Identification of novel bioactive metabolites of 5-demethylnobiletin in mice

Mol Nutr Food Res. 2013 Nov;57(11):1999-2007. doi: 10.1002/mnfr.201300211. Epub 2013 Jul 26.

Abstract

Scope: Biotransformation of dietary components is important for their in vivo biological activities after oral ingestion. Herein, we investigated biotransformation of 5-demethylnobiletin (a polymethoxyflavone found in citrus fruits) in mice, and its implication in the inhibition of human colon cancer cells.

Methods and results: Urine samples were collected from mice fed with 5-demethylnobiletin (1) and analyzed by LC-ESI-MS and HPLC coupled with an electrochemical detector. Three major metabolites were identified as 5,3'-didemethylnobiletin (2), 5,4'-didemethylnobiletin (3), and 5,3',4'-tridemethylnobiletin (4) by comparing their ESI-MS and HPLC profiles with those of authentic standards synthesized by a multistep route. Cell viability assay in human colon cancer cells demonstrated that all three metabolites of 5-demethylnobiletin, especially 5,3'-didemethylnobiletin (2), showed much stronger inhibitory effects on cancer cell growth than 5-demethylnobiletin. For example, metabolites (2-4) showed IC50 of 0.12, 5.5, and 4.2 μM in SW620 cells, respectively, while 5-demethylnobiletin at 10 μM only caused 37% inhibition after 72 h of treatment.

Conclusion: Three novel metabolites were identified in mice after oral administration of 5-demethylniobiletin. These metabolites exhibited strong inhibitory effects against human colon cancer cells. Our results provide a first report on these bioactive metabolites and warrant further investigation on their molecular mechanism of actions.

Keywords: 5,3′-Didemethylnobiletin; 5-Demethylnobiletin; Cancer inhibition; Metabolites.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Administration, Oral
  • Animals
  • Biotransformation
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Chromatography, High Pressure Liquid
  • Chromatography, Liquid
  • Citrus / chemistry
  • Colonic Neoplasms / drug therapy
  • Flavones / administration & dosage
  • Flavones / pharmacokinetics*
  • Flavones / urine*
  • Humans
  • Male
  • Mass Spectrometry
  • Mice
  • Plant Extracts / administration & dosage
  • Plant Extracts / chemistry

Substances

  • Flavones
  • Plant Extracts
  • 5-O-demethylnobiletin