Liu, S.; Xiao, P.; Elgeti, M.; Fine, E. J.; Lucero, E. Y.; Vestergaard, M.; Wang, J.; Jyothidasan, A.; Li, A.; Qu, C.; Olsen, E.; Mazis, G.; Madsen, J. K.; Suomivuori, C.-M.; Kim, J.; Pakharukova, N.; Rahman, R.; Kereliuk, S. M.; Koch, W. J.; Strachan, R. T.; Staus, D. P.; Masoudi, A.; Hubbell, W. L.; Kahsai, A. W.; Dror, R. O.; Rockman, H. A.; Sun, J.; Ahn, S.; Lefkowitz, R. J.

Blockade of signaling through the angiotensin II type 1 receptor (AT1R), a prototypical G protein-coupled receptor (GPCR), by angiotensin receptor blockers (ARBs) is a major therapeutic approach to treating a wide variety of cardiovascular and renal diseases. Like most GPCRs, the AT1R signals through two transducers, G proteins and beta-arrestins. Previous reports have described beta-arrestin-biased peptide orthosteric agonists for the AT1R with potential therapeutic advantages over currently available unbiased ARBs. Here we report the DNA-encoded library screening-guided isolation and pharmacological characterization of the first small molecule AT1R allosteric ligands. We use cryo-electron microscopy, double electron-electron resonance spectroscopy, molecular dynamics simulations, and targeted mutagenesis to determine their binding sites, binding modes and conformational mechanisms driving their unique and divergent modulatory effects on G protein and beta-arrestin pathways. Our findings uncover new mechanisms for precisely controlling the dynamic behavior of the AT1R with implications for drug development targeting this pathophysiologically important receptor family.