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. 1992 Mar;33(3):313–317. doi: 10.1111/j.1365-2125.1992.tb04042.x

Human and mouse liver coumarin 7-hydroxylases do not metabolize warfarin in vitro.

P Honkakoski 1, P Arvela 1, R Juvonen 1, M A Lang 1, M Kairaluoma 1, O Pelkonen 1
PMCID: PMC1381282  PMID: 1576054

Abstract

1. The contribution of human P450 2A6 and mouse P450 2a-5 isoenzymes, both highly active in coumarin 7-hydroxylation, to the metabolism of warfarin was studied in several in vitro systems with human and mouse liver preparations. 2. The reconstituted P450 2a-5 purified from DBA/2 mouse liver did not metabolize warfarin. 3. An anti-P450 2a-5 antibody did not consistently inhibit any of the warfarin biotransformation reactions catalyzed by human or mouse liver microsomes, although coumarin 7-hydroxylation was inhibited by over 90%. 4. In some human microsomal samples, 4- and 8-hydroxylations of warfarin were inhibited to some extent by the anti-P450 2a-5 antibody. 5. Warfarin (less than 1 mM) did not inhibit coumarin 7-hydroxylation by human or mouse liver microsomes in vitro. 6. We conclude that mouse and human coumarin 7-hydroxylases do not oxidise warfarin.

