Development of CoV inhibitors against non-enzymatic targets

总结 使用病毒酶作为抗病毒药物开发的靶标已经导致许多抗病毒疗法用于治疗癌症。 诊所已经开发了几种用于HIV、HCV和HSV的病毒聚合酶和/或蛋白酶抑制剂，其中 他人不太常见的是针对具有未知酶特性的病毒产物的抗病毒药物的开发， 尽管一些例外包括用于治疗HCV感染的NS 5 b抑制剂。许多病毒 产品缺乏可用于开发抑制剂的体外筛选测定的明确的酶活性， 病毒酶作为治疗干预的靶点的局限性限制了可用的病毒靶点的数量。在项目4中- 开发针对非酶靶点的CoV抑制剂在这个抗病毒开发财团中，我们 探索具有未知酶活性且未被其他SARS-CoV-2涵盖的SARS-CoV-2靶标 这个提案的项目。我们选择了毒力所需的病毒蛋白Nsp 1以及 病毒RNA。病毒Nsp 1和病毒RNA都具有特定的独特序列和结构， 将它们与宿主分子区分开来。在aim 1中，我们将生物化学，遗传学，结构学和计算机技术相结合， 鉴定防止Nsp 1对宿主应答的抑制的治疗性分子的方法 被感染的细胞在aim 2中，我们使用先进的反义策略来靶向特定的保守功能序列 参与复制、转录和/或翻译的病毒RNA。在aim 3中，我们计划识别分子 其结合并阻止引导病毒RNA复制和转录的保守RNA结构的功能。 为此，我们组建了一个研究团队，他们在使用基于RNA的 治疗、病毒学、细胞生物学、RNA生物学和宿主-病毒相互作用。该团队与 该联盟的核心受益于结构、ADME和PK专业知识以及抗病毒检测， 动物模型和化学库。虽然项目主要是探索性的， 阶段，已经鉴定了靶向RNA序列（Aim 2）或RNA结构（Aim 3）的几种命中 并将在我们联盟的抗病毒产品线上继续推进。对于其他人，如Nsp 1， 生物化学和细胞生物学可用的信息，其中许多信息是由 我们的团队，这证明了Nsp 1抑制剂的筛选。这类抑制剂的进展 非常规的病毒靶点将为开发针对其他病毒的新疗法打开大门， 具有未知酶功能的病毒产品，扩大了我们治疗病毒性疾病的可制药空间 感染.

SUMMARY The use of viral enzymes as targets for antiviral development has led to many antiviral therapies of use in the clinic. Several viral polymerase and/or protease inhibitors have been developed for HIV, HCV and HSV, among others. Less common is the development of antivirals against viral products with unknown enzymatic properties, although some exceptions include the NS5b inhibitors used for the treatment of HCV infections. As many viral products lack clear enzymatic activities that can be used to develop in vitro screen assays for inhibitors, the use of viral enzymes as targets for therapeutic intervention limits the number of available viral targets. In project 4 – Development of CoV Inhibitors Against Non-Enzymatic Targets of this antiviral development consortium, we explore SARS-CoV-2 targets with unknown enzymatic activities and not covered by the additional SARS-CoV-2 projects of this proposal. We have chosen as targets both a viral protein required for virulence, Nsp1, as well as the viral RNA. Both the viral Nsp1 and the viral RNA have specific unique sequences and structures that distinguish them from host molecules. In aim 1, we combine biochemical, genetic, structural and in silico approaches to identify therapeutic molecules that prevent the inhibition of host responses exerted by Nsp1 in infected cells. In aim 2 we use advanced antisense strategies to target specific conserved functional sequences of the viral RNA involved in replication, transcription and/or translation. In aim 3, we plan to identify molecules that bind and prevent the function of conserved RNA structures that guide viral RNA replication and transcription. For this purpose, we have assembled a team of investigators with ample expertise in the use of RNA-based therapies, virology, cell biology, RNA biology and host-virus interactions. This team interacts closely with the Cores of this consortium to benefit of the structural, ADME and PK expertise, as well as of the antiviral assays, animal models and chemical libraries provided by these Cores. While the Project is mainly in an exploratory phase, several hits that target RNA sequences (Aim 2) or RNA structures (Aim 3), have already been identified and will move forward in the antiviral pipeline of our consortium. For others, like Nsp1, there is enough biochemical and cell biology available information, with many of this information generated by the members of our team, that justifies the proposed screen for Nsp1 inhibitors. Advancement of such classes of inhibitors against unconventional viral targets will open the door for the development of new therapies against other viruses and viral products with unknown enzymatic function, expanding our druggable space for the treatment of viral infections.