Advertisement

Lipid nanoparticles as vehicles for oral delivery of insulin and insulin analogs: preliminary ex vivo and in vivo studies

  • Elisabetta Muntoni
  • Elisabetta Marini
  • Nahid Ahmadi
  • Paola Milla
  • Corrado Ghè
  • Alessandro Bargoni
  • Maria Teresa Capucchio
  • Elena Biasibetti
  • Luigi BattagliaEmail author
Original Article
  • 47 Downloads

Abstract

Aims

Subcutaneous administration of insulin in patients suffering from diabetes is associated with the distress of daily injections. Among alternative administration routes, the oral route seems to be the most advantageous for long-term administration, also because the peptide undergoes a hepatic first-pass effect, contributing to the inhibition of the hepatic glucose output. Unfortunately, insulin oral administration has so far been hampered by degradation by gastrointestinal enzymes and poor intestinal absorption. Loading in lipid nanoparticles should allow to overcome these limitations.

Methods

Entrapment of peptides into such nanoparticles is not easy, because of their high molecular weight, hydrophilicity and thermo-sensitivity. In this study, this objective was achieved by employing fatty acid coacervation method: solid lipid nanoparticles and newly engineered nanostructured lipid carriers were formulated. Insulin and insulin analog—glargine insulin—were entrapped in the lipid matrix through hydrophobic ion pairing.

Results

Bioactivity of lipid entrapped peptides was demonstrated through a suitable in vivo experiment. Ex vivo and in vivo studies were carried out by employing fluorescently labelled peptides. Gut tied up experiments showed the superiority of glargine insulin-loaded nanostructured lipid carriers, which demonstrated significantly higher permeation (till 30% dose/mL) compared to free peptide. Approximately 6% absolute bioavailability in the bloodstream was estimated for the same formulation through in vivo pharmacokinetic studies in rats. Consequently, a discrete blood glucose responsivity was noted in healthy animals.

Conclusions

Given the optimized ex vivo and in vivo intestinal uptake of glargine insulin from nanostructured lipid carriers, further studies will be carried out on healthy and diabetic rat models in order to establish a glargine insulin dose–glucose response relation.

Keywords

Insulin Glargine insulin Solid lipid nanoparticles (SLN) Nanostructured lipid carriers (NLC) 

Notes

Acknowledgements

The authors thank Italian MIUR (Ricerca Locale 2016–2017 and FFABR 2018) for funding.

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

Ethical approval

Animal experiments were performed owing to Italian and International Guidelines (DL 26/2014 implementation of directive 2010/63 UE). An experimental protocol approved by the Turin University Bioethical Committee and the Italian Ministry of Health (Aut. N. 32/2016-PR) was employed.

Informed consent

For this type of study no informed consent is required.

Supplementary material

592_2019_1403_MOESM1_ESM.doc (208 kb)
Supplementary material 1 (DOC 208 kb)

