Site of action of beta-ligands at the human beta1-adrenoceptor.

Antagonist affinity measurements have traditionally been considered important in defining the receptor or receptor subtypes present within cells or tissues. Any change in this value has normally been taken as evidence for the presence of a second receptor. However, highly efficacious ligands induce a time and phosphorylation-dependent change in the beta2-adrenoceptor resulting in 10-fold lower affinity for antagonists. Also the beta1-adrenoceptor is now considered to exist in two different active conformations which are distinguished by their pharmacological properties. In this study, the site of action of a range of beta-agonists and beta-antagonists was determined using the human beta1-adrenoceptor stably expressed in Chinese hamster ovary cells with cyclic AMP response element reporter genes. Adrenaline and noradrenaline were confirmed as having agonist actions via the catecholamine site, whereas all antagonists had higher affinity for the catecholamine rather than secondary site. However, the rank order of affinity for the two sites was different suggesting that they are indeed separate entities. The measurements of antagonist affinity at the catecholamine site, however, were found to depend upon the agonist present. For example, xamoterol, cimaterol, terbutaline, and formoterol agonist responses were more readily antagonized by CGP 20712A[2-hydroxy-5-(2-[{hydroxy-3-(4-[1-methyl-4-trifluoromethyl-2-imidazolyl]phenoxy)propyl}amino]ethoxy)benzamide] than the catecholamine responses themselves. This, however, was not related to agonist efficacy as has previously been reported for the human beta2-adrenoceptor. Therefore, it may be that some agonists (e.g., cimaterol) purely activate the catecholamine site and others purely activate the secondary site (e.g., CGP 12177 [(-)-4-(3-tert-butylamino-2-hydroxypropoxy)-benzimidazol-2-one]), whereas the others (e.g., catecholamines) activate both sites to differing degrees.