PO.ET02.11 · 实验与分子治疗
一种用于特异性检测的新方法学（NAM）：膜蛋白质组阵列的ISTAND资格认证以评估基于单克隆抗体疗法的脱靶结合
A new approach methodology (NAM) for specificity testing: ISTAND qualification of the membrane proteome array to evaluate off-target binding of MAb-based therapies
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：经科学验证的新方法学（NAMs）为更好地预测人体药物安全性提供了机会，同时也加速了药物开发和监管审查。FDA已建立多种途径，通过包括新药创新科学与技术方法（ISTAND）计划在内的各种机制，将NAMs认证为药物开发工具（DDTs）。
方法：我们开发了膜蛋白质组阵列（MPA）平台，用于抗体类治疗药物的特异性检测。MPA评估约6,000种膜蛋白的结合相互作用，每种膜蛋白均以其天然结构构象单独表达于活细胞或未固定细胞中。该筛选采用高通量流式细胞术，可实现定量和高灵敏度检测，任何鉴定出的靶点均通过滴定分析进行验证。MPA正通过FDA的ISTAND计划被认证为一种新型DDT。
结果：作为我们DDT资格认证的一部分，我们对35种抗体治疗药物进行了比较研究，使用我们的MPA和常规组织交叉反应性（TCR）研究评估其特异性。我们发现，其中17%的药物存在根据TCR结果无法预料的脱靶。总体而言，MPA和TCR结果仅在约一半受试分子中一致。在许多情况下，TCR结果因技术变异性、非特异性或胞浆染色、或假阳性染色而无法定论。此类结果凸显了使用TCR数据来决定可采取的后续步骤的困难。相比之下，MPA的脱靶结果通过统计学量化相对亲和力和结合水平、将结合定位于细胞外与细胞内表位、以及确定人体组织表达模式以评估体内安全风险，从而降低了风险。通过MPA资格认证过程，我们还量化了该平台的灵敏度和可重复性，并实施了额外的质量控制程序和数据分析。
结论：MPA数据已常规用于抗体类生物治疗药物的监管申请，并可用于替代或补充其他交叉反应性研究。MPA目前正处于通过FDA ISTAND计划进行DDT资格认证的最后审查阶段。
查看英文原文 English abstract
Background: Scientifically validated New Approach Methodologies (NAMs) provide an opportunity to better predict drug safety in humans while also accelerating drug development and regulatory review. The FDA has established pathways to qualify NAMs as Drug Development Tools (DDTs) through various mechanisms that now include the Innovative Science and Technology Approaches for New Drugs (ISTAND) program.
Methods: We developed the Membrane Proteome Array (MPA) platform for specificity testing of antibody-based therapeutics. The MPA assesses binding interactions across ~6,000 membrane proteins, each individually expressed in their native structural configuration within live or unfixed cells. The screen uses high-throughput flow cytometry, allowing for quantitative and high-sensitivity detection, and any identified targets are validated by titration analysis. The MPA is being qualified as a novel DDT through FDA's ISTAND program.
Results: As part of our DDT qualification, we performed a comparison study of 35 antibody therapeutics evaluated for specificity using both our MPA and conventional tissue cross-reactivity (TCR) studies. We found that 17% of these drugs had an off-target that was not expected based on TCR results. Overall, the MPA and TCR results were consistent for only about half of the molecules tested. In numerous instances, TCR results were inconclusive based on technical variability, nonspecific or cytoplasmic staining, or false-positive staining. Such results highlight the difficulty of using TCR data for deciding actionable next steps. In contrast, off-targets from the MPA are de-risked by statistically quantifying relative affinity and binding levels, localizing binding to extracellular vs. intracellular epitopes, and determining tissue expression patterns in humans to assess the in vivo safety risk. Through the MPA qualification process, we have also quantified the sensitivity and reproducibility of the platform and implemented additional quality control procedures and data analyses.
Conclusions: MPA data is already routinely used in regulatory applications for antibody-based biotherapeutics and may be used to replace or complement other cross-reactivity studies. The MPA is currently in the last stages of review for DDT qualification through FDA's ISTAND program.
利益披露 Disclosure
D. Norden,
Integral Molecular Employment.
C. Navia,
Integral Molecular Employment.
J. Richards,
Integral Molecular Employment.
A. Hobby, None.
B. Doranz,
Integral Molecular Employment.