Inhibition of flavivirus replication and assembly

抑制黄病毒复制和组装

基本信息

批准号：
7672018
负责人：
Richard J. Kuhn
金额：
$ 24.75万
依托单位：
UNIVERSITY OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-03-24 至 2010-02-28
项目状态：
已结题

项目摘要

This project will use a multidisciplinary team of molecular, structural, and computational biologists together with medicinal chemists to develop and evaluate compounds that have inhibitory activity against the flavivirus genus of plus strand RNA viruses. These compounds will be used to probe the function of their target proteins in the life cycle of the virus. This project builds on existing collaborations and extends previous observations by the Purdue group. The flaviviruses contain members of MAID Priority Pathogens that are found in all three categories A-C. The focus will be primarily on dengue virus, a category A pathogen, because of both its medical importance and the substantial structural data and reagents that we and others have compiled over the last several years. We will combine both foundational basic research, together with a translational component that involves in vitro and in vivo compound evaluation to identify small molecules suitable for pre-clinical antiviral development. The approaches used to identify such compounds will also yield insight into the assembly and replication of flaviviruses. We will determine the conformational states of the flavivirus structural proteins as they progress in the immature virion to the mature state, and evaluate the ability of compounds to interfere with these transitions and the process of viral entry. We will evaluate the dynamic states of the flavivirus particle, the structural transitions that occur during maturation and entry, and the binding of small molecule inhibitors to the envelope proteins. Two sets of inhibitory molecules have been developed based on predicted binding to 1) an E protein n-octyl-b-Dglucoside (BOG) binding pocket; and 2) sites on E that are involved in protein - protein interactions. These two sets serve as starting reagents in compound evaluation and further development. We will also identify compounds that inhibit flavivirus genome replication, identify their mechanism of inhibition, and evaluate their efficacy in an animal model. A high throughput flavivirus replicon screen was used to identify small molecule inhibitors of replication. Targets of these inhibitors will be identified by genetic selection, initial hits will be expanded by successive rounds of medicinal chemistry, and the compounds will be evaluated both in vitro using cell culture models and in vivo using small animal models.

该项目将使用分子，结构和计算生物学家的多学科团队一起用药物学家开发和评估具有抑制活性的化合物加链RNA病毒的黄病毒属。这些化合物将用于探测其功能在病毒生命周期中靶向蛋白质。该项目以现有协作为基础，并扩展普渡组的先前观察。黄病毒包含女仆优先病原体的成员在所有三个类别A-C中都可以找到。重点将主要放在登革热病毒上，一个类别病原体，由于其医学重要性以及我们的实质性结构数据和试剂在过去的几年中，其他人进行了编译。我们将结合两个基础基础研究，以及涉及体外和体内化合物评估的翻译成分适用于临床前抗病毒发育的小分子。用于识别这样的方法化合物还将洞悉黄病毒的组装和复制。我们将确定黄病毒结构蛋白在未成熟病毒中的构象状态成熟状态，并评估化合物干扰这些过渡的能力和病毒进入。我们将评估黄素粒子的动态状态，即发生的结构转变在成熟和进入过程中，小分子抑制剂与包膜蛋白的结合。两套已经基于预测的结合1）E蛋白N-辛基-B-葡萄糖苷开发了抑制性分子（沼泽）装订口袋； 2）E上与蛋白质相互作用有关的位点。这些两套是复合评估和进一步开发的起始试剂。我们还将确定抑制黄病毒基因组复制，确定其抑制机制的化合物并评估其动物模型的功效。使用高吞吐量黄病毒复制子屏幕来识别小分子复制抑制剂。这些抑制剂的靶标将通过遗传选择确定，初始命中将是通过连续的药物化学扩展，化合物将在体外评估使用细胞培养模型和使用小动物模型的体内。