Characterization of a non-approved selective androgen receptor modulator drug candidate sold via the Internet and identification of in vitro generated phase-I metabolites for human sports drug testing

Rapid Commun Mass Spectrom. 2015 Jun 15;29(11):991-9. doi: 10.1002/rcm.7189.

Abstract

Rationale: Potentially performance-enhancing agents, particularly anabolic agents, are advertised and distributed by Internet-based suppliers to a substantial extent. Among these anabolic agents, a substance referred to as LGD-4033 has been made available, comprising the core structure of a class of selective androgen receptor modulators (SARMs).

Methods: In order to provide comprehensive analytical data for doping controls, the substance was obtained and characterized by nuclear magnetic resonance spectroscopy (NMR) and liquid chromatography/electrospray ionization high resolution/high accuracy tandem mass spectrometry (LC/ESI-HRMS). Following the identification of 4-(2-(2,2,2-trifluoro-1-hydroxyethyl)pyrrolidin-1-yl)-2-(trifluoromethyl)benzonitrile, the substance was subjected to in vitro metabolism studies employing human liver microsomes and Cunninghamella elegans (C. elegans) preparations as well as electrochemical metabolism simulations.

Results: By means of LC/ESI-HRMS, five main phase-I metabolites were identified as products of liver microsomal preparations including three monohydroxylated and two bishydroxylated species. The two most abundant metabolites (one mono- and one bishydroxylated product) were structurally confirmed by LC/ESI-HRMS and NMR. Comparing the metabolic conversion of 4-(2-(2,2,2-trifluoro-1-hydroxyethyl)pyrrolidin-1-yl)-2-(trifluoromethyl)benzonitrile observed in human liver microsomes with C. elegans and electrochemically derived metabolites, one monohydroxylated product was found to be predominantly formed in all three methodologies.

Conclusions: The implementation of the intact SARM-like compound and its presumed urinary phase-I metabolites into routine doping controls is suggested to expand and complement existing sports drug testing methods.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anabolic Agents / chemistry*
  • Anabolic Agents / economics
  • Anabolic Agents / metabolism*
  • Androgens / chemistry*
  • Androgens / metabolism*
  • Chromatography, Liquid
  • Cunninghamella / drug effects
  • Cunninghamella / metabolism
  • Doping in Sports / economics
  • Humans
  • Internet
  • Magnetic Resonance Spectroscopy
  • Microsomes, Liver / chemistry
  • Microsomes, Liver / metabolism
  • Molecular Structure
  • Receptors, Androgen / metabolism*
  • Substance Abuse Detection
  • Tandem Mass Spectrometry

Substances

  • AR protein, human
  • Anabolic Agents
  • Androgens
  • Receptors, Androgen