A high-affinity, cis-on photoswitchable beta blocker to optically control β2-adrenergic receptors in vitro and in vivo

Shi, Shuang; Zheng, Yang; Goulding, Joëlle; Marri, Silvia; Lucarini, Laura; Konecny, Benjamin; Sgambellone, Silvia; Villano, Serafina; Bosma, Reggie; Wijtmans, Maikel; Briddon, Stephen J.; Zarzycka, Barbara A.; Vischer, Henry F.; Leurs, Rob

Abstract

This study introduces (S)-Opto-prop-2, a second-generation photoswitchable ligand designed for precise modulation of β -adrenoceptor (β AR). Synthesised by incorporating an azobenzene moiety with propranolol, (S)-Opto-prop-2 exhibited a high PSS (photostationary state for cis isomer) percentage (∼90 %) and a favourable half-life (>10 days), facilitating diverse bioassay measurements. In vitro, the cis-isomer displayed substantially higher β AR binding affinity than the trans-isomer (1000-fold), making (S)-Opto-prop-2 one of the best photoswitchable GPCR (G protein-coupled receptor) ligands reported so far. Molecular docking of (S)-Opto-prop-2 in the X-ray structure of propranolol-bound β AR followed by site-directed mutagenesis studies, identified D113 , N312 and F289 as crucial residues that contribute to ligand-receptor interactions at the molecular level. In vivo efficacy was assessed using a rabbit ocular hypertension model, revealing that the cis isomer mimicked propranolol's effects in reducing intraocular pressure, while the trans isomer was inactive. Dynamic optical modulation of β AR by (S)-Opto-prop-2 was demonstrated in two different cAMP bioassays and using live-cell confocal imaging, indicating reversible and dynamic control of β AR activity using the new photopharmacology tool. In conclusion, (S)-Opto-prop-2 emerges as a promising photoswitchable ligand for precise and reversible β AR modulation with light. The new tool shows superior cis-on binding affinity, one of the largest reported differences in affinity (1000-fold) between its two configurations, in vivo efficacy, and dynamic modulation. This study contributes valuable insights into the evolving field of photopharmacology, offering a potential avenue for targeted therapy in β AR-associated pathologies.

Citation

Shi, S., Zheng, Y., Goulding, J., Marri, S., Lucarini, L., Konecny, B., Sgambellone, S., Villano, S., Bosma, R., Wijtmans, M., Briddon, S. J., Zarzycka, B. A., Vischer, H. F., & Leurs, R. (2024). A high-affinity, cis-on photoswitchable beta blocker to optically control β2-adrenergic receptors in vitro and in vivo. Biochemical Pharmacology, 226, Article 116396. https://doi.org/10.1016/j.bcp.2024.116396

• Journal Article Type: Article
• Acceptance Date: Jun 25, 2024
• Online Publication Date: Jun 26, 2024
• Publication Date: 2024-08
• Deposit Date: Feb 17, 2025
• Publicly Available Date: Feb 18, 2025
• Journal: Biochemical Pharmacology
• Electronic ISSN: 1873-2968
• Publisher: Elsevier
• Peer Reviewed: Peer Reviewed
• Volume: 226
• Article Number: 116396
• DOI: https://doi.org/10.1016/j.bcp.2024.116396
• Keywords: Photoswitchable β-blockers, β(2)AR, Molecular modelling, GPCR signalling, Site-directed mutagenesis, Rabbit intraocular pressure
• Public URL: https://nottingham-repository.worktribe.com/output/37300477
• Publisher URL: https://www.sciencedirect.com/science/article/pii/S0006295224003794?via%3Dihub