High-Throughput, Multispecies, Parallelized Plasma Stability Assay for the Determination and Characterization of Antibody-Drug Conjugate Aggregation and Drug Release.
High-Throughput, Multispecies, Parallelized Plasma Stability Assay for the Determination and Characterization of Antibody-Drug Conjugate Aggregation and Drug Release.
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DOI：
10.1021/acsomega.7b00452
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发表时间：
2017-08-31
期刊：
影响因子：
4.1
通讯作者：
Jenkins GJ
中科院分区：
文献类型：
--
作者：
Durbin KR;Nottoli MS;Catron ND;Richwine N;Jenkins GJ
抗体-药物偶联物（adc）在循环中的稳定性对最大功效和最小毒性至关重要。完整到达预期靶点的ADC可以向肿瘤输送尽可能高的药物负荷，并减少血液中游离药物的脱靶毒性。因此，ADC稳定性的评估是开发过程中至关重要的数据。然而，传统的ADC稳定性分析可能是人工密集型的，低通量的，并且需要大量的ADC材料。在这里，我们介绍了一种自动化的高通量血浆稳定性检测方法，用于同时在5种基质中筛选多达40种adc的144小时内药物释放和聚集。在早期药物开发过程中，ADC材料的数量通常是有限的，因此该检测采用384孔格式，以最大限度地减少每种ADC的材料需求<100 μg，每种物种类型的血浆需求<100 μL。使用非线性回归方程对药物释放和聚集输出进行建模，以计算每种数据类型的形成率。我们测试了一组15种具有不同抗体和相同缬氨酸-瓜氨酸-对氨基氨基氨基甲酸酯-单甲基lauristatin E连接物的adc，并比较了adc之间和物种之间的形成率，揭示了几个值得注意的趋势。特别是，当仅改变抗体时，发现了大范围的聚集，这表明在开发过程的早期进行血浆稳定性筛选，以发现和去除可能产生不稳定adc的候选抗体的关键作用。本文介绍的分析可用于提供新化学和抗体筛选计划的稳定性数据，为体内研究选择最佳候选药物，并提供在ADC开发的所有阶段中突出特定ADC设计固有的稳定性问题的结果。
The stability of antibody–drug conjugates (ADCs) in circulation is critical for maximum efficacy and minimal toxicity. An ADC reaching the intended target intact can deliver the highest possible drug load to the tumor and reduce off-target toxicity from free drug in the blood. As such, assessment of ADC stability is a vital piece of data during development. However, traditional ADC stability assays can be manually intensive, low-throughput, and require large quantities of ADC material. Here, we introduce an automated, high-throughput plasma stability assay for screening drug release and aggregation over 144 h for up to 40 ADCs across five matrices simultaneously. The amount of ADC material during early drug development is often limited, so this assay was implemented in 384-well format to minimize material requirements to <100 μg of each ADC and 100 μL of plasma per species type. Drug release and aggregation output were modeled using nonlinear regression equations to calculate formation rates for each data type. A set of 15 ADCs with different antibodies and identical valine–citrulline–p-aminobenzylcarbamate–monomethylauristatin E linker-drug payloads was tested and formation rates were compared across ADCs and between species, revealing several noteworthy trends. In particular, a wide range in aggregation was found when altering only the antibody, suggesting a key role for plasma stability screening early in the development process to find and remove antibody candidates with the potential to create unstable ADCs. The assay presented here can be leveraged to provide stability data on new chemistry and antibody screening initiatives, select the best candidate for in vivo studies, and provide results that highlight stability issues inherent to particular ADC designs throughout all stages of ADC development.
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