title: A kinetic model for the metabolic interaction of two substrates at the active site of cytochrome P450 3A4.
abstract: In many cases, CYP3A4 exhibits unusual kinetic characteristics that result from the metabolism of multiple substrates that coexist at the active site. In the present study, we observed that alpha-naphthoflavone (alpha-NF) exhibited a differential effect on CYP3A4-mediated product formation as shown by an increase and decrease, respectively, of the carboxylic acid (P(2)) and omega-3-hydroxylated (P(1)) metabolites of losartan, while losartan was found to be an inhibitor of the formation of the 5,6-epoxide of alpha-NF. Thus, to address this problem, a kinetic model was developed on the assumption that CYP3A4 can accommodate two distinct and independent binding domains for the substrates within the active site, and the resulting velocity equations were employed to predict the kinetic parameters for all possible enzyme-substrate species. Our results indicate that the predicted values had a good fit with the experimental observations. Therefore, the kinetic constants can be used to adequately describe the nature of the metabolic interaction between the two substrates. Applications of the model provide some new insights into the mechanism of drug-drug interactions at the level of CYP3A4.
authors.affiliation: Department of Drug Metabolism, Merck Research Laboratories, West Point, Pennsylvania 19486, USA. magang_shou@merck.com
authors.forename: M
authors.initials: M
authors.lastname: Shou
authors.forename: R
authors.initials: R
authors.lastname: Dai
authors.forename: D
authors.initials: D
authors.lastname: Cui
authors.forename: K R
authors.initials: KR
authors.lastname: Korzekwa
authors.forename: T A
authors.initials: TA
authors.lastname: Baillie
authors.forename: T H
authors.initials: TH
authors.lastname: Rushmore
mesh_terms: D001571:Benzoflavones / Q000378:metabolism; D001665:Binding Sites; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D007700:Kinetics; D019808:Losartan / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D008954:Models, Biological; D013379:Substrate Specificity
chemical_list: D001571:Benzoflavones; C011512:alpha-naphthoflavone; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; C104464:CYP3A protein, human; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human; D019808:Losartan