Discovery and Structure–Activity Relationship Studies of Novel Adenosine A₁ Receptor-Selective Agonists

A series of benzyloxy and phenoxy derivatives of the adenosine receptor agonists N 6 -cyclopentyl adenosine (CPA) and N 6 -cyclopentyl 5-N-ethylcarboxamidoadenosine (CP-NECA) were synthesized, and their potency and selectivity were assessed. We observed that the most potent were the compounds with a halogen in the meta position on the aromatic ring of the benzyloxy-or phenoxycyclopentyl substituent. In general, the NECA-based compounds displayed greater A 1 R selectivity than the adenosine-based compounds, with N 6 -2-(3-bromobenzyloxy)cyclopentyl-NECA and N 6 -2-(3-methoxyphenoxy)cyclopentyl-NECA showing 1500fold improved A 1 R selectivity compared to NECA. In addition, we quantified the compounds' affinity and kinetics of binding at both human and rat A 1 R using a NanoBRET binding assay and found that the halogen substituent in the benzyloxy-or phenoxycyclopentyl moiety seems to confer high affinity for the A 1 R. Molecular modeling studies suggested a hydrophobic subpocket as contributing to the A 1 R selectivity displayed. We believe that the identified selective potent A 1 R agonists are valuable tool compounds for adenosine receptor research.