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.

项目概要寨卡病毒 (ZIKV) 和登革热病毒 (DENV) 是蚊媒黄病毒，可引起以下疾病：严重程度从轻微症状到致命的出血性登革热。五年前 ZIKV 迅速传播南美洲和中美洲 20 多个国家的 ZIKV 感染已达到大流行水平，威胁要扩展到美国南部。过去两年来，登革热病毒的威胁急剧增加几十年来一直是热带地区最严重的蚊媒病原体之一。治疗 DENV 暴发 ZIKV 流行地区，反之亦然，针对蚊媒黄病毒的广谱抑制剂药物将提供一种有效的治疗方法，降低免疫相关恶化的风险，例如先前感染或接种过疫苗的患者。 DENV 和 ZIKV 临床分离株的序列比对与二级结构预测相结合使我们发现了一个保守的RNA三路连接，它作为复制启动子黄病毒。在这里，我们建议鉴定通过靶向启动子 RNA 抑制病毒复制的化合物作为未来开发 DENV 和 ZIKV 广谱疗法的主要候选者。该提案的具体目标是 1) 开发基于 FRET 的结合测定来评估小分子作为 DENV 和 ZIKV 复制启动子 RNA 的选择性配体，2) 建立体外复制启动测定来鉴定启动子 RNA 结合配体作为 ZIKV 复制的抑制剂，并且 3) 发现小的通过靶向作为病毒的 RNA 3WJ 基序来抑制细胞培养中的 DENV 和 ZIKV 的分子复制启动子。