2-and 8-alkynyladenosines:: conformational studies and docking to human adenosine A3 receptor can explain their different biological behavior

Adenosine (Ado) derivatives substituted at the, C2 position with an alkynyl chain are endowed with high affinity for A(1), A(2A) and A(3) human adenosine receptors, while being less active at the low affinity A(2B) subtype. On the other hand, the introduction of an alkynyl chain at the C8 position of adenosine is detrimental for the affinity and potency at A,, A(2A), and A(2B) receptors, while is more tolerated by the A3 receptor. The evaluation of the stimulation of [S-35]GTPgammaS binding revealed that 2-alkynyladenosines behave as adenosine receptors agonists while, on the contrary, 8-alkynyladenosines behave as antagonists. With this work we demonstrated, by means of an NMR-based and a computational conformational analysis, that 8-alkynyladenosines, differently from 2-alkynyladenosines, cannot adopt the sugar-base anti conformation required for adenosine receptor activation. Furthermore, using the recently reported X-ray crystal structure of bovine rhodopsin as template, we built a 3D model of the seven transmembrane domains of the human adenosine A(3) receptor with the homology modeling. After identification of the binding site we carried out docking experiments, demonstrating that the two class of molecules have different binding modes that explain their different degree of affinity and the shift of their activity from agonism to antagonism. (C) 2002 Elsevier Science Inc. All rights reserved.