Detection of cannabinoid receptor type 2 in native cells and zebrafish with a highly potent, cell-permeable fluorescent probe

Gazzi, T.; Brennecke, B.; Atz, K.; Korn, C.; Sykes, D.; Forn-Cuni, G.; ... ; Nazaré, M. (2022)

Article / Letter to editor

Despite its essential role in the (patho)physiology of several diseases, CB₂R tissue expression profiles and signaling mechanisms are not yet fully understood. We report the development of a highly potent, fluorescent CB₂R agonist probe employing structure-based reverse design. It commences with a highly potent, preclinically validated ligand, which is conjugated to a silicon-rhodamine fluorophore, enabling cell permeability. The probe is the first to preserve interspecies affinity and selectivity for both mouse and human CB₂R. Extensive cross-validation (FACS, TR-FRET and confocal microscopy) set the stage for CB₂R detection in endogenously expressing living cells along with zebrafish larvae. Together, these findings will benefit clinical translatability of CB₂R based drugs.

• All authors: Gazzi, T.; Brennecke, B.; Atz, K.; Korn, C.; Sykes, D.; Forn-Cuni, G.; Pfaff, P.; Sarott, R.C.; Westphal, M.V.; Mostinski, Y.; Mach, L.; Wasinska-Kalwa, M.; Weise, M.; Hoare, B.L.; Miljuš, T.; Mexi, M.; Roth, N.; Koers, E.J.; Guba, W.; Alker, A.; Rufer, A.C.; Kusznir, E.A.; Huber, S.; Raposo, C.; Zirwes, E.A.; Osterwald, A.; Pavlovic, A.; Moes, S.; Beck, J.; Nettekoven, M.; Benito-Cuesta, I.; Grande, T.; Drawnel, F.; Widmer, G.; Holzer, D.; Wel, T. van der; Mandhair, H.; Honer, M.; Fingerle, J.; Scheffel, J.; Broichhagen, J.; Gawrisch, K.; Romero, J.; Hillard, C.J.; Varga, Z.V.; Stelt, M. van der; Pacher, P.; Gertsch, J.; Ullmer, C.; McCormick, P.J.; Oddi, S.; Spaink, H.P.; Maccarrone, M.; Veprintsev, D.B.; Carreira, E.M.; Grether, U.; Nazaré, M.
• Date: 2022-04-01
• Journal: Chemical Science
• Volume: 13
• Issue: 19
• Pages: 5539 - 5545
• DOI: doi:10.1039/d1sc06659e
• Link: https://pubs.rsc.org/en/content/articlelanding/2022/SC/D1SC06659E