Back to Search Start Over

Molecular-interaction and signaling profiles of AM3677, a novel covalent agonist selective for the cannabinoid 1 receptor.

Authors :
Janero DR
Yaddanapudi S
Zvonok N
Subramanian KV
Shukla VG
Stahl E
Zhou L
Hurst D
Wager-Miller J
Bohn LM
Reggio PH
Mackie K
Makriyannis A
Source :
ACS chemical neuroscience [ACS Chem Neurosci] 2015 Aug 19; Vol. 6 (8), pp. 1400-10. Date of Electronic Publication: 2015 May 29.
Publication Year :
2015

Abstract

The cannabinoid 1 receptor (CB1R) is one of the most abundant G protein-coupled receptors (GPCRs) in the central nervous system. CB1R involvement in multiple physiological processes, especially neurotransmitter release and synaptic function, has made this GPCR a prime drug discovery target, and pharmacological CB1R activation has been demonstrated to be a tenable therapeutic modality. Accordingly, the design and profiling of novel, drug-like CB1R modulators to inform the receptor's ligand-interaction landscape and molecular pharmacology constitute a prime contemporary research focus. For this purpose, we report utilization of AM3677, a designer endocannabinoid (anandamide) analogue derivatized with a reactive electrophilic isothiocyanate functionality, as a covalent, CB1R-selective chemical probe. The data demonstrate that reaction of AM3677 with a cysteine residue in transmembrane helix 6 of human CB1R (hCB1R), C6.47(355), is a key feature of AM3677's ligand-binding motif. Pharmacologically, AM3677 acts as a high-affinity, low-efficacy CB1R agonist that inhibits forskolin-stimulated cellular cAMP formation and stimulates CB1R coupling to G protein. AM3677 also induces CB1R endocytosis and irreversible receptor internalization. Computational docking suggests the importance of discrete hydrogen bonding and aromatic interactions as determinants of AM3677's topology within the ligand-binding pocket of active-state hCB1R. These results constitute the initial identification and characterization of a potent, high-affinity, hCB1R-selective covalent agonist with utility as a pharmacologically active, orthosteric-site probe for providing insight into structure-function correlates of ligand-induced CB1R activation and the molecular features of that activation by the native ligand, anandamide.

Details

Language :
English
ISSN :
1948-7193
Volume :
6
Issue :
8
Database :
MEDLINE
Journal :
ACS chemical neuroscience
Publication Type :
Academic Journal
Accession number :
25978068
Full Text :
https://doi.org/10.1021/acschemneuro.5b00090