Manufacturing and Characterization of Potent mRNA Lipid Nanoparticle Vaccines at Multiple Scales

多尺度有效 mRNA 脂质纳米颗粒疫苗的制造和表征

• 批准号: 10491863
• 负责人: MIKELL PAIGE
• 金额: $ 49.14万
• 依托单位: GEORGE MASON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-15 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10491863
• 关键词: Manufacturing; Characterization; Potent; mRNA; Lipid

PROJECT SUMMARY mRNA vaccines have received emergency authorization approval for Covid-19 and are under development for several other infectious diseases. The mRNA-encoded immunogen in these vaccines is delivered inside a lipid nanoparticle (LNP) that comprises four lipids, the most important being the ionizable lipid that is responsible for releasing the mRNA from intracellular endosomes through protonation of amine groups that interact with the endosomal membrane. Clinical studies showed these vaccines are highly efficacious with over 94% of patients protected from SARS-CoV-2 infection. Although the successful development of mRNA LNP vaccines for SARS- Cov-2 constitutes a major breakthrough for a novel medical modality, it has also highlighted several unanswered questions surrounding this platform that require urgent investigation including : 1) What are the critical features of mRNA LNP manufacturing that need to be respected to create potent systems ? 2) What is functional consequence of truncated mRNA transcripts produced by in vitro translation (IVT) or by degradation and how can they be minimized through IVT optimization ? 3) What causes degradation of mRNA LNPs during manufacturing and storage and how can this be measured and minimized ? We recently discovered a new process to rationally design ionizable lipids for increased potency. We also discovered a novel manufacturing process to assemble the mRNA LNP such that the same lipid and mRNA components produce a much more efficient and potent delivery system. This allows more of the mRNA in the LNP to be translated thereby reducing dose and reactogenicity and increasing the number of people that can be vaccinated with the same quantity of vaccine. We have initiated studies to characterize and understand mRNA LNP stability that can affect the quality and performance of these vaccines. In this proposal we will perform studies to further the understanding of how mRNA LNPs are assembled and manufactured at different manufacturing scales from laboratory scale to commercial scale. The lack of public information in this area is a major impediment to improving and broadening the use of this new vaccine modality. We will also illuminate the presence and importance of the heterogeneity of different transcripts in the mRNA drug substance and how that heterogeneity could impact product performance. And finally, the stability of mRNA LNPs during manufacturing and storage will be studied to understand the degradation mechanisms causing loss of bioactivity and structural integrity of the mRNA and lipids, how to measure these properties, and develop formulations that have extended stability profiles compared to current products.

项目摘要 mRNA疫苗已获得COVID-19的紧急授权批准，并正在开发 其他几种传染病。这些疫苗中的mRNA编码的免疫原被输送到脂质中 包括四个脂质的纳米颗粒（LNP），最重要的是电离脂质，负责的脂质 通过与胺的质子化释放mRNA从细胞内内体释放 内体膜。临床研究表明，这些疫苗高度有效，超过94％的患者 免受SARS-COV-2感染的保护。尽管成功开发了mRNA LNP疫苗用于SARS- COV-2构成了一种新型医学方式的重大突破，它还突出了几个未回答的 围绕需要紧急调查的该平台的问题，包括：1）关键功能是什么 需要尊重建立有效系统的mRNA LNP制造业？ 2）什么是功能 通过体外翻译（IVT）或通过降解产生的截短的mRNA转录本的结果 可以通过IVT优化最小化它们吗？ 3）是什么导致mRNA LNP降解期间 制造和存储，如何测量和最小化？ 最近，我们发现了一个新的过程，以合理设计可电离的脂质，以提高效力。我们也是 发现了一个新的制造过程来组装mRNA LNP，使得相同的脂质和mRNA 组件产生更有效和有效的输送系统。这允许更多的mRNA LNP要翻译，从而降低剂量和反应生成性，并增加可以是的人数 用相同数量的疫苗接种。我们已经开始研究以表征和理解mRNA LNP稳定性会影响这些疫苗的质量和性能。在这个建议中，我们将执行 研究以进一步了解mRNA LNP的组装和制造方式 制造规模从实验室量表到商业规模。该领域缺乏公共信息是 改善和扩大这种新疫苗模式的使用的主要障碍。我们还将照亮 mRNA药物中不同转录本的异质性的存在和重要性以及如何 异质性可能会影响产品性能。最后，制造过程中mRNA LNP的稳定性 将研究存储以了解导致生物活性和结构性丧失的降解机制 mRNA和脂质的完整性，如何测量这些特性并开发已扩展的制剂 与当前产品相比，稳定性概况。