References

  1. 1.
    Peterson GE (2006) Intermediate and long-acting insulins: a review of NPH insulin, insulin glargine and insulin detemir. Curr Med Res Opin 22(12):2613–2619CrossRefGoogle Scholar
  2. 2.
    Alleman E, Leroux JC, Gurny R (1998) Polymeric nano- and microparticles for the oral delivery of peptides and peptidomimetics. Adv Drug Del Rev 34:171–187CrossRefGoogle Scholar
  3. 3.
    Mutalik M (2011) Long awaited dream of oral insulin: Where did we reach? Asian J Pharm Clin Res 4(S2):15–20Google Scholar
  4. 4.
    Hussain NH, Jaitley V, Florence A (2001) Transcytosis of nanoparticle and dendrimer delivery systems: evolving vistas. Adv Drug Del Rev 50:107–142CrossRefGoogle Scholar
  5. 5.
    Florence A (2004) Issues in oral nanoparticle drug carrier uptake and targeting. J Drug Target 12(2):65–70CrossRefGoogle Scholar
  6. 6.
    Porter CJ, Charman WN (2001) Intestinal lymphatic drug transport: an update. Adv Drug Del Rev 50:61–80CrossRefGoogle Scholar
  7. 7.
    Humberstone AJ, Charman WN (1997) Lipid-based vehicles for the oral delivery of poorly water soluble drugs. Adv Drug Del Rev 25:103–128CrossRefGoogle Scholar
  8. 8.
    Müller RH, Mäder K, Gohla S (2000) Solid lipid nanoparticles (SLN) for controlled drug delivery—a review of the state of the art. Eur J Pharm Biopharm 50:161–177CrossRefGoogle Scholar
  9. 9.
    Jawahar N, Meyyanathan SN, Reddy G, Sood S (2012) Solid lipid nanoparticles for oral delivery of poorly soluble drugs. J Pharm Sci & Res 4(7):1848–1855Google Scholar
  10. 10.
    Muranishi S (1991) Drug targeting towards the lymphatics. In: Testa B (ed) Advances in drug research, vol 21. Academic Press, London, pp 1–38Google Scholar
  11. 11.
    Yuan H, Chen J, Du Y-Z, Hu FQ, Zeng S, Zhao HL (2007) Studies on oral absorption of stearic acid SLN by a novel fluorometric method. Colloids Surf B 58:157–164CrossRefGoogle Scholar
  12. 12.
    Müller RH, Radtke M, Wissing A (2002) Solid lipid nanoparticles (SLN) and nanostructured lipid carriers (NLC) in cosmetic and dermatological preparations. Adv Drug Del Rev 54:S131–S155CrossRefGoogle Scholar
  13. 13.
    Battaglia L, Gallarate M (2012) Lipid nanoparticles: state of the art, new preparation methods and challenges in drug delivery. Expert Opin Drug Del 9(5):497–508CrossRefGoogle Scholar
  14. 14.
    Almeida AJ, Souto E (2007) Solid lipid nanoparticles as a drug delivery system for peptides and proteins. Adv Drug Del Rev 59:478–490CrossRefGoogle Scholar
  15. 15.
    Battaglia L, Gallarate M, Cavalli R, Trotta M (2010) Solid lipid nanoparticles produced through a coacervation method. J Microencapsul 27:78–85CrossRefGoogle Scholar
  16. 16.
    Battaglia L, D’Addino I, Peira E, Trotta M, Gallarate M (2012) Solid lipid nanoparticles prepared by coacervation method as vehicles for ocular cyclosporine. J Drug Del Sci Technol 22(2):125–130CrossRefGoogle Scholar
  17. 17.
    Battaglia L, Trotta M, Cavalli R PCT n. WO2008 149215 A2Google Scholar
  18. 18.
    Powers ME, Matsuura J, Brassell J, Manning MC, Shefter E (1993) Enhanced solubility of proteins and peptides in nonpolar solvents through hydrophobic ion pairing. Biopolymers 33:927–932CrossRefGoogle Scholar
  19. 19.
    Gallarate M, Battaglia L, Peira E, Trotta M (2011) Peptide-loaded solid lipid nanoparticles prepared through coacervation technique. Int J Chem Eng.  https://doi.org/10.1155/2011/132435 Google Scholar
  20. 20.
    Battaglia L, Trotta M, Gallarate M, Chirio D (2007) Solid lipid nanoparticles formed by solvent-in-water emulsion-diffusion technique: development and influence on insulin stability. J Microencapsul 14:672–684CrossRefGoogle Scholar
  21. 21.
    Silva CM, Ribeiro AJ, Figuereido IV, Gonçalves AR, Veiga F (2006) Alginate microspheres prepared by internal gelation: development and effect on insulin stability. Int J Pharm 311:1–10CrossRefGoogle Scholar
  22. 22.
    Cocco M, Pellegrini C, Martínez-Banaclocha H et al (2017) Development of an acrylate derivative targeting the NLRP3 inflammasome for the treatment of inflammatory bowel disease. J Med Chem 60(9):3656–3671CrossRefGoogle Scholar
  23. 23.
    Battaglia L, Serpe L, Muntoni E, Zara G, Trotta M, Gallarate M (2011) Methotrexate-loaded SLNs prepared by coacervation technique: in vitro cytotoxicity and in vivo pharmacokinetics and biodistribution. Nanomedicine (London) 6(9):1561–1573CrossRefGoogle Scholar
  24. 24.
    Trotta M, Carlotti ME, Gallarate M, Zara GP, Muntoni E, Battaglia L (2011) Insulin-loaded SLN prepared with the emulsion dilution technique. in vivo tracking of nanoparticles after oral administration to rats. J Disp Sci Technol 32:1041–1045CrossRefGoogle Scholar
  25. 25.
    Damgè C, Michel C, Aprahamian M, Couvreur P (1988) New approach for oral administration of insulin with polyalkylcyanoacrylate nanocapsules as drug carrier. Diabetes 37:246–250CrossRefGoogle Scholar
  26. 26.
    Quintanar-Guerrero D, Allemann E, Fessi H, Doelker E (1997) Applications of the ion-pair concept to hydrophilic substances with special emphasis on peptides. Pharm Res 14:119–127CrossRefGoogle Scholar
