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.

摘要 单克隆抗体和其他生物治疗剂经过各种修饰， 活性和稳定性，因此必须作为QC和可比性的一部分进行分析。质谱 (MS)由于其检测蛋白质的能力， 在分子水平上进行修饰。在过去的几年里，多属性方法（MAM）已经获得了 在整个药物开发和QC实验室的牵引，与几个开发人员实现了一些 MAM在表征或释放方面的形式。虽然更换多个QC测试提供了机会， 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中实施MAM，但MAM在 生物仿制药和小型生物制药公司。这项建议涉及的一个主要考虑领域是 如FDA工作人员2019年出版物所述，在QC中实施MAM：MAM的性能 与传统方法相比。收集数据以支持从传统技术到MAM的过渡是一个挑战。 可能阻止或延迟生物仿制药开发的重大投资。 这项工作的目的是评估基于MS的MAM与传统QC的性能 确定强制降解后产品质量属性（PQA）变化的方法， 这些PQA的生物活性、结合亲和力和结构发生变化。研究结果将有助于支持 通过创建知识库，从传统技术过渡到MAM， 采用MAM，并使生物仿制药制造商能够更广泛地使用MAM。 本文拟定的工作将评估和比较单克隆抗体（阿达木单抗）和Fc的PQA 融合蛋白（依那西普）使用常规QC方法和 基于MAM的方法。每种技术对强制降解期间变化的敏感性将 评价并与使用基于细胞的测定检测到的功能和结构变化进行比较， 测定和圆二色性。我们建议首先进行阿达木单抗的强制降解研究， 来自多个来源的依那西普（具体目标1）。将使用常规方法评价PQA， 评估电荷变体、糖基化和大小变体（特定目标2），并与特定 使用基于MS的MAM工作流程（特定目标3）检测分子修饰，以评估可比性 和对PQA变化的敏感性。最后，根据具体目标4，我们将评估生物活性和结构 在PQA方面表现出显著差异的生物制品， 结构和功能的变化。