Eakins Julie, Bauch Caroline, Woodhouse Heather, Park Benjamin, Bevan Samantha, Dilworth Clive, Walker Paul
Cyprotex Discovery Ltd, BioHub at Alderley Park, Alderley Edge, Cheshire, SK10 4TG, UK.
Cyprotex Discovery Ltd, BioHub at Alderley Park, Alderley Edge, Cheshire, SK10 4TG, UK.
Toxicol In Vitro. 2016 Aug;34:161-170. doi: 10.1016/j.tiv.2016.03.016. Epub 2016 Apr 12.
Drug induced mitochondrial dysfunction has been implicated in organ toxicity and the withdrawal of drugs or black box warnings limiting their use. The development of highly specific and sensitive in vitro assays in early drug development would assist in detecting compounds which affect mitochondrial function. Here we report the combination of two in vitro assays for the detection of drug induced mitochondrial toxicity. The first assay measures cytotoxicity after 24h incubation of test compound in either glucose or galactose conditioned media (Glu/Gal assay). Compounds with a greater than 3-fold toxicity in galactose media compared to glucose media imply mitochondrial toxicity. The second assay measures mitochondrial respiration, glycolysis and a reserve capacity with mechanistic responses observed within one hour following exposure to test compound. In order to assess these assays a total of 72 known drugs and chemicals were used. Dose-response data was normalised to 100× Cmax giving a specificity, sensitivity and accuracy of 100%, 81% and 92% respectively for this combined approach.
药物诱导的线粒体功能障碍与器官毒性有关，并且药物的撤市或黑框警告限制了它们的使用。在药物早期研发中开发高度特异且灵敏的体外试验将有助于检测影响线粒体功能的化合物。在此，我们报告两种用于检测药物诱导的线粒体毒性的体外试验的联合应用。第一种试验在测试化合物于葡萄糖或半乳糖条件培养基中孵育24小时后测量细胞毒性（葡萄糖/半乳糖试验）。与葡萄糖培养基相比，在半乳糖培养基中具有大于3倍毒性的化合物意味着线粒体毒性。第二种试验测量线粒体呼吸、糖酵解以及在暴露于测试化合物后一小时内观察到的具有机制性反应的储备能力。为了评估这些试验，总共使用了72种已知药物和化学品。剂量-反应数据被归一化为100×Cmax，这种联合方法的特异性、灵敏度和准确度分别为100%、81%和92%。
