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：
10181523
负责人：
Rong Hai
金额：
$ 25.33万
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/10181523
关键词：
Address Adult Antiviral Agents Arboviruses 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 ZIKA ZIKV infection Zika Virus arthropod-borne 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.

Zika NS5 蛋白的机制理解和抑制抽象的寨卡病毒 (ZIKV) 属于含有单链 RNA 的黄病毒科。它最近的爆发和对人类疾病（例如神经系统疾病）的影响已引起全球健康警报，迫切需要采取措施制定针对 ZIKV 感染的治疗策略。然而，目前尚无批准的抗病毒药物对抗 ZIKV 的方法尚未推出。该应用程序旨在开发针对非结构性病毒的抗病毒策略 ZIKV 的蛋白 5 (NS5)，负责病毒特异性基因组复制。一方面，当前将评估已确定的黄病毒抑制剂抑制 ZIKV 的效率。另一方面，将研究 ZIKV NS5 介导的 RNA 复制的机制细节，从而为开发针对 ZIKV NS5 各种酶促步骤的协同抑制策略。在目标 1 中，将采用结构、生化和细胞方法来评估 ZIKV NS5 介导的抑制作用由非核苷抑制剂噻吩基炔丙醇 (TPA) 化合物从头合成 RNA (NNI) 已被鉴定为体外和离体登革热病毒 (DENV) NS5 的抑制剂。我们最近的 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，本申请将解决 TPA化合物是否可以作为ZIKV NS5的抑制剂，更重要的是，为研究提供基础 ZIKV NS5 基于结构的药物优化。目标 2，ZIKV NS5 介导的 RNA 的机制基础复制将通过复制起始和延伸的结构阐明来确定 ZIKV NS5 复合物，结合突变和酶分析。关于结构的知识 ZIKV NS5 从复制起始到延伸的构象转变将为基于结构的药物设计，用于全面抑制 ZIKV NS5 活性。总之，拟议的研究将为 NS5 介导的基因组复制提供关键的机制见解，并为开发针对 ZIKV 的有效抑制剂。