagram for clear nanoemulsion. The Design of Experiment approach was used for the optimization of the formulation. Blank self-nanoemulsifying formulations containing ethyl oleate as oil phase, Cremophor EL®, and Labrasol® as surfactant, absolute ethanol, and propylene glycol as co-solvent in various proportions were approximately 18–50 nm, 0.08–0.204 and − 3 to − 23 mV in droplet size, polydispersity index, and zeta potential, respectively. When all formulations were compared by statistical analysis, five of them with smaller droplet sizes were selected for further studies. The physical stability test was performed for 1 month at 5 °C ± 3 °C and 25 °C ± 2 °C/60% RH ± 5% RH storage conditions. As a result of the characterization and physical stability test results, ethyl oleate: Cremophor EL®:absolute ethanol (30:52.5:17.5) formulation and four formulations containing ethyl oleate: Cremophor EL®:Labrasol®:propylene glycol:absolute ethanol at varying concentrations were considered for peptide encapsulation efficiency. Formulation having the highest encapsulation efficiency of exendin-4 containing ethyl oleate: Cremophor EL®:Labrasol®:propylene glycole:absolute ethanol (15:42.5:21.25:15.94:5.31) was selected for in vitro Caco-2 intestinal permeability study. The permeabiliy coefficient was increased by 1.5-folds by exendin-4-loaded self-nanoemulsifying formulation as compared to the exendin-4 solution. It can be concluded that intestinal permeability has been improved by self-nanoemulsifying formulation.
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Abbreviations
- Cr EL® :
-
Cremophor EL®
- DoE:
-
Design of experiment
- DS:
-
Droplet Size
- ELISA:
-
Enzyme-linked immuno sorbent assay
- Eq:
-
Equation
- Ex-4:
-
Exendin-4
- GI:
-
Gastrointestinal
- HLB:
-
Hydrophilic–lipophilic balance
- Lab® :
-
Labrasol®
- Papp:
-
Permeability coefficient
- PDI:
-
Polydispersity index
- PG:
-
Propylene glycol
- RH:
-
Relative humidity
- SNEDDS:
-
Self-nanoemulsifying drug delivery system
- ZP:
-
Zeta potential
References
American Diabetes Association (2013) Diagnosis and classification of diabetes mellitus. Diabetes Care 36(Suppl 1):67–74. https://doi.org/10.2337/dc13-S067
Balakrishnan P, Lee BJ, Oh DH, Kim JO, Hong MJ, Jee JP, Kim JA, Yoo BK, Woo JS, Yong CS, Choi HG (2009) Enhanced oral bioavailability of Coenzyme Q10 by self-emulsifying drug delivery systems. Int J Pharm 374(1–2):66–72. https://doi.org/10.1016/j.ijpharm.2009.03.008 (Epub 2009 Mar 19)
Celik-Tekeli M, Celebi N, Aktas Y (2017) Development and characterization of self nanoemulsifiying systems for oral delivery of peptides. In: Noszái B, Zelkó R, Hankó Z. (Eds). Proceedings of the 7th BBBB International Conference on Pharmaceutical Sciences, Balatonfüred, Hungary, October 05–07, 2017. Acta pharmaceutica Hungarica 87 (3–4):112–112.
Chakraborti CK (2010) Exenatide: a new promising antidiabetic agent. Indian J Pharm Sci 72(1):1
Chuang EY, Nguyen GT, Su FY, Lin KJ, Chen CT, Mi FL, Yen TC, Juang JH, Sung HW (2013) Combination therapy via oral co-administration of insulin and exendin-4 loaded nanoparticles to treat type 2 diabetic rats undergoing OGTT. Biomaterials 34(32):7994–8001. https://doi.org/10.1016/j.biomaterials.2013.07.021 (Epub 2013 Jul 24)
Cilek A, Celebi N, Tirnaksiz F, Tay A (2005) A lecithin-based microemulsion of rh-insulin with aprotinin for oral administration: Investigation of hypoglycemic effects in non-diabetic and STZ-induced diabetic rats. Int J Pharm 298:176–185. https://doi.org/10.1016/j.ijpharm.2005.04.016
Constantinides PP (1995) Lipid microemulsions for improving drug dissolution and oral absorption: physical and biopharmaceutical aspects. Pharm Res 12(11):1561–1572. https://doi.org/10.1023/a:1016268311867
Deutel B, Greindl M, Thaurer M, Bernkop-Schnürch A (2008) Novel insulin thiomer nanoparticles: in vivo evaluation of an oral drug delivery system. Biomacromol 9(1):278–285. https://doi.org/10.1021/bm700916h (Epub 2007 Dec 27)
FDA (2009) Guidance for industry, Q8(R2) Pharmaceutical development, US Department of Health and Human Services, Food and Drug Administration, CDER, Rockville. https://www.fda.gov/downloads/drugs/guidances/ucm073507.pdf Accessed 9 July 2020
