Identifying GSK-3β kinase inhibitors of Alzheimer's disease: Virtual screening, enzyme, and cell assays

• Published in: European journal of pharmaceutical sciences 2016-06, Vol.89, p.11-19
• Main Authors: Lin, Chih-Hsin, Hsieh, Yu-Shao, Wu, Yih-Ru, Hsu, Chia-Jen, Chen, Hsuan-Chiang, Huang, Wun-Han, Chang, Kuo-Hsuan, Hsieh-Li, Hsiu Mei, Su, Ming-Tsan, Sun, Ying-Chieh, Lee, Guan-Chiun, Lee-Chen, Guey-Jen
• Format: Article
• Language: English
• Subjects: Alzheimer Disease - drug therapy; Alzheimer Disease - metabolism; Alzheimer's disease; Cell assay; Cell Line; Enzyme assay; Enzyme Inhibitors - pharmacology; Glycogen Synthase Kinase 3 beta - antagonists & inhibitors; GSK-3β kinase inhibitor; HEK293 Cells; Humans; Neurites - drug effects; Phosphorylation - drug effects; tau Proteins - metabolism; Virtual screening
• ISSN: 0928-0987; 1879-0720
• DOI: 10.1016/j.ejps.2016.04.012

Glycogen synthase kinase 3β (GSK-3β) is widely known as a critical target protein for treating Alzheimer's disease (AD). We utilized virtual screening to search databases for compounds with the potential to be used in drugs targeting GSK-3β kinase, and kinase as well as cell assays to investigate top-scored, selected compounds. Virtual screening of >1.1 million compounds in the ZINC and in-house databases was conducted using an optimized computational protocol in the docking program GOLD. Of the top-ranked compounds, 16 underwent a luminescent kinase assay and a cell assay using HEK293 cells expressing DsRed-tagged ΔK280 in the repeat domain of tau (tauRD). The compounds VB-003 (a potent GSK-3β inhibitor) and VB-008 (AM404, an anandamide transport inhibitor), with determined IC50 values of 0.25 and 5.4μM, respectively, were identified as reducing tau aggregation. Both compounds increased expression of phospho-GSK-3β (Ser9) and reduced endogenous tau phosphorylation at the sites of Ser202, Thr231, and Ser396. In the ∆K280 tauRD-DsRed SH-SY5Y cells, VB-008, but not VB-003, enhanced HSPB1 and GRP78 expression, increased ∆K280 tauRD-DsRed solubility, and promoted neurite outgrowth. Thus VB-008 performed best to the end of the present study. The identified compound VB-008 may guide the identification and synthesis of potential inhibitors analogous to this compound. [Display omitted]