Vol. 58, Issue 6, 1461-1469, December 2000

Allosteric Inhibition of Endothelin ETA Receptors by 3,5-Dibromosalicylic Acid

Virginie Blandin, Paul Vigne, Jean Philippe Breittmayer, and Christian Frelin

Institut de Pharmacologie Moléculaire et Cellulaire du Centre National de la Recherche Scientifique (V.B., P.V., C.F.), Valbonne, France; and Institut National de la Santé et de la Recherche Médicale U343 (J.P.B.), Hôpital de l'Archet, BP 79, Nice, France

Derivatives of salicylic acid (SA) and benzoic acid prevent endothelin-1 (ET-1) binding to ETA receptors. This study analyzed actions of 30 derivatives of benzoic acid and salicylic acid on ¹²⁵I-ET-1 binding to recombinant rat ETA receptors. The most active compounds were 3,5-dibromosalicylic acid (Br2SA, K_i = 0.5 mM) and 3,5-diiodosalicylic acid (K_i = 0.3 mM). They were about 50 times more potent than SA and aspirin. Br2SA inhibited equilibrium ¹²⁵I-ET-1 binding in an apparently competitive manner. It accelerated 8-fold the dissociation of ¹²⁵I-ET-1 receptor complexes and did not modify the second order rate constant of association of ¹²⁵I-ET-1 to its receptors. Br2SA also decreased the affinity of ETA receptors for receptor antagonists BQ-123 and bosentan. Br2SA accelerated dissociation of ¹²⁵I-ET-1-solubilized ETA receptor complexes and decreased the apparent molecular size of solubilized receptors. Br2SA and 3,5-diiodosalicylic acid inhibited two cellular actions of ET-1: the mobilization of intracellular Ca²⁺ stores in isolated cells and contractions of rat aortic rings. They accelerated the relaxing action of BQ-123 and bosentan in ET-1-treated aortic rings. The results suggest the existence of an allosteric modifier site on ETA receptors that recognizes selected derivatives of SA. SA derivatives might be of therapeutic interest to relieve tight ET-1 binding and to favor actions of receptor antagonists.