Design and in Vivo Characterization of A₁ Adenosine Receptor Agonists in the Native Ribose and Conformationally Constrained (N)-Methanocarba Series

Abstract

(N)-Methanocarba ([3.1.0]bicyclohexyl) adenosines and corresponding ribosides were synthesized to identify novel A₁ adenosine receptor (A₁AR) agonists for CNS or peripheral applications. Human and mouse AR binding was determined to assess the constrained ring system's A₁AR compatibility. N⁶-Dicyclobutylmethyl ribose agonist (9, MRS7469, >2000-fold selective for A₁AR) and known truncated N⁶-dicyclopropylmethyl methanocarba 7 (MRS5474) were drug-like. The pure diastereoisomer of known riboside 4 displayed high hA₁AR selectivity. Methanocarba modification reduced A₁AR selectivity of N⁶-dicyclopropylmethyl and endo-norbornyladenosines but increased ribavirin selectivity. Most analogues tested (ip) were inactive or weak in inducing mouse hypothermia, despite mA₁AR full agonism and variable mA₃AR efficacy, but strong hypothermia by 9 depended on A₁AR, which reflects CNS activity (determined using A₁AR or A₃AR null mice). Conserved hA₁AR interactions were preserved in modeling of 9 and methanocarba equivalent 24 (∼400-fold A₁AR-selective). Thus, we identified, and characterized in vivo, ribose and methanocarba nucleosides, including with A₁AR-enhancing N⁶-dicyclobutylmethyl-adenine and 1,2,4-triazole-3-carboxamide (40, MRS7451) nucleobases.