Functional assay for assessment of agonistic or antagonistic activity of angiotensin AT2 receptor ligands reveals that EMA401 and PD123319 have agonistic properties

• DOI: 10.1016/j.bcp.2023.115793
• Publication Date: 2023-09-07T21:08:29Z
• Authors: A. Augusto Peluso; Igor M. Souza-Silva; Daniel C. Villela; Pernille B.L. Hansen; Anders Hallberg; Michael Bader; Robson Santos; Colin Sumners; U. Muscha Steckel...
• Keywords: 290; Angiotensin II; Endothelial Cells; Nitric oxide; Pharmacology and Toxicology; PD123319; Farmakologi och toxikologi; Receptor, Angiotensin, Type 2; Receptor, Angiotensin, Type 1; 3. Good health; Angiotensin AT 2-receptor; Cricetulus; HEK293 Cells; EMA401; Cardiovascular and Metabolic Diseases; Cricetinae; Integrative Biomedicine [Topic 3]; Animals; Humans; Angiotensin AT-receptor

With the discovery of protective arm renin-angiotensin system (RAS), interest has grown in RAS-related receptors such as angiotensin AT2-receptor [AT2R] potential new drug targets. While it is known that AT2R couple to Gi, also apparent they do not signal via inhibition adenylyl cyclase/decrease cAMP, many Gi-coupled receptors. Thus, standard commercially-available assays cannot be applied test for agonistic or antagonistic properties ligands. This lack hampered development drugs targeting AT2R. Therefore, we aimed at developing a reliable, technically easy assay determination intrinsic activity ligands, primarily distinguishing between agonists and antagonists. We found measurement NO release by DAF-FM fluorescence primary human aortic endothelial cells (HAEC) AT2R-transfected CHO reliable characterization testing assay, made several novel findings, including: a) C21 full agonist (with same efficacy II); b) no receptor Mas; c) HEK-293 are unresponsive stimulation; d) EMA401 PD123319, which commonly regarded antagonists, partial Collectively, have developed tested an based on quantification HAEC AT2R-CHO suitable characterisation established