title: Chlorpropamide 2-hydroxylation is catalysed by CYP2C9 and CYP2C19 in vitro: chlorpropamide disposition is influenced by CYP2C9, but not by CYP2C19 genetic polymorphism.
abstract: OBJECTIVE
  We evaluated the involvement of cytochrome P450 (CYP) isoforms 2C9 and 2C19 in chlorpropamide 2-hydroxylation in vitro and in chlorpropamide disposition in vivo.
  
  
  METHODS
  To identify CYP isoforms(s) that catalyse 2-hydroxylation of chlorpropamide, the incubation studies were conducted using human liver microsomes and recombinant CYP isoforms. To evaluate whether genetic polymorphisms of CYP2C9 and/or CYP2C19 influence the disposition of chlorpropamide, a single oral dose of 250 mg chlorpropamide was administered to 21 healthy subjects pregenotyped for CYP2C9 and CYP2C19.
  
  
  RESULTS
  In human liver microsomal incubation studies, the formation of 2-hydroxychlorpropamide (2-OH-chlorpropamide), a major chlorpropamide metabolite in human, has been best described by a one-enzyme model with estimated K(m) and V(max) of 121.7 +/- 19.9 microm and 16.1 +/- 5.0 pmol min(-1) mg(-1) protein, respectively. In incubation studies using human recombinant CYP isoforms, however, 2-OH-chlorpropamide was formed by both CYP2C9 and CYP2C19 with similar intrinsic clearances (CYP2C9 vs. CYP2C19: 0.26 vs. 0.22 microl min(-1) nmol(-1) protein). Formation of 2-OH-chlorpropamide in human liver microsomes was significantly inhibited by sulfaphenazole, but not by S-mephenytoin, ketoconazole, quinidine, or furafylline. In in vivo clinical trials, eight subjects with the CYP2C9*1/*3 genotype exhibited significantly lower nonrenal clearance [*1/*3 vs.*1/*1: 1.8 +/- 0.2 vs. 2.4 +/- 0.1 ml h(-1) kg(-1), P < 0.05; 95% confidence interval (CI) on the difference 0.2, 1.0] and higher metabolic ratios (of chlorpropamide/2-OH-chlorpropamide in urine: *1/*3 vs.*1/*1: 1.01 +/- 0.19 vs. 0.56 +/- 0.08, P < 0.05; 95% CI on the difference - 0.9, - 0.1) than did 13 subjects with CYP2C9*1/*1 genotype. In contrast, no differences in chlorpropamide pharmacokinetics were observed for subjects with the CYP2C19 extensive metabolizer vs. poor metabolizer genotypes.
  
  
  CONCLUSIONS
  These results suggest that chlorpropamide disposition is principally determined by CYP2C9 activity in vivo, although both CYP2C9 and CYP2C19 have a catalysing activity of chlorpropamide 2-hydroxylation pathway.
authors.affiliation: Department of Pharmacology and PharmacoGenomics Research Centre, Inje University College of Medicine and Clinical Pharmacology Centre, Busan Paik Hospital, Busan 641-735, Korea.
authors.forename: Ji-Hong
authors.initials: JH
authors.lastname: Shon
authors.forename: Young-Ran
authors.initials: YR
authors.lastname: Yoon
authors.forename: Min-Jung
authors.initials: MJ
authors.lastname: Kim
authors.forename: Kyoung-Ah
authors.initials: KA
authors.lastname: Kim
authors.forename: Young-Chae
authors.initials: YC
authors.lastname: Lim
authors.forename: Kwang-Hyeon
authors.initials: KH
authors.lastname: Liu
authors.forename: Dong-Hoon
authors.initials: DH
authors.lastname: Shin
authors.forename: Chung Han
authors.initials: CH
authors.lastname: Lee
authors.forename: In-June
authors.initials: IJ
authors.lastname: Cha
authors.forename: Jae-Gook
authors.initials: JG
authors.lastname: Shin
mesh_terms: D000284:Administration, Oral; D000328:Adult; D001189:Aryl Hydrocarbon Hydroxylases / Q000235:genetics* / Q000378:metabolism; D002747:Chlorpropamide / Q000378:metabolism* / Q000493:pharmacokinetics; D065731:Cytochrome P-450 CYP2C19; D065729:Cytochrome P-450 CYP2C9; D005260:Female; D006801:Humans; D006900:Hydroxylation; D007004:Hypoglycemic Agents / Q000378:metabolism* / Q000493:pharmacokinetics; D008099:Liver / Q000201:enzymology*; D008297:Male; D008861:Microsomes / Q000201:enzymology*; D006899:Mixed Function Oxygenases / Q000235:genetics* / Q000378:metabolism; D011110:Polymorphism, Genetic
chemical_list: D007004:Hypoglycemic Agents; D006899:Mixed Function Oxygenases; C450260:CYP2C9 protein, human; D065729:Cytochrome P-450 CYP2C9; D001189:Aryl Hydrocarbon Hydroxylases; C045793:CYP2C19 protein, human; D065731:Cytochrome P-450 CYP2C19; D002747:Chlorpropamide