Assessment of the performance of MAM vs conventional QC methods for evaluation of Product Quality Attributes of adalimumab and etanercept

评估 MAM 与传统 QC 方法的性能，以评估阿达木单抗和依那西普的产品质量属性

• 批准号: 10619721
• 负责人: Diane McCarthy
• 金额: $ 153.07万
• 依托单位: U.S. PHARMACOPEIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10619721
• 关键词: Assessment; performance; MAM; vs; conventional

ABSTRACT Monoclonal antibodies and other biotherapeutics are subject to a variety of modifications that can impact activity and stability and therefore must be analyzed as part of QC and comparability. Mass spectrometry (MS) has become a workhorse for biopharmaceutical analytical laboratories due to its ability to detect protein modifications at a molecular level. Over the past few years, the Multi-Attribute Method (MAM) has gained traction throughout pharmaceutical development and QC labs, with several developers implementing some form of MAM in characterization or release. While replacing multiple QC tests provides an opportunity to streamline lab work and decrease development time and post-approval costs, several challenges remain. While some large biopharma companies are implementing MAM in QC, MAM is not as commonly used in biosimilar and small biopharma companies. This proposal addresses one of the key areas of consideration for implementation of MAM in QC as outlined in a 2019 publication from FDA staff: the performance of MAM vs conventional methods. Collecting data to support transitioning from conventional techniques to MAM is a significant investment that can prevent or delay development of biosimilars. The objective of this work is to assess the performance of the MS-based MAM versus conventional QC methods to identify changes in product quality attributes (PQAs) upon forced degradation and to correlate changes in those PQAs with bioactivity, binding affinity, and structure. Results of this study will help support transitioning from conventional techniques to MAM by creating a knowledge base that can lower the barrier to adoption of MAM and enable wider use of MAM by biosimilar manufacturers. The work proposed here will assess and compare PQAs of a monoclonal antibody (adalimumab) and Fc fusion protein (etanercept) acquired from three different sources using both conventional QC methods and MAM-based approaches. Sensitivity of each technique to changes during forced degradation will be evaluated and compared to functional and structural changes detected using cell-based assays, binding assays and circular dichroism. We propose to first conduct forced degradation studies of adalimumab and etanercept from multiple sources (Specific Aim 1). PQAs will be evaluated using conventional methods to assess charge variants, glycosylation, and size variants, (Specific Aim 2) and compared to the specific molecular modifications detected using an MS-based MAM workflow (Specific Aim 3) to assess comparability and sensitivity to changes in PQAs. Lastly, under Specific Aim 4, we will assess the bioactivity and structure of biotherapeutic products exhibiting significant differences in PQAs to correlate molecular changes to changes in structure and function.

抽象的 单克隆抗体和其他生物疗法受到各种修饰，可能会影响 活动和稳定性，因此必须作为QC的一部分和可比性进行分析。质谱法 （MS）由于能够检测蛋白质的能力而成为生物药物分析实验室的主力军 分子水平的修饰。在过去的几年中，多属性方法（MAM）已获得 整个药品开发和QC实验室的吸引力，几位开发人员实施了一些 MAM的表征或释放形式。在替换多个质量控制测试的同时，为 简化实验室工作并减少开发时间和批准后成本，仍然存在一些挑战。 虽然一些大型生物制药公司正在QC实施MAM，但MAM并不常用于 生物仿制药和小型生物制药公司。该提案涉及考虑关键领域之一 如FDA员工2019年出版物中概述的QC中MAM的实施：MAM的表现 vs常规方法。收集数据以支持从常规技术过渡到MAM是一个 可以防止或延迟生物仿制药的发展的大量投资。 这项工作的目的是评估基于MS的MAM与常规QC的性能 在强制降解后识别产品质量属性（PQA）变化并关联的方法 具有生物活性，结合亲和力和结构的那些PQA的变化。这项研究的结果将有助于支持 通过创建可以降低障碍的知识基础，从传统技术过渡到MAM 为了采用MAM，并可以通过生物仿制药制造商更广泛地使用MAM。 这里提出的工作将评估和比较单克隆抗体（Adalimumab）和FC的PQA 使用常规QC方法和 基于MAM的方法。每种技术对强制降解过程中的变化的敏感性将是 评估并与使用基于细胞的测定的功能和结构变化进行了比较，结合 测定和循环二分性能。我们建议首先对Adalimumab和 来自多个来源的依那感（特定目标1）。 PQA将使用常规方法进行评估 评估电荷变体，糖基化和尺寸变体（特定目标2），并与特定 使用基于MS的MAM工作流（特定目标3）检测到的分子修饰以评估可比性 对PQA的变化的敏感性。最后，在特定目标4下，我们将评估生物活性和结构 在PQA上表现出显着差异以将分子变化与 结构和功能的变化。