Abstract
The sedative clomethiazole (CMZ) has been used in Europe since the mid-1960s to treat insomnia and alcoholism. It has been previously demonstrated in clinical studies to reversibly inhibit human CYP2E1 in vitro and decrease CYP2E1-mediated elimination of chlorzoxazone. We have investigated the selectivity of CMZ inhibition of CYP2E1 in pooled human liver microsomes (HLMs). In a reversible inhibition assay of the major drug-metabolizing cytochrome P450 (P450) isoforms, CYP2A6 and CYP2E1 exhibited IC50 values of 24 µM and 42 µM, respectively with all other isoforms exhibiting values >300 µM. When CMZ was preincubated with NADPH and liver microsomal protein for 30 minutes before being combined with probe substrates, however, more potent inhibition was observed for CYP2E1 and CYP2B6 but not CYP2A6 or other P450 isoforms. The substantial increase in potency of CYP2E1 inhibition upon preincubation enables the use of CMZ to investigate the role of human CYP2E1 in xenobiotic metabolism and provides advantages over other chemical inhibitors of CYP2E1. The KI and kinact values obtained with HLM-catalyzed 6-hydroxylation of chlorzoxazone were 40 µM and 0.35 minute−1, respectively, and similar to values obtained with recombinant CYP2E1 (41 µM, 0.32 minute−1). The KI and kinact values, along with other parameters, were used in a mechanistic static model to explain earlier observations of a profound decrease in the rate of chlorzoxazone elimination in volunteers despite the absence of detectable CMZ in blood.
Footnotes
- Received February 23, 2016.
- Accepted May 4, 2016.
↵1 Current affiliation: AbbVie Inc., 1 N. Waukegan Rd, North Chicago, Illinois.
↵2 Current address: Somerville, Massachusetts.
↵3 Current affiliation: Department of Biomedical Engineering, Rensselaer Polytechnic Institute, Troy, New York.
↵4 Current affiliation: Mylan Institutional, Sugar Land, Texas.
↵5 Current affiliation: Cardinal Health, Woburn, Massachusetts.
↵6 Current affiliation: AstraZeneca, Waltham, Massachusetts.
Portions of this work were previously presented at the 2010 International Society of Study of Xenobiotics (Istanbul, Turkey).
Clomethiazole is also known as [5-(2-chloroethyl) 4-methylthiazole], chlormethiazole, Distraneurin, Hemineurin, Heminevrin).
E.S.P., A.K.M., A.P.B., S.S.D., and E.T.G. were employed by Discovery Labware, Inc., a business unit of BD Biosciences, Woburn, Massachusetts, at the time of their experimental contributions to the work. Discovery Labware, Inc., was subsequently acquired by Corning Life Sciences.
↵This article has supplemental material available at dmd.aspetjournals.org.
- Copyright © 2016 by The American Society for Pharmacology and Experimental Therapeutics
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