Identifying GSK-3β kinase inhibitors of Alzheimer's disease: Virtual screening, enzyme, and cell assays.
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.
Department of Neurology, Chang Gung Memorial Hospital, Chang Gung University, Taipei 10507, Taiwan.
Chih-Hsin
CH
Lin
Department of Chemistry, National Taiwan Normal University, 88, Ting Chow Road Section 4, Taipei 11677, Taiwan.
Yu-Shao
YS
Hsieh
Department of Neurology, Chang Gung Memorial Hospital, Chang Gung University, Taipei 10507, Taiwan.
Yih-Ru
YR
Wu
Department of Chemistry, National Taiwan Normal University, 88, Ting Chow Road Section 4, Taipei 11677, Taiwan.
Chia-Jen
CJ
Hsu
Department of Life Science, National Taiwan Normal University, 88, Ting Chow Road Section 4, Taipei 11677, Taiwan.
Hsuan-Chiang
HC
Chen
Department of Life Science, National Taiwan Normal University, 88, Ting Chow Road Section 4, Taipei 11677, Taiwan.
Wun-Han
WH
Huang
Department of Neurology, Chang Gung Memorial Hospital, Chang Gung University, Taipei 10507, Taiwan.
Kuo-Hsuan
KH
Chang
Department of Life Science, National Taiwan Normal University, 88, Ting Chow Road Section 4, Taipei 11677, Taiwan.
Hsiu Mei
HM
Hsieh-Li
Department of Life Science, National Taiwan Normal University, 88, Ting Chow Road Section 4, Taipei 11677, Taiwan.
Ming-Tsan
MT
Su
Department of Chemistry, National Taiwan Normal University, 88, Ting Chow Road Section 4, Taipei 11677, Taiwan. Electronic address: sun@ntnu.edu.tw.
Ying-Chieh
YC
Sun
Department of Life Science, National Taiwan Normal University, 88, Ting Chow Road Section 4, Taipei 11677, Taiwan. Electronic address: gclee@ntnu.edu.tw.
Guan-Chiun
GC
Lee
Department of Life Science, National Taiwan Normal University, 88, Ting Chow Road Section 4, Taipei 11677, Taiwan. Electronic address: t43019@ntnu.edu.tw.
Guey-Jen
GJ
Lee-Chen
D000544:Alzheimer Disease / Q000188:drug therapy* / Q000378:metabolism; D002460:Cell Line; D000091342:Endoplasmic Reticulum Chaperone BiP; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D000071679:Glycogen Synthase Kinase 3 beta / Q000037:antagonists & inhibitors*; D057809:HEK293 Cells; D006801:Humans; D016501:Neurites / Q000187:drug effects; D010766:Phosphorylation / Q000187:drug effects; D016875:tau Proteins / Q000378:metabolism
D000091342:Endoplasmic Reticulum Chaperone BiP; D004791:Enzyme Inhibitors; C000717247:HSPA5 protein, human; D016875:tau Proteins; D000071679:Glycogen Synthase Kinase 3 beta
Alzheimer's disease; Cell assay; Enzyme assay; GSK-3β kinase inhibitor; Virtual screening