HepG2 as promising cell-based model for biosynthesis of long-term metabolites: Exemplified for metandienone

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

  • Annette Zschiesche - , Institute of Doping Analysis and Sports Biochemistry Dresden (Author)
  • Zdenek Chundela - , Institute of Doping Analysis and Sports Biochemistry Dresden (Author)
  • Detlef Thieme - , Institute of Doping Analysis and Sports Biochemistry Dresden (Author)
  • Annekathrin Martina Keiler - , Environmental Monitoring and Endocrinology (Research Group), Institute of Doping Analysis and Sports Biochemistry Dresden (Last author)

Abstract

In order to detect the abuse of substances in sports, the knowledge of their metabolism is of undisputable importance. As in vivo administration of compounds faces ethical problems and might even not be applicable for nonapproved compounds, cell-based models might be a versatile tool for biotransformation studies. We coincubated HepG2 cells with metandienone and D3 -epitestosterone for 14 days. Phase I and II metabolites were analyzed by high-performance liquid chromatography (HPLC)-tandem mass spectrometry and confirmed by gas chromatography-mass spectrometry (GC-MS). The metandienone metabolites formed by HepG2 cells were comparable with those renally excreted by humans. HepG2 cells also generated the two long-term metabolites 17β-hydroxymethyl-17α-methyl-18-nor-androst-1,4,13-trien-3-one and 17α-hydroxymethyl-17β-methyl-18-nor-androst-1,4,13-trien-3-one used in doping analyses, though in an inverse ratio compared with that observed in human urine. In conclusion, we showed that HepG2 cells are suitable as model for the investigation of biotransformation of androgens, especially for the anabolic androgenic steroid metandienone. They further proved to cover phase I and II metabolic pathways, which combined with a prolonged incubation time with metandienone resulted in the generation of its respective long-term metabolites known from in vivo metabolism. Moreover, we showed the usability of D3 -epitestosterone as internal standard for the incubation. The method used herein appears to be suitable and advantageous compared with other models for the investigation of doping-relevant compounds, probably enabling the discovery of candidate metabolites for doping analyses.

Details

Original languageEnglish
Pages (from-to)298-306
Number of pages9
JournalDrug testing and analysis
Volume14
Issue number2
Publication statusPublished - 1 Feb 2022
Peer-reviewedYes

External IDs

Scopus 85118312225
Mendeley 8222eaa0-ef73-327f-9e18-89eb6e3c624f
unpaywall 10.1002/dta.3184
ORCID /0000-0002-2157-4711/work/142251648

Keywords

DFG Classification of Subject Areas according to Review Boards

Sustainable Development Goals

Keywords

  • GC–MS, HPLC–MS, HepG2, in vitro metabolism, metandienone