On the high affinity of 8-cyclohexylcaffeine for the presynaptic inhibitory adenosine receptor present in rat motor nerve terminals.

Rat neuromuscular junction was used to study the characteristics of presynaptic A1 adenosine receptors. We investigated the ability of the 8-substituted caffeine, 8-cyclohexylcaffeine (CHC), as well as of 1,3,8-substituted xanthines, 1,3-dipropyl-8-p-sulfophenylxanthine (DPSPX) and 8-p-sulfophenyl-1-isoamyl-3-isobutylxanthine (SPIIBX) to antagonize the inhibitory effect of 2-chloroadenosine on the amplitude of nerve-evoked twitches of the rat phrenic-hemidiaphragm, and we compared the affinity of these xanthines with that of 1,3-dipropyl-8-cyclopenthylxanthine (DPCPX). CHC, DPSPX and SPIIBX in a near parallel manner shifted to the right the log concentration-response curve for the inhibitory effect of 2-chloroadenosine on nerve-evoked twitch amplitude. Linear Schild plots with slopes near to unity were obtained for all these xanthines. The order of potency of the xanthines was DPCPX (Ki = 0.53 nM) > DPSPX (38 nM) = CHC (41 nM) > SPIIBX (404 nM). The affinities of DPSPX and SPIIBX for the A1 receptor at the rat neuromuscular junction are in agreement with the affinities described for A1 receptors at brain membranes. The now reported affinity of CHC for the presynaptic A1 receptor is 683 times higher than that obtained in binding studies in rat brain membranes, and is only 49 times higher than that obtained in functional assays (adenylate cyclase activity) in non-neuronal preparations (rat fat cells).