Abstract
The oral anticoagulant acenocoumarol is given as a racemic mixture. The (S)-enantiomer is rapidly cleared and is the reason why only (R)-acenocoumarol contributes to the pharmacological effect. The objective of the study was to establish the cytochrome P450 (CYP) enzymes catalyzing the hydroxylations of the acenocoumarol enantiomers. Of various cDNA-expressed human CYPs, only CYP2C9 hydroxylated (S)-acenocoumarol. Hydroxylation occurred at the 6-, 7-, and 8-position with equalKm values and a ratio of 0.9:1:0.1 forVmax. CYP2C9 also mediated the 6-, 7-, and 8-hydroxylations of (R)-acenocoumarol withKm values three to four times andVmax values one-sixth times those of (S)-acenocoumarol. (R)-Acenocoumarol was also metabolized by CYP1A2 (6-hydroxylation) and CYP2C19 (6-, 7-, and 8-hydroxylation). In human liver microsomes one enzyme only catalyzed (S)-acenocoumarol hydroxylations withKm values < 1 μM. In most of the samples tested the 7-hydroxylation of (R)-acenocoumarol was also catalyzed by one enzyme only. The 6-hydroxylation was catalyzed by at least two enzymes. Sulfaphenazole could completely inhibit in a competitive way the hydroxylations of (S)-acenocoumarol and the 7-hydroxylation of (R)-acenocoumarol. The 6-hydroxylation of (R)-acenocoumarol could be partially inhibited by sulfaphenazole, 40 to 50%, and by furafylline, 20 to 30%. Significant mutual correlations were obtained between the hydroxylations of (S)-acenocoumarol, the 7-hydroxylation of (R)-acenocoumarol, the 7-hydroxylation of (S)-warfarin, and the methylhydroxylation of tolbutamide. The results demonstrate that (S)-acenocoumarol is hydroxylated by a single enzyme, namely CYP2C9. CYP2C9 is also the main enzyme in the 7-hydroxylation of (R)-acenocoumarol. Other enzymes involved in (R)-acenocoumarol hydroxylation reactions are CYP1A2 and CYP2C19. Drug interactions must be expected, particularly for drugs interfering with CYP2C9. Also, drugs interfering with CYP1A2 and CYP2C19 may potentiate acenocoumarol anticoagulant therapy.
Footnotes
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Send reprint requests to: H.H.W. Thijssen, Dept. of Pharmacology, University of Maastricht, P.O. Box 616, 6200 MD Maastricht, the Netherlands. E-mail: h.thijssen{at}farmaco.unimaas.nl
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Part of the work was supported by the BioAvenir program (to P.H.B.) and by region Ile de France.
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↵1 The abbreviation used is: CYP, cytochrome P450.
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↵2 The clinical plasma concentrations of acenocoumarol range between 50 and 150 ng/ml and are mainly (R)-acenocoumarol. Assuming that the free hepatic concentration equals free plasma concentration (unbound fraction, 1–2%), the substrate concentration would be 2 to 10 nmol/l.
- Received May 1, 2000.
- Accepted August 15, 2000.
- The American Society for Pharmacology and Experimental Therapeutics
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