Yao Ming, Zhu Mingshe, Sinz Michael W, Zhang Hongjian, Humphreys W Griffith, Rodrigues A David, Dai Renke
Department of Biotransformation, Bristol-Myers Squibb Pharmaceutical Research Institute, P.O. Box 4000, Princeton, NJ 08543, USA.
J Pharm Biomed Anal. 2007 May 9;44(1):211-23. doi: 10.1016/j.jpba.2007.02.034. Epub 2007 Mar 3.
Substrate inhibition assays for five of the major CYP enzymes (phenacetin for CYP1A2, diclofenac for CYP2C9, (S)-mephenytoin for CYP2C19, dextromethorphan for CYP2D6 and midazolam and testosterone for CYP3A4) in human liver microsomes were developed. Fully automated incubations were conducted in a 96-well format under optimized enzyme kinetic conditions. Metabolites of probe substrates were analyzed with rapid LC-MS/MS methods. The assays were fully validated following the procedure for validating bioanalytical methods recommended by regulatory agencies. Quality control samples and a positive control CYP inhibitor were included in each assay. The IC(50) values determined for typical CYP inhibitors were reproducible and consistent with those reported in the literature. The high quality and throughput of these assays make them ideally suited for providing information for decision making in late drug discovery and early development and for providing labeling input for new drug registrations.
开发了用于人肝微粒体中五种主要细胞色素P450（CYP）酶的底物抑制试验（CYP1A2用非那西丁、CYP2C9用双氯芬酸、CYP2C19用(S)-美芬妥因、CYP2D6用右美沙芬以及CYP3A4用咪达唑仑和睾酮）。在优化的酶动力学条件下，以96孔板形式进行全自动孵育。用快速液相色谱 - 串联质谱法分析探针底物的代谢产物。按照监管机构推荐的生物分析方法验证程序对试验进行了全面验证。每次试验都包含质量控制样品和阳性对照CYP抑制剂。为典型CYP抑制剂测定的IC50值具有可重复性，且与文献报道的值一致。这些试验的高质量和高通量使其非常适合为药物发现后期和早期开发中的决策提供信息，以及为新药注册提供标签信息。
