Norovirus Capsid: a Novel Drug Target

描述(申请人提供)：诺如病毒(NV)是急性胃肠炎非细菌性流行的最重要原因，它是全世界发病率和死亡率的常见原因。在美国，NV相关胃肠炎的暴发估计每年影响2300万人，对公共健康构成重大威胁。为了开发治疗NV感染的新的治疗方法，这项建议旨在识别能够有效破坏NV衣壳组装和/或破坏NV衣壳组装的小分子。靶向NV衣壳蛋白是一种新的策略，近年来已成功应用于其他病毒。建议的方法源于我们之前的合作努力，并建立在NV研究和蛋白质及其相互作用的计算研究方面的专业知识。我们提出了一种集成策略，它独特地结合了：i)一个新的计算框架，用于对NV衣壳蛋白中的热点进行建模和排序；ii)虚拟筛选以这些位置为靶点的小分子；iii)使用新建立的功能和结构分析以及不同的模型系统对Top Hit进行实验验证。绘制功能和结构热点图将利用关于NV的丰富的序列和结构数据。通过将进化、结构和功能信息与与衣壳蛋白齐聚相关的构象变化和变构效应的建模相结合，将进一步提高对预测热点的排名和评估。这些已确定的热点将通过突变、体外结合和体内功能分析进行验证和进一步表征。将使用最先进的对接模拟、化学信息学方法和类药物化合物的综合库进行虚拟筛选，以确定针对排名靠前的位置的候选分子。TOP HITS的实验验证将使用我们最近开发的用于评估NV衣壳组装的体外结合和体内功能分析来进行。为了方便这个和未来对其他病毒的应用，一个用于自动分析内部结构和功能热点的通用界面 衣壳蛋白将被整合到我们的PolyView-MM门户中。

DESCRIPTION (provided by applicant): Noroviruses (NVs) are the most important cause of non-bacterial epidemics of acute gastroenteritis, which is a common cause of morbidity and mortality worldwide. In US, outbreaks of NV-related gastroenteritis are estimated to affect 23 million people annually, representing a significant threat to public health. With the goal of developing novel therapeutic approaches for the treatment of NV infection, this proposal aims to identify small molecules that can effectively destabilize and/or disrupt the assembly of the NV capsid. Targeting NV capsid represents a novel strategy that has been successfully applied to other viruses in recent years. The proposed approach stems from our previous collaborative efforts and builds on expertise in both NV research and computational studies of proteins and their interactions. We propose an integrated strategy that uniquely combines: i) a novel computational framework for modeling and ranking of hot spots within NV capsid proteins; ii) virtual screening for small molecules targeting such sites; iii) experimental validation of top hit using newly established functional and structural assays and different model systems. Mapping of functional and structural hot spots will utilize the wealth of sequence and structural data on NVs. Ranking and assessment of the predicted hot spots will be further enhanced by integrating evolutionary, structural and functional information with modeling of conformational changes and allosteric effects associated with oligomerization of capsid proteins. Validation and further characterization of such identified hot spots will be performed by using mutagenesis, in vitro binding and in vivo functional assays. Virtual screening using state-of-art docking simulations, cheminformatic methods and comprehensive libraries of drug-like compounds will be used to identify candidate molecules targeting top ranking sites. Experimental validation of top hits will be performed using in vitro binding and in vivo functional assays developed by us recently for the assessment of NV capsid assembly. In order to facilitate this and future applications to other viruses, a versatile interface for automated analysis of structural and functional hot spots within capsid proteins will be integrated into our Polyview-MM portal.