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Selected References

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  1. Aitio A. A simple and sensitive assay of 7-ethoxycoumarin deethylation. Anal Biochem. 1978 Apr;85(2):488–491. doi: 10.1016/0003-2697(78)90245-2. [DOI] [PubMed] [Google Scholar]
  2. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  3. Honkakoski P., Lang M. A. Mouse liver phenobarbital-inducible P450 system: purification, characterization, and differential inducibility of four cytochrome P450 isozymes from D2 mouse. Arch Biochem Biophys. 1989 Aug 15;273(1):42–57. doi: 10.1016/0003-9861(89)90160-4. [DOI] [PubMed] [Google Scholar]
  4. Juvonen R. O., Autio S., Lang M. A. Pyrazole as a modifier of liver microsomal monooxygenase in DBA/2N and AKR/J mice. Biochem Pharmacol. 1987 Nov 15;36(22):3993–3997. doi: 10.1016/0006-2952(87)90469-2. [DOI] [PubMed] [Google Scholar]
  5. Juvonen R. O., Shkumatov V. M., Lang M. A. Purification and characterization of a liver microsomal cytochrome P-450 isoenzyme with a high affinity and metabolic capacity for coumarin from pyrazole-treated D2 mice. Eur J Biochem. 1988 Jan 15;171(1-2):205–211. doi: 10.1111/j.1432-1033.1988.tb13777.x. [DOI] [PubMed] [Google Scholar]
  6. Kaminsky L. S., Dannan G. A., Guengerich F. P. Composition of cytochrome P-450 isozymes from hepatic microsomes of C57BL/6 and DBA/2 mice assessed by warfarin metabolism, immunoinhibition, and immunoelectrophoresis with anti-(rat cytochrome P-450). Eur J Biochem. 1984 May 15;141(1):141–148. doi: 10.1111/j.1432-1033.1984.tb08168.x. [DOI] [PubMed] [Google Scholar]
  7. Kling L., Legrum W., Netter K. J. Induction of liver cytochrome P-450 in mice by warfarin. Comparison of warfarin-, phenobarbitone-, and cobalt-induced hepatic microsomal protein patterns by PAGE after partial purification on octyl-sepharose CL-4B. Biochem Pharmacol. 1985 Jan 1;34(1):85–91. doi: 10.1016/0006-2952(85)90104-2. [DOI] [PubMed] [Google Scholar]
  8. Lang M. A., Juvonen R., Järvinen P., Honkakoski P., Raunio H. Mouse liver P450Coh: genetic regulation of the pyrazole-inducible enzyme and comparison with other P450 isoenzymes. Arch Biochem Biophys. 1989 May 15;271(1):139–148. doi: 10.1016/0003-9861(89)90264-6. [DOI] [PubMed] [Google Scholar]
  9. Lush I. E., Arnold C. J. High coumarin 7-hydroxylase activity does not protect mice against Warfarin. Heredity (Edinb) 1975 Oct;35(2):279–281. doi: 10.1038/hdy.1975.92. [DOI] [PubMed] [Google Scholar]
  10. Miles J. S., McLaren A. W., Forrester L. M., Glancey M. J., Lang M. A., Wolf C. R. Identification of the human liver cytochrome P-450 responsible for coumarin 7-hydroxylase activity. Biochem J. 1990 Apr 15;267(2):365–371. doi: 10.1042/bj2670365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Nebert D. W., Nelson D. R., Coon M. J., Estabrook R. W., Feyereisen R., Fujii-Kuriyama Y., Gonzalez F. J., Guengerich F. P., Gunsalus I. C., Johnson E. F. The P450 superfamily: update on new sequences, gene mapping, and recommended nomenclature. DNA Cell Biol. 1991 Jan-Feb;10(1):1–14. doi: 10.1089/dna.1991.10.1. [DOI] [PubMed] [Google Scholar]
  12. Negishi M., Lindberg R., Burkhart B., Ichikawa T., Honkakoski P., Lang M. Mouse steroid 15 alpha-hydroxylase gene family: identification of type II P-450(15)alpha as coumarin 7-hydroxylase. Biochemistry. 1989 May 16;28(10):4169–4172. doi: 10.1021/bi00436a007. [DOI] [PubMed] [Google Scholar]
  13. Pelkonen O., Sotaniemi E. A., Ahokas J. T. Coumarin 7-hydroxylase activity in human liver microsomes. Properties of the enzyme and interspecies comparisons. Br J Clin Pharmacol. 1985 Jan;19(1):59–66. doi: 10.1111/j.1365-2125.1985.tb02613.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Raunio H., Syngelmä T., Pasanen M., Juvonen R., Honkakoski P., Kairaluoma M. A., Sotaniemi E., Lang M. A., Pelkonen O. Immunochemical and catalytical studies on hepatic coumarin 7-hydroxylase in man, rat, and mouse. Biochem Pharmacol. 1988 Oct 15;37(20):3889–3895. doi: 10.1016/0006-2952(88)90070-6. [DOI] [PubMed] [Google Scholar]
  15. Raunio H., Valtonen J., Honkakoski P., Lang M. A., Ståhlberg M., Kairaluoma M. A., Rautio A., Pasanen M., Pelkonen O. Immunochemical detection of human liver cytochrome P450 forms related to phenobarbital-inducible forms in the mouse. Biochem Pharmacol. 1990 Dec 1;40(11):2503–2509. doi: 10.1016/0006-2952(90)90092-y. [DOI] [PubMed] [Google Scholar]
  16. Rettie A. E., Eddy A. C., Heimark L. D., Gibaldi M., Trager W. F. Characteristics of warfarin hydroxylation catalyzed by human liver microsomes. Drug Metab Dispos. 1989 May-Jun;17(3):265–270. [PubMed] [Google Scholar]
  17. Sutcliffe F. A., MacNicoll A. D., Gibson G. G. Aspects of anticoagulant action: a review of the pharmacology, metabolism and toxicology of warfarin and congeners. Rev Drug Metab Drug Interact. 1987;5(4):225–272. doi: 10.1515/dmdi.1987.5.4.225. [DOI] [PubMed] [Google Scholar]
  18. Yamano S., Tatsuno J., Gonzalez F. J. The CYP2A3 gene product catalyzes coumarin 7-hydroxylation in human liver microsomes. Biochemistry. 1990 Feb 6;29(5):1322–1329. doi: 10.1021/bi00457a031. [DOI] [PubMed] [Google Scholar]

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