PO.CH02.02 · 化学
整合的质谱工作流程助力癌症研究中的下一代偶联疗法
Integrated mass spectrometric workflows enabling next-generation conjugate therapeutics in cancer research
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
抗体-药物偶联物（ADCs）以及更广泛的抗体-X偶联物（Antibody-X Conjugates, AXCs）——包括新兴的抗体-寡核苷酸偶联物（Antibody-Oligonucleotide Conjugates, AOCs）——正作为肿瘤学和自身免疫领域的治疗手段迅速发展。它们日益扩大的影响力增加了对稳健生物分析策略的需求，这些策略需能够定量总抗体和偶联抗体、释放的载荷（payload）以及相关的生物转化途径。LC-MS/MS已成为应对AXCs分析复杂性的重要平台，支持用于总抗体和偶联抗体的混合LBA-LC-MS工作流程，以及游离载荷和载荷代谢物的直接测定。为肿瘤选择性活化而设计的点击化学（click-chemistry）药物系统也带来了类似的分析挑战，需要高灵敏度和特异性。本文所述平台通过增强的灵敏度、选择性和全面的生物转化表征来满足这些需求。我们开发了一套针对ADC、AXC和AOC项目量身定制的整合LC-MS/MS工作流程。经优化的受控还原和选择性提取方法能够准确定量多种载荷类别——包括auristatin类（MMAE/MMAF）、拓扑异构酶I抑制剂（DXd、SN-38、exatecan）以及相关的连接子-药物（linker-drug）种类——同时最大限度减少人为的载荷释放。这些检测在血浆、肿瘤、组织和基于细胞的系统中均表现出高灵敏度、线性和基质稳健性，支持可靠的药代动力学、稳定性和生物转化评估。我们还建立了一套采用叠氮-炔烃和TCO-四嗪反应的互补生物正交点击化学工作流程，以选择性标记前体和产物种类，增强质谱可检测性并实现对活化行为的定量分析。这些基于LC-MS/MS的方法共同为ADCs、AOCs、AXCs和点击活化疗法提供了统一的分析解决方案。该平台支持跨血浆和组织基质的全面体外、离体和体内药代动力学研究，促进这些新兴治疗手段的稳健PK/PD相关性分析和暴露量分析。
查看英文原文 English abstract
Antibody-drug conjugates (ADCs) and the broader class of Antibody-X Conjugates (AXCs)-including emerging Antibody-Oligonucleotide Conjugates (AOCs)-are rapidly advancing as therapeutic modalities in oncology and autoimmunity. Their expanding impact increases the need for robust bioanalytical strategies capable of quantifying total and conjugated antibodies, released payloads, and associated biotransformation pathways.LC-MS/MS has become an essential platform for addressing the analytical complexity of AXCs, supporting hybrid LBA-LC-MS workflows for total and conjugated antibodies, as well as direct measurement of free payloads and payload metabolites. Click-chemistry-enabled drug systems, designed for tumor-selective activation, present similar analytical challenges that require high sensitivity and specificity. The platform described here meets these demands through enhanced sensitivity, selectivity, and comprehensive biotransformation characterization.We developed an integrated LC-MS/MS workflow tailored for ADC, AXC, and AOC programs. Optimized controlled-reduction and selective-extraction methods enabled accurate quantification of diverse payload classes-including auristatins (MMAE/MMAF), topoisomerase-I inhibitors (DXd, SN-38, exatecan), and associated linker-drug species-while minimizing artificial payload release. These assays demonstrated high sensitivity, linearity, and matrix robustness across plasma, tumor, tissue, and cell-based systems, supporting reliable pharmacokinetic, stability, and biotransformation assessments.A complementary bioorthogonal click-chemistry workflow using azide-alkyne and TCO-tetrazine reactions was also established to selectively tag precursor and product species, enhancing MS detectability and enabling quantitative analysis of activation behavior.Together, these LC-MS/MS-based approaches provide a unified analytical solution for ADCs, AOCs, AXCs, and click-activated therapeutics. The platform supports comprehensive in vitro, ex vivo, and in vivo pharmacokinetic studies across plasma and tissue matrices, facilitating robust PK/PD correlation and exposure profiling for these emerging modalities.
利益披露 Disclosure
M. Fang, None..
N. Sonawane, None..
W. Ma, None..
B. Wei, None..
Q. Lin, None.