Larson Brad, Banks Peter, Cali James J, Sobol Mary, Shultz Sarah
BioTek Instruments, Inc., Highland Park, Winooski, VT 05404, USA.
J Lab Autom. 2011 Feb;16(1):47-55. doi: 10.1016/j.jala.2010.01.005. Epub 2010 Mar 19.
The determination of inhibitory effects that lead compounds have on cytochrome P450 (CYP) enzymes is an important part of today's drug discovery process. Assays can be performed early in the discovery process to predict adverse drug-drug interactions caused by CYP inhibition and to minimize the costs associated with terminating candidates in late stage development or worse, removing a drug from the market after launch. For early discovery work, testing substantial numbers of compounds is desirable, thus automated "mix and read" assays are beneficial. Here, we demonstrate the automation of the CYP profiling process using a simple, yet robust robotic platform. Compound titration, as well as transfer of compounds and assay components was performed by the same automated pipetting system. IC(50)s of small molecule drugs were determined using recombinant CYP enzymes, CYP3A4, -2C9, and -2D6 and luminogenic substrates specific to each. Compounds were profiled against all three enzymes on the same 384-well assay plate.
确定先导化合物对细胞色素P450（CYP）酶的抑制作用是当今药物发现过程的重要组成部分。在发现过程的早期就可以进行检测，以预测由CYP抑制引起的不良药物相互作用，并将与在后期开发中终止候选药物相关的成本降至最低，更糟糕的是，在药物上市后将其撤出市场。对于早期发现工作，需要测试大量化合物，因此自动化的“混合并读取”检测方法很有帮助。在这里，我们展示了使用一个简单但强大的机器人平台实现CYP分析过程的自动化。化合物滴定以及化合物和检测组件的转移均由同一自动移液系统完成。使用重组CYP酶CYP3A4、-2C9和-2D6以及每种酶特异性的发光底物测定小分子药物的半数抑制浓度（IC50）。在同一384孔检测板上针对所有三种酶对化合物进行分析。
