Identification of the human liver cytochrome P450 enzymes involved in the in vitro metabolism of a novel 5-lipoxygenase inhibitor.
In vitro studies were conducted to identify the hepatic cytochrome P450 (CYP) forms involved in the oxidative metabolism of [14C]ABT-761 and its N-dehydroxylated metabolite, [14C]ABT-438, by human liver microsomes. The two compounds were metabolized by parallel pathways, to form the corresponding methylene bridge hydroxy metabolites. There was no evidence of sulfoxidation and/or ring hydroxylation. Over the ABT-761 and ABT-438 concentration ranges studied (1-300 microM), the rate of NADPH-dependent hydroxylation was linear with respect to substrate concentration ([S]) and did not conform to saturable Michaelis-Menten kinetics. Under these conditions ([S] < KM), the intrinsic clearance (Vmax/KM) of ABT-438 was 10-fold higher than that of ABT-761 (1.7 +/- 0.8 vs. 0.17 +/- 0.06 microl/min/mg, mean +/- SD, N = 3 livers). The hydroxylation of both compounds was shown to be highly correlated (r = 0.83, p < 0.01, N = 11 different human livers) with CYP3A-selective erythromycin N-demethylase activity, and the correlation between ABT-761 hydroxylation and tolbutamide hydroxylase (CYP2C9-selective) activity (r = 0.63, p < 0.05, N = 10) was also statistically significant. Ketoconazole (2.0 microM), a CYP3A-selective inhibitor, inhibited the hydroxylation of both compounds by 53-67%, and sulfaphenazole (CYP2C9-selective) decreased activity by 10-20%. By comparison, alpha-naphthoflavone, a known activator of CYP3A, stimulated the hydroxylation of ABT-761 (8-fold) and ABT-438 (4-fold). In addition, the abundance-normalized rates of cDNA-expressed CYP-dependent metabolism indicated that hydroxylation was largely mediated (66-86%) by CYP3A(4). Therefore, it is concluded that the hydroxylation of ABT-761 and ABT-438 (</=10 microM) is primarily mediated by CYP3A, although CYP2C9 may play an ancillary role.
Machinist
J M
JM
Drug Metabolism Department, Pharmaceutical Products Division, Abbott Laboratories, Abbott Park, IL 60064, USA.
Mayer
M D
MD
Roberts
E M
EM
Surber
B W
BW
Rodrigues
A D
AD
D002384:Catalysis; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D018076:DNA, Complementary; D005464:Fluorobenzenes / Q000494:pharmacology*; D006801:Humans; D006918:Hydroxyurea / Q000031:analogs & derivatives* / Q000493:pharmacokinetics; D016859:Lipoxygenase Inhibitors / Q000493:pharmacokinetics*; D008862:Microsomes, Liver / Q000201:enzymology*; D013876:Thiophenes / Q000494:pharmacology*
D018076:DNA, Complementary; D005464:Fluorobenzenes; D016859:Lipoxygenase Inhibitors; C115398:N-3-(5-(4-fluorobenzyl)thiophen-2-yl)-1-methyl-2-propynylurea; D013876:Thiophenes; D003577:Cytochrome P-450 Enzyme System; C096921:atreleuton; D006918:Hydroxyurea