ABSTRACT

Ligand-based pharmacophore modelling and virtual screening along with in vitro screening were performed as a rational strategy for the identification of novel compounds as apoptosis inducers and anticancer agents from the chemical database. Known apoptosis inducers were selected from the literature for generation of pharmacophore models, which were subjected to validation using Receiver operating characteristic (ROC) and Günere-Henry (GH) scoring methods. Based on highest fitness score of 4680.61, ROC value of 0.872 and GH score of 0.758, pharmacophore model-2 was selected as the best model. Model-2 as 3D search query was searched against the IBS database to find novel compounds as hits. Three hits were selected with a Q_FIT value more than 82 for in vitro screening as apoptosis inducers and anticancer agents. In vitro anticancer activity was performed using resazurin cell variability assay, and apoptosis inducing activity was determined using caspase-3 activation and annexin-FITC assays. One of the retrieved hit, STOCK5S-44056 demonstrated IC₅₀ value of 23.56 µM in cell variability assay, and had EC₅₀ value of 26.95 µM in caspase-3 activation assay. STOCK5S-44056 also indicated late stage induction of apoptosis in annexin assay. The results of in vitro activity revealed that STOCK5S-44056 has a potential to become anticancer agents.

摘要

基于配体的药效团建模和虚拟筛选以及体外筛选是从化学数据库中鉴定新化合物作为凋亡诱导剂和抗癌药的合理策略。从文献中选择已知的凋亡诱导剂以产生药效团模型，使用接收器操作特征（ROC）和Günere-Henry（GH）评分方法对其进行验证。根据最高健身得分4680.61，ROC值0.872和GH得分0.758，选择药效团模型2作为最佳模型。针对IBS数据库搜索Model-2作为3D搜索查询，以查找新化合物作为命中。Q _FIT选择了三首在体外筛选凋亡诱导剂和抗癌剂的价值超过82。使用刃天青细胞变异性测定法进行体外抗癌活性，并使用caspase-3激活和膜联蛋白-FITC测定法测定凋亡诱导活性。检索到的命中之一，STOCK5S-44056在细胞变异性测定中显示IC ₅₀值为23.56 µM，在caspase-3激活测定中显示EC ₅₀值为26.95 µM。STOCK5S-44056还表明在膜联蛋白测定中凋亡的晚期诱导。体外活性的结果表明，STOCK5S-44056有潜力成为抗癌药。