title: Evaluation of induction potency of new drug candidates on CYP1A2 and CYP3A4 using real-time one-step RT-PCR in primary cultures of cryopreserved human hepatocytes.
abstract: This study evaluates the induction potency of new drug candidates on mRNA levels of CYP1A2 and CYP3A4 in primary cultures of cryopreserved human hepatocytes. Analysis was performed by quantitative real-time RT-PCR using primers and TaqMan probes. Positive controls for CYP1A2 and CYP3A4 used beta-naphthoflavone (beta-NF) and rifampicin (Rif), respectively. In the first stage of the study, the lot showing the best induction of mRNA expression CYP1A2 and CYP3A4 from among eight lots of hepatocytes was selected. In the second stage, we evaluated the levels of CYP1A2 and CYP3A4 gene expression in hepatocytes after exposure to eight NO-1886 (ibrolipim) derivatives. A combination of real-time one-step RT-PCR and primary culture of cryopreserved human hepatocytes is suitable for evaluating of induction potency of a large number of new drug candidates.
authors.affiliation: Research and Development Center, Otsuka Pharmaceutical Factory, Inc, Tokushima, Japan.
authors.forename: Masuhiro
authors.initials: M
authors.lastname: Nishimura
authors.forename: Shizuo
authors.initials: S
authors.lastname: Narimatsu
authors.forename: Shinsaku
authors.initials: S
authors.lastname: Naito
mesh_terms: D000328:Adult; D000368:Aged; D001549:Benzamides / Q000494:pharmacology*; D002478:Cells, Cultured; D002675:Child, Preschool; D015925:Cryopreservation; D019388:Cytochrome P-450 CYP1A2 / Q000096:biosynthesis*; D051544:Cytochrome P-450 CYP3A / Q000096:biosynthesis*; D004790:Enzyme Induction / Q000187:drug effects; D005260:Female; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D009943:Organophosphorus Compounds / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
chemical_list: D001549:Benzamides; D009943:Organophosphorus Compounds; C082137:4-diethoxyphosphorylmethyl-N-(4-bromo-2-cyanophenyl)benzamide; D019388:Cytochrome P-450 CYP1A2; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human