Abstract
CYP2A5, a mouse cytochrome P450 monooxygenase that shows high similarities to human CYP2A6 and CYP2A13 in protein sequence and substrate specificity, is expressed in multiple tissues, including the liver, kidney, lung, and nasal mucosa. Heterologously expressed CYP2A5 is active in the metabolism of both endogenous substrates, such as testosterone, and xenobiotic compounds, such as nicotine and cotinine. To determine the biological and pharmacological functions of CYP2A5 in vivo, we have generated a Cyp2a5-null mouse. Homozygous Cyp2a5-null mice are viable and fertile; they show no evidence of embryonic lethality or developmental deficits; and they have normal circulating levels of testosterone and progesterone. The Cyp2a5-null mouse and wild-type mouse were then used for determination of the roles of CYP2A5 in the metabolism of nicotine and its major circulating metabolite, cotinine. The results indicated that the Cyp2a5-null mouse has lower hepatic nicotine 5′-hydroxylation activity in vitro, and slower systemic clearance of both nicotine and cotinine in vivo. For both compounds, a substantially longer plasma half-life and a greater area under the concentration-time curve were observed for the Cyp2a5-null mice, compared with wild-type mice. Further pharmacokinetics analysis confirmed that the brain levels of nicotine and cotinine are also influenced by the Cyp2a5 deletion. These findings provide direct evidence that CYP2A5 is the major nicotine and cotinine oxidase in mouse liver. The Cyp2a5-null mouse will be valuable for in vivo studies on the role of CYP2A5 in drug metabolism and chemical toxicity, and for future production of CYP2A6- and CYP2A13-humanized mouse models.
Footnotes
This work was supported in part by the National Institutes of Health National Cancer Institute [Grant CA092596]; and the National Institutes of Health National Institute of Environmental Health Sciences [Grant ES007462].
Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.
doi:10.1124/jpet.109.162610
↵ The online version of this article (available at http://jpet.aspetjournals.org) contains supplemental material.
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ABBREVIATIONS:
- ES
- embryonic stem
- P450
- cytochrome P450
- UGT
- UDP-glucuronosyltransferase
- PCR
- polymerase chain reaction
- LC
- liquid chromatography
- HPLC
- high-performance liquid chromatography
- kb
- kilobase pair
- MS
- mass spectrometer
- AUC
- area under the concentration-time curve
- CL
- clearance
- F
- bioavailability
- cotinine-d3
- (±)-cotinine-methyl-d3
- G418
- (2R,3S,4R,5R,6S)-5-amino-6-[(1R,2S,3S,4R,6S)-4,6-diamino-3-[(2R,3R,4R,5R)-3,5-dihydroxy-5-methyl-4-methylaminooxan-2-yl]oxy-2-hydroxycyclohexyl]oxy-2-(1-hydroxyethyl)oxane-3,4-diol
- WT
- wild type
- psig
- pound-force per square inch gauge.
- Received October 11, 2009.
- Accepted November 13, 2009.
- Copyright © 2010 by The American Society for Pharmacology and Experimental Therapeutics
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