Inhibition of flavivirus replication and assembly

这个项目将使用一个由分子、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作用的化合物 黄病毒正链RNA病毒属。这些化合物将被用来探索它们的功能 病毒生命周期中的靶蛋白。该项目建立在现有协作的基础上，并扩展 普渡集团之前的观察结果。黄病毒含有MAID优先病原体成员 在所有三个类别A-C中都有。重点将主要放在A类登革热病毒上 病原体，因为它的医学重要性和大量的结构数据和试剂，我们 而其他人则是在过去几年里汇编的。我们将把基础基础研究和 与涉及体外和体内化合物评估的翻译成分一起鉴定 适合临床前抗病毒开发的小分子药物。用来识别这类 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向 成熟状态，并评估化合物干扰这些转变和过程的能力 病毒入侵。我们将评估黄病毒颗粒的动态，即发生的结构转变 在成熟和进入的过程中，以及小分子抑制剂与包膜蛋白的结合。两套 基于预测与1)E蛋白N-辛基-b-D葡萄糖苷的结合，已经开发出许多抑制分子 (BOG)结合口袋；以及2)E上参与蛋白质-蛋白质相互作用的位点。这些 两套作为化合物评价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 在动物模型中的疗效。高通量黄病毒复制子筛选用于鉴定小分子 复制的抑制因子。这些抑制剂的靶点将通过基因选择来确定，最初的打击将是 通过连续几轮的药物化学膨胀，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。

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.