Lyophilization Provides Long-term Stability for a Lipid Nanoparticle-formulated, Nucleoside-modified MRNA Vaccine

Overview

Journal Mol Ther

Publisher Cell Press

Specialties Molecular Biology
Pharmacology

Date 2022 Feb 8

PMID 35131437

Authors

Hiromi Muramatsu

Kieu Lam

Csaba Bajusz

Dorottya Laczko

Katalin Kariko

Petra Schreiner

Alan Martin

Peter Lutwyche

James Heyes

Norbert Pardi

Affiliations

Soon will be listed here.

Abstract

Lipid nanoparticle (LNP)-formulated nucleoside-modified mRNA vaccines have proven to be very successful in the fight against the coronavirus disease 2019 (COVID-19) pandemic. They are effective, safe, and can be produced in large quantities. However, the long-term storage of mRNA-LNP vaccines without freezing is still a challenge. Here, we demonstrate that nucleoside-modified mRNA-LNPs can be lyophilized, and the physicochemical properties of the lyophilized material do not significantly change for 12 weeks after storage at room temperature and for at least 24 weeks after storage at 4°C. Importantly, we show in comparative mouse studies that lyophilized firefly luciferase-encoding mRNA-LNPs maintain their high expression, and no decrease in the immunogenicity of a lyophilized influenza virus hemagglutinin-encoding mRNA-LNP vaccine was observed after 12 weeks of storage at room temperature or for at least 24 weeks after storage at 4°C. Our studies offer a potential solution to overcome the long-term storage-related limitations of nucleoside-modified mRNA-LNP vaccines.

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References

Jeffs L, Palmer L, Ambegia E, Giesbrecht C, Ewanick S, MacLachlan I . A scalable, extrusion-free method for efficient liposomal encapsulation of plasmid DNA. Pharm Res. 2005; 22(3):362-72. DOI: 10.1007/s11095-004-1873-z. View

Buschmann M, Carrasco M, Alishetty S, Paige M, Alameh M, Weissman D . Nanomaterial Delivery Systems for mRNA Vaccines. Vaccines (Basel). 2021; 9(1). PMC: 7836001. DOI: 10.3390/vaccines9010065. View

Baiersdorfer M, Boros G, Muramatsu H, Mahiny A, Vlatkovic I, Sahin U . A Facile Method for the Removal of dsRNA Contaminant from In Vitro-Transcribed mRNA. Mol Ther Nucleic Acids. 2019; 15:26-35. PMC: 6444222. DOI: 10.1016/j.omtn.2019.02.018. View

Hou X, Zaks T, Langer R, Dong Y . Lipid nanoparticles for mRNA delivery. Nat Rev Mater. 2021; 6(12):1078-1094. PMC: 8353930. DOI: 10.1038/s41578-021-00358-0. View

Patel S, Nail S, Pikal M, Geidobler R, Winter G, Hawe A . Lyophilized Drug Product Cake Appearance: What Is Acceptable?. J Pharm Sci. 2017; 106(7):1706-1721. DOI: 10.1016/j.xphs.2017.03.014. View

Crommelin D, Anchordoquy T, Volkin D, Jiskoot W, Mastrobattista E . Addressing the Cold Reality of mRNA Vaccine Stability. J Pharm Sci. 2020; 110(3):997-1001. PMC: 7834447. DOI: 10.1016/j.xphs.2020.12.006. View

Geisbert T, Lee A, Robbins M, Geisbert J, Honko A, Sood V . Postexposure protection of non-human primates against a lethal Ebola virus challenge with RNA interference: a proof-of-concept study. Lancet. 2010; 375(9729):1896-905. PMC: 7138079. DOI: 10.1016/S0140-6736(10)60357-1. View

Geisbert T, Hensley L, Kagan E, Yu E, Geisbert J, Daddario-DiCaprio K . Postexposure protection of guinea pigs against a lethal ebola virus challenge is conferred by RNA interference. J Infect Dis. 2006; 193(12):1650-7. PMC: 7110204. DOI: 10.1086/504267. View

Freyn A, Silva J, Rosado V, Bliss C, Pine M, Mui B . A Multi-Targeting, Nucleoside-Modified mRNA Influenza Virus Vaccine Provides Broad Protection in Mice. Mol Ther. 2020; 28(7):1569-1584. PMC: 7335735. DOI: 10.1016/j.ymthe.2020.04.018. View

10.

Chen C, Han D, Cai C, Tang X . An overview of liposome lyophilization and its future potential. J Control Release. 2009; 142(3):299-311. DOI: 10.1016/j.jconrel.2009.10.024. View

11.

Kasper J, Winter G, Friess W . Recent advances and further challenges in lyophilization. Eur J Pharm Biopharm. 2013; 85(2):162-9. DOI: 10.1016/j.ejpb.2013.05.019. View

12.

Polack F, Thomas S, Kitchin N, Absalon J, Gurtman A, Lockhart S . Safety and Efficacy of the BNT162b2 mRNA Covid-19 Vaccine. N Engl J Med. 2020; 383(27):2603-2615. PMC: 7745181. DOI: 10.1056/NEJMoa2034577. View

13.

Hogan M, Pardi N . mRNA Vaccines in the COVID-19 Pandemic and Beyond. Annu Rev Med. 2021; 73:17-39. DOI: 10.1146/annurev-med-042420-112725. View

14.

Ball R, Bajaj P, Whitehead K . Achieving long-term stability of lipid nanoparticles: examining the effect of pH, temperature, and lyophilization. Int J Nanomedicine. 2017; 12:305-315. PMC: 5221800. DOI: 10.2147/IJN.S123062. View

15.

Semple S, Akinc A, Chen J, Sandhu A, Mui B, Cho C . Rational design of cationic lipids for siRNA delivery. Nat Biotechnol. 2010; 28(2):172-6. DOI: 10.1038/nbt.1602. View

16.

Schoenmaker L, Witzigmann D, Kulkarni J, Verbeke R, Kersten G, Jiskoot W . mRNA-lipid nanoparticle COVID-19 vaccines: Structure and stability. Int J Pharm. 2021; 601:120586. PMC: 8032477. DOI: 10.1016/j.ijpharm.2021.120586. View

17.

Hassett K, Higgins J, Woods A, Levy B, Xia Y, Hsiao C . Impact of lipid nanoparticle size on mRNA vaccine immunogenicity. J Control Release. 2021; 335:237-246. DOI: 10.1016/j.jconrel.2021.05.021. View

18.

Baden L, El Sahly H, Essink B, Kotloff K, Frey S, Novak R . Efficacy and Safety of the mRNA-1273 SARS-CoV-2 Vaccine. N Engl J Med. 2020; 384(5):403-416. PMC: 7787219. DOI: 10.1056/NEJMoa2035389. View

19.

Heyes J, Hall K, Tailor V, Lenz R, MacLachlan I . Synthesis and characterization of novel poly(ethylene glycol)-lipid conjugates suitable for use in drug delivery. J Control Release. 2006; 112(2):280-90. DOI: 10.1016/j.jconrel.2006.02.012. View

20.

Zhao P, Hou X, Yan J, Du S, Xue Y, Li W . Long-term storage of lipid-like nanoparticles for mRNA delivery. Bioact Mater. 2020; 5(2):358-363. PMC: 7078456. DOI: 10.1016/j.bioactmat.2020.03.001. View