Production of certified reference materials for the sports doping control of the REV‐ERB agonist SR9009

Abstract

Two human metabolites of the REV-ERB agonist SR9009, identified by researchers with an interest in sports doping control, have been synthesized and assessed for purity. The synthesis employed was a modification of published procedures for the parent SR9009, careful attention to the purification of intermediates and the final product ensuring materials of the highest purity were available for certification. For each candidate material impurities of related structure were identified and quantified as a relative mass fraction using high performance liquid chromatography-ultraviolet (HPLC-UV) detection and proton nuclear magnetic resonance ( H NMR) spectroscopy. The quantification of water, occluded solvent, and inorganic residue was assessed using Karl Fischer, H NMR, and thermogravimetric analysis, thereby completing the assessment of all impurities typically characterized by the mass balance approach. Summation and subtraction from 1000 mg/g afforded the mass fraction of the main component, the associated uncertainty ensuring certified reference material status can be applied to the resulting pure substance calibration standards. The availability of these standards to the sports doping control community will facilitate delivery of metrological traceability to the SI unit for mass (kg) to routine testing results and aid method development for the detection and quantification of SR9009 abuse.

SR9009 has REV-ERB–independent effects on cell proliferation and metabolism

“…Like other NRs, REV-ERBs can be regulated by ligands, including naturally occurring heme (22,23), which modulate their repressive activity. Attempts to pharmacologically target REV-ERBs by the use of putatively specific synthetic agonists, particularly SR9009 (24), have suggested a wide range of beneficial effects in healthy as well as diseased animal models and cell systems (11,(24)(25)(26)(27)(28)(29)(30)(31)(32)(33)(34). The beneficial effects of SR9009 on weight and exercise in mice (24,27) have led to an interest in the use of the compound by humans, with online advertisements selling SR9009 highlighting REV-ERBs as the molecular target (e.g., https://www.simplyanabolics.com/sarms/sr9009-stenabolic/).…”

“…The beneficial effects of SR9009 on weight and exercise in mice (24,27) have led to an interest in the use of the compound by humans, with online advertisements selling SR9009 highlighting REV-ERBs as the molecular target (e.g., https://www.simplyanabolics.com/sarms/sr9009-stenabolic/). This, in turn, has prompted newly developed assays that detect its surreptitious use (31,33,35).…”

The Role of Circadian Clock Genes in Critical Illness: The Potential Role of Translational Clock Gene Therapies for Targeting Inflammation, Mitochondrial Function, and Muscle Mass in Intensive Care

“…The unlicensed use of these drugs as performance enhancers ( Davies et al, 2019 ) raises the issue of how to enhance “performance” in the critically ill. The early phase of clinical trials of new, circadian clock therapeutics is now urgently needed.…”

“…REVERB agonists can affect skeletal muscle oxidative and functional capacity ( Woldt et al, 2013 ; Amador et al, 2018 ) and so may find an application in this context. Indeed, REVERB doping has been proposed in elite sports ( Davies et al, 2019 ). REVERB may prevent muscle atrophy, so preserving muscle mass ( Mayeuf-Louchart et al, 2017 ).…”

Annual banned‐substance review – Analytical approaches in human sports drug testing

“…While sex did not appear to affect the metabolism of SR9009, a significant influence of genetic polymorphism (especially concerning CYP2D6 and CYP2C19) as well as the presence of antifungal therapeutics or antidepressants was recorded. Overall, the targeting of three metabolites resulting from dealkylation reactions (Figure A) was recommended, two of which were recently synthesized and commercialized as phase‐I metabolites for sports drug testing purposes . Of note, in vivo metabolism data indicated a significant share of glucuronic acid conjugation of these metabolites, and hence hydrolysis and/or inclusion of intact phase‐II metabolites into routine doping controls is recommended.…”