Identification of Flavonoids-based HITS to Inhibit GSK3β Activity: An <i>In Silico</i> and <i>In Vitro</i> Analysis

Abstract

Background: Glycogen synthase kinase-3 (GSK-3) is a protein kinase with two isoforms, alpha and beta. GSK-3β, specifically, is found in nervous tissue and has been linked to the development of various human diseases. Aim: This study aimed to investigate the potential of flavonoids as inhibitors of GSK-3β using in silico and in vitro analyses. Methods: Schrodinger Maestro software was used for in silico molecular docking simulations to assess the interaction between flavonoids (ligands) and GSK-3β (target protein). In vitro cytotoxicity studies were performed on Rat L6 skeletal muscle cells to determine the safety of the selected flavonoids. Additionally, the neuroprotective effects of the flavonoids were evaluated using an oxygen-glucose deprivation model on SHSY 5Y neuroblastoma cells. Results: The in silico analysis revealed that all the studied flavonoids interacted with valine 135, a key amino acid for GSK-3β inhibition. Furthermore, the in vitro studies showed that the selected flavonoids were safe up to a concentration of 30 μM in rat L6 cells. Among the tested flavonoids, hesperetin, a flavanone, demonstrated the most potent neuroprotective effect in the SHSY 5Y cell line, with an IC50 value of 13.30 μM. Conclusion: Hesperetin, a flavanone, emerged as the most promising candidate among the tested flavonoids for further investigation as a potential GSK-3β inhibitor with neuroprotective properties.