TY - JOUR
T1 - Flipping the GPCR Switch
T2 - Structure-Based Development of Selective Cannabinoid Receptor 2 Inverse Agonists
AU - Kosar, Miroslav
AU - Sarott, Roman C.
AU - Sykes, David A.
AU - Viray, Alexander E.G.
AU - Vitale, Rosa Maria
AU - Tomašević, Nataša
AU - Li, Xiaoting
AU - Ganzoni, Rudolf L.Z.
AU - Kicin, Bilal
AU - Reichert, Lisa
AU - Patej, Kacper J.
AU - Gómez-Bouzó, Uxía
AU - Guba, Wolfgang
AU - McCormick, Peter J.
AU - Hua, Tian
AU - Gruber, Christian W.
AU - Veprintsev, Dmitry B.
AU - Frank, James A.
AU - Grether, Uwe
AU - Carreira, Erick M.
N1 - Publisher Copyright:
© 2024 The Authors. Published by American Chemical Society
PY - 2023
Y1 - 2023
N2 - We report a blueprint for the rational design of G protein coupled receptor (GPCR) ligands with a tailored functional response. The present study discloses the structure-based design of cannabinoid receptor type 2 (CB2R) selective inverse agonists (S)-1 and (R)-1, which were derived from privileged agonist HU-308 by introduction of a phenyl group at the gem-dimethylheptyl side chain. Epimer (R)-1 exhibits high affinity for CB2R with Kd = 39.1 nM and serves as a platform for the synthesis of a wide variety of probes. Notably, for the first time these fluorescent probes retain their inverse agonist functionality, high affinity, and selectivity for CB2R independent of linker and fluorophore substitution. Ligands (S)-1, (R)-1, and their derivatives act as inverse agonists in CB2R-mediated cAMP as well as G protein recruitment assays and do not trigger β-arrestin-receptor association. Furthermore, no receptor activation was detected in live cell ERK1/2 phosphorylation and Ca2+-release assays. Confocal fluorescence imaging experiments with (R)-7 (Alexa488) and (R)-9 (Alexa647) probes employing BV-2 microglial cells visualized CB2R expressed at endogenous levels. Finally, molecular dynamics simulations corroborate the initial docking data in which inverse agonists restrict movement of toggle switch Trp2586.48 and thereby stabilize CB2R in its inactive state.
AB - We report a blueprint for the rational design of G protein coupled receptor (GPCR) ligands with a tailored functional response. The present study discloses the structure-based design of cannabinoid receptor type 2 (CB2R) selective inverse agonists (S)-1 and (R)-1, which were derived from privileged agonist HU-308 by introduction of a phenyl group at the gem-dimethylheptyl side chain. Epimer (R)-1 exhibits high affinity for CB2R with Kd = 39.1 nM and serves as a platform for the synthesis of a wide variety of probes. Notably, for the first time these fluorescent probes retain their inverse agonist functionality, high affinity, and selectivity for CB2R independent of linker and fluorophore substitution. Ligands (S)-1, (R)-1, and their derivatives act as inverse agonists in CB2R-mediated cAMP as well as G protein recruitment assays and do not trigger β-arrestin-receptor association. Furthermore, no receptor activation was detected in live cell ERK1/2 phosphorylation and Ca2+-release assays. Confocal fluorescence imaging experiments with (R)-7 (Alexa488) and (R)-9 (Alexa647) probes employing BV-2 microglial cells visualized CB2R expressed at endogenous levels. Finally, molecular dynamics simulations corroborate the initial docking data in which inverse agonists restrict movement of toggle switch Trp2586.48 and thereby stabilize CB2R in its inactive state.
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U2 - 10.1021/acscentsci.3c01461
DO - 10.1021/acscentsci.3c01461
M3 - Article
AN - SCOPUS:85187570798
SN - 2374-7943
JO - ACS Central Science
JF - ACS Central Science
ER -