The fungusCunninghamella eleganscan produce human and equine metabolites of selective androgen receptor modulators (SARMs)

Abstract

1. Selective androgen receptor modulators (SARMs) are a group of substances that have potential to be used as doping agents in sports. Being a relatively new group not available on the open market means that no reference materials are commercially available for the main metabolites. In the presented study, the in vitro metabolism of SARMs by the fungus Cunninghamella elegans has been investigated with the purpose of finding out if it can produce relevant human and equine metabolites. 2. Three different SARMs, S1, S4 and S24, were incubated for 5 days with C. elegans. The samples were analysed both with and without sample pretreatment using ultra performance liquid chromatography coupled to high resolution mass spectrometry. 3. All the important phase I and some phase II metabolites from human and horse were formed by the fungus. They were formed through reactions such as hydroxylation, deacetylation, O-dephenylation, nitro-reduction, acetylation and sulfonation. 4. The study showed that the fungus produced relevant metabolites of the SARMs and thus can be used to mimic mammalian metabolism. Furthermore, it has the potential to be used for future production of reference material.

Metabolic Profile of Synthetic Cannabinoids 5F-PB-22, PB-22, XLR-11 and UR-144 by Cunninghamella elegans

“…This finding agrees with the study on the synthetic cannabinoid AM2201 that glucosidation and sulfation are the only phase II reactions observed for C. elegans metabolism while glucuronidation is the major phase II transformation in humans [33]. Similarly, for some other drugs, glucosidation and sulfation are reported to be the phase II reactions of C. elegans while the same drugs are glucuronidated and sulfated in humans [38,48]. The latter experiments did not involve extraction step as a sample preparation, supporting that poor recovery of glucuronides in dichloromethane is not likely a reason for the absence of glucuronides in this study.…”

Investigation of the selective androgen receptor modulators S1, S4 and S22 and their metabolites in equine plasma using high‐resolution mass spectrometry

“…Its elemental composition was determined to be C 11 H 10 N 2 O 5 F 3 for the negatively charged ion and it fragmented to m/z 205, by a loss of C 4 H 6 O 3 . The reaction that had occurred could not be elucidated, but both the retention time [14] and fragment [11] corresponded with previous findings which supports the identification of M6b. The intensities of the metabolites were comparable for the three horses administered with SARM S1 and all metabolites observed in plasma have been previously described in urine.…”

“…[7] Furthermore, the identification of metabolites also corroborates that a substance has passed through a metabolic system and that an adverse finding is not caused by contamination of the sample. The metabolism of different SARMs of the arylpropionamide class has been studied in vitro in a variety of systems such as human liver microsomes, [8] equine liver microsomes, [9] bovine liver microsomes [10] and fungal incubates, [11] as well as in vivo in both human [12,13] and equine [14] urine. The three aforementioned in vivo studies showed that the SARMs were metabolized rapidly and extensively.…”

Detection of the selective androgen receptor modulator andarine (S‐4) in a routine equine blood doping control sample

“…[7] Analytical methods utilizing the sensitivity and specificity of liquid chromatography-high resolution accurate mass (LC-HRAM) spectrometry to detect andarine (S-4) have been reported since 2006. [10] For the benefit of urine analysis in equine [11][12][13] and human [13,14] anti-doping, in vitro and in vivo metabolic studies of andarine (S-4) have been performed. [10] For the benefit of urine analysis in equine [11][12][13] and human [13,14] anti-doping, in vitro and in vivo metabolic studies of andarine (S-4) have been performed.…”