Mechanistic understanding and inhibition of Zika NS5 protein

Zika NS5 蛋白的机制理解和抑制

基本信息

批准号：
9806591
负责人：
Rong Hai
金额：
$ 19.44万
依托单位：
UNIVERSITY OF CALIFORNIA RIVERSIDE
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-01 至 2021-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9806591
关键词：
Address Adult Antiviral Agents Arboviruses Arthropods Binding Sites Biochemical Biological Assay Complex Crystallization Dengue Virus Development Disease Outbreaks Drug Design Evaluation Family Fetus Flaviviridae Flavivirus Foundations Genome Genomics Goals Guillain-Barré Syndrome Health In Vitro Infection Knowledge Lead Ligands Link Mediating Methods Molecular Conformation Mutation Neurologic Nonstructural Protein Pharmaceutical Preparations Phase Proteins Protocols documentation RNA RNA chemical synthesis RNA replication RNA-Directed RNA Polymerase Replication Initiation Research Resistance Resolution South America Structure Therapeutic Titrations Vaccines Viral Virus Virus Diseases Virus Inhibitors Virus Replication ZIKV infection Zika Virus base conformational conversion design experimental study global health human disease inhibitor/antagonist insight lead optimization member mutant nervous system disorder novel novel therapeutics propargyl alcohol small molecule structural biology virology

项目摘要

Mechanistic understanding and inhibition of Zika NS5 protein ABSTRACT Zika virus (ZIKV) belongs to the single-stranded RNA-containing flavivirus family. Its recent outbreak and implication in human diseases (e.g. neurological disorders) have raised a global health alarm, and urgency to develop a therapeutic strategy against ZIKV infection. However, there are no currently approved antivirals against ZIKV available yet. This application seeks to develop an antiviral strategy against the non-structural protein 5 (NS5) of ZIKV, which is responsible for virus-specific genomic replication. On one hand, the currently identified flavivirus inhibitors will be evaluated for their efficiency on ZIKV inhibition. On the other hand, mechanistic details of ZIKV NS5-mediated RNA replication will be investigated, thereby providing a basis for development of synergistic inhibition strategies targeting various enzymatic steps of ZIKV NS5. In Aim 1, structural, biochemical and cellular approaches will be taken to evaluate the inhibition of ZIKV NS5-mediated de novo RNA synthesis by the thiophenyl propargyl alcohol (TPA) compounds, the non-nucleoside inhibitors (NNIs) that have been identified as inhibitors for Dengue virus (DENV) NS5, in vitro and ex vivo. Our recent structural study of ZIKV NS5 revealed that the TPA-binding site of DENV NS5 is conserved in ZIKV NS5. Through evaluation of the inhibitory effects of the TPA compounds on ZIKV NS5, this application will address whether the TPA compounds can serve as inhibitors to ZIKV NS5, and more importantly, to provide a basis for structure-based drug optimization for ZIKV NS5. In Aim 2, the mechanistic basis of ZIKV NS5-mediated RNA replication will be determined through structure elucidation of the replication initiation and elongation complexes of ZIKV NS5, combined with mutational and enzymatic analyses. The structural knowledge on the conformational transition of ZIKV NS5 from replication initiation to elongation will then provide a framework for structure-based drug design for comprehensive inhibition of ZIKV NS5 activity. Together, the proposed studies will provide key mechanistic insights into the NS5-mediated genome replication and establish a foundation for development of effective inhibitors against ZIKV.

机械理解和抑制Zika NS5蛋白抽象的 Zika病毒（ZIKV）属于单链RNA的黄病毒家族。它最近的爆发和对人类疾病的影响（例如神经系统疾病）引发了全球健康警报，并紧迫制定针对ZIKV感染的治疗策略。但是，目前尚无批准的抗病毒药针对ZIKV可用。该应用程序旨在制定针对非结构性的抗病毒策略 ZIKV的蛋白5（NS5），该蛋白质是病毒特异性基因组复制的原因。一方面，目前将评估确定的黄素抑制剂在ZIKV抑制方面的效率。另一方面，将研究ZIKV NS5介导的RNA复制的机械细节，从而为针对ZIKV NS5的各种酶促步骤的协同抑制策略的发展。在AIM 1中，将采用结构，生化和细胞方法来评估ZIKV NS5介导的抑制硫代苯基丙醇（TPA）化合物，非核苷抑制剂的从头RNA合成（NNI）已被鉴定为登革热病毒（DENV）NS5的抑制剂，体外和ex vivo。我们最近 ZIKV NS5的结构研究表明，DENV NS5的TPA结合位点在ZIKV NS5中是保守的。通过评估TPA化合物对ZIKV NS5的抑制作用，此应用将解决 TPA化合物是否可以作为对ZIKV NS5的抑制剂，更重要的是，基于结构的药物优化ZIKV NS5。在AIM 2中，ZIKV NS5介导的RNA的机械基础复制将通过结构阐明复制启动和伸长来确定 ZIKV NS5的复合物，结合突变和酶促分析。关于然后，ZIKV NS5从复制启动到伸长的构象过渡将为框架提供一个框架基于结构的药物设计，用于全面抑制ZIKV NS5活性。共同提出的研究将为NS5介导的基因组复制提供关键的机理见解，并为开发针对ZIKV的有效抑制剂。