Skip to main content
SpringerLink
Log in

Haplotype structures of EPHX1 and their effects on the metabolism of carbamazepine-10,11-epoxide in Japanese epileptic patients

  • Pharmacogenetics
  • Published:
European Journal of Clinical Pharmacology Aims and scope Submit manuscript

Abstract

Objective

Microsomal epoxide hydrolase (mEH) is an enzyme that detoxifies reactive epoxides and catalyzes the biotransformation of carbamazepine-10,11-epoxide (CBZ-epoxide) to carbamazepine-10,11-diol (CBZ-diol). Utilizing single nucleotide polymorphisms (SNPs) of the EPHX1 gene encoding mEH, we identified the haplotypes of EPHX1 blocks and investigated the association between the block haplotypes and CBZ-epoxide metabolism.

Methods

SNPs of EPHX1 were analyzed by means of polymerase chain reaction amplification and DNA sequencing using DNA extracted from the blood leukocytes of 96 Japanese epileptic patients, including 58 carbamazepine-administered patients. The plasma concentrations of CBZ and its four metabolites were determined using high-performance liquid chromatography.

Results

From sequencing all 9 exons and their surrounding introns, 29 SNPs were found in EPHX1. The SNPs were separated into three blocks on the basis of linkage disequilibrium, and the block haplotype combinations (diplotypes) were assigned. Using plasma CBZ-diol/CBZ-epoxide ratios (diol/epoxide ratios) indicative of the mEH activity, the effects of the diplotypes in each EPHX1 block were analyzed on CBZ-epoxide metabolism. In block 2, the diol/epoxide ratios increased significantly depending on the number of haplotype *2 bearing Y113H (P=0.0241). In block 3, the ratios decreased depending on the number of haplotype *2 bearing H139R (P=0.0351). Also, an increasing effect of a *1 subtype, *1c, was observed on the ratio.

Conclusion

These results show that some EPHX1 haplotypes are associated with altered CBZ-epoxide metabolism. This is the first report on the haplotype structures of EPHX1 and their potential in vivo effects.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Similar content being viewed by others

References

  1. Weinshilboum R (2003) Inheritance and drug response. N Engl J Med 348:529–537

    Google Scholar 

  2. Hassett C, Aicher L, Sidhu JS, Omiecinski CJ (1994) Human microsomal epoxide hydrolase: genetic polymorphism and functional expression in vitro of amino acid variants. Hum Mol Genet 3:421–428

    Google Scholar 

  3. Maekawa K, Itoda M, Hanioka N, Saito Y, Murayama N, Nakajima O, Soyama A, Ishida S, Ozawa S, Ando M, Sawada J (2003) Non-synonymous single nucleotide alterations in the microsomal epoxide hydrolase gene and their functional effects. Xenobiotica 33:277–287

    Google Scholar 

  4. Tomson T, Tybring G, Bertilsson L (1983) Single-dose kinetics and metabolism of carbamazepine-10,11-epoxide. Clin Pharmacol Ther 33:58–65

    Google Scholar 

  5. Kitteringham NR, Davis C, Howard N, Pirmohamed M, Park BK (1996) Interindividual and interspecies variation in hepatic microsomal epoxide hydrolase activity: studies with cis-stilbene oxide, carbamazepine 10,11-epoxide and napthalene. J Pharmacol Exp Ther 278:1018–1027

    Google Scholar 

  6. Kitamura Y, Moriguch M, Kaneko H, Morisaki H, Morisaki T, Toyama K, Kamatani N (2002) Determination of probability distribution of diplotype configuration (diplotype distribution) for each subject from genotypic data using the EM algorithm. Ann Hum Genet 66:183–193

    Google Scholar 

  7. Seidegard J, DePierre JW (1983) Microsomal epoxide hydrolase. Properties, regulation and function. Biochim Biophys Acta 695:251–270

    Google Scholar 

  8. Fretland AJ, Omiecinski CJ (2000) Epoxide hydrolases: biochemistry and molecular biology. Chem Biol Interact 129:41–59

    Google Scholar 

  9. Laurenzana EM, Hassett C, Omiecinski CJ (1998) Post-transcriptional regulation of human microsomal epoxide hydrolase. Pharmacogenetics 8:157–167

    Google Scholar 

  10. Mandrioli R, Albani F, Casamenti G, Sabbioni C, Raggi M (2001) Simultaneous high-performance liquid chromatography determination of carbamazepine and five of its metabolites in plasma of epileptic patients. J Chromatogr B 762:109–116

    Google Scholar 

  11. Raggi MA, Pucci V, Maurizio A, Muzikar J, Kenndler E (2002) Separation of carbamazepine and five metabolites, and analysis in human plasma by micellar electrokinetic capillary chromatography. J Chromatogr B 770:217–225

    Google Scholar 

  12. Judson R, Stephens JC, Windemuth A (2000) The predictive power of haplotypes in clinical response. Pharmacogenomics 1:15–26

    Google Scholar 

  13. Zhang K, Calabrese P, Nordborg M, Sun F (2002) Haplotype block structure and its applications to association studies: power and study designs. Am J Hum Genet 71:1386–1394

    Google Scholar 

  14. Yoshikawa M, Hiyama K, Ishioka S, Maeda H, Maeda A, Yamakido M (2000) Microsomal epoxide hydrolase genotypes and chronic obstructive pulmonary disease in Japanese. Int J Mol Med 51:49–53

    Google Scholar 

  15. Shen LX, Basilion JP, Stanton VP Jr (1999) Single-nucleotide polymorphisms can cause different structural folds of mRNA. Proc Natl Acad Sci U S A 96:7871–7876

