Broad-spectrum inhibitors targeting the viral replication promoter conserved in dengue and Zika viruses

针对登革热和寨卡病毒中保守的病毒复制启动子的广谱抑制剂

基本信息

批准号：
10194075
负责人：
Thomas C Hermann
金额：
$ 23.3万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2023-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10194075
关键词：
3&apos Untranslated Regions 5&apos Untranslated Regions Address Affect Affinity Antiviral Agents Antiviral Therapy Binding Binding Sites Biological Assay Cell Culture Techniques Cells Central America Clinical Country Dengue Fever Dengue Hemorrhagic Fever Dengue Virus Development Disease Disease Outbreaks Dose Flavivirus Fluorescence Fluorescence Resonance Energy Transfer Fluorescent Dyes Future Genome Goals Hepatitis C virus Human In Vitro Label Lead Ligand Binding Ligands Liver Microsomes Medical Metabolic Modeling Molecular Molecular Conformation Monitor Open Reading Frames Outcome Patients Permeability Pharmaceutical Preparations Polymerase RNA RNA Binding RNA Conformation RNA Sequences RNA replication RNA-Directed RNA Polymerase Replication Initiation Research Risk Sequence Alignment Serotyping Severities Signal Transduction South America Structure Symptoms Testing Therapeutic Titrations Toxic effect Translations Untranslated RNA Vaccinated Viral Viral Genome Virus Replication ZIKV infection Zika Virus base design expression cloning high throughput screening inhibitor/antagonist lead candidate mosquito-borne mosquito-borne pathogen pandemic disease pathogen promoter recruit replicase response screening small molecule small molecule inhibitor tissue culture tool viral RNA

项目摘要

Project Summary Zika virus (ZIKV) and dengue virus (DENV) are mosquito-borne flaviviruses that cause disease ranging in severity from mild symptoms to deadly hemorrhagic dengue fever. ZIKV was rapidly spreading five years ago to over 20 countries in South and Central America, where infections by ZIKV reached pandemic levels and threatened to expand to the southern US. The threat by DENV has increased dramatically over the last two decades as one of the worst mosquito-borne pathogens in tropical regions. To treat outbreaks of DENV in ZIKV-endemic regions, and vice versa, broad-spectrum inhibitor drugs against mosquito-borne flaviviruses would provide an efficient therapeutic approach that reduces the risk of immuno-associated exacerbation, for example in previously infected or vaccinated patients. Sequence alignment of DENV and ZIKV clinical isolates in combination with secondary structure prediction have led us to identify a conserved RNA three-way junction that serves as the replication promoter in flaviviruses. Here, we propose to identify compounds that inhibit viral replication by targeting the promoter RNA as lead candidates for the future development of DENV and ZIKV broad-spectrum therapeutics. The Specific Aims of this proposal are to 1) develop a FRET-based binding assay to assess small molecules as selective ligands of the DENV and ZIKV replication promoter RNA, 2) establish an in vitro replication initiation assay to identify promoter RNA-binding ligands as inhibitors of ZIKV replication, and 3) discover small molecules that inhibit DENV and ZIKV in cell culture by targeting the RNA 3WJ motif that serves as the viral replication promoter.

项目摘要 Zika病毒（ZIKV）和登革热病毒（DENV）是蚊子传播的黄病毒，引起疾病的范围从轻度症状到致命的出血登革热。 Zikv五年前迅速蔓延在南美洲和中美洲的20多个国家，ZIKV感染达到大流行水平，威胁要扩展到美国南部。 DENV的威胁在过去两个方面急剧增加数十年是热带地区最严重的蚊子传播病原体之一。治疗DENV的爆发 ZIKV流行区，反之亦然，针对蚊子传播的黄病毒的广谱抑制剂药物将提供有效的治疗方法，以降低免疫相关加重的风险以前感染或接种疫苗的患者的示例。 DENV和ZIKV临床分离株与二级结构预测的序列比对使我们确定了保守的RNA三向连接点，该连接处是复制启动子黄病毒。在这里，我们建议通过靶向启动子RNA来识别抑制病毒复制的化合物作为DENV和ZIKV广谱治疗学的未来发展的主要候选人。该提案的具体目的是1）开发基于FRET的结合测定法以评估小分子作为DENV和ZIKV复制启动子RNA的选择性配体，2）建立体外复制启动分析以鉴定启动子RNA结合配体为ZIKV复制的抑制剂，3）发现小通过靶向用作病毒的RNA 3WJ基序，抑制细胞培养中DENV和ZIKV的分子复制启动子。