PDF(Pubmed)
Abstract：
The use of computer-aided methods have continued to propel accelerated drug discovery across various disease models, interestingly allowing the specific inhibition of pathogenic targets. Chloride Intracellular Channel Protein 4 (CLIC4) is a novel class of intracellular ion channel highly implicated in tumor and vascular biology. It regulates cell proliferation, apoptosis and angiogenesis; and is involved in multiple pathologic signaling pathways. Absence of specific inhibitors however impedes its advancement to translational research. Here, we integrate structural bioinformatics and experimental research approaches for the discovery and validation of small-molecule inhibitors of CLIC4. High-affinity allosteric binders were identified from a library of 1615 Food and Drug Administration (FDA)-approved drugs via a high-performance computing-powered blind-docking approach, resulting in the selection of amphotericin B and rapamycin. NMR assays confirmed the binding and conformational disruptive effects of both drugs while they also reversed stress-induced membrane translocation of CLIC4 and inhibited endothelial cell migration. Structural and dynamics simulation studies further revealed that the inhibitory mechanisms of these compounds were hinged on the allosteric modulation of the catalytic glutathione (GSH)-like site loop and the extended catalytic β loop which may elicit interference with the catalytic activities of CLIC4. Structure-based insights from this study provide the basis for the selective targeting of CLIC4 to treat the associated pathologies.
摘要：
计算机辅助方法的使用继续推动各种疾病模型加速药物发现，有趣的是，允许特异性抑制致病靶标。氯化物细胞内通道蛋白4（CLIC4）是一类与肿瘤和血管生物学密切相关的新型细胞内离子通道。它调节细胞增殖，细胞凋亡和血管生成；并参与多种病理信号通路。然而，缺乏特异性抑制剂阻碍了其向转化研究的发展。这里,我们整合了结构生物信息学和实验研究方法,以发现和验证CLIC4的小分子抑制剂.通过高性能计算驱动的盲对接方法，从1615个食品和药物管理局（FDA）批准的药物库中鉴定出高亲和力变构结合剂，导致选择两性霉素B和雷帕霉素。NMR测定证实了两种药物的结合和构象破坏作用，同时它们还逆转了应激诱导的CLIC4的膜易位并抑制了内皮细胞迁移。结构和动力学模拟研究进一步表明，这些化合物的抑制机制取决于催化谷胱甘肽（GSH）样位点环和延伸的催化β环的变构调节，这可能引起对CLIC4催化活性的干扰。来自本研究的基于结构的见解为CLIC4的选择性靶向治疗相关病理提供了基础。
