In Vitro Metabolism of a New Neuroprotective Agent, KR-31543 in the Human Liver Microsomes : Identification of Human Cytochrome P450

  • Ji, Hye-Young (Drug Metabolism and Bioanalysis Laboratory, College of Pharmacy, Wonkwang University) ;
  • Lee, Seung-Seok (Drug Metabolism and Bioanalysis Laboratory, College of Pharmacy, Wonkwang University) ;
  • Yoo, Sung-Eun (Korea Research Institute of Chemical Technology) ;
  • Kim, Hosoon (AgroPharma Research Institute, Dongbu Hannong Chemical Company) ;
  • Lee, Dong-Ha (AgroPharma Research Institute, Dongbu Hannong Chemical Company) ;
  • Lim, Hong (AgroPharma Research Institute, Dongbu Hannong Chemical Company) ;
  • Lee, Hye-Suk (Drug Metabolism and Bioanalysis Laboratory, College of Pharmacy, Wonkwang University)
  • Published : 2004.02.01

Abstract

KR-31543, (2S,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(2 -methyl-2H-tetrazol-5-ylmethyl) amino]-3,4-dihydro-2-dimethoxymethyl-3-hydroxy-2-methyl-2H-1-benzopyran, is a new neuroprotective agent for preventing ischemia-reperfusion damage. This study was performed to identify the metabolic pathway of KR-31543 in human liver microsomes and to characterize cytochrome P450 (CYP) enzymes that are involved in the metabolism of KR-31543. Human liver microsomal incubation of KR-31543 in the presence of NADPH resulted in the formation of two metabolites, M1 and M2. M1 was identified as N-(4-chlorophenyl)-N-(2-methyl-2H-tetrazol-5-ylmethyl)amine on the basis of LC/MS/MS analysis with a synthesized authentic standard, and M2 was suggested to be hydroxy-KR-31543. Correlation analysis between the known CYP enzyme activities and the rates of the formation of M 1 and M2 in the 12 human liver microsomes have showed significant correlations with testosterone 6$\beta$-hydroxylase activity (a marker of CYP3A4). Ketoconazole, a selective inhibitor of CYP3A4, and anti-CYP3A4 monoclonal antibodies potently inhibited both N-hydrolysis and hydroxylation of KR-31543 in human liver microsomes. These results provide evidence that CYP3A4 is the major isozyme responsible for the metabolism of KR-31543 to M1 and M2.

Keywords

References

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