Pharmacophore-Based Virtual Screening and Experimental Validation of Novel Inhibitors against Cyanobacterial Fructose-1, 6-/Sedoheptulose-1, 7-bisphosphatase
Pharmacophore-Based Virtual Screening and Experimental Validation of Novel Inhibitors against Cyanobacterial Fructose-1, 6-/Sedoheptulose-1, 7-bisphosphatase
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DOI：
10.1021/ci4007529
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发表时间：
2014-02
影响因子：
5.6
通讯作者：
Yao Sun;Rui Zhang;Ding Li;Lingling Feng;Di Wu;Lina Feng;Peipei Huang;Yanliang Ren;JiangTao Feng;San Xiao;Jian Wan
中科院分区：
文献类型：
--
作者：
Yao Sun;Rui Zhang;Ding Li;Lingling Feng;Di Wu;Lina Feng;Peipei Huang;Yanliang Ren;JiangTao Feng;San Xiao;Jian Wan
蓝藻果糖-1，6-/景天庚酮糖-1，7-二磷酸酶（cy-FBP/SBH 4）是筛选新型抑制剂的潜在靶标。在本研究中，我们靶向cy-FBP/SBB 1的底物结合口袋。通过基于药效团的虚拟筛选策略，从SPECs数据库中筛选出一系列新的热门化合物。大多数受试化合物对cy-FBP/SBF 2表现出中等抑制活性（IC 50 = 20.7-176.9 μM）。化合物2及其类似物10和11表现出较强的抑制活性，体外对cy-FBP/SBB 2的IC 50值分别为20.7、13.4和19.0 μM，体内对蓝藻集胞藻PCC 6803的EC 50值分别为12.3、10.9和2.9 ppm。选择化合物10以进行精细对接研究，以研究抑制剂与cy-FBP/SBR 2的合理结合模式。此外，可能的相互作用的残基与抑制剂进行了检查，通过定点诱变，酶测定，和荧光光谱分析。结果提供洞察抑制剂和底物结合口袋之间的结合模式。所观察到的理论和实验结果是一致的，表明所采用的建模策略和筛选方法是适当的，以寻找新的先导化合物，具有结构的多样性和高抑制活性对cy-FBP/SBB 12。
Cyanobacterial fructose-1,6-/sedoheptulose-1,7-bisphoshatase (cy-FBP/SBPase) is a potential enzymatic target for screening of novel inhibitors that can combat harmful algal blooms. In the present study, we targeted the substrate binding pocket of cy-FBP/SBPase. A series of novel hit compounds from the SPECs database were selected by using a pharmacophore-based virtual screening strategy. Most of the compounds tested exhibited moderate inhibitory activities (IC50 = 20.7-176.9 μM) against cy-FBP/SBPase. Compound 2 and its analogues 10 and 11 exhibited strong inhibitory activities, with IC50 values of 20.7, 13.4, and 19.0 μM against cy-FBP/SBPase in vitro and EC50 values of 12.3, 10.9, and 2.9 ppm against cyanobacteria Synechocystis PCC6803 in vivo, respectively. The compound 10 was selected in order to perform a refined docking study to investigate the rational binding mode of inhibitors with cy-FBP/SBPase. Furthermore, possible interactions of the residues with inhibitors were examined by site-directed mutagenesis, enzymatic assays, and fluorescence spectral analyses. The results provide insight into the binding mode between the inhibitors and the substrate binding pocket. The observed theoretical and experimental results are in concert, indicating that the modeling strategies and screening methods employed are appropriate to search for novel lead compounds having both structural diversity and high inhibitory activity against cy-FBP/SBPase.