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.

项目概要 mRNA疫苗已获得针对Covid-19的紧急授权批准，并正在开发中 其他几种传染病。这些疫苗中的 mRNA 编码的免疫原在脂质内传递 纳米颗粒（LNP）包含四种脂质，最重要的是可电离脂质，它负责 通过与胞内体相互作用的胺基的质子化从细胞内释放 mRNA 内体膜。临床研究表明这些疫苗对超过 94% 的患者非常有效 免受 SARS-CoV-2 感染。尽管SARS的mRNA LNP疫苗研发成功 Cov-2 是一种新型医疗方式的重大突破，同时也凸显了一些尚未解答的问题 围绕该平台需要紧急调查的问题包括：1）有哪些关键功能 需要尊重 mRNA LNP 制造来创建有效的系统？ 2）什么是功能性的 体外翻译 (IVT) 或降解产生的 mRNA 转录物截短的后果以及如何 可以通过 IVT 优化将它们最小化吗？ 3) 是什么原因导致 mRNA LNP 降解 制造和储存以及如何测量和最小化这种情况？ 我们最近发现了一种合理设计可电离脂质以提高效力的新工艺。我们也 发现了一种组装 mRNA LNP 的新制造工艺，使得相同的脂质和 mRNA 组件产生更高效、更有效的输送系统。这使得更多的 mRNA LNP 被转化，从而降低剂量和反应原性，并增加可治疗的人数 接种相同数量的疫苗。我们已经启动研究来表征和理解 mRNA LNP 稳定性会影响这些疫苗的质量和性能。在这个提案中我们将执行 研究进一步了解 mRNA LNP 如何在不同的环境中组装和制造 生产规模从实验室规模到商业规模。缺乏该领域的公共信息是一个问题 改进和扩大这种新疫苗方式的使用的主要障碍。我们还将照亮 mRNA 药物中不同转录物异质性的存在和重要性以及如何实现这一点 异质性可能会影响产品性能。最后，mRNA LNP 在制造过程中的稳定性 将研究和储存，以了解导致生物活性和结构损失的降解机制 mRNA 和脂质的完整性，如何测量这些特性，并开发具有延长寿命的配方 与当前产品相比的稳定性概况。