RT Journal Article
SR Electronic
T1 CYP3A4 Is the Major CYP Isoform Mediating the in Vitro Hydroxylation and Demethylation of Flunitrazepam
JF Drug Metabolism and Disposition
JO Drug Metab Dispos
FD American Society for Pharmacology and Experimental Therapeutics
SP 133
OP 140
VO 29
IS 2
A1 Leah M. Hesse
A1 Karthik Venkatakrishnan
A1 Lisa L. von Moltke
A1 Richard I. Shader
A1 David J. Greenblatt
YR 2001
UL http://dmd.aspetjournals.org/content/29/2/133.abstract
AB The kinetics of flunitrazepam (FNTZ) N-demethylation to desmethylflunitrazepam (DM FNTZ), and 3-hydroxylation to 3-hydroxyflunitrazepam (3-OH FNTZ), were studied in human liver microsomes and in microsomes containing heterologously expressed individual human CYPs. FNTZ was N-demethylated by cDNA-expressed CYP2A6 (Km = 1921 μM), CYP2B6 (Km = 101 μM), CYP2C9 (Km = 50 μM), CYP2C19 (Km = 60 μM), and CYP3A4 (Km = 155 μM), and 3-hydroxylated by cDNA-expressed CYP2A6 (Km = 298 μM) and CYP3A4 (Km = 286 μM). The 3-hydroxylation pathway was predominant in liver microsomes, accounting for more than 80% of intrinsic clearance compared with theN-demethylation pathway. After adjusting for estimated relative abundance, CYP3A accounted for the majority of intrinsic clearance via both pathways. This finding was supported by chemical inhibition studies in human liver microsomes. Formation of 3-OH FNTZ was reduced to 10% or less of control values by ketoconazole (IC50 = 0.11 μM) and ritonavir (IC50 = 0.041 μM). Formation of DM FNTZ was inhibited to 40% of control velocity by 2.5 μM ketoconazole and to 30% of control by 2.5 μM ritonavir. Neither 3-OH FNTZ nor DM FNTZ formation was inhibited to less than 85% of control activity by α-naphthoflavone (CYP1A2), sulfaphenazole (CYP2C9), omeprazole (CYP2C19), or quinidine (CYP2D6). Thus, CYP-dependent FNTZ biotransformation, like that of many benzodiazepine derivatives, is mediated mainly by CYP3A. Clinical interactions of FNTZ with CYP3A inhibitors can be anticipated. The American Society for Pharmacology and Experimental Therapeutics