A comprehensive assay for nine major cytochrome P450 enzymes activities with 16 probe reactions on human liver microsomes by a single LC/MS/MS run to support reliablein vitroinhibitory drug–drug interaction evaluation

• DOI: 10.3109/00498254.2015.1036954
• Publication Date: 2015-05-26T12:24:33Z
• Authors: Ying Peng; Hui Wu; Xueyuan Zhang; Fengyi Zhang; Huanhuan Qi; Yunxi Zhong; Yu Wang; Hua Sang; Guangji Wang; Jianguo Sun
• Keywords: Microsoma; Human liver

1. A comprehensive method for the simultaneous characterization of xenobiotic compound inhibition nine major CYP enzymes in human liver microsomes was established by using 16 CYP-catalyzed reactions 14 probe substrates with three cocktail incubation sets and a single LC/MS/MS analysis. 2. The subgroups were developed to minimize effects organic solvents, polyunsaturated fatty acids mutual substrate interactions: Group I composed tolbutamide (CYP2C9), S-mephenytoin (CYP2C19), testosterone (CYP3A4), dextromethorphan (CYP2D6); II nifedipine midazolam coumarin (CYP2A6), bupropion (CYP2B6), diclofenac (CYP2C9); III phenacetin (CYP1A2), chlorzoxazone (CYP2E1), omeprazole (CYP2C19 CYP3A4), paclitaxel (CYP2C8), (+)-bufuralol (CYP2D6). In case CYP2C9, CYP2C19, CYP2D6 CYP3A4, multiple used due phenomenon substrate-binding pockets substrate-dependent inhibition. All metabolites simultaneously analyzed polarity switching mode run. 3. This validated against assay 12 well-characterized inhibitors two new entities (GT0918, MDV3100). IC50 values each inhibitor agreed well that individual drug as reported previous literatures.