Identification of the First Synthetic Allosteric Modulator of the CB₂ Receptors and Evidence of Its Efficacy for Neuropathic Pain Relief

The direct activation of cannabinoid receptors (CBRs) results in several beneficial effects; therefore several CBRs ligands have been synthesized and tested in vitro and in vivo. However, none of them reached an advanced phase of clinical development due mainly to side effects on the CNS. Medicinal chemistry approaches are now engaged to develop allosteric modulators that might offer a novel therapeutic approach to achieve potential therapeutic benefits avoiding inherent side effects of orthosteric ligands. Here we identify the first ever synthesized positive allosteric modulator (PAM) that targets CB₂Rs. The evidence for this was obtained using [³H]­CP55940 and [³⁵S]­GTPγS binding assays. This finding will be useful for the characterization of allosteric binding site(s) on CB₂Rs which will be essential for the further development of CB₂R allosteric modulators. Moreover, the new CB₂R PAM displayed antinociceptive activity in vivo in an experimental mouse model of neuropathic pain, raising the possibility that it might be a good candidate for clinical development

Identification of the First Synthetic Allosteric Modulator of the CB₂ Receptors and Evidence of Its Efficacy for Neuropathic Pain Relief

The direct activation of cannabinoid receptors (CBRs) results in several beneficial effects; therefore several CBRs ligands have been synthesized and tested in vitro and in vivo. However, none of them reached an advanced phase of clinical development due mainly to side effects on the CNS. Medicinal chemistry approaches are now engaged to develop allosteric modulators that might offer a novel therapeutic approach to achieve potential therapeutic benefits avoiding inherent side effects of orthosteric ligands. Here we identify the first ever synthesized positive allosteric modulator (PAM) that targets CB₂Rs. The evidence for this was obtained using [³H]­CP55940 and [³⁵S]­GTPγS binding assays. This finding will be useful for the characterization of allosteric binding site(s) on CB₂Rs which will be essential for the further development of CB₂R allosteric modulators. Moreover, the new CB₂R PAM displayed antinociceptive activity in vivo in an experimental mouse model of neuropathic pain, raising the possibility that it might be a good candidate for clinical development

Enantiospecific Allosteric Modulation of Cannabinoid 1 Receptor

The cannabinoid 1 receptor (CB1R) is one of the most widely expressed metabotropic G protein-coupled receptors in brain, and its participation in various (patho)­physiological processes has made CB1R activation a viable therapeutic modality. Adverse psychotropic effects limit the clinical utility of CB1R orthosteric agonists and have promoted the search for CB1R positive allosteric modulators (PAMs) with the promise of improved drug-like pharmacology and enhanced safety over typical CB1R agonists. In this study, we describe the synthesis and <i>in vitro</i> and <i>ex vivo</i> pharmacology of the novel allosteric CB1R modulator GAT211 (racemic) and its resolved enantiomers, GAT228 (<i>R</i>) and GAT229 (<i>S</i>). GAT211 engages CB1R allosteric site(s), enhances the binding of the orthosteric full agonist [³H]­CP55,490, and reduces the binding of the orthosteric antagonist/inverse agonist [³H]­SR141716A. GAT211 displayed both PAM and agonist activity in HEK293A and Neuro2a cells expressing human recombinant CB1R (hCB1R) and in mouse-brain membranes rich in native CB1R. GAT211 also exhibited a strong PAM effect in isolated vas deferens endogenously expressing CB1R. Each resolved and crystallized GAT211 enantiomer showed a markedly distinctive pharmacology as a CB1R allosteric modulator. In all biological systems examined, GAT211’s allosteric agonist activity resided with the <i>R-</i>(+)-enantiomer (GAT228), whereas its PAM activity resided with the <i>S</i>-(−)-enantiomer (GAT229), which lacked intrinsic activity. These results constitute the first demonstration of enantiomer-selective CB1R positive allosteric modulation and set a precedent whereby enantiomeric resolution can decisively define the molecular pharmacology of a CB1R allosteric ligand