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Long-term stability of influenza vaccine in a dissolving microneedle patch

Abstract

This study tested the hypothesis that optimized microneedle patch formulations can stabilize trivalent subunit influenza vaccine during long-term storage outside the cold chain and when exposed to potential stresses found during manufacturing and storage. Formulations containing combinations of trehalose/sucrose, sucrose/arginine, and arginine/heptagluconate were successful at retaining most or all vaccine activity during storage at 25 °C for up to 24 months as determined by ELISA assay. The best formulation of microneedle patches contained arginine/heptagluconate, which showed no significant loss of vaccine activity during the study. To validate these in vitro findings, mice were immunized using trivalent influenza vaccine stored in microneedle patches for more than 1 year at 25 °C, which elicited antibody titers greater than or equal to fresh liquid vaccine delivered by intradermal injection, indicating the retention of immunogenicity during storage. Finally, influenza vaccine in microneedle patches lost no significant activity during exposure to 60 °C for 4 months, multiple freeze-thaw cycles, or electron beam irradiation. We conclude that optimally formulated microneedle patches can retain influenza vaccine activity during extended storage outside the cold chain and during other environmental stresses, which suggests the possibility of microneedle patch storage on pharmacy shelves without refrigeration.

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References

  1. 1.

    World Health Organization. Influenza (seasonal) fact sheet 2014 [10/24/15]. Available from: http://www.who.int/mediacentre/factsheets/fs211/en/index.html.

  2. 2.

    Grohskopf L et al. Prevention and control of seasonal influenza with vaccines: recommendations of the Advisory Committee on Immunization Practices (ACIP)—United States, 2014–15 influenza season. Morb Mortal Wkly Rep. 2014;63:691–7.

  3. 3.

    CDC. Flu vaccination coverage, United States, 2014–15 influenza season: United States centers for disease control and prevention,; 2015 [cited 2015 December]. Available from: http://www.cdc.gov/flu/fluvaxview/coverage-1415estimates.htm.

  4. 4.

    Das P. Revolutionary vaccine technology breaks the cold chain. Lancet Infect Dis. 2004;4(12):719.

  5. 5.

    Patois E, Capelle MAH, Gurny R, Arvinte T. Stability of seasonal influenza vaccines investigated by spectroscopy and microscopy methods. Vaccine. 2011;29(43):7404–13.

  6. 6.

    Cicerone MT, Soles CL. Fast dynamics and stabilization of proteins: binary glasses of trehalose and glycerol. Biophys J. 2004;86(6):3836–45.

  7. 7.

    Chang L, Pikal MJ. Mechanisms of protein stabilization in the solid state. J Pharm Sci. 2009;98(9):2886–908.

  8. 8.

    Lee VHL. Changing needs in drug delivery in the era of peptide and protein drugs. In: Lee VHL, editor. Peptide and protein drug delivery. New York: Marcel Dekker, Inc.; 1991. p. 1–56.

  9. 9.

    Chen X, Fernando GJP, Crichton ML, Flaim C, Yukiko SR, Fairmaid EJ, et al. Improving the reach of vaccines to low-resource regions, with a needle-free vaccine delivery device and long-term thermostabilization. J Control Release. 2011;152(3):349–55.

  10. 10.

    Pearton M, Kang SM, Song JM, Kim YC, Quan FS, Ivory M, et al. Immune stimulation following microneedle delivery of influenza virus-like particle (VLP) vaccines to human skin. Drug Discov Today. 2010;15(23–24):A87.

  11. 11.

    Kommareddy S, Baudner BC, Bonificio A, Gallorini S, Palladino G, Determan AS, et al. Influenza subunit vaccine coated microneedle patches elicit comparable immune responses to intramuscular injection in guinea pigs. Vaccine. 2013;31(34):3435–41.

  12. 12.

    Donnelly RF, Singh TRR, Woolfson AD. Microneedle-based drug delivery systems: microfabrication, drug delivery, and safety. Drug Deliv. 2010;17(4):187–207.

  13. 13.

