Abstract
CYP2E1 metabolizes compounds, including clinical drugs, organic solvents, and tobacco-specific carcinogens. Chlorzoxazone (CZN) is a probe drug used to phenotype for CYP2E1 activity. Smokers have increased CZN clearance during smoking compared with nonsmoking periods; however, it is unclear which cigarette smoke component is causing the increased activity. The relationships between in vivo CZN disposition, in vitro CZN metabolism, and hepatic CYP2E1 have not been investigated in a within-animal design. In control-treated monkeys (Cercopithecus aethiops), the in vivo CZN area under the curve extrapolated to infinity (AUCinf) was 19.7 ± 4.5 μg μ h/ml, t1/2 was 0.57 ± 0.07 h, and terminal disposition rate constant calculated from last three to four points on the log-linear end of the concentration versus time curve was 1.2 ± 0.2 /h. In vitro, the apparent Vmax was 3.48 ± 0.02 pmol/min/μg microsomal protein, and the Km was 95.4 ± 1.8 μM. Chronic nicotine treatment increased in vivo CZN disposition, as indicated by a 52% decrease in AUCinf (p < 0.01) and 52% decrease in Tmax (p < 0.05) compared with control-treated monkeys. The log metabolic ratios at 0.5, 1, 2, and 4 h significantly negatively correlated with CZN AUCinf (p = 0.01–0.0001). Monkey hepatic CYP2E1 levels significantly correlated with both in vivo AUCinf (p = 0.03) and in vitro (p = 0.004) CZN metabolism. Together, the data indicated that nicotine induction of in vivo CZN disposition is related to the rates of in vitro CZN metabolism and hepatic microsomal CYP2E1 protein levels. Nicotine is one component in cigarette smoke that can increase in vivo CZN metabolism via induction of hepatic CYP2E1 levels. Thus, nicotine exposure may affect the metabolism of CYP2E1 substrates such as acetaminophen, ethanol, and benzene.
Footnotes
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This work was supported by Canadian Institutes of Health Research (CIHR) Grant MT14173, as well as a CIHR Tobacco Use in Special Populations Fellowship (A.M.L.) and a Canada Research Chair (R.F.T.).
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Conflict of Interest Statement: R.F.T. is a shareholder in Nicogen, a company focused on novel treatment approaches. No support from Nicogen was used, and no benefit to the company was obtained.
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Article, publication date, and citation information can be found at http://dmd.aspetjournals.org.
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doi:10.1124/dmd.106.010363.
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ABBREVIATIONS: CZN, chlorzoxazone; 6OHCZN, 6-hydroxychlorzoxazone; PBS, phosphate-buffered saline; BSA, bovine serum albumin; TBS, Tris-buffered saline; AUCT, area under the curve to the last quantifiable concentration; Kel, terminal disposition rate constant calculated from last three to four points on the log-linear end of the concentration versus time curve; AUCinf, area under the curve extrapolated to infinity; P450, cytochrome(s) P450.
- Received March 31, 2006.
- Accepted June 8, 2006.
- The American Society for Pharmacology and Experimental Therapeutics
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