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.

该项目将使用由分子、结构和计算生物学家组成的多学科团队与药物化学家合作开发和评估具有抑制活性的化合物正链RNA病毒的黄病毒属。这些化合物将用于探测其功能病毒生命周期中的目标蛋白。该项目建立在现有合作的基础上，并扩展了普渡大学小组之前的观察结果。黄病毒包含 MAID 优先病原体成员在 A-C 所有三个类别中都可以找到。重点将主要集中在 A 类登革热病毒上病原体，因为其医学重要性以及我们所掌握的大量结构数据和试剂以及其他人在过去几年中编制的。我们将结合基础基础研究，以及涉及体外和体内化合物评估的翻译组件，以识别适合临床前抗病毒开发的小分子。用于识别此类的方法化合物还将深入了解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粒子中进展到病毒颗粒时的构象状态成熟状态，并评估化合物干扰这些转变和过程的能力病毒进入。我们将评估黄病毒颗粒的动态以及发生的结构转变成熟和进入过程中，以及小分子抑制剂与包膜蛋白的结合。两套抑制分子的开发基于预测与 1) E 蛋白正辛基-b-D 葡萄糖苷的结合 (BOG) 装订袋； 2) E 上参与蛋白质-蛋白质相互作用的位点。这些两套作为化合物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的化合物，确定其抑制机制，并评估其作用在动物模型中的功效。使用高通量黄病毒复制子筛选来鉴定小分子复制抑制剂。这些抑制剂的靶点将通过基因选择来确定，最初的命中将是通过连续几轮药物化学进行扩展，并且这些化合物将在体外进行评估使用细胞培养模型和体内使用小动物模型。