  27. 27.
    Ruan LP, Chen S, Yu BY, Zhu DN, Cordell GA, Qiu SX (2006) Prediction of human absorption of natural compounds by the non-everted rat intestinal sac model. Eur J Med Chem 41:605–610CrossRefGoogle Scholar
  28. 28.
    Luo Z, Liu Y, Zhao B et al (2013) Ex vivo and in situ approaches used to study intestinal absorption. J Pharm Toxicol Method 68:208–216CrossRefGoogle Scholar
  29. 29.
    Sawai T, Drongowski RA, Lampman RW, Coran AG, Harmon CM (2001) The effect of phospholipids and fatty acids on tight junction permeability and bacterial translocation. Pediatr Surg Int 17:269–274CrossRefGoogle Scholar
  30. 30.
    Sandri G, Bonferoni MC, Rossi S, Ferrari F, Boselli C, Caramella C (2010) Insulin-loaded nanoparticles base on N-Trimethyl Chitosan. In Vitro (CaCo-2 Model) and Ex Vivo (Excised Rat Jejunum, Duodenum, and Ileum) evaluation of penetration enhancement properties. AAPS J 11:362–371Google Scholar
  31. 31.
    Iiboshi Y, Nezu R, Khan J et al (1996) Development changes in distribution of the mucous gel layer in rat small intestine. J Parenter Enteral Nutr 20(6):406–411CrossRefGoogle Scholar
  32. 32.
    Atuma C, Strugala V, Allen A, Holm L (2001) The adherent gastrointestinal mucous gel layer; thickness and physical state in vivo. Am J Physiol Gastrointest Liver Physiol 280(5):G922–G929CrossRefGoogle Scholar
  33. 33.
    Miyazaki M, Mukai H, Iwanaga K, Morimoto K, Kakemi M (2001) Pharmacokinetic–pharmacodynamic modelling of human insulin: validity of pharmacological availability as a substitute for extent of bioavailability. J Pharm Pharmacol 53:1235–1246CrossRefGoogle Scholar
  34. 34.
    Taraghdari ZB, Imani R, Mohabatpour F (2019) A review on bioengineering approaches to insulin delivery: a pharmaceutical and engineering perspective. Macromol Biosci 19:1800458CrossRefGoogle Scholar
  35. 35.
    Wong CY, Al-Salamia H, Dass CR (2017) Potential of insulin nanoparticle formulations for oral delivery and diabetes treatment. J Control Release 264:247–275CrossRefGoogle Scholar
  36. 36.
    Xia CQ, Wang J, Shen WC (2000) Hypoglycemic effect of insulin-transferrin conjugate in streptozotocin-induced diabetic rats. J Pharmacol Exp Ther 295(2):594–600Google Scholar
  37. 37.
    Clement S, Still JG, Kosutic G, McAllister RG (2002) Oral insulin product hexyl-insulin monoconjugate 2 (HIM2) in Type 1 Diabetes Mellitus: the glucose stabilization effects of HIM2. Diabetes Technol Ther 4:459CrossRefGoogle Scholar
  38. 38.
    Sarmento B, Ribeiro A, Veiga F, Ferreira D, Neufeld R (2007) Oral bioavailability of insulin contained in polysaccharide nanoparticles. Biomacromolecules 8:3054–3060CrossRefGoogle Scholar
  39. 39.
    Hurkat P, Jain A, Jain A, Shilpi S, Gulbake A, Jain SK (2012) Concanavalin A conjugated biodegradable nanoparticles for oral insulin delivery. J Nanopart Res 14:1219CrossRefGoogle Scholar
  40. 40.
    Pridgen EM, Alexis F, Kuo TT et al (2013) Transepithelial transport of Fc-targeted nanoparticles by the neonatal fc receptor for oral delivery. Sci Transl Med 5(213):213ra167CrossRefGoogle Scholar
  41. 41.
    Gordin D, Saraheimo M, Tuomikangas J et al (2019) Insulin exposure mitigates the increase of arterial stiffness in patients with type 2 diabetes and albuminuria: an exploratory analysis. Acta Diabetol.  https://doi.org/10.1007/s00592-019-01351-4 Google Scholar
  42. 42.
    Sarmento B, Martins S, Ferreira D, Souto EB (2007) Oral insulin delivery by means of solid lipid nanoparticles. Int J Nanomed 2(4):743–749Google Scholar
  43. 43.
    Tang S, Wu W, Tang W et al (2017) Suppression of Rho-kinase 1 is responsible for insulin regulation of the AMPK/SREBP-1c pathway in skeletal muscle cells exposed to palmitate. Acta Diabetol 54:635–644CrossRefGoogle Scholar
  44. 44.
    Zhang XW, Zhang XL, Xu B, Kang LN (2018) Comparative safety and efficacy of insulin degludec with insulin glargine in type 2 and type 1 diabetes: a meta-analysis of randomized controlled trials. Acta Diabetol 55:429–441CrossRefGoogle Scholar
  45. 45.
    Wang J, Yu J, Zhang Y et al (2019) Glucose transporter inhibitor-conjugated insulin mitigates hypoglycaemia. PNAS 116(22):10744–10748CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Italia S.r.l., part of Springer Nature 2019

Authors and Affiliations

  • Elisabetta Muntoni
    • 1
  • Elisabetta Marini
    • 1
  • Nahid Ahmadi
    • 2
  • Paola Milla
    • 1
  • Corrado Ghè
    • 1
  • Alessandro Bargoni
    • 3
  • Maria Teresa Capucchio
    • 4
  • Elena Biasibetti
    • 5
  • Luigi Battaglia
    • 1
    Email author
  1. 1.Dipartimento di Scienza e Tecnologia del FarmacoUniversità degli Studi di TorinoTurinItaly
  2. 2.Department of ChemistryUniversity of Sistan and BaluchistanZahedanIran
  3. 3.Dipartimento di Scienze della Sanità Pubblica e PediatricheUniversità degli Studi di TorinoTurinItaly
  4. 4.Dipartimento di Scienze VeterinarieUniversità degli Studi di TorinoGrugliascoItaly
  5. 5.Histopathology Department CIBAIstituto Zooprofilattico Sperimentale del PiemonteTurinItaly

Personalised recommendations