    Google Scholar 

  16. Duan J, Wainwright MS, Comeron JM, Saitou N, Sanders AR, Gelernter J, Gejman PV (2003) Synonymous mutations in the human dopamine receptor D2 (DRD2) affect mRNA stability and synthesis of the receptor. Hum Mol Genet 12:205–216

    Article  CAS  PubMed  Google Scholar 

  17. Frittitta L, Ercolino T, Bozzali M, Argiolas A, Graci S, Santagati MG, Spampinato D, Di Paola R, Cisternino C, Tassi V, Vigneri R, Pizzuti A, Trischitta V (2001) A cluster of three single nucleotide polymorphisms in the 3′-untranslated region of human glycoprotein PC-1 gene stabilizes PC-1 mRNA and is associated with increased PC-1 protein content and insulin resistance-related abnormalities. Diabetes 50:1952–1955

    Google Scholar 

  18. Spina E, Martines C, Fazio A, Trio R, Pisani F, Tomson T (1991) Effect of phenobarbital on the pharmacokinetics of carbamazepine-10,11-epoxide, an active metabolite of carbamazepine. Ther Drug Monit 13:109–112

    Google Scholar 

  19. Kroetz DL, Kerr BM, McFarland LV, Loiseau P, Wilensky AJ, Levy RH (1993) Measurement of in vivo microsomal epoxide hydrolase activity in white subjects. Clin Pharmacol Ther 53:306–315

    Google Scholar 

  20. Pisani F, Fazio A, Oteri G, Spina E, Perucca E, Bertilsson L (1988) Effect of valpromide on the pharmacokinetics of carbamazepine-10,11-epoxide. Br J Clin Pharmacol 25:611–613

    Google Scholar 

  21. Bertilsson L, Tomson T (1986) Clinical pharmacokinetics and pharmacological effects of carbamazepine and carbamazepine-10,11-epoxide. An update. Clin Pharmacokinet 11:177–198

    Google Scholar 

  22. Green VJ, Pirmohamed M, Kitteringham NR, Gaedigk A, Grant DM, Boxer M, Burchell B, Park BK (1995) Genetic analysis of microsomal epoxide hydrolase in patients with carbamazepine hypersensitivity. Biochem Pharmacol 50:1353–1359

    Google Scholar 

  23. Shiseki K, Itoda M, Saito Y, Nakajima Y, Maekawa K, Kimura H, Goto Y, Saitoh O, Katoh M, Ohnuma T, Kawai M, Sugai K, Ohtsuki T, Suzuki C, Minami N, Ozawa S, Sawada J (2003) Five novel single nucleotide polymorphisms in the EPHX1 gene encoding microsomal epoxide hydrolase. Drug Metab Pharmacokinet 18:150–153

    Google Scholar 

  24. Gaedigk A, Spielberg SP, Grant DM (1994) Characterization of the microsomal epoxide hydrolase gene in patients with anticonvulsant adverse drug reactions. Pharmacogenetics 4:142–153

    Google Scholar 

  25. Saito S, Iida A, Sekine A, Eguchi C, Miura Y, Nakamura Y (2001) Seventy genetic variations in human microsomal and soluble epoxide hydrolase genes (EPHX1 and EPHX2) in the Japanese population. J Hum Genet 46:325–329

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported in part by the Program for Promotion of Fundamental Studies in Health Sciences (MPJ-2 and −6) of the Pharmaceuticals and Medical Devices Agency (PMDA) of Japan. We thank Ms. Chie Knudsen for her secretarial assistance.

Author information

Authors and Affiliations

  1. Project team for Pharmacogenetics, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo, 158-8501, Japan

    Yukiko Nakajima, Yoshiro Saito, Kisho Shiseki, Hiromi Fukushima-Uesaka, Shogo Ozawa, Nahoko Kaniwa & Jun-ichi Sawada

  2. Division of Medicinal Safety Science, National Institute of Health Sciences, Tokyo, Japan

    Ryuichi Hasegawa & Nahoko Kaniwa

  3. National Center Hospital for Mental, Nervous and Muscular Disorders, National Center of Neurology and Psychiatry, Tokyo, Japan

    Kenji Sugai, Masaaki Katoh, Osamu Saitoh, Teiichi Ohnuma, Mitsuru Kawai, Taisuke Ohtsuki, Chieko Suzuki & Narihiro Minami

  4. National Institute of Neuroscience, National Center of Neurology and Psychiatry, Tokyo, Japan

    Narihiro Minami, Hideo Kimura & Yu-ichi Goto

  5. Division of Genomic Medicine, Department of Advanced Biomedical Engineering and Science, Tokyo Women’s Medical University, Tokyo, Japan

    Naoyuki Kamatani

Authors
  1. Yukiko Nakajima
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yoshiro Saito
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kisho Shiseki
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hiromi Fukushima-Uesaka
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ryuichi Hasegawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Shogo Ozawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Kenji Sugai
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Masaaki Katoh
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Osamu Saitoh
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Teiichi Ohnuma
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Mitsuru Kawai
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Taisuke Ohtsuki
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Chieko Suzuki
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Narihiro Minami
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Hideo Kimura
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. Yu-ichi Goto
    View author publications

    You can also search for this author in PubMed Google Scholar

  17. Naoyuki Kamatani
    View author publications

    You can also search for this author in PubMed Google Scholar

  18. Nahoko Kaniwa
    View author publications

    You can also search for this author in PubMed Google Scholar

  19. Jun-ichi Sawada
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Nahoko Kaniwa.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Nakajima, Y., Saito, Y., Shiseki, K. et al. Haplotype structures of EPHX1 and their effects on the metabolism of carbamazepine-10,11-epoxide in Japanese epileptic patients. Eur J Clin Pharmacol 61, 25–34 (2005). https://doi.org/10.1007/s00228-004-0878-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00228-004-0878-1

Keywords

Search

Navigation