Products Cited in Publications Worldwide Synthesis of Novel Chloro-Benzo [d] imidazole Regioisomers as Selective CB2 Receptor Agonists: Indirect Functional Evaluation and Molecular Insights

Synthesis of Novel Chloro-Benzo [d] imidazole Regioisomers as Selective CB2 Receptor Agonists: Indirect Functional Evaluation and Molecular Insights

Salazar, Valeria Zuñiga; Ravanal, Renato Burgos; Soto-Flores, Jonathan; Sabadini, Gianfranco; González, José Vicente; Mella, Jaime; Romero-Parra, Javier

DOI:

Abstract

Background/Objectives: The cannabinoid type 2 receptor (CB2 receptor) has been extensively studied in recent years due to the benefits associated with its modulation, including the regulation of the inflammatory response, neuroimmunomodulatory properties, and antitumor effects, all with the advantage of lacking significant psychoactive effects. Herein, we report the design, synthesis, characterization, biological assays, and molecular modelling analyses of novel (5/6-chloro-2-aryl-1H-benzo [d]imidazol-1-yl)(4-methoxyphenyl)methanone and 5/6-chloro-1-(4-methoxybenzyl)-2-aryl-1H-benzo [d]imidazole regioisomers as potential cannabinoid type 2 receptor ligands. Methods: The compounds were evaluated for their presumed CB2 agonist activity using an indirect receptor-dependent apoptotic cell death assay exerted by cannabinoids, using the cell lines HEK293 (low CB1/CB2 expression), U-87 MG (high CB1 expression), and HL-60 (exclusive CB2 expression), and including the known cannabinoid ligands WIN-55,212-2 and AM630 as reference ligands. Flow cytometry was performed to assess apoptosis. Molecular docking and molecular dynamics simulations were used to explore ligand-receptor interactions at the CB2 active site. Results: Compounds 3a, 3b’, 3c, and 4b selectively reduced HL-60 cell viability, similar to WIN-55,212-2, while showing no toxicity toward HEK293 or U-87 MG cells. Flow cytometry indicated that compounds 3a and 3c induced apoptosis in HL-60 cells comparable to WIN-55,212-2. Computational studies suggested that both compounds bind within the CB2 receptor active site predominantly through π–π and hydrophobic interactions involving their benzo [d]imidazole cores, 2-aryl moieties, and 4-methoxybenzoyl scaffolds, resembling the binding patterns of established CB2 ligands. Conclusions: Compounds 3a and 3c exert selective cytotoxicity against HL-60 cells, likely via a CB2 agonist-mediated apoptotic mechanism. The applied combined experimental and computational approach provides a rapid, informative strategy for preliminary evaluation of CB2 ligands and guides subsequent detailed pharmacological studies.

Keywords

cannabinoids ; benzo [d]imidazole ; cytotoxic studies ; flow cytometry ; indirectagonism ; docking ; molecular dynamics

Purchased from AmBeed

Propidium Iodide

A79595025535-16-495%

Propidium Iodide

110.0/100mg

Detail