mRNA治疗的临床潜力分析与应用
2022-04-11 来源:本站 点击次数:1484
信使RNA（mRNA）是一类新兴的生物治疗药物。虽然它们有相当大的前景，但这一领域仍处于起步阶段。mRNA治疗为挑战性疾病的靶向治疗和灵活生产提供了新的机遇，抗COVID-19的mRNA疫苗的快速发展就证明了这一点。
mRNA的临床潜力已经超出了疫苗的范畴，扩展到其他先进的治疗方法，例如蛋白质替代、基因治疗和癌症免疫治疗。
由于mRNA是脆弱的，不稳定的，开发 mRNA 疫苗的最重要的挑战仍然是其固有的不稳定性，因为它很有可能在冰点以上分解。
mRNA 构建模块的修饰以及能够相对安全地包裹它颗粒的开发帮助了mRNA 疫苗候选者。但是这种新型疫苗仍然需要前所未有的冷冻条件存储、运输和管理。
WHO ：
在用 LNP 包封期间以及制造条件下应特别注意尽量减少 mRNA 的降解，这个包封和生产添加会影响 LNP 及最终 mRNA-LNP 疫苗产品的稳定性，例如：产生的影响条件包括mRNA解冻和LNPs或mRNA LNPs的冷冻速率。
Assessment report---COVID-19 Vaccine（根据某上市的mRNA疫苗的评估报告---EMA）
Characterisation 特征 ---提供了冻融降解的数据
The applicant provided a detailed characterisation of the mRNA active substance including detailed information on the structural properties of the mRNA CX-024414. Additionally, data on the chromatographic profile, the thermal denaturation as well as freeze thaw degradation and their influence on protein expression have been provided.
申请人提供了mRNA活性物质的详细特征，包括mRNA CX-024414结构特性的详细信息。此外，还提供了色谱图、热变性和冻融降解及其对蛋白质表达的影响数据。
The composition of the finished product
最终产品的成分---采用蔗糖来提高冻融和存储稳定性
Sucrose is added to promote product stability through freeze/thaw and long-term storage.
添加蔗糖是为了通过冷冻/解冻和长期储存来提高产品的稳定性。
Manufacturing process development
生产工艺开发---增加了放大和大体积冷冻步骤
mRNA-loaded LNP intermediate 载有 mRNA 的 LNP 中间体
The product consists of an mRNA-lipid complex dispersion that contains the mRNA (CX-024414) that encodes for the pre-fusion stabilised Spike protein of 2019-novel Coronavirus (SARS-CoV-2).Comprehensive development data have been provided. Changes made during development have been described. They included scale-up, addition of a bulk freezing step, change of some equipment and revision of the lipid molar ratios to allow for harmonisation with the platform process.
该产品由一种mRNA-脂质复合分散体组成，其中包含编码2019年新型冠状病毒（SARS-CoV-2）融合前稳定 Spike 蛋白的mRNA（CX-024414）。已提供全面的开发数据。已经描述了在开发过程中所做的变更。它们包括放大、增加批量冷冻步骤、改变一些设备和修改脂质摩尔比以与平台工艺匹配。
Manufacture of the product and process controls
产品生产和工艺控制---涵盖了冷冻工艺
The LNP manufacturing process comprises lipid stock solution (LSS) preparation, nanoprecipitation mixing, tangential flow filtration (TFF), dilution and cryoprotectant addition, clarification, fill, and freezing and storage.