Furman BL (2012) The development of Byetta (exenatide) from the venom of the Gila monster as an anti-diabetic agent. Toxicon 59(4):464–471. https://doi.org/10.1016/j.toxicon.2010.12.016
Green BD, Flatt PR (2007) Incretin hormone mimetics and analogues in diabetes therapeutics. Best Pract Res Clin Endocrinol Metab 21(4):497–516. https://doi.org/10.1016/j.beem.2007.09.003
Gupta V, Doshi N, Mitragotri S (2013) Permeation of insulin, calcitonin and exenatide across Caco-2 monolayers: measurement using a rapid, 3-day system. PLoS ONE 8(2):571–536. https://doi.org/10.1371/journal.pone.0057136 (Epub 2013 Feb 27)
Gursoy RN, Benita S (2004) Self-emulsifying drug delivery systems (SEDDS) for improved oral delivery of lipophilic drugs. Biomed Pharmacother 58:173–182. https://doi.org/10.1016/j.biopha.2004.02.001
Holm R, Jensen IH, Sonnergaard J (2006) Optimization of self-microemulsifying drug delivery systems (SMEDDS) using a D-optimal design and the desirability function. Drug Dev Ind Pharm 32(9):1025–1032
Ismail R, Csóka I (2017) Novel strategies in the oral delivery of antidiabetic peptide drugs –Insulin, GLP 1 and its analogs. Eur J Pharm Biopharm 115:257–267. https://doi.org/10.1016/j.ejpb.2017.03.015 (Epub 2017 Mar 21)
Kohli K, Chopra S, Dhar D, Arora S, Khar RK (2010) Self-emulsifying drug delivery systems: an approach to enhance oral bioavailability. Drug Discov Today 15(21–22):958–965. https://doi.org/10.1016/j.drudis.2010.08.007
Kumar A, Sharma S, Kamble R (2010) Self emulsifying drug delivery system (Sedds): future aspects. Int J Pharm 2:7–13
Leonaviciute G, Bernkop-Schnurch A (2015) Self-emulsifying drug delivery systems in oral (poly)peptide drug delivery. Expert Opin Drug Deliv 12(11):1703–1716. https://doi.org/10.1517/17425247.2015.1068287 (Epub 2015 Aug 24)
Li P, Nielsen HM, Müllertz A (2012) Oral delivery of peptides and proteins using lipid-based drug delivery systems. Expert Opin Drug Deliv 9(10):1289–1304. https://doi.org/10.1517/17425247.2012.717068 (Epub 2012 Aug 17)
Li P, Tan A, Prestidge CA, Nielsen HM, Müllertz A (2014) Self-nanoemulsifying drug delivery systems for oral insulin delivery: in vitro and in vivo evaluations of enteric coating and drug loading. Int J Pharm 477(1–2):390–398. https://doi.org/10.1016/j.ijpharm.2014.10.039
Li X, Wang C, Liang R, Sun F, Shi Y, Wang A, Liu W, Sun K, Li Y (2015) The glucose-lowering potential of exenatide delivered orally via goblet cell-targeting nanoparticles. Pharm Res 32(3):1017–1027. https://doi.org/10.1007/s11095-014-1513-1 (Epub 2014 Oct 1)
Mason RL, Gunst RF, Hess JL (2003) Statistical principles in experimental design. In: Mason RL, Gunst RF, Hess JL (eds) Statistical design and analysis of experiments: with applications to engineering and science, 2nd edn. John Wiley Sons Inc, New Jersey, pp 107–139
Morcol T, Nagappan P, Nerenbaum L, Mitchell A, Bell SJ (2004) Calcium phosphate-PEG-insulin-casein (CAPIC) particles as oral delivery systems for insulin. Int J Pharm 277:91–97. https://doi.org/10.1016/j.ijpharm.2003.07.015
Mu H, Holm R, Müllertz A (2013) Lipid-based formulations for oral administration of poorly water-soluble drugs. Int J Pharm 453(1):215–224. https://doi.org/10.1016/j.ijpharm.2013.03.054 (Epub 2013 Apr 8)
Oh KS, Kim JY, Yoon BD, Lee M, Kim H, Kim M, Seo JH, Yuk SH (2014) Sol–gel transition of nanoparticles/polymer mixtures for sustained delivery of exenatide to treat type 2 diabetes mellitus. Eur J Pharm and Biopharm 88(3):664–669. https://doi.org/10.1016/j.ejpb.2014.08.004 (Epub 2014 Aug 23)
Park K, Kwon CI, Park K (2011) Oral protein delivery: current status and future prospect. React Funct Polym 71:280–287. https://doi.org/10.1016/j.reactfunctpolym.2010.10.002
Parmar B, Patel U, Bhimani B, Sanghavi K, Patel G, Daslaniya D (2012) SMEDDS: a dominant dosage form which improve bioavailability. Am J PharmTech Res 2(4):55–72
Politis SN, Colombo P, Colombo G, Rekkas DM (2017) Design of experiments (DoE) in pharmaceutical development. Drug Dev Ind Pharm 43(6):889–901. https://doi.org/10.1080/03639045.2017.1291672
