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Pharmaceutical Research

, Volume 28, Issue 5, pp 1157–1166 | Cite as

Inhalable Sustained-Release Formulation of Glucagon: In Vitro Amyloidogenic and Inhalation Properties, and In Vivo Absorption and Bioactivity

  • Satomi Onoue
  • Kazuki Kuriyama
  • Atsushi Uchida
  • Takahiro Mizumoto
  • Shizuo Yamada
Research Paper

ABSTRACT

Purpose

The present study aimed to develop novel glucagon-loaded PLGA nanospheres without cytotoxic fibril formation for chronic glucagon replacement therapy.

Methods

Glucagon-loaded nanospheres (GLG/NS) were prepared by an emulsion solvent diffusion method in oil, and a respirable powder formulation (GLG/NS-RP) was prepared with a jet mill. Physicochemical and inhalation properties of GLG/NS-RP were characterized, and pharmacokinetic behavior and hyperglycemic effect of intratracheally instilled GLG/NS-RP were evaluated in rats.

Results

Although preparation of GLG/NS using glucagon solution at concentrations over 10 mg/mL led to significant formation of cytotoxic glucagon aggregates, glucagon solution at less than 5 mg/mL did not cause structural changes. Drug release behavior of GLG/NS showed a biphasic pattern with an initial burst and slow diffusion. Laser diffraction and cascade impactor analyses of GLG/NS-RP suggested high dispersion and deposition in the respiratory organs with a fine particle fraction of 20.5%. After the intratracheal administration of the GLG/NS-RP (200 μg glucagon/kg) in rats, glucagon was released in a sustained manner, leading to sustained hyperglycemic effects compared with those of normal glucagon powder.

Conclusion

These data would suggest a therapeutic benefit of the newly developed GLG/NS-RP as an alternative to the injection form of glucagon currently used.

KEY WORDS

cytotoxic fibril dry powder inhaler glucagon PLGA sustained release 

Notes

ACKNOWLEDGMENTS

This work was supported in part by a Grant-in-Aid for Young Scientists (B) (No.22790043; S. Onoue) from the Ministry of Education, Culture, Sports, Science and Technology, and Project of Shizuoka Prefecture and Shizuoka City Collaboration of Regional Entities for the Advancement of Technological Excellence, Japan Science and Technology Agency (JST).

