Wedelolactone metabolism in rats through regioselective glucuronidation catalyzed by uridine diphosphate-glucuronosyltransferases 1As (UGT1As)

Phytomedicine. 2016 Apr 15;23(4):340-9. doi: 10.1016/j.phymed.2016.01.007. Epub 2016 Feb 10.

Abstract

Background: Wedelolactone (WEL), a medicinal plant-derived coumestan, has been reported to exhibit a diverse range of pharmacological activities. However, the metabolism and disposition of WEL remain unexplored.

Purpose: The present study aims to investigate the metabolism of WEL in rats and identify the enzymes responsible for forming major WEL metabolites.

Methods: Plasma, urine, feces, and bile samples were collected before and after 50 mg/kg WEL was orally administered to rats. Metabolites were profiled by ultrahigh performance liquid chromatography/quadrupole time-of-flight mass spectrometry and identified by high-performance liquid chromatography-solid-phase extraction-nuclear magnetic resonance spectroscopy. The in vitro WEL glucuronidation activities of human liver microsomes, human kidney microsomes, human intestine microsomes, and 12 recombinant human uridine diphosphate-glucuronosyltransferase (UGT) isoforms were screened. Molecular docking simulation of the interaction between WEL and UGT1A9 was conducted.

Results: WEL underwent extensive metabolism, and 17 metabolites were identified. The major metabolic pathways observed were glucuronidation and methylation. Glucuronic acid was preferentially introduced into 5-OH, whereas no obvious regioselectivity was observed in the methylation of 11-OH and 12-OH. Multiple UGTs, including UGT1A1, UGT1A3, UGT1A6, UGT1A7, UGT1A8, UGT1A9, and UGT1A10, were involved in forming WEL glucuronides and O-methylated WEL glucuronides.

Conclusion: The extensive glucuronidation and methylation is responsible for the low oral bioavailability of WEL in rats. UGT1A1 and UGT1A9 were the major enzymes involved in the glucuronidation of WEL and O-methylated WEL. Molecular docking studies revealed that 5-OH was accessible to the catalytic domain of UGT1As; therefore, 5-OH exhibited a high probability of glucuronidation.

Keywords: Metabolism; Molecular docking; Uridine diphosphate-glucuronosyltransferase (UGT); Wedelolactone.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Asteraceae / chemistry
  • Biological Availability
  • Coumarins / metabolism
  • Coumarins / pharmacokinetics*
  • Glucuronic Acid / metabolism
  • Glucuronides / metabolism*
  • Glucuronosyltransferase / metabolism*
  • Humans
  • Intestinal Mucosa / metabolism*
  • Kidney / metabolism*
  • Liver / metabolism*
  • Male
  • Mass Spectrometry
  • Methylation
  • Microsomes / metabolism
  • Molecular Docking Simulation
  • Plant Extracts / metabolism
  • Protein Isoforms
  • Rats
  • UDP-Glucuronosyltransferase 1A9
  • UGT1A1 Enzyme
  • Uridine Diphosphate / metabolism*

Substances

  • Coumarins
  • Glucuronic Acid
  • Glucuronides
  • Glucuronosyltransferase
  • Plant Extracts
  • Protein Isoforms
  • UDP-Glucuronosyltransferase 1A9
  • Uridine Diphosphate
  • UGT1A1 Enzyme
  • UGT1A9 protein, human
  • wedelolactone
  • coumestan