LNP 制造过程包括脂质储备溶液 (LSS) 制备、纳米沉淀混合、切向流过滤 (TFF)、稀释和冷冻保护剂添加、澄清、填充以及冷冻和储存。
Stability of the product
产品的稳定性--现有的冻融工艺对产品的稳定性没有影响
mRNA-loaded LNP intermediate载有 mRNA 的 LNP 中间体
Stability studies in support of mRNA-loaded LNP intermediate freeze-thaw cycling were performed using the development lot. The results show that the mRNA-1273 LNP biophysical attributes (particle size, polydispersity, encapsulation) are not impacted by the different freeze-thaw processes (room temperature or 2°C to 8°C) or 5 cycles of freeze-thaw
使用开发批次进行了支持负载 mRNA 的 LNP 中间冻融循环的稳定性研究。结果表明，mRNA-1273 LNP 生物物理属性（粒径、多分散性、封装）不受不同冻融过程（室温或 2°C 至 8°C）或 5 个冻融循环的影响，还进行了支持成品冻融循环的稳定性研究。冻融循环最多重复五次。所有结果都在规格范围内，表明最终产品在重复冻融步骤方面相对稳定，支持五个冻融循环。然而，批准的产品信息中不允许在上市时反复冻融小瓶。
Finished product
最终成品---仍然需要做冻融测试
Stability studies in support of finished product freeze-thaw cycling were also performed. The freeze-thaw cycling was repeated up to five times. All results were within the specification and indicate that the final product is relatively stable with regard to repeated freeze-thaw steps support five freeze/thaw cycles. Repeated freeze-thaw of vials upon marketing is however, not permitted in the approved product information
Design and conduct of clinical studies
临床研究的设计和执行---仍需要评估冻融的稳定性，涵盖运输和存储
The RT-PCR assay is acceptably validated and additional validation studies to evaluate stability of clinical samples at various temperatures and freeze/thawing cycles to cover shipping and storage conditions were provided.
RT-PCR 测定得到了可接受的验证，并提供了额外的验证研究，以评估临床样品在各种温度和冷冻/解冻循环中的稳定性，以涵盖运输和储存条件。
Why not impacted by the different freeze-thaw processes
为什么冻融工艺对产品没有影响?
冻融工艺的开发
特殊要求的冻融曲线降低了对mRNA产品的影响
The goal must be to reduce STRESSES DURING FREEZING
目标必须是在冻结过程中减少压力
Stress is coursed by:
压力主要这几方面
• CRYOCONCENTRATION冷冻浓缩
• PH SHIFT pH pH漂移
• PROTEIN INTERACTION WITH THE ICE SURFACE蛋白冰液界面的变性
mRNA Freezing experience
mRNA冷冻实验
mRNA experienced companies are freezing as fast as possible, so with rates from + to -60° C in < 5hours
经验丰富的mRNA公司会尽可能快地冻结mRNA（在< 5小时内从+到-60°C冻结）
mRNA的临床潜力已经超出了疫苗的范畴，扩展到其他先进的治疗方法，例如蛋白质替代、基因治疗和癌症免疫治疗。
由于mRNA是脆弱的，不稳定的，开发 mRNA 疫苗的最重要的挑战仍然是其固有的不稳定性，因为它很有可能在冰点以上分解。
mRNA 构建模块的修饰以及能够相对安全地包裹它颗粒的开发帮助了mRNA 疫苗候选者。但是这种新型疫苗仍然需要前所未有的冷冻条件存储、运输和管理。
WHO ：
在用 LNP 包封期间以及制造条件下应特别注意尽量减少 mRNA 的降解，这个包封和生产添加会影响 LNP 及最终 mRNA-LNP 疫苗产品的稳定性，例如：产生的影响条件包括mRNA解冻和LNPs或mRNA LNPs的冷冻速率。
Assessment report---COVID-19 Vaccine（根据某上市的mRNA疫苗的评估报告---EMA）
Characterisation 特征 ---提供了冻融降解的数据
The applicant provided a detailed characterisation of the mRNA active substance including detailed information on the structural properties of the mRNA CX-024414. Additionally, data on the chromatographic profile, the thermal denaturation as well as freeze thaw degradation and their influence on protein expression have been provided.
申请人提供了mRNA活性物质的详细特征，包括mRNA CX-024414结构特性的详细信息。此外，还提供了色谱图、热变性和冻融降解及其对蛋白质表达的影响数据。
The composition of the finished product
最终产品的成分---采用蔗糖来提高冻融和存储稳定性
Sucrose is added to promote product stability through freeze/thaw and long-term storage.
