title: Metabolic characterization of the major human small intestinal cytochrome p450s.
abstract: Human small intestine epithelial cells (enterocytes) provide the first site for cytochrome P450 (CYP)-catalyzed metabolism of orally ingested xenobiotics. CYP3A4 is the major form of CYP expressed in enterocytes and CYP2C is also expressed at a significant level. In this study, we further characterized the expression of CYP3A4 and CYP2C in human enterocytes and their interindividual variations by examining the metabolic activities from 10 individuals. CYP3A4 in human jejunum microsomes, as determined by 6beta-testosterone hydroxylase activity, varied from 0.36 to 2.46 nmol/min/mg. The apparent average K(m) and V(max) values from two representative individuals were 54 microM and 3.2 nmol/min/mg, respectively. CYP2C9 and CYP2C19 in human jejunum microsomes, as determined by diclofenac 4'-hydroxylase and mephenytoin 4'-hydroxylase activities, varied over an 18-fold range (7.3-129 pmol/min/mg) and 17-fold range (0.8-13.1 pmol/min/mg), respectively. The mean apparent K(m) for diclofenac 4'-hydroxylase was 9.9 microM , whereas the apparent mean K(m) for S-mephenytoin 4'-hydroxylase was 79.3 microM . The mean intrinsic clearance (V(max)/K(m)) was approximately 130-fold greater for diclofenac 4'-hydroxylase than for mephenytoin 4'-hydroxylase. The metabolic activities of CYP2C9 and CYP2C19 were confirmed by inhibition by sulfaphenazole for CYP2C9 and ticlopidine for CYP2C19. In addition, CYP2C9 activities did not correlate with CYP3A4 activities, while CYP2C19 activities had a significant but poor correlation with those of CYP3A4. Thus the major CYP activities in human enterocytes have large interindividual variabilities that are not strongly related.
authors.affiliation: Pfizer Inc., Department of Drug Metabolism, Central Research Division, Groton, Connecticut, USA.
authors.forename: R S
authors.initials: RS
authors.lastname: Obach
authors.forename: Q Y
authors.initials: QY
authors.lastname: Zhang
authors.forename: D
authors.initials: D
authors.lastname: Dunbar
authors.forename: L S
authors.initials: LS
authors.lastname: Kaminsky
mesh_terms: D000328:Adult; D000368:Aged; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004008:Diclofenac / Q000378:metabolism; D020895:Enterocytes / Q000166:cytology / Q000201:enzymology / Q000378:metabolism; D005260:Female; D006801:Humans; D006900:Hydroxylation; D015151:Immunoblotting; D007421:Intestine, Small / Q000201:enzymology* / Q000378:metabolism; D007527:Isoenzymes / Q000378:metabolism; D007700:Kinetics; D008297:Male; D008617:Mephenytoin / Q000378:metabolism; D008861:Microsomes / Q000201:enzymology / Q000378:metabolism; D008875:Middle Aged; D006899:Mixed Function Oxygenases / Q000378:metabolism; D012044:Regression Analysis; D013426:Sulfaphenazole / Q000494:pharmacology; D013739:Testosterone / Q000378:metabolism; D013988:Ticlopidine / Q000494:pharmacology
chemical_list: D007527:Isoenzymes; C400322:cytochrome P-450 CYP2C subfamily; D013426:Sulfaphenazole; D004008:Diclofenac; D013739:Testosterone; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; C104464:CYP3A protein, human; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human; D013988:Ticlopidine; D008617:Mephenytoin