Nucleotide P2Y1 receptor agonists are in vitro and in vivo prodrugs of A1/A3 adenosine receptor agonists: implications for roles of P2Y1 and A1/A3 receptors in physiology and pathology

Theodore E. Liston, Sonja Hinz, Christa E. Müller, Deborah M. Holstein, Jay Wendling, Roger J. Melton, Mary Campbell, William S. Korinek, R. Rama Suresh, Dane A. Sethre-Hofstad, Zhan Guo Gao, Dilip K. Tosh, Kenneth A. Jacobson, James D. Lechleiter

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Rapid phosphoester hydrolysis of endogenous purine and pyrimidine nucleotides has challenged the characterization of the role of P2 receptors in physiology and pathology. Nucleotide phosphoester stabilization has been pursued on a number of medicinal chemistry fronts. We investigated the in vitro and in vivo stability and pharmacokinetics of prototypical nucleotide P2Y1 receptor (P2Y1R) agonists and antagonists. These included the riboside nucleotide agonist 2-methylthio-ADP and antagonist MRS2179, as well as agonist MRS2365 and antagonist MRS2500 containing constrained (N)-methanocarba rings, which were previously reported to form nucleotides that are more slowly hydrolyzed at the α-phosphoester compared with the ribosides. In vitro incubations in mouse and human plasma and blood demonstrated the rapid hydrolysis of these compounds to nucleoside metabolites. This metabolism was inhibited by EDTA to chelate divalent cations required by ectonucleotidases for nucleotide hydrolysis. This rapid hydrolysis was confirmed in vivo in mouse pharmacokinetic studies that demonstrate that MRS2365 is a prodrug of the nucleoside metabolite AST-004 (MRS4322). Furthermore, we demonstrate that the nucleoside metabolites of MRS2365 and 2-methylthio-ADP are adenosine receptor (AR) agonists, notably at A3 and A1ARs. In vivo efficacy of MRS2365 in murine models of traumatic brain injury and stroke can be attributed to AR activation by its nucleoside metabolite AST-004, rather than P2Y1R activation. This research suggests the importance of reevaluation of previous in vitro and in vivo research of P2YRs and P2XRs as there is a potential that the pharmacology attributed to nucleotide agonists is due to AR activation by active nucleoside metabolites.

Original languageEnglish (US)
Pages (from-to)543-559
Number of pages17
JournalPurinergic Signalling
Volume16
Issue number4
DOIs
StatePublished - Dec 2020

Keywords

  • A receptor
  • A receptor
  • Adenosine
  • Ectonucleotidase
  • P2Y receptor
  • Prodrug

ASJC Scopus subject areas

  • Molecular Biology
  • Cellular and Molecular Neuroscience
  • Cell Biology

Fingerprint Dive into the research topics of 'Nucleotide P2Y<sub>1</sub> receptor agonists are in vitro and in vivo prodrugs of A<sub>1</sub>/A<sub>3</sub> adenosine receptor agonists: implications for roles of P2Y<sub>1</sub> and A<sub>1</sub>/A<sub>3</sub> receptors in physiology and pathology'. Together they form a unique fingerprint.

Cite this