title: Multiple P450 substrates in a single run: rapid and comprehensive in vitro interaction assay.
abstract: The dramatically increased number of new chemical entities (NCE) used in drug discovery has raised a demand for efficient and rapid drug metabolism screening techniques. The aim of this study was to develop a global in vitro metabolic interaction screening test utilising the N-in-1 approach. A cocktail consisting of 10 CYP-selective probes with known kinetic, metabolic and interaction properties in vivo was incubated in a pool of human liver microsomes, and metabolites of melatonin (CYP1A2), coumarin (CYP2A6), bupropion (CYP2B6), amodiaquine (CYP2C8), tolbutamide (CYP2C9), omeprazole (CYP2C19 and CYP3A4), dextromethorphan (CYP2D6), chlorzoxazone (CYP2E1), midazolam (CYP3A4) and testosterone (CYP3A4) were analysed simultaneously using LC/TOF-MS. Performance of the method was assessed with cDNA expressed P450s and diagnostic CYP-specific inhibitors. The results were in good accordance with literature and our previous studies. The cocktail developed is suitable for fast and reliable in vitro screening of the interaction potential and characteristics of NCEs.
authors.affiliation: Department of Pharmacology and Toxicology, 90014 University of Oulu, P.O.Box 5000, Oulu, Finland. miia.turpeinen@oulu.fi
authors.forename: Miia
authors.initials: M
authors.lastname: Turpeinen
authors.forename: Jouko
authors.initials: J
authors.lastname: Uusitalo
authors.forename: Uusitalo
authors.initials: U
authors.lastname: Jouko
authors.forename: Jorma
authors.initials: J
authors.lastname: Jalonen
authors.forename: Jalonen
authors.initials: J
authors.lastname: Jorma
authors.forename: Olavi
authors.initials: O
authors.lastname: Pelkonen
authors.forename: Pelkeonen
authors.initials: P
authors.lastname: Olavi
mesh_terms: D002626:Chemistry, Pharmaceutical / Q000379:methods*; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D008862:Microsomes, Liver / Q000187:drug effects / Q000378:metabolism; D013379:Substrate Specificity / Q000187:drug effects / Q000502:physiology
chemical_list: D065607:Cytochrome P-450 Enzyme Inhibitors; D004791:Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System