In-silico and In-vitro Evaluation of Acetylcholinesterase Activity of Methanolic Extract of Vitex negundo

Introduction: Vitex negundo has a myriad of medicinal uses, such as antioxidant, analgesic and anthelminthic properties, and it is used for dysmenorrhoea. The plant contains various phytochemicals, such as flavonoids, vitamins and casticin. All components of the plant are utilised medicinally due to the minimal occurrence of adverse drug reactions, making it a potential drug target for various chronic diseases. However, the role of this plant in neurodegenerative disorders, such as Alzheimer’s Disease (AD), has not yet been extensively studied. Aim: To evaluate the in-silico and in-vitro activity of the antiAlzheimer properties of the Methanolic Extract of Vitex negundo (MEVN) leaves. Materials and Methods: The present in-silico docking study and in-vitro study were conducted in the Department of Pharmacology at Sri Ramachandra Institute of Higher Education and Research, Porur, Chennai, Tamil Nadu, India, over a period of two months. The in-silico analysis was performed to determine the intermolecular interactions between the ligand and protein of the top five scored molecules using PyMol software. To assess the Acetylcholinesterase (AChE) inhibitory property, the findings were presented as docking scores. Cell viability and cytotoxicity were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay, with results expressed as percentages for various concentrations: 6.25 μg/mL, 12.5 μg/ mL, 25 μg/mL, 50 μg/mL, 75 μg/mL, and 100 μg/mL of MEVN. Results: The MEVN exhibited competitive inhibition of the AChE enzyme. There were active interactions between Agnuside, Isochlorogenic Acid B, Isochlorogenic Acid C, Kaempferol-3- O-rutinoside, and Quercetin found in the MEVN with AChE. The percentage of cell viability for the concentrations of 6.25 μg/ mL, 12.5 μg/mL, 25 μg/mL, 50 μg/mL, 75 μg/mL, and 100 μg/ mL, as determined by the MTT assay, was 98.62%, 98.86%, 97.19%, 96.66%, 91.86%, and 78.82%, respectively. The results indicated that MEVN does not exhibit any toxicity. Conclusion: The findings demonstrated that MEVN possesses AChE inhibitory properties and maintains cell viability, both of which are indicative of anti-Alzheimer activity. Therefore, MEVN can be further evaluated for its potential in preventing cell cytotoxicity and treating AD.