Glycogen synthase kinase 3β (GSK-3β) is a widely investigated molecular target for numerous diseases including Alzheimer's disease, cancer, and diabetes mellitus. The present study was aimed to discover new scaffolds for GSK-3β inhibition, through protein structure-guided virtual screening approach. With the availability of large number of GSK-3β crystal structures with varying degree of RMSD in protein backbone and RMSF in side chain geometry, herein appropriate crystal structures were selected based on the characteristic ROC curve and percentage enrichment of actives. The validated docking protocol was employed to screen a library of 50,000 small molecules using molecular docking and binding affinity calculations. Based on the GLIDE docking score, Prime MMGB/SA binding affinity, and interaction pattern analysis, the top 50 ligands were selected for GSK-3β inhibition. (Z)-2-(3-chlorobenzylidene)-3,4-dihydro-N-(2-methoxyethyl)-3-oxo-2H-benzo[b][1,4]oxazine-6-carboxamide (F389-0663, 7) was identified as a potent inhibitor of GSK-3β with an IC50 value of 1.6 μm. Further, GSK-3β inhibition activity was then investigated in cell-based assay. The treatment of neuroblastoma N2a cells with 12.5 μm of F389-0663 resulted in the significant increase in GSK-3β Ser9 levels, which is indicative of the GSK-3β inhibitory activity of a compound. The molecular dynamic simulations were carried out to understand the interactions of F389-0663 with GSK-3β protein.

中文翻译：

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（Z）-2-（3-氯亚苄基）-3,4-二氢-N-（2-甲氧基乙基）-3-氧代-2H-苯并[b] [1,4]恶嗪-6-羧酰胺为GSK-3β抑制剂：通过虚拟筛选进行鉴定，并在基于酶和细胞的测定中进行验证。

糖原合酶激酶3β（GSK-3β）是许多疾病的分子靶标，包括阿尔茨海默氏病，癌症和糖尿病。本研究旨在通过蛋白质结构指导的虚拟筛选方法发现抑制GSK-3β的新支架。由于可获得大量的GSK-3β晶体结构，其中蛋白质骨架中的RMSD的程度不同，而侧链几何结构中的RMSF的程度不同，此处根据特征ROC曲线和活性物质的富集百分比选择合适的晶体结构。使用分子对接和结合亲和力计算，采用经过验证的对接方案筛选50,000个小分子的文库。根据GLIDE对接得分，Prime MMGB / SA绑定亲和力和交互模式分析，选择前50个配体用于GSK-3β抑制。（Z）-2-（3-氯亚苄基）-3,4-二氢-N-（2-甲氧基乙基）-3-氧代-2H-苯并[b] [1,4]恶嗪-6-羧酰胺（F389-0663 ，7）被鉴定为有效的GSK-3β抑制剂，IC50值为1.6μm。此外，然后在基于细胞的测定中研究了GSK-3β的抑制活性。用12.5μmF389-0663处理神经母细胞瘤N2a细胞导致GSK-3βSer9水平显着增加，这表明该化合物具有GSK-3β抑制活性。进行了分子动力学模拟，以了解F389-0663与GSK-3β蛋白的相互作用。6微米 此外，然后在基于细胞的测定中研究了GSK-3β抑制活性。用12.5μmF389-0663处理神经母细胞瘤N2a细胞导致GSK-3βSer9水平显着增加，这表明该化合物具有GSK-3β抑制活性。进行了分子动力学模拟，以了解F389-0663与GSK-3β蛋白的相互作用。6微米 此外，然后在基于细胞的测定中研究了GSK-3β抑制活性。用12.5μmF389-0663处理神经母细胞瘤N2a细胞导致GSK-3βSer9水平显着增加，这表明该化合物具有GSK-3β抑制活性。进行了分子动力学模拟，以了解F389-0663与GSK-3β蛋白的相互作用。