Identification and characterization of human UDP-glucuronosyltransferases responsible for the in vitro glucuronidation of salvianolic acid A

Drug Metab Pharmacokinet. 2012;27(6):579-85. doi: 10.2133/dmpk.dmpk-12-rg-023. Epub 2012 Apr 27.

Abstract

Glucuronidation is an important pathway in the elimination of salvianolic acid A (Sal A); however the mechanism of UDP-glucuronosyltransferases (UGTs) in this process remains to be investigated. In this study, the kinetics of Sal A glucuronidation by pooled human liver microsomes (HLMs), pooled human intestinal microsomes (HIMs) and 12 recombinant UGT isozymes were investigated. The glucuronidation of Sal A can be shown both in HLMs and HIMs with K(m) values of 39.84 ± 3.76 and 54.04 ± 4.36 µM, respectively. Among the 12 human UGTs investigated, UGT1A1 and UGT1A9 were the major isoforms that catalyzed the glucuronidation of Sal A (K(m) values of 29.72 ± 2.20 and 24.40 ± 2.60 µM). UGT1A9 showed the highest affinity of Sal A glucuronidation. Furthermore, a significant correlation between Sal A glucuronidation and propofol glucuronidation (a typical UGT1A9 substrate) was observed. The chemical inhibition study showed that the IC(50) for phenylbutazone inhibition of Sal A glucuronidation was 50.3 ± 4.3 and 39.4 ± 2.9 µM by HLMs and UGT1A9, respectively. Mefenamic acid inhibited Sal A glucuronidation in UGT1A1 and HLMs with IC(50) values of >200 and 12.4 ± 2.2 µM, respectively.

MeSH terms

  • Caffeic Acids / metabolism
  • Caffeic Acids / pharmacokinetics*
  • Glucuronidase / metabolism
  • Glucuronosyltransferase / antagonists & inhibitors
  • Glucuronosyltransferase / metabolism*
  • Humans
  • Hydrolysis / drug effects
  • Intestinal Mucosa / metabolism
  • Intestines / drug effects
  • Intestines / enzymology
  • Isoenzymes / metabolism
  • Kinetics
  • Lactates / metabolism
  • Lactates / pharmacokinetics*
  • Mefenamic Acid / pharmacology
  • Microsomes / drug effects
  • Microsomes / enzymology
  • Microsomes / metabolism
  • Microsomes, Liver / drug effects
  • Microsomes, Liver / enzymology
  • Microsomes, Liver / metabolism
  • Propofol / metabolism
  • Propofol / pharmacokinetics
  • Statistics as Topic
  • UDP-Glucuronosyltransferase 1A9

Substances

  • Caffeic Acids
  • Isoenzymes
  • Lactates
  • UGT1A9 protein, human
  • Mefenamic Acid
  • salvianolic acid A
  • UGT1A1 enzyme
  • Glucuronosyltransferase
  • UDP-Glucuronosyltransferase 1A9
  • Glucuronidase
  • Propofol