Relationships between Dapsone Metabolic Activity and Polymorphism of Arylamine N-acetyltransferase 2 in the F2 Hybrid Rats

잡종 2세대(Fischer 계: Wistar-Kyoto 계) 흰쥐에서 Arylamine N-acetyltransferase 2의 다형성과 Dapsone의 대사능과의 연관성에 대한 연구

  • 신인철 (한양대학교 의과대학 약리학교실 및 의과학연구소) ;
  • 강주섭 (한양대학교 의과대학 약리학교실 및 의과학연구소) ;
  • 고현철 (한양대학교 의과대학 약리학교실 및 의과학연구소) ;
  • 이창호 (한양대학교 의과대학 약리학교실 및 의과학연구소) ;
  • 안동춘 (강원대학교 동물지원과학대학 수의학과) ;
  • 백두진 (한양대학교 의과대학 해부학교실) ;
  • 심성한 (한양대학교 의과대학 유전학교실) ;
  • 조율희 (한양대학교 의과대학 유전학교실)
  • Published : 2002.09.01


The arylamine N-acetyltransferases (NATs) are a family of enzymes that N-acetylate mylhydrazines and arylamines through transfer of an acetyl group from acetyl coenzyme A. This activity was found to vary among individuals as a Mendalian trait and the basis of the genetic differences in human NAT activity is one of the best of the genetic studied examples of pharmacogenetic variation. The classical N-acetylation polymorphism is regulated at the NAT2 locus, which segregates individuals into rapid, intermediate, and slow acetylator phenotypes. In this study, the relationship between NAT2 activity phenotype using HPLC:UV assay for the determination of dapsone and monoacetyldapsone in plasma and NAT2 genotype by PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) was investigated in the F2 hybrid (Fischer 344 vs Wistar-Kyoto) rats. Three Common mutant alleles at the NAT2 gene locus have been identified in the F2 generation progeny of Fischer 344 rats as raid acetylator and Wistar-Kyoto rats as slow acetylator segregated into three modes (low, intermediates, and high) with simple Mendelian inheritance. The metabolic activity of NAT2 of the intermediate and rapid acetylators is significant1y greater than slow acetylator, but the metabolic activity of rapid acetylator is not significantly different from Intermediate type. Therefore, we could observe that complete trimodal NAT2 genotypic alleles and incomplete trimodal NAT2 metabolic phenotypic distribution in tile F2 hybrid rats. These observations suggest that the relationships between NAT2 genotype and metabolic phenotype exists and F2 hybrid (Fischer 344: Wistar-Kyoto) animal models about NAT2 polymorphism might be applied in the toxicity and pharmacogenetic studies of arylamine drugs and carcinogens.



