Abstract
The elucidation of metabolic pathways and the detection of emerging therapeutics potentially enhancing athletic performance are of paramount importance to doping control authorities to protect the integrity of elite sports. A new drug candidate belonging to the family of the peroxisome proliferator-activated receptor-delta agonists termed GW1516 (also referred to as GW501516) has been prohibited by the World Anti-Doping Agency in 2009 due to its potential to artificially increase endurance. Consequently, sports drug testing laboratories need to establish detection methods enabling the identification of the intact substance and/or its metabolite(s) that unambiguously prove the presence or absence of the target substances in doping control specimens. Simulating human metabolic reactions using liver microsomal preparations, minute amounts of possible urinary metabolites were obtained that were characterized by mass spectrometry-based methods. Subsequently, the most abundant metabolic products were chemically synthesized and as well characterized by mass spectrometry and nuclear magnetic resonance spectroscopy. Finally, GW1516 and two oxidized metabolites were implemented in a routine doping control analytical assay based on liquid chromatography-(tandem) mass spectrometry (LC-MS/MS), which was tested for its fitness-for-purpose using spiked urine samples.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Thevis M (2010) Mass spectrometry in sports drug testing—characterization of prohibited substances and doping control analytical assays. Wiley, Hoboken
WADA (2011) World Anti-Doping Agency, vol 2010
Thevis M, Kuuranne T, Geyer H, Schänzer W (2011) Annual banned-substance review: analytical approaches in human sports drug testing. Drug Test Anal 3:1–14
Kazlauskas R (2010) Designer steroids. Handb Exp Pharmacol 195:155–185
Thevis M, Thomas A, Kohler M, Beuck S, Schänzer W (2009) Emerging drugs: mechanism of action, mass spectrometry and doping control analysis. J Mass Spectrom 44:442–460
Narkar VA, Downes M, Yu RT, Embler E, Wang YX, Banayo E, Mihaylova MM, Nelson MC, Zou Y, Juguilon H, Kang H, Shaw RJ, Evans RM (2008) AMPK and PPARdelta agonists are exercise mimetics. Cell 134:405–415
US National Institutes of Health (2009) Clinical trials including GW501516. http://www.clinicaltrials.gov/ct2/results?term=gw501516. Accessed 20 Jun 2009
Wang YX, Lee CH, Tiep S, Yu RT, Ham J, Kang H, Evans RM (2003) Peroxisome-proliferator-activated receptor delta activates fat metabolism to prevent obesity. Cell 113:159–170
Krämer DK, Al-Khalili L, Guigas B, Leng Y, Garcia-Roves PM, Krook A (2007) Role of AMP kinase and PPARdelta in the regulation of lipid and glucose metabolism in human skeletal muscle. J Biol Chem 282:19313–19320
Luquet S, Lopez-Soriano J, Holst D, Gaudel C, Jehl-Pietri C, Fredenrich A, Grimaldi PA (2004) Roles of peroxisome proliferator-activated receptor delta (PPARdelta) in the control of fatty acid catabolism. A new target for the treatment of metabolic syndrome. Biochimie 86:833–837
Brunmair B, Staniek K, Dorig J, Szocs Z, Stadlbauer K, Marian V, Gras F, Anderwald C, Nohl H, Waldhausl W, Furnsinn C (2006) Activation of PPAR-delta in isolated rat skeletal muscle switches fuel preference from glucose to fatty acids. Diabetologia 49:2713–2722
Wang YX, Zhang CL, Yu RT, Cho HK, Nelson MC, Bayuga-Ocampo CR, Ham J, Kang H, Evans RM (2004) Regulation of muscle fiber type and running endurance by PPARdelta. PLoS Biol 2:e294
Thevis M, Möller I, Thomas A, Beuck S, Rodchenkov G, Bornatsch W, Geyer H, Schänzer W (2010) Characterization of two major urinary metabolites of the PPARdelta-agonist GW1516 and implementation of the drug in routine doping controls. Anal Bioanal Chem 396:2479–2491
Acknowledgments
The presented work was supported by Antidoping Switzerland (Berne, Switzerland) and the Manfred-Donike-Institute for Doping Analysis (Cologne, Germany).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media New York
About this protocol
Cite this protocol
Thevis, M., Möller, I., Beuck, S., Schänzer, W. (2013). Synthesis, Mass Spectrometric Characterization, and Analysis of the PPARδ Agonist GW1516 and Its Major Human Metabolites: Targets in Sports Drug Testing. In: Badr, M., Youssef, J. (eds) Peroxisome Proliferator-Activated Receptors (PPARs). Methods in Molecular Biology, vol 952. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-155-4_22
Download citation
DOI: https://doi.org/10.1007/978-1-62703-155-4_22
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-154-7
Online ISBN: 978-1-62703-155-4
eBook Packages: Springer Protocols