In silico and in vitro study to identify neuroprotective compounds against factors implicated in Alzheimers disease

Abstract

To screen the neuroprotective compounds from the existing database based on molecular docking and ADME studies. The drug targets selected in the present study are cholinesterase enzyme (AChE and BuChE) and BACE 1 enzyme. Further in vitro screening the shortlisted compounds for their anti-cholinesterase, antiBACE 1, anti-amyloidogenic, anti-MAO B, antioxidant activities, protective effect against Aand#946; induced toxicity and H2O2 induced oxidative stress in PC12 cell lines. Methodology: The chemical database, Pubchem was screened against various drug targets AChE (4EY6), BuChE (1P0M) and BACE 1 (4D8C) of AD using the Glide module of Schrodinger suite. All the shortlisted compounds were then subjected to ADME studies using the QikProp module of Schrodinger suite. Shortlisted compounds were then checked for their anti-cholinesterase, anti-BACE 1, antiamyloidogenic, anti-MAO B, antioxidant activities, protective effect against Aand#946; induced toxicity and H2O2 induced oxidative stress in PC12 cell lines. The in vitro assays used in this study were Ellman s assay for anticholinesterase activity and for the mechanism of action, kinetics was performed which was analysed using Lineweaver-Burk (LB) plot method. Molecular dynamics of the lead compounds was also performed using Desmond module of Schrodinger suite. FRET assay kits were used for both BACE 1 and MAO B inhibition. Thioflavin T (ThT) assay, transmission electron microscopy (TEM) studies, circular dichroism (CD) spectroscopy was used for the anti-amyloidogenic activity. The protective activity of the shortlisted test compounds was examined using MTT assay against Aand#946; induced toxicity in the PC12 cell line. Antioxidant capacities of the shortlisted test compounds were determined using TEAC and FRAP assays. The protective activity of the shortlisted test compounds was assessed using MTT and nitric oxide assay against H2O2 induced oxidative stress in PC12 cell lines