A high content screening assay to predict human drug-induced liver injury during drug discovery
A high content screening assay to predict human drug-induced liver injury during drug discovery
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DOI：
10.1016/j.vascn.2013.08.001
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发表时间：
2013-11-01
影响因子：
1.9
通讯作者：
Hornberg, Jorrit J.
中科院分区：
文献类型：
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作者：
Persson, Mikael;Loye, Anni F.;Hornberg, Jorrit J.
简介：药物不良反应是导致药物开发计划失败、药物撤回和使用限制的主要原因。因此，早期危险识别和勤勉的风险规避策略至关重要。对于药物性肝损伤（DILI），使用传统的安全测试很难做到这一点。为了降低 DILI 的风险，需要识别和取消选择具有高风险的候选药物。而且，为了生产没有相关风险的候选药物，需要在药物发现过程中尽早评估风险因素，以便可以根据安全参数优化先导系列。这需要能够进行中到高通量化合物分析并生成适合在先导化合物优化程序期间建立结构-活性关系的定量结果的方法。方法：我们对这种方法进行了验证，这是一种基于六个参数（细胞核计数、细胞核面积、质膜完整性、溶酶体活性、线粒体膜电位 (MMP) 和线粒体面积）的新型高内涵筛选测定，使用类似于 100 种临床肝毒性特征已知的药物。结果讨论：我们发现最低毒性浓度和治疗 Cmax 之间的 100 倍 TI 是根据各个参数将化合物分类为肝毒性或非肝毒性的最佳选择。大多数参数的敏感性约为 50%，特异性约为 90%。在低于 Cmax 100 倍的浓度下达到 >= 2 个参数的药物通常具有肝毒性，而非肝毒性药物通常会达到
Introduction: Adverse drug reactions are a major cause for failures of drug development programs, drug withdrawals and use restrictions. Early hazard identification and diligent risk avoidance strategies are therefore essential. For drug-induced liver injury (DILI), this is difficult using conventional safety testing. To reduce the risk for DILI, drug candidates with a high risk need to be identified and deselected. And, to produce drug candidates without that risk associated, risk factors need to be assessed early during drug discovery, such that lead series can be optimized on safety parameters. This requires methods that allow for medium-to-high throughput compound profiling and that generate quantitative results suitable to establish structure-activity-relationships during lead optimization programs. Methods: We present the validation of such a method, a novel high content screening assay based on six parameters (nuclei counts, nuclear area, plasma membrane integrity, lysosomal activity, mitochondrial membrane potential (MMP), and mitochondrial area) using similar to 100 drugs of which the clinical hepatotoxicity profile is known. Results discussion: We find that a 100-fold TI between the lowest toxic concentration and the therapeutic Cmax is optimal to classify compounds as hepatotoxic or non-hepatotoxic, based on the individual parameters. Most parameters have similar to 50% sensitivity and similar to 90% specificity. Drugs hitting >= 2 parameters at a concentration below 100-fold their Cmax are typically hepatotoxic, whereas non-hepatotoxic drugs typically hit