Defining the affinity and receptor sub-type selectivity of four classes of endothelin antagonists in clinically relevant human cardiovascular tissues

AIMS: We have compared the endothelin receptor subtype affinity (K(D)) and selectivity of four structural classes of antagonists (peptide, sulphonamide-based, carboxylic acid-based, myceric acid-based) in human cardiovascular tissues to determine whether these are predicted by values reported for human cloned receptors. Additionally, affinities (K(B)) for these antagonists, determined in ET-1-mediated vasoconstriction assays in human blood vessels, were used to identify discrepancies between K(B) and K(D) determined in the same tissues. MAIN METHODS: Competition binding experiments were carried out in sections of human left ventricle, coronary artery and homogenates of saphenous vein to determine K(D) values for structurally different ET(A)-selective (FR139317, BMS 182874, S97-139, sitaxentan, ambrisentan) and mixed (PD142893, Ro462005, bosentan, L-749329, SB209670) antagonists. Schild-derived values of antagonist affinity were obtained in vascular functional studies. KEY FINDINGS: When compared with previously reported data in human cloned endothelin receptors, those antagonists reported to be ET(A)-selective exhibited even greater ET(A) selectivity in human ventricle (BMS 182874, sitaxentan, ambrisentan) that expressed both receptor subtypes. Those antagonists reported to have <100 fold selectivity in cloned receptors (PD142893, Ro-462005, bosentan, SB209670, L-749329) did not distinguish between receptor subtypes in human left ventricle. For antagonists where we determined affinity in vascular functional and binding assays (Ro462005, bosentan, BMS 182874, L-749329, SB209670) there was no correlation between the degree of discrepancy in K(B) and K(D) and structural class. SIGNIFICANCE: For an antagonist to retain ET(A)-selectivity in vivo it may be necessary to identify those compounds that have at least 1000 fold ET(A):ET(B) selectivity in in vitro assays.