Effect of N-phenylbenzenesulfonamide targeting CNR2 over human bone marrow mesenchymal stem cell population and osteoblast/adipocyte differentiation

Abstract

Cannabinoids receptor type 2 (CNR2) signaling using selective receptor compounds encourages the expression of osteogenic genes and enhances mineralization. The effect of six newly synthesized Cannabinoid receptors type 2 (CB2) selective compounds over the viability of human bone marrow mesenchymal stem cells (h-BMMSCs) and their osteogenic and adipogenic potential study was carried out. Human mesenchymal stem cells (MSCs) were treated with 0.5 μM of compounds 1–6 to induce lineage differentiation (adipogenic, osteogenic). The cells were analyzed using differentiation assays, lipid droplets and mineralized matrix was stained with “Niel red and Alizarin Red, further performed reverse transcription-polymerase chain reaction (RT-PCR) for lineage-specific markers expression.” Treatment with synthesized compounds N-(4-(dimethyl amino) benzyl)− 4-methyl-N-phenylbenzenesulfonamide (1), N-(4-(dimethyl amino) benzyl)-N-phenyl benzamide (3) and N-(4-tert-butylbenzyl)− 4-methyl-N-phenylbenzenesulfonamide (5), increased both lipid droplets formation and Alkaline Phosphatase (ALP) activity. Treatment of N-(4-(dimethylamino) benzyl)-N-phenylbenzene sulfonamide (2), and 4-methyl-N-phenyl-N-(4-(trifluoromethyl) benzyl) benzene sulfonamide (4) was found to enhance the adipogenic differentiation of (h-BMSCs) and inhibit osteogenic differentiation. A Reduction in adipocyte differentiation upon exposure to N-(4-tert-butylbenzyl)-N-phenylbenzene sulfonamide (6) was noted. Conversely, enhanced mineral deposition was accompanied by increased alkaline phosphatase (ALP) activity in osteogenic differentiation. Taken together, these newly synthesized selective compounds work in favor of adipocyte formation except for 6 which reduces lipid droplets accumulation and enhanced calcium deposition. “In conclusion, compounds 1, 3, 5 and 6 demonstrated a positive effect over osteoblast formation compared to control.” As per molecular docking study, compounds 1, 2 and 3 are the most active derivative will serve as anti-osteoporosis drug discovery.