    McGrath MG, Vrdoljak A, O’Mahony C, Oliveira JC, Moore AC, Crean AM. Determination of parameters for successful spray coating of silicon microneedle arrays. Int J Pharm. 2011;415(1–2):140–9.

  14. 14.

    van der Maaden K, Jiskoot W, Bouwstra J. Microneedle technologies for (trans)dermal drug and vaccine delivery. J Control Release. 2012;161(2):645–55.

  15. 15.

    Lee JW, Park JH, Prausnitz MR. Dissolving microneedles for transdermal drug delivery. Biomaterials. 2008;29(13):2113–24.

  16. 16.

    Zhang J, Pritchard E, Hu X, Valentin T, Panilaitis B, Omenetto FG, et al. Stabilization of vaccines and antibiotics in silk and eliminating the cold chain. Proc Natl Acad Sci U S A. 2012;109(30):11981–6.

  17. 17.

    Kim YC, Quan FS, Compans RW, Kang SM, Prausnitz MR. Stability kinetics of influenza vaccine coated onto microneedles during drying and storage. Pharm Res. 2011;28(1):135–44.

  18. 18.

    Norman JJ, Arya JM, McClain MA, Frew PM, Meltzer MI, Prausnitz MR. Microneedle patches: usability and acceptability for self-vaccination against influenza. Vaccine. 2014;32(16):1856–62.

  19. 19.

    Kim YC, Quan FS, Yoo DG, Compans RW, Kang SM, Prausnitz MR. Enhanced memory responses to seasonal H1N1 influenza vaccination of the skin with the use of vaccine-coated microneedles. J Infect Dis. 2010;201(2):190–8.

  20. 20.

    Vassilieva EV, Kalluri H, McAllister D, Taherbhai MT, Esser ES, Pewin WP, et al. Improved immunogenicity of individual influenza vaccine components delivered with a novel dissolving microneedle patch stable at room temperature. Drug Deliv Transl Res. 2015;5(4):360–71.

  21. 21.

    Carpenter JF, Pikal MJ, Chang BS, Randolph TW. Rational design of stable lyophilized protein formulations: some practical advice. Pharm Res. 1997;14(8):969–75.

  22. 22.

    Wang W. Lyophilization and development of solid protein pharmaceuticals. Int J Pharm. 2000;203(1–2):1–60.

  23. 23.

    Brandau DT, Jones LS, Wiethoff CM, Rexroad J, Middaugh CR. Thermal stability of vaccines. J Pharm Sci. 2003;92(2):218–31.

  24. 24.

    Cicerone MT, Pikal MJ, Qian KK. Stabilization of proteins in solid form. Adv Drug Deliv Rev. 2015;93:14–24.

  25. 25.

    Crowe LM, Reid DS, Crowe JH. Is trehalose special for preserving dry biomaterials? Biophys J. 1996;71(4):2087–93.

  26. 26.

    Grasmeijer N, Stankovic M, de Waard H, Frijlink HW, Hinrichs WLJ. Unraveling protein stabilization mechanisms: vitrification and water replacement in a glass transition temperature controlled system. Biochim Biophys Acta Protein Proteomics. 2013;1834(4):763–9.

  27. 27.

    Monkos K. Studies of protein solution conformations using viscometric measurements. In: Uversky VN, Permyakov EA, editors. Conformational stability, size, shape and surface of protein molecules. Molecular Anatomy and Physiology of Proteins: Nova Science Publishers; 2007. p. 355–87.

  28. 28.

    Geeraedts F, Saluja V, ter Veer W, Amorij J-P, Frijlink H, Wilschut J, et al. Preservation of the immunogenicity of dry-powder influenza H5N1 whole inactivated virus vaccine at elevated storage temperatures. AAPS J. 2010;12(2):215–22.

  29. 29.

    Ohtake S, Kita Y, Arakawa T. Interactions of formulation excipients with proteins in solution and in the dried state. Adv Drug Deliv Rev. 2011;63(13):1053–73.

  30. 30.