  1. Blum, M., Grant, D.M., Mcbride, W., Hein, M. and Meyer, U.A. (1990). Human arylamine N-acetyltransferase genes; isolation, chromosomal localization, and functional expres- sion. DNA Cell. Biol., 9, 193-203
  2. Doll, M.A. and Hein, D.W. (1995). Cloning, sequencing and expression of NAATl and NAT2 encoding genes from rapid and slow acetylator inbred rats. Pharmacogenetics, 5, 247- 251
  3. Fiala, E.S., Weisburger, J.H., Katayama, S., Chandrasekaran, V. and Williams, G.M. (1981). The effect of disulfiram on the carcinogenicity of 3,2'-dimethyl-4-aminobiphenyl in Syrian golden hamsters and rats. Carcinogenesis, 2, 965-969
  4. Grant, D.M., Blum, M., Beer, M. and Meyer, U.A. (1991). Monomorphic and polymorphic human arylamine N-acetyl- transferases: a comparison of liver isozymes and expressed products of two cloned genes. Mol. Pharmacol., 39, 184-191
  5. Hein, D.W. (1988). Acetylator genotype and arylamine-induced carcinogenesis. Biochim. Biophys. Acta., 948, 37-66
  6. Hein, D.W., Doll, M.A., Fretland, A.J., Gray, K., Deitz, A.C., Feng, Y., Jiang, W., Rustan, T.D., Satran, S.L. and Wilkie, T.R. $J_I$. (1997). Rodent models of the human acetyla-tion polymorphism: comparisons of recombinant acetyltrans- ferases. Mutat. Res., 376, 101-106
  7. Hein, D.W., Kirlin, W.G., Ferguson, R.J., Thompson, L.K. and Ogolla, F. (1986). Identification and inheritance of inbred hamster N-acetyltransferase isozyme in peripheral blood. J. Pharmacol. Exp. Ther., 239, 823-828
  8. Hein, D.W., Kirlin, W.G., Ferguson, R.J. and Weber, W.W. (1985). Inheritance of liver N-acetyltransferase activity in the rapid and slow acetylator inbred hamster. J, Pharmacol. Exp. Ther., 233, 584-587
  9. Hein, D.W., Rustan, T.D., Bucher, K.D. and Miller, L. S. (1991). Polymorphic and monomorphic expression of arylamine carcinogen N-acetyltransferase isozymes in tumor target organ cytosols of Syrian hamsters cogenic at the polymorphic acetlytransferase locus. J. Pharmacol. Exp. Ther., 259, 699-704
  10. Hein, D.W., Smolen, T.N., Fox, R.R. and Weber, W.W. (1982). Identification of genetically homozygous rapid and slow acetylators of drugs and environmental carcinogens among established inbred rabbit strains. J. Pharmacol. Exp. Ther., 223, 40-44
  11. Hein, D.W. and Weber, W.W. (1984). Relationship between N- acetylator phenotype and susceptibility toward hydrazine- induced lethal central nervous system toxicity in the rabbit. J. Pharmacol. Exp. Ther., 228, 588-592
  12. Ito, N., Shirai, T., Tagawa, Y., Nakamura, A. and Fukushima, S. (1988). Variation in tumor yield in the prostate and other target organs of the rat in response to varied dosage and duration of administration of 3,2'-dimethy1-4-aminopheno1. Cancer Res., 48, 4629-4632
  13. Martell, R.T. and Weber, W.W. (1993). N-acetylation poly morphism in liver and pancreas of inbred rats. Drug Metab. Disposit., 21, 965-966
  14. McDonald, M.M. and Boorman, G.A. (1989). Pancreatic hepatocytes associated with chronic 2,6-dichloro-p-pheny1- enediamine admmistrations in Fischer 344 rats. Toxicol. Pathol., 17, 1-6
  15. Newton, J.F., Yoshimoto, M., Bernstein, J., Rush, G.F. and Hook, J.B. (1983). Acetaminophen nephrotoxicity in the rat. II. Strain differences in nephrotoxicity and metabolism of p- aminophenol, a metabolite of acetaminophen. Toxicol. Appl. Pharmacol., 69, 307-318
  16. Ogolla, F., Ferguson, R.J., Kirlin, W.G., Trinidad, A., Andrews, A.F., Mpezo, M. and Hein, D.W. (1990). Acetylator genotype-dependent expression of arylamine N-acetylt- ransferase and N-hydroxylamine O-acetyltransferase in Syrian inbred hamster intestine and colon. Identity with the hepatic acetylation polymorphism. Drug Metab. Dispos., 18, 680-685
  17. Ozawa, S., Abu-Zeid, M., Kawakubo, Y., Toyama, S., Yamazoe, Y. and Kato, R. (1990). Monomorphic and polymorphic isozymes of arylamine N-acetyltransferases in hamster liver: purification of the isozymes and genetic basis of N- acetylation polymorphism. Carcinogenesis, 11, 2137-2144
  18. Queiroz, R.H., Dreossi, S.A. and Carvalho, D. (1997). A rapid, specific, and sensitive method for the determination of acetylation phenotype using dapsone. J. Anal. Toxicol., 21, 203-207
  19. Roberts, S.M., Budinsky, R.A., Adams, L.E., Litwin, A. and Hess, E. V. (1985). Procainamide acetylation in strains of rat and mouse. Drug. Metab. Dispos., 13, 517-519
  20. Schneck, D.W., Grove, K., Dewitt, F.O., Shiroff, R.A. and Hayes, A.H. $J_I$. (1978). The quantative disposition of procainamide and N-acetylprocainamide in the rat. J. Pharmacol. Exp. Ther., 204, 219-225
  21. Shirai, T., Nakamura, A., Fukushima, S., Wang, C.Y., Yamada, H. and Ito, N. (1990). Selective induction of prostate carcinoma in F344 rats treated with intraperitoneal injections of N-hydroxy-3, 2'-dimethyl-4-aminobiphenyl. Jpn. J. Cancer Res., 81, 320-323
  22. Shirai, T, Tamano, S., Kato, T., Iwasaki, S., Takahashi, S. and Ito, N. (1991). Induction of invasive carcinoma in the acces- sory sex organs other than the ventral prostate of rats given 3,2'-dimethyl biphenyl-4-aminobiphenyl and testoster-one propionate. Cancer Res., 51, 1264-1269, 1991
  23. Takayama, S., Nakatsuru, Y., Masuda, M., Ohgaki, H., Sato, S. and Sugimura, T. (1984). Demonstration of carcinogenicity in Fischer 344 rats of 2-amino-3-methyl-imidazo1e '4,5-F' qu- inoline from broiled sardine, fried beef and beef extract. Gann., 75, 467-470
  24. Tannen, R.H., Weber, W.W. (1980). Inheritance of acetylator phenotype in mice. J. Pharmacol. Exp. Ther., 213, 480-484
  25. Weber, W.W. and Hein, D.W. (1985). N-acetylation pharmaco- genetics. Pharmacol. Rev. 37, 25-79
  26. Wick, M.J. and Hanna, P.E. (1990). Bioactivation of N- arylhydroxamic acids by rat hepatic N-acetyltransferase: De- tection of multiple enzymes forms by mechanism-based inactivation. Biochem. Pharmcol., 39, 991-1003