S-0139 (27-O-3-[2-(3-carboxy-acryloylamino)-5-hydroxyphenyl]-acryloylo xy myricerone, sodium salt) is a highly specific nonpeptide endothelin ET(A) receptor antagonist. The binding of [3H]S-0139 was compared to that of [125I]endothelin-1 to characterize the binding of the antagonist in porcine aortic smooth muscle membranes. Scatchard analysis revealed a single class of [3H]S-0139 binding sites with a Kd value of 0.61 +/- 0.10 nM and a Bmax of 0.72 +/- 0.16 pmol/mg protein. These sites were saturable and reversible. [125I]Endothelin-1 also showed binding with high affinity (Kd = 0.12 +/- 0.02 nM) to a homogeneous population of binding sites, whose Bmax (0.71 +/- 0.20 pmol/mg protein) was almost the same as that for [3H]S-0139. In both cases, the binding could be displaced by known endothelin receptor ligands and their IC50 values in each case showed a very close correlation (r = 0.986). The potency of seven endothelin receptor antagonists to displace [3H]S-0139 binding also correlated highly to the potency for inhibiting the endothelin-1-induced increase in cytosolic Ca2+ concentration (r = 0.949). Myriceric acid A showed a more potent functional activity than expected from its binding affinity, but this seemed to result from the different assay conditions, such as incubation time. Together, the results suggest that S-0139 labels only endothelin ET(A) receptor binding sites in porcine aortic smooth muscle.
As the first non-peptide endothelin receptor antagonist from a higher plant, a new triterpenoid, myriceric acid A (50-235) (1) was isolated from the bayberry, Myrica cerifera. Myriceric acid A (1) inhibited not only an endothelin-1-induced increase in cytosolic free Ca2+ concentration (IC50 = 11 +/- 2 nM) but [125I]endothelin-1 binding in rat aortic smooth muscle cells (Ki = 66 +/- 15 nM). Two new related triterpenoids, myriceric acid C (6), and myriceric acid D (8), were also isolated. Furthermore, the chemical modification of these natural products led to the synthesis of sulfated derivatives (13, 14, 15) which showed 1.5 to 20 times higher affinity for endothelin receptors. The structure activity relationships of myriceric acids and their derivatives are discussed.
1994-06-01·Journal of Hypertension2区 · 医学
Myricerone caffeoyl ester (50-235) is a non-peptide antagonist selective for human ETA receptors
2区 · 医学
作者: Maguire, Janet J. ; Bacon, Caragh R. ; Fujimoto, Masafumi ; Davenport, Anthony P.
To assess the pharmacological profile of a novel non-peptide endothelin antagonist (50-235) at endothelin receptors in human vascular smooth muscle preparations using radiolabelled binding techniques and in vitro pharmacological assays.
The antagonist was investigated for its ability to inhibit specific [125I]-endothelin-1 binding to ETA and ETB receptors using cryostat sections of media of human coronary artery. Antagonism by 50-235 (1-30 mumol/l) of endothelin-1-induced vasoconstriction in isolated preparations of human coronary artery, saphenous vein and left internal mammary artery was also determined.
In coronary artery 50-235 (10(-11) to 10(-4) mol/l) inhibited specifically bound [125I]-endothelin-1 (0.1 nmol/l) in a biphasic manner. The ratio of ETA:ETB receptor was 79:21. Increasing concentrations of 50-235 produced progressive rightwards displacements of the endothelin-1 dose-response curve in each of the three types of blood vessel. The dose-response curves were parallel and no attenuation of the maximum endothelin-1 response was observed suggesting that 50-235 was antagonizing endothelin-1 vasoconstriction in a competitive manner. The pA2 values determined by analysis of the Schild regression lines were 6.05 in coronary artery, 6.12 in saphenous vein and 6.18 in left internal mammary artery, and the slopes were not significantly different from unity.
The antagonist 50-235 exhibits nanomolar affinity for human ETA receptors and 500-fold selectivity for ETA compared with ETB receptors. Its novel non-peptide structure demonstrates that the carbon-nitrogen bond is not crucial for endothelin antagonist activity, and might provide important information for the development of therapeutic agents for conditions in which endothelins may be pathophysiologically relevant.