title: A fluorescent cell-based assay for cytochrome P-450 isozyme 1A2 induction and inhibition.
abstract: High throughput lead optimization requires simple, homogeneous cell-based assays capable of defining the druglike properties of first-round screening hits. Induction and inhibition of the Phase I drug-metabolizing enzymes are central to this process. We report here an assay for induction and inhibition of cytochrome P-450 (CYP) isozyme 1A2 that meets these requirements. It utilizes HepG2/C3A, a human liver cell line, and ethoxyresorufin. Using methylcholanthrene, CYP1A2 can be induced dramatically, and it is inhibited by furafylline, a mechanism-based inhibitor of this enzyme.
authors.affiliation: Amphioxus Cell Technologies, Inc, Houston, TX 77082, USA. jkelly@amphioxus.com
authors.forename: J H
authors.initials: JH
authors.lastname: Kelly
authors.forename: N L
authors.initials: NL
authors.lastname: Sussman
mesh_terms: D002460:Cell Line; D019388:Cytochrome P-450 CYP1A2 / Q000032:analysis* / Q000096:biosynthesis; D065609:Cytochrome P-450 CYP1A2 Inhibitors; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004790:Enzyme Induction / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005453:Fluorescence; D006801:Humans; D008748:Methylcholanthrene / Q000494:pharmacology; D010078:Oxazines; D013806:Theophylline / Q000031:analogs & derivatives / Q000494:pharmacology
chemical_list: D065609:Cytochrome P-450 CYP1A2 Inhibitors; D004791:Enzyme Inhibitors; D010078:Oxazines; D008748:Methylcholanthrene; C007649:ethoxyresorufin; D013806:Theophylline; C050131:furafylline; D019388:Cytochrome P-450 CYP1A2