Capsaicin from <i>Capsicum annuum</i> var. Cayenne: Isolation, GC-MS Analysis, Anticancer Activity on MG63 Osteosarcoma Cells, and <i>In Silico</i> Interaction with 6T2W

Background: Capsicum annuum (cayenne pepper), a member of the Solanaceae family, is known for its bioactive compounds, particularly capsaicin, which have demonstrated anticancer potential. This study aimed to explore the cytotoxic effects of cayenne pepper’s bioactive components on osteosarcoma cells and investigate the molecular mechanisms underlying their activity using advanced computational tools. Aim: This study’s main goal was to identify and isolate bioactive substances from C. annuum, evaluate their cytotoxic effects on MG-63 osteosarcoma cells, and explore the binding interactions between capsaicin and the 6T2W protein using molecular docking. Methods: Dried cayenne pepper fruit was powdered and subjected to ethanol extraction to isolate its bioactive components. Gas Chromatography-Mass Spectrometry (GC-MS) analysis identified seven compounds, including capsaicin, dihydrocapsaicin, linolelaidic acid, and hexadecanoic acid. The cytotoxicity of Capsaicin was evaluated using the MTT assay on MG-63 osteosarcoma cells. Molecular docking studies, conducted using ChemDraw Ultra 8.0, AutoDock Vina, and Pyrx Virtual Screening Tools, examined capsaicin’s interaction with the active site of the 6T2W protein. Results: The IC50 values determined from the MTT assay showed capsaicin to be the most potent compound with an IC50 of 50 μg/ml. Molecular docking revealed strong interactions between capsaicin and the 6T2W protein, suggesting a potential mechanism for its anticancer activity. Conclusion: Capsaicin and the mother extract from cayenne pepper demonstrated the highest cytotoxicity against MG-63 osteosarcoma cells, positioning them as potential candidates for osteosarcoma therapy. The molecular docking studies offer insights into the binding mechanisms of capsaicin with the 6T2W protein, paving the way for further investigation into its therapeutic applications. Major Findings: Capsaicin was successfully isolated from C. annuum var. Cayenne and confirmed through GC-MS analysis. It exhibited significant cytotoxic activity against MG63 osteosarcoma cells and showed strong binding affinity to the 6T2W protein target in molecular docking studies.