Development of fluorinated and methoxylated benzothiazole derivatives as highly potent and selective cannabinoid CB2 receptor ligands

Abstract

The upregulation of the CB₂ receptors in neuroinflammation and cancer and their potential visualization with PET (positron emission tomography) could provide a valuable diagnostic and therapy-monitoring tool in such disorders. However, the availability of reliable CB₂-selective imaging probes is still lacking in clinical practice. We have recently identified a benzothiazole-2-ylidine amide hit (6a) as a highly potent CB₂ ligand. With the aim of enhancing its CB₂ over CB₁ selectivity and introducing structural sites suitable for radiolabeling, we herein describe the development of fluorinated and methoxylated benzothiazole derivatives endowed with extremely high CB₂ binding affinity and an exclusive selectivity to the CB₂ receptor. Compounds 14, 15, 18, 19, 21, 24 and 25 displayed subnanomolar CB₂K_i values (ranging from 0.16 nM to 0.68 nM) and interestingly, all of the synthesized compounds completely lacked affinity at the CB₁ receptor (K_i > 10,000 nM for all compounds), indicating their remarkably high CB₂ over CB₁ selectivity factors. The fluorinated analogs, 15 and 21, were evaluated for their in vitro metabolic stability in mouse and human liver microsomes (MLM and HLM). Both 15 and 21 displayed an exceptionally high stability (98% and 91% intact compounds, respectively) after 60 min incubation with MLM. Contrastingly, a 5- and 2.8-fold lower stability was demonstrated for compounds 15 and 21, respectively, upon incubation with HLM for 60 min. Taken together, our data present extremely potent and selective CB₂ ligands as credible leads that can be further exploited for ¹⁸F- or ¹¹C-radiolabeling and utilization as PET tracers.

Keywords: Benzothiazole; CB(2) ligands; Cannabinoid receptor type 2; Fluorine; Metabolic studies; PET.