Putative role of the adenosine A(3) receptor in the antiproliferative action of N(6)-(2-isopentenyl)adenosine

We tested a panel of naturally occurring nucleosides for their affinity towards adenosine receptors. Both N (6)-(2-isopentenyl)adenosine (IPA) and racemic zeatin riboside were shown to be selective human adenosine A(3) receptor (hA(3)R) ligands with affinities in the high nanomolar range (K (i) values of 159 and 649 nM, respectively). These values were comparable to the observed K (i) value of adenosine on hA(3)R, which was 847 nM in the same radioligand binding assay. IPA also bound with micromolar affinity to the rat A(3)R. In a functional assay in Chinese hamster ovary cells transfected with hA(3)R, IPA and zeatin riboside inhibited forskolin-induced cAMP formation at micromolar potencies. The effect of IPA could be blocked by the A(3)R antagonist VUF5574. Both IPA and reference A(3)R agonist 2-chloro-N (6)-(3-iodobenzyl)adenosine-5′-N-methylcarboxamide (Cl-IB-MECA) have known antitumor effects. We demonstrated strong and highly similar antiproliferative effects of IPA and Cl-IB-MECA on human and rat tumor cell lines LNCaP and N1S1. Importantly, the antiproliferative effect of low concentrations of IPA on LNCaP cells could be fully blocked by the selective A(3)R antagonist MRS1523. At higher concentrations, IPA appeared to inhibit cell growth by an A(3)R-independent mechanism, as was previously reported for other A(3)R agonists. We used HPLC to investigate the presence of endogenous IPA in rat muscle tissue, but we could not detect the compound. In conclusion, the antiproliferative effects of the naturally occurring nucleoside IPA are at least in part mediated by the A(3)R. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11302-011-9244-9) contains supplementary material, which is available to authorized users.