添加蔗糖是为了通过冷冻/解冻和长期储存来提高产品的稳定性。
Manufacturing process development
生产工艺开发---增加了放大和大体积冷冻步骤
mRNA-loaded LNP intermediate 载有 mRNA 的 LNP 中间体
The product consists of an mRNA-lipid complex dispersion that contains the mRNA (CX-024414) that encodes for the pre-fusion stabilised Spike protein of 2019-novel Coronavirus (SARS-CoV-2).Comprehensive development data have been provided. Changes made during development have been described. They included scale-up, addition of a bulk freezing step, change of some equipment and revision of the lipid molar ratios to allow for harmonisation with the platform process.
该产品由一种mRNA-脂质复合分散体组成，其中包含编码2019年新型冠状病毒（SARS-CoV-2）融合前稳定 Spike 蛋白的mRNA（CX-024414）。已提供全面的开发数据。已经描述了在开发过程中所做的变更。它们包括放大、增加批量冷冻步骤、改变一些设备和修改脂质摩尔比以与平台工艺匹配。
Manufacture of the product and process controls
产品生产和工艺控制---涵盖了冷冻工艺
The LNP manufacturing process comprises lipid stock solution (LSS) preparation, nanoprecipitation mixing, tangential flow filtration (TFF), dilution and cryoprotectant addition, clarification, fill, and freezing and storage.
LNP 制造过程包括脂质储备溶液 (LSS) 制备、纳米沉淀混合、切向流过滤 (TFF)、稀释和冷冻保护剂添加、澄清、填充以及冷冻和储存。
Stability of the product
产品的稳定性--现有的冻融工艺对产品的稳定性没有影响
mRNA-loaded LNP intermediate载有 mRNA 的 LNP 中间体
Stability studies in support of mRNA-loaded LNP intermediate freeze-thaw cycling were performed using the development lot. The results show that the mRNA-1273 LNP biophysical attributes (particle size, polydispersity, encapsulation) are not impacted by the different freeze-thaw processes (room temperature or 2°C to 8°C) or 5 cycles of freeze-thaw
使用开发批次进行了支持负载 mRNA 的 LNP 中间冻融循环的稳定性研究。结果表明，mRNA-1273 LNP 生物物理属性（粒径、多分散性、封装）不受不同冻融过程（室温或 2°C 至 8°C）或 5 个冻融循环的影响，还进行了支持成品冻融循环的稳定性研究。冻融循环最多重复五次。所有结果都在规格范围内，表明最终产品在重复冻融步骤方面相对稳定，支持五个冻融循环。然而，批准的产品信息中不允许在上市时反复冻融小瓶。
Finished product
最终成品---仍然需要做冻融测试
Stability studies in support of finished product freeze-thaw cycling were also performed. The freeze-thaw cycling was repeated up to five times. All results were within the specification and indicate that the final product is relatively stable with regard to repeated freeze-thaw steps support five freeze/thaw cycles. Repeated freeze-thaw of vials upon marketing is however, not permitted in the approved product information
Design and conduct of clinical studies
临床研究的设计和执行---仍需要评估冻融的稳定性，涵盖运输和存储
The RT-PCR assay is acceptably validated and additional validation studies to evaluate stability of clinical samples at various temperatures and freeze/thawing cycles to cover shipping and storage conditions were provided.
RT-PCR 测定得到了可接受的验证，并提供了额外的验证研究，以评估临床样品在各种温度和冷冻/解冻循环中的稳定性，以涵盖运输和储存条件。
Why not impacted by the different freeze-thaw processes
为什么冻融工艺对产品没有影响?
冻融工艺的开发
特殊要求的冻融曲线降低了对mRNA产品的影响
The goal must be to reduce STRESSES DURING FREEZING
目标必须是在冻结过程中减少压力
Stress is coursed by:
压力主要这几方面
• CRYOCONCENTRATION冷冻浓缩
• PH SHIFT pH pH漂移
• PROTEIN INTERACTION WITH THE ICE SURFACE蛋白冰液界面的变性
mRNA Freezing experience
mRNA冷冻实验
mRNA experienced companies are freezing as fast as possible, so with rates from + to -60° C in < 5hours
经验丰富的mRNA公司会尽可能快地冻结mRNA（在< 5小时内从+到-60°C冻结）
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