    Mistilis MJ, Bommarius AS, Prausnitz MR. Development of a thermostable microneedle patch for influenza vaccination. J Pharm Sci. 2015;104(2):740–9.

  31. 31.

    Chu LY, Choi SO, Prausnitz MR. Fabrication of dissolving polymer microneedles for controlled drug encapsulation and delivery: bubble and pedestal microneedle designs. J Pharm Sci. 2010;99(10):4228–38.

  32. 32.

    Koutsonanos DG, Vassilieva EV, Stavropoulou A, Zarnitsyn VG, Esser ES, Taherbhai MT, et al. Delivery of subunit influenza vaccine to skin with microneedles improves immunogenicity and long-lived protection. Sci Rep. 2012;2:357.

  33. 33.

    Crowe JH, Spargo BJ, Crowe LM. Preservation of dry liposomes does not require retention of residual water. Proc Natl Acad Sci U S A. 1987;84(6):1537–40.

  34. 34.

    Costantino HR, Carrasquillo KG, Cordero RA, Mumenthaler M, Hsu CC, Griebenow K. Effect of excipients on the stability and structure of lyophilized recombinant human growth hormone. J Pharm Sci. 1998;87(11):1412–20.

  35. 35.

    Choi HJ, Bondy BJ, Yoo DG, Compans RW, Kang SM, Prausnitz MR. Stability of whole inactivated influenza virus vaccine during coating onto metal microneedles. J Control Release. 2013;166(2):159–71.

  36. 36.

    Mattern M, Winter G, Kohnert U, Lee G. Formulation of proteins in vacuum-dried glasses. II. Process and storage stability in sugar-free amino acid systems. Pharm Dev Technol. 1999;4(2):199–208.

  37. 37.

    Hai TT, Nelson DJ. Stabilization of therapeutic hemoglobin compositions Patent WO1997039027 A1. 1997.

  38. 38.

    Roe KD, Labuza TP. Glass transition and crystallization of amorphous trehalose-sucrose mixtures. Int J Food Prop. 2005;8(3):559–74.

  39. 39.

    Wolfe J, Bryant G, Koster KL. What is ‘unfreezable water’, how unfreezable is it, and how much is there? Cryo Lett. 2002;23(3):157–66.

  40. 40.

    Ameri M, Wang XM, Maa YF. Effect of irradiation on parathyroid hormone PTH(1–34) coated on a novel transdermal microprojection delivery system to produce a sterile product-adhesive compatibility. J Pharm Sci. 2010;99(4):2123–34.

  41. 41.

    Williams MS. Single-radial-immunodiffusion as an in vitro potency assay for human inactivated viral vaccines. Vet Microbiol. 1993;37(3–4):253–62.

  42. 42.

    Skountzou I, Compans RW. Skin immunization with influenza vaccines. Curr Top Microbiol Immunol. 2015;386:343–69.

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Acknowledgments

We thank Novartis for generously providing monovalent influenza vaccine stock. This work was supported in part by the National Institutes of Health. We thank Polo Gaputan and Miraj Desai for their work on this project. The work was carried out in the Center for Drug Design, Development, and Delivery and the Institute for Bioengineering and Bioscience at the Georgia Institute of Technology. Matthew Mistilis, Andreas Bommarius, and Mark Prausnitz are inventors of patent(s) that have been or may be licensed to companies developing microneedle-based products, and Mark Prausnitz is a paid advisor to companies developing microneedle-based products and is a founder/shareholder of companies developing microneedle-based products. This potential conflict of interest has been disclosed and is overseen by Georgia Tech and Emory University.

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Correspondence to Mark R. Prausnitz.

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Mistilis, M.J., Joyce, J.C., Esser, E.S. et al. Long-term stability of influenza vaccine in a dissolving microneedle patch. Drug Deliv. and Transl. Res. 7, 195–205 (2017). https://doi.org/10.1007/s13346-016-0282-2

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Keywords

  • Vaccine delivery
  • Transdermal delivery
  • Vaccine stability
  • Vaccine formulation
  • Solid dosage form
  • Microneedle
  • Influenza vaccine