Metabolic mapping of A3 adenosine receptor agonist MRS5980

Abstract

(1S,2R,3S,4R,5S)-4-(2-((5-Chlorothiophen-2-yl)ethynyl)-6-(methylamino)-9H-purin-9-yl)-2,3-dihydroxy-N-methylbicyclo[3.1.0]hexane-1-carboxamide (MRS5980) is an A3AR selective agonist containing multiple receptor affinity- and selectivity-enhancing modifications and a therapeutic candidate drug for many inflammatory diseases. Metabolism-related poor pharmacokinetic behavior and toxicities are a major reason of drug R&D failure. Metabolomics with UPLC-MS was employed to profile the metabolism of MRS5980 and MRS5980-induced disruption of endogenous compounds. Recombinant drug-metabolizing enzymes screening experiment were used to determine the enzymes involved in MRS5980 metabolism. Analysis of lipid metabolism-related genes was performed to investigate the reason for MRS5980-induced lipid metabolic disorders. Unsupervised principal components analysis separated the control and MRS5980 treatment group in feces, urine, and liver samples, but not in bile and serum. The major ions mainly contributing to the separation for feces and urine were oxidized MRS5980, glutathione (GSH) conjugates and cysteine conjugate (degradation product of the GSH conjugates) of MRS5980. The major ions contributing to the group separation of liver samples were phosphatidylcholines. In vitro incubation experiments showed the major involvement of CYP3A enzymes in the oxidative metabolism of MRS5980 and direct GSH reactivity of MRS5980. The electrophilic attack by MRS5980 is a minor pathway and did not alter GSH levels in liver or liver histology, and thus may be of minor clinical consequence. Gene expression analysis further showed decreased expression of PC biosynthetic genes choline kinase a and b, which further accelerated conversion of lysophosphatidylcholine to phosphatidylcholines through increasing the expression of lysophosphatidylcholine acyltransferase 3. These data will be useful to guide rational design of drugs targeting A3AR, considering efficacy, metabolic elimination, and electrophilic reactivity.

A3 Adenosine Receptors as Modulators of Inflammation: From Medicinal Chemistry to Therapy

“…C2‐triazole substitution (two positional isomers) was previously found to be compatible with A 3 AR binding in the riboside series, as in 15a and 15b . As a postscript to that effort to replace the C2‐arylethynyl group, this ethynyl group was found to be relatively unreactive toward thiols such as glutathione, and the risk of such compounds depleting liver glutathione was shown to be very small …”

“…132 As a postscript to that effort to replace the C2-arylethynyl group, this ethynyl group was found to be relatively unreactive toward thiols such as glutathione, and the risk of such compounds depleting liver glutathione was shown to be very small. 133 The surprising finding that substitution of the exocylic NH of adenine with H or CH 3 in MRS5919 32 allowed full activation of the A 3 AR emphasized that the loss of otherwise important recognition elements in a ligand can be compensated by other affinity enhancing moieties on the nucleoside. 134 Moreover, the ribose moiety is the main effector of receptor activation, while adenine modifications tend to change the subtype selectivity but usually do not have a major effect on the agonist efficacy.…”

Metabolic mapping of Schisandra sphenanthera extract and its active lignans using a metabolomic approach based on ultra high performance liquid chromatography with high‐resolution mass spectrometry

“…Drug metabolism is significant in explaining and predicting efficacy and toxicity [11]. The metabolic profile of drugs and drug candidates can be clearly elucidated through combining in vitro incubation mixtures and in vivo animal models [12]. The pharmacokinetics of the six bioactive lignans in WZ were also measured and we found that most of…”

“…Furthermore, our previous studies reveal that WZ and its active lignans significantly protect against liver injury including acetaminophen-induced liver injury [7,8], partial hepatectomy [9] and lithocholic acid-induced cholestasis [10].Drug metabolism is significant in explaining and predicting efficacy and toxicity [11]. The metabolic profile of drugs and drug candidates can be clearly elucidated through combining in vitro incubation mixtures and in vivo animal models [12]. The pharmacokinetics of the six bioactive lignans in WZ were also measured and we found that most of…”

Historical and Current Adenosine Receptor Agonists in Preclinical and Clinical Development

“…MRS5980 37 is a highly selective C2-arylethynyl (N)-methanocarba A 3 AR agonist, which has been demonstrated to be highly efficacious in in vivo pain models following administered by oral gavage, with a protective effect lasting up to 3 h (Tosh et al, 2014; Janes et al, 2016), and its metabolomics has been studied (Fang et al, 2015). The 2-chlorothienyl group is stable in vivo , and the aryl alkyne group was shown to be not highly reactive.…”