Rao SV, Shao J (2008) Self-nanoemulsifying drug delivery system (SNEDDS) for oral delivery of protein drugs: I Formulation Development. Int J Pharm 362(1–2):2–9. https://doi.org/10.1016/j.ijpharm.2008.05.018 (Epub 2008 May 27)
Renukuntla J, Vadlapudi AD, Patel A, Boddu SH, Mitra AK (2013) Approaches for enhancing oral bioavailability of peptides and proteins. Int J Pharm 447(1–2):75–93. https://doi.org/10.1016/j.ijpharm.2013.02.030 (Epub 2013 Feb 18)
Sakloetsakun D, Dünnhaupt S, Barthelmes J, Perera G, Bernkop-Schnürch A (2013) Combining two technologies: multifunctional polymers and self-nanoemulsifying drug delivery system (SNEDDS) for oral insulin administration. Int J Biol Macromol 61:363–372. https://doi.org/10.1016/j.ijbiomac.2013.08.002 (Epub 2013 Aug 8)
Shafiq S, Shakeel F, Talegaonkar S, Ahmad FJ, Khar RK, Ali M (2007) Development and bioavailability assessment of ramipril nanoemulsion formulation. Eur J Pharm Biopharm 66:227–243. https://doi.org/10.1016/j.ejpb.2006.10.014 (Epub 2006 Oct 24)
Soudry-Kochavi L, Naraykin N, Nassar T, Benita S (2015) Improved oral absorption of exenatide using an original nanoencapsulation and microencapsulation approach. J Control Release 217:202–210. https://doi.org/10.1016/j.jconrel.2015.09.012 (Epub 2015 Sep 14)
Tan ML, Choong PF, Dass CR (2010) Recent developments in liposomes, microparticles and nanoparticles for protein and peptide drug delivery. Peptides 31(1):184–193. https://doi.org/10.1016/j.peptides.2009.10.002 (Epub 2009 Oct 9)
Wang M, Zhang Y, Feng J, Gu T, Dong Q, Yang X, Sun Y, Wu Y, Chen Y, Kong W (2013) Preparation, characterization, and in vitro and in vivo investigation of chitosan-coated poly (d, l-lactide-co glycolide) nanoparticles for intestinal delivery of exendin-4. Int J Pharm 8:1141–1154. https://doi.org/10.2147/IJN.S41457 (Epub 2013 Mar 15)
Wilcox G (2005) Insulin and insulin resistance. Clin Biochem Rev 26(2):19
Yu LX (2008) Pharmaceutical quality by design: product and process development, understanding and control. Pharm Res 25:781–791. https://doi.org/10.1007/s11095-007-9511-1 (Epub 2008 Jan 10)
Zhang ZH, Zhang YL, Zhou JP, Lv HX (2012a) Solid lipid nanoparticles modified with stearic acid–octaarginine for oral administration of insulin. Int J Pharm 7:3333–3339. https://doi.org/10.2147/IJN.S31711 (Epub 2012 Jul 2)
Zhang Q, He N, Zhang L, Zhu F, Chen Q, Qin Y, Zhang Z, Zhang Q, Wang S, He Q (2012b) The in vitro and in vivo study on self-nanoemulsifying drug delivery system (SNEDDS) based on insulin-phospholipid complex. J Biomed Nanotechnol 8(1):90–97. https://doi.org/10.1166/jbn.2012.1371
Zhang Y, Wu X, Meng L, Zhang Y, Ai R, Qi N, He H, Xu H, Tang X (2012c) Thiolated Eudragit nanoparticles for oral insulin delivery: preparation, characterization and in vivo evaluation. Int J Pharm 436(1–2):341–350. https://doi.org/10.1016/j.ijpharm.2012.06.054 (Epub 2012 Jul 3)
Zupančič O, Grieβinger JA, Rohrer J, Pereira de Sousa I, Danninger L, Partenhauser A, Sündermann NE, Laffleur F, Bernkop-Schnürch A (2016) Development, in vitro and in vivo evaluation of a self-emulsifying drug delivery system (SEDDS) for oral enoxaparin administration. Eur J Pharm Biopharm 109:113–121. https://doi.org/10.1016/j.ejpb.2016.09.013 (Epub 2016 Sep 28)
Acknowledgements
This work was supported by the [Research Fund of the Erciyes] under Grant [TCD-2017-7087]; [Scientific and Technological Research Council of Turkey (TUBİTAK)] under Grant [217S416]. We would like to thank Dr. Alptug Eren Karakucuk for his contributions to Design of Experiment studies which is examined in this research paper.
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Tekeli, M.C., Aktas, Y. & Celebi, N. Oral self-nanoemulsifying formulation of GLP-1 agonist peptide exendin-4: development, characterization and permeability assesment on Caco-2 cell monolayer. Amino Acids 53, 73–88 (2021). https://doi.org/10.1007/s00726-020-02926-0
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DOI: https://doi.org/10.1007/s00726-020-02926-0