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

Chih Hsin Lin, Yu Shao Hsieh, Yih Ru Wu, Chia Jen Hsu, Hsuan Chiang Chen, Wun Han Huang, Kuo Hsuan Chang, Hsiu Mei Hsieh-Li, Ming-Tsan Su, Ying-Chieh Sun, Guan-Chiun Lee, Guey-Jen Lee

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

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 5.4 μM, 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 ΔK280 tauRD-DsRed SH-SY5Y cells, VB-008, but not VB-003, enhanced HSPB1 and GRP78 expression, increased ΔK280 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.

Original languageEnglish
Pages (from-to)11-19
Number of pages9
JournalEuropean Journal of Pharmaceutical Sciences
Volume89
DOIs
Publication statusPublished - 2016 Jun 30

Fingerprint

Glycogen Synthase Kinase 3
Enzyme Assays
Alzheimer Disease
Phosphotransferases
Databases
Luminescent Measurements
HEK293 Cells
Drug Delivery Systems
Solubility
Inhibitory Concentration 50
Phosphorylation
fluorescent protein 583
Proteins

Keywords

  • Alzheimer's disease
  • Cell assay
  • Enzyme assay
  • GSK-3β kinase inhibitor
  • Virtual screening

ASJC Scopus subject areas

  • Pharmaceutical Science

Cite this

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

In: European Journal of Pharmaceutical Sciences, Vol. 89, 30.06.2016, p. 11-19.

Research output: Contribution to journalArticle

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, Guey-Jen. / Identifying GSK-3β kinase inhibitors of Alzheimer's disease : Virtual screening, enzyme, and cell assays. In: European Journal of Pharmaceutical Sciences. 2016 ; Vol. 89. pp. 11-19.
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AU - Hsu, Chia Jen

AU - Chen, Hsuan Chiang

AU - Huang, Wun Han

AU - Chang, Kuo Hsuan

AU - Hsieh-Li, Hsiu Mei

AU - Su, Ming-Tsan

AU - Sun, Ying-Chieh

AU - Lee, Guan-Chiun

AU - Lee, Guey-Jen

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AB - 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 5.4 μM, 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 ΔK280 tauRD-DsRed SH-SY5Y cells, VB-008, but not VB-003, enhanced HSPB1 and GRP78 expression, increased ΔK280 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.

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