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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
Author(s) -
Thevis Mario,
Lagojda Andreas,
Kuehne Dirk,
Thomas Andreas,
Dib Josef,
Hansson Annelie,
Hedeland Mikael,
Bondesson Ulf,
Wigger Tina,
Karst Uwe,
Schänzer Wilhelm
Publication year - 2015
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.7189
Subject(s) - chemistry , drug , pharmacology , in vitro , androgen receptor , drug discovery , computational biology , biochemistry , medicine , cancer , prostate cancer , biology
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. Copyright © 2015 John Wiley & Sons, Ltd.