Samuel Liu, Peng Xiao, Matthias Elgeti, Eve J. Fine, Emilio Y. Lucero, Mikkel Vestergaard, Junyan Wang, Arun Jyothidasan, Angus Li, Changxiu Qu, Eva Olsen, Georgios Mazis, Josephine K. Madsen, Carl-Mikael Suomivuori, Jihee Kim, Natalia Pakharukova, Rashad Rahman, Stephanie M. Kereliuk, Walter J. Koch, Ryan T. Strachan, Dean P. Staus, Ali Masoudi, Wayne L. Hubbell, Alem W. Kahsai, Ron O. Dror, Howard A. Rockman, Jin-Peng Sun, Seungkirl Ahn, Robert J. Lefkowitz

bioRxiv - Biochemistry

DOI: 10.64898/2026.07.15.736838

Abstract

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 diseases¹. Like most GPCRs, the AT1R signals through two transducers, G proteins and β-arrestins^2,3. Previous reports have described β-arrestin-biased peptide orthosteric agonists for the AT1R with potential therapeutic advantages over currently available unbiased ARBs^4–6. 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 β-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.