A₃ adenosine receptor agonists containing dopamine moieties for enhanced interspecies affinity

Following our study of 4'-truncated (N)-methanocarba-adenosine derivatives that displayed unusually high mouse (m) A₃AR affinity, we incorporated dopamine-related N⁶ substituents in the full agonist 5'-methylamide series. N⁶-(2-(4-Hydroxy-3-methoxy-phenyl)ethyl) derivative MRS7618 11 displayed K_i (nM) 0.563 at hA₃AR (∼20,000-fold selective) and 1.54 at mA₃AR. 2-Alkyl ethers maintained A₃ affinity, but with less selectivity than 2-alkynes. Parallel functional assays of G protein-dependent and β-arrestin 2 (βarr2)-dependent pathways indicate these are full agonists but not biased. Through use of computational modeling, we hypothesized that phenyl OH/OMe groups interact with polar residues, particularly Gln261, on the mA₃AR extracellular loops as the basis for the affinity enhancement. Although the pharmacokinetics indicated facile clearance of parent O-methyl catechol nucleosides 21 and 31, prolonged mA₃AR activation in vivo was observed in a hypothermia model, suggested potential formation of active metabolites through demethylation. Selected analogues induced mouse hypothermia following i.p. injection, indicative of peripheral A₃AR agonism in vivo.