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全面优化冻干粉化工艺,提高冻干 mRNA-LNPs 的长期稳定性和体内性能。

Comprehensive Optimization of a Freeze-Drying Process Achieving Enhanced Long-Term Stability and In Vivo Performance of Lyophilized mRNA-LNPs.

机构信息

CerTest Biotec S.L., 50840 San Mateo de Gállego, Spain.

出版信息

Int J Mol Sci. 2024 Oct 1;25(19):10603. doi: 10.3390/ijms251910603.


DOI:10.3390/ijms251910603
PMID:39408932
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11476828/
Abstract

The success of mRNA vaccines against SARS-CoV-2 has prompted interest in mRNA-based pharmaceuticals due to their rapid production, adaptability, and safety. Despite these advantages, the inherent instability of mRNA and its rapid degradation in vivo underscores the need for an encapsulation system for the administration and delivery of RNA-based therapeutics. Lipid nanoparticles (LNPs) have proven the most robust and safest option for in vivo applications. However, the mid- to long-term storage of mRNA-LNPs still requires sub-zero temperatures along the entire chain of supply, highlighting the need to develop alternatives to improve mRNA vaccine stability under non-freezing conditions to facilitate logistics and distribution. Lyophilization presents itself as an effective alternative to prolong the shelf life of mRNA vaccines under refrigeration conditions, although a complex optimization of the process parameters is needed to maintain the integrity of the mRNA-LNPs. Recent studies have demonstrated the feasibility of freeze-drying LNPs, showing that lyophilized mRNA-LNPs retain activity and stability. However, long-term functional data remain limited. Herein, we focus on obtaining an optimized lyophilizable mRNA-LNP formulation through the careful selection of an optimal buffer and cryoprotectant and by tuning freeze-drying parameters. The results demonstrate that our optimized lyophilization process maintains LNP characteristics and functionality for over a year at refrigerated temperatures, offering a viable solution to the logistical hurdles of mRNA vaccine distribution.

摘要

mRNA 疫苗在对抗 SARS-CoV-2 方面的成功,促使人们对基于 mRNA 的药物产生了兴趣,因为它们具有快速生产、适应性强和安全性高的特点。尽管有这些优势,但 mRNA 的内在不稳定性及其在体内的快速降解,凸显了需要一种封装系统来管理和输送基于 RNA 的治疗药物。脂质纳米粒(LNPs)已被证明是最稳健和最安全的体内应用选择。然而,mRNA-LNP 的中至长期储存仍然需要在整个供应链中保持零下温度,这凸显了需要开发替代品来提高 mRNA 疫苗在非冻结条件下的稳定性,以便利物流和配送。冷冻干燥是一种有效的延长冷藏条件下 mRNA 疫苗保质期的替代方法,尽管需要对工艺参数进行复杂的优化,以保持 mRNA-LNP 的完整性。最近的研究已经证明了冻干 LNPs 的可行性,表明冻干的 mRNA-LNPs 保持活性和稳定性。然而,长期功能数据仍然有限。本文中,我们专注于通过仔细选择最佳缓冲液和冷冻保护剂,并调整冷冻干燥参数,获得优化的可冻干 mRNA-LNP 配方。结果表明,我们优化的冷冻干燥工艺可在冷藏温度下保持 LNP 特性和功能超过一年,为 mRNA 疫苗配送的物流障碍提供了可行的解决方案。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb49/11476828/713f9f1f45ae/ijms-25-10603-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb49/11476828/7557c8f0e52d/ijms-25-10603-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb49/11476828/9190773633a4/ijms-25-10603-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb49/11476828/fcec8180aff8/ijms-25-10603-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb49/11476828/713f9f1f45ae/ijms-25-10603-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb49/11476828/7557c8f0e52d/ijms-25-10603-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb49/11476828/9190773633a4/ijms-25-10603-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb49/11476828/fcec8180aff8/ijms-25-10603-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb49/11476828/713f9f1f45ae/ijms-25-10603-g004.jpg

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引用本文的文献

[1]
Freeze-Drying of mRNA-LNPs Vaccines: A Review.

Vaccines (Basel). 2025-8-12

[2]
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[3]
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[4]
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Int J Mol Sci. 2025-5-27

[6]
Revolutionizing mRNA Vaccines Through Innovative Formulation and Delivery Strategies.

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本文引用的文献

[1]
Purification of linearized template plasmid DNA decreases double-stranded RNA formation during IVT reaction.

Front Mol Biosci. 2023-9-29

[2]
Lyophilization of Nanoparticles, Does It Really Work? Overview of the Current Status and Challenges.

Int J Mol Sci. 2023-9-13

[3]
Induction of Bleb Structures in Lipid Nanoparticle Formulations of mRNA Leads to Improved Transfection Potency.

Adv Mater. 2023-8

[4]
Continuous freeze-drying of messenger RNA lipid nanoparticles enables storage at higher temperatures.

J Control Release. 2023-5

[5]
The Effect of Cryoprotectants and Storage Conditions on the Transfection Efficiency, Stability, and Safety of Lipid-Based Nanoparticles for mRNA and DNA Delivery.

Adv Healthc Mater. 2023-7

[6]
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Langmuir. 2023-2-21

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