In vitro metabolism of beta-lapachone (ARQ 501) in mammalian hepatocytes and cultured human cells

Rapid Commun Mass Spectrom. 2009 Jan;23(1):12-22. doi: 10.1002/rcm.3835.


ARQ 501 (3,4-dihydro-2,2-dimethyl-2H-naphthol[1,2-b]pyran-5,6-dione, beta-lapachone) is an anticancer agent, currently in multiple phase II clinical trials as monotherapy and in combination with other cytotoxic drugs. This study focuses on in vitro metabolism in cryopreserved hepatocytes from mice, rats, dogs and humans using [(14)C]-labeled ARQ 501. Metabolite profiles were characterized using liquid chromatography/mass spectrometry combined with an accurate radioactivity counter. Ion trap mass spectrometry was employed for further structural elucidation. A total of twelve metabolites were detected in the mammalian hepatocytes studied; all of which but one were generated from phase II conjugation reactions. Ten of the observed metabolites were produced by conjugations occurring at the reduced ortho-quinone carbonyl groups of ARQ 501. The metabolite profiles revealed that glucuronidation was the major biotransformation pathway in mouse and human hepatocytes. Monosulfation was the major pathway in dog, while, in rat, it appears glucuronidation and sulfation pathways contributed equally. Three major metabolites were found in rats: monoglucuronide M1, monosulfate M6, and glucuronide-sulfate M9. Two types of diconjugation metabolites were formed by attachment of the second glycone to an adjacent hydroxyl or to an existing glycone. Of the diconjugation metabolites, glucosylsulfate M10, diglucuronide M5, and glucuronide-glucoside M11 represent rarely observed phase II metabolites in mammals. The only unconjugated metabolite was generated through hydrolysis and was observed in rat, dog and human hepatocytes. ARQ 501 appeared less stable in human hepatocytes than in those of other species. To further elucidate the metabolism of ARQ 501 in extrahepatic sites, its metabolism in human kidney, lung and intestine cells was also studied, and only monoglucuronide M1 was observed in all the cell types examined.

MeSH terms

  • Animals
  • Antineoplastic Agents / metabolism*
  • Cells, Cultured
  • Chromatography, Liquid / methods*
  • Dogs
  • Glucosides / chemistry
  • Glucosides / metabolism
  • Glucuronates / chemistry
  • Glucuronates / metabolism
  • Hepatocytes / metabolism*
  • Humans
  • Hydrolysis
  • Intestinal Mucosa / metabolism
  • Isotope Labeling
  • Kidney / metabolism
  • Lung / metabolism
  • Mass Spectrometry / methods*
  • Metabolomics / methods
  • Mice
  • Naphthoquinones / metabolism*
  • Rats
  • Sulfates / chemistry
  • Sulfates / metabolism


  • Antineoplastic Agents
  • Glucosides
  • Glucuronates
  • Naphthoquinones
  • Sulfates
  • beta-lapachone