REFERENCES

  1. 1.
    Unger RH, Dobbs RE, Orci L. Insulin, glucagon, and somatostatin secretion in the regulation of metabolism. Annu Rev Physiol. 1978;40:307–43.PubMedCrossRefGoogle Scholar
  2. 2.
    Mochiki E, Suzuki H, Takenoshita S, Nagamachi Y, Kuwano H, Mizumoto A, et al. Mechanism of inhibitory effect of glucagon on gastrointestinal motility and cause of side effects of glucagon. J Gastroenterol. 1998;33:835–41.PubMedCrossRefGoogle Scholar
  3. 3.
    Slezak LA, Andersen DK. Pancreatic resection: effects on glucose metabolism. World J Surg. 2001;25:452–60.PubMedCrossRefGoogle Scholar
  4. 4.
    Hirota M, Ikei S, Mishima M, Mori K, Sakamoto K, Yamane T, et al. Glucagon in the metabolic and nutritional management after total pancreatectomy–a case report. Jpn J Surg. 1989;19:586–92.PubMedCrossRefGoogle Scholar
  5. 5.
    Teshima D, Yamauchi A, Makino K, Kataoka Y, Arita Y, Nawata H, et al. Nasal glucagon delivery using microcrystalline cellulose in healthy volunteers. Int J Pharm. 2002;233:61–6.PubMedCrossRefGoogle Scholar
  6. 6.
    Endo K, Amikawa S, Matsumoto A, Sahashi N, Onoue S. Erythritol-based dry powder of glucagon for pulmonary administration. Int J Pharm. 2005;290:63–71.PubMedCrossRefGoogle Scholar
  7. 7.
    Onoue S, Yamamoto K, Kawabata Y, Hirose M, Mizumoto T, Yamada S. Novel dry powder inhaler formulation of glucagon with addition of citric acid for enhanced pulmonary delivery. Int J Pharm. 2009;382:144–50.PubMedCrossRefGoogle Scholar
  8. 8.
    Okada H. One- and three-month release injectable microspheres of the LH-RH superagonist leuprorelin acetate. Adv Drug Deliv Rev. 1997;28:43–70.PubMedCrossRefGoogle Scholar
  9. 9.
    Houchin ML, Topp EM. Chemical degradation of peptides and proteins in PLGA: a review of reactions and mechanisms. J Pharm Sci. 2008;97:2395–404.PubMedCrossRefGoogle Scholar
  10. 10.
    Onoue S, Ohshima K, Debari K, Koh K, Shioda S, Iwasa S, et al. Mishandling of the therapeutic peptide glucagon generates cytotoxic amyloidogenic fibrils. Pharm Res. 2004;21:1274–83.PubMedCrossRefGoogle Scholar
  11. 11.
    Maji SK, Perrin MH, Sawaya MR, Jessberger S, Vadodaria K, Rissman RA, et al. Functional amyloids as natural storage of peptide hormones in pituitary secretory granules. Science. 2009;325:328–32.PubMedCrossRefGoogle Scholar
  12. 12.
    Merrifield RB. Solid-phase peptide synthesis. Adv Enzymol Relat Areas Mol Biol. 1969;32:221–96.PubMedGoogle Scholar
  13. 13.
    Kawashima Y, Yamamoto H, Takeuchi H, Hino T, Niwa T. Properties of a peptide containing DL-lactide/glycolide copolymer nanospheres prepared by novel emulsion solvent diffusion methods. Eur J Pharm Biopharm. 1998;45:41–8.PubMedCrossRefGoogle Scholar
  14. 14.
    Onoue S, Waki Y, Nagano Y, Satoh S, Kashimoto K. The neuromodulatory effects of VIP/PACAP on PC-12 cells are associated with their N-terminal structures. Peptides. 2001;22:867–72.PubMedCrossRefGoogle Scholar
  15. 15.
    LeVine 3rd H. Thioflavine T interaction with synthetic Alzheimer’s disease beta- amyloid peptides: detection of amyloid aggregation in solution. Protein Sci. 1993;2:404–10.PubMedCrossRefGoogle Scholar
  16. 16.
    Ikegami K, Kawashima Y, Takeuchi H, Yamamoto H, Isshiki N, Momose D, et al. Improved inhalation behavior of steroid KSR-592 in vitro with Jethaler by polymorphic transformation to needle-like crystals (beta-form). Pharm Res. 2002;19:1439–45.PubMedCrossRefGoogle Scholar
  17. 17.
    Onoue S, Yajima T. Fibrillar aggregates of amyloidogenic peptides: new Insight into the neuroprotective effects of neuropeptides on fibril-induced neuronal cell death. NY: Nova Science Publishers; 2006.Google Scholar
  18. 18.
    Bucciantini M, Giannoni E, Chiti F, Baroni F, Formigli L, Zurdo J, et al. Inherent toxicity of aggregates implies a common mechanism for protein misfolding diseases. Nature. 2002;416:507–11.PubMedCrossRefGoogle Scholar
  19. 19.
    Musumeci T, Ventura CA, Giannone I, Ruozi B, Montenegro L, Pignatello R, et al. PLA/PLGA nanoparticles for sustained release of docetaxel. Int J Pharm. 2006;325:172–9.PubMedCrossRefGoogle Scholar
  20. 20.
    Zheng XM, Martin GP, Marriott C. Particulate interactions in dry powder formulations for inhalation. London: Taylor&Francis; 2001.CrossRefGoogle Scholar
  21. 21.
    Larhrib H, Martin GP, Marriott C, Prime D. The influence of carrier and drug morphology on drug delivery from dry powder formulations. Int J Pharm. 2003;257:283–96.PubMedCrossRefGoogle Scholar
  22. 22.
    Maeda H, Okabayashi T, Yatabe T, Yamashita K, Hanazaki K. Perioperative intensive insulin therapy using artificial endocrine pancreas in patients undergoing pancreatectomy. World J Gastroenterol. 2009;15:4111–5.PubMedCrossRefGoogle Scholar
  23. 23.
    Cryan SA. Carrier-based strategies for targeting protein and peptide drugs to the lungs. AAPS J. 2005;7:E20–41.PubMedCrossRefGoogle Scholar
  24. 24.
    Patton JS. Pulmonary delivery of drugs for bone disorders. Adv Drug Deliv Rev. 2000;42:239–48.PubMedCrossRefGoogle Scholar
  25. 25.
    Agu RU, Ugwoke MI, Armand M, Kinget R, Verbeke N. The lung as a route for systemic delivery of therapeutic proteins and peptides. Respir Res. 2001;2:198–209.PubMedCrossRefGoogle Scholar
  26. 26.
    Patton JS, Byron PR. Inhaling medicines: delivering drugs to the body through the lungs. Nat Rev Drug Discov. 2007;6:67–74.PubMedCrossRefGoogle Scholar
  27. 27.
    Huypens P, Ling Z, Pipeleers D, Schuit F. Glucagon receptors on human islet cells contribute to glucose competence of insulin release. Diabetologia. 2000;43:1012–9.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Satomi Onoue
    • 1
  • Kazuki Kuriyama
    • 1
  • Atsushi Uchida
    • 1
  • Takahiro Mizumoto
    • 2
    • 3
  • Shizuo Yamada
    • 1
  1. 1.Department of Pharmacokinetics and Pharmacodynamics and Global Center of Excellence (COE) ProgramSchool of Pharmaceutical Sciences, University of ShizuokaShizuokaJapan
  2. 2.Department of Product DevelopmentIto Life Sciences Inc.MoriyaJapan
  3. 3.American Peptide CompanySunnyvaleUSA

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