Inverse agonism and neutral antagonism at a constitutively active alpha‐1a adrenoceptor

We have studied the antagonist action of prazosin and KMD‐3213 in a constitutively active mutant of the human alpha‐1a adrenoceptor in which Ala 271 was substituted to Thr and was expressed in CHO cells. Inverse agonism was characterized by up‐regulation of receptor density, a decrease in basal GTPγS binding, and a reduction in basal inositol‐1,4,5‐trisphosphate (IP 3 ) level. According to the above criteria, prazosin acted as an inverse agonist, whilst KMD‐3213 behaved as a neutral antagonist. Compared with the wild‐type receptor, mutant receptor exhibited single affinity sites for [ 3 H]‐prazosin, [ 3 H]‐KMD and the non‐radioactive ligands tested, and displayed significantly higher affinities for several agonists but not for the two antagonists. Administration of KMD‐3213 to prazosin‐treated CHO cells expressing the mutant receptor reversed the inverse agonism of prazosin resulting in rapid increases in cellular IP 3 , in intracellular [Ca 2+ ] and in the rate of extracellular acidification. These results indicated that a neutral antagonist can reverse the action of an inverse agonist at the receptor site. The distinct properties of inverse agonist and neutral antagonist in affecting receptor function may be important for the clinical use of such antagonists.British Journal of Pharmacology (2000) 131 , 546–552; doi: 10.1038/sj.bjp.0703584