The specific binding sites for S-145, a novel thromboxane A2/prostaglandin H2 (TXA2/PGH2) receptor antagonist with weak partial agonistic activity, were studied in human platelet membranes. [3H]S-145 displayed high affinity and specificity, as well as saturable and displaceable binding, to a single class of recognition sites with the same maximum number of sites (2100 fmol/mg protein) as the other two TXA2/PGH2 receptor antagonists, [3H]SQ29,548 and [3H]ONO3708. Binding of S-145 to the platelet membranes was enhanced by divalent cations (Mg2+ and Ca2+), and the binding affinity in the presence of 20 mM MgCl2 was 0.75 nM, a value which was smaller than those of SQ29,548 (8.7 nM) and ONO3708 (3.7 nM). The rank order of potency (Ki) for a series of TXA2/PGH2 receptor antagonists to displace [3H]S-145 binding to the membranes was correlated with those determined from [3H]SQ29,548 or [3H]ONO3708 binding to the same preparations. Kinetic analysis for the binding of the above radiolabeled antagonist to the crude platelet membranes revealed that the dissociation rate constant (K-1) for S-145 was much smaller than that for other ligands in human, rat and rabbit platelets. The extremely slow dissociation of S-145 from the receptors may explain the long-lasting characteristic of this compound in vivo as well as the abolishment of partial agonistic activity.