Validating the Flavivirus Envelope Protein as an Antiviral Target

验证黄病毒包膜蛋白作为抗病毒靶点

基本信息

批准号：
10578759
负责人：
NATHANAEL Schiander GRAY
金额：
$ 96.12万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-03-01 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10578759
关键词：
ADME Study Address Animal Model Antiviral Agents Automobile Driving Binding Binding Proteins Binding Sites Biochemical Biological Assay Biology Chemicals Complex Computational Biology Computer Analysis Computer Models Culicidae Dengue Infection Dengue Virus Disease Docking Drug Design Drug Kinetics Drug Targeting E protein Engineering Enzymes Evaluation Flavivirus Foundations Future Glucosides Goals Immune response Immunity In Vitro Infection Japanese encephalitis virus Kinetics Lead Libraries Liposomes Maximum Tolerated Dose Measures Mediating Membrane Fusion Microsomes Minority Modeling Peptide Hydrolases Pharmaceutical Chemistry Pharmaceutical Preparations Phenotype Plasma Proteins Polymerase Property Proteins Public Health Recombinants Resistance Resistance profile Resources Serial Passage Series Serotyping Site Site-Directed Mutagenesis Solubility Structural Models Structure Therapeutic Time Viral Viral Envelope Proteins Viremia Virion Virus Virus Inhibitors West Nile virus Work X-Ray Crystallography ZIKA Zika Virus anti-viral efficacy antiviral drug development aqueous combat cytotoxicity design efficacy testing env Gene Products experimental study extracellular flavivirus envelope protein E high throughput screening human pathogen in silico in vivo in vivo Model in vivo evaluation inhibitor lead optimization lead series mouse model mutant novel therapeutics particle pathogen process optimization prophylactic resistance mutation screening small molecule small molecule inhibitor structural biology transmission process viral entry inhibitor viral resistance

项目摘要

PROJECT SUMMARY Dengue virus (DENV) and other flaviviruses are major human pathogens that cause significant disease. Transmitted by widespread mosquito species, many of these viruses spread rapidly and can have a devastating impact on public health where prior immunity does not exist. There is thus a significant need for countermeasures to combat both current and future flavivirus threats. Major limitations in current antivirals development are the relatively small number of validated antiviral targets, most of which are viral enzymes (e.g., polymerases, proteases); the low barrier to resistance when direct-acting antivirals are used as monotherapies; and the narrow spectrum activity of most of these agents (“one bug, one drug”). New classes of targets that can mediate broad-spectrum activity against related viruses and that have high barriers to resistance are particularly needed to combat emerging viruses since we generally lack sufficient time and resources to develop new drugs on a useful time scale once these viruses pose significant threats. Small molecules targeting the flavivirus envelope protein, E, have the potential to mimic the humoral immune response by engaging their target extracellularly and blocking viral entry early in the replication cycle. We have identified multiple small molecule inhibitor series that bind to the DENV envelope protein, E, and inhibit E-mediated membrane fusion during viral entry even when only a minority of copies of E on the particle are inhibitor-bound. These compounds bind in a pocket between domains I and II and inhibit West Nile, Zika, and Japanese encephalitis viruses due to at least partial conservation of this site. We recently established a target-based assay and validated its use in the identification of new inhibitors of DENV and Zika E proteins that bind in the conserved pocket and that have more drug-like properties than our original inhibitors. Building on this work, we now propose a comprehensive plan to rationally optimize small molecule inhibitors of the DENV E protein as a potential anti-viral strategy. Towards this end, we will combine modeling and structure- guided drug design with an efficient screening cascade using complementary target-based biochemical, cellular and mechanistic assays to enable efficient optimization of two chemically distinct lead series. Our primary goal in this work is to demonstrate antiviral efficacy in a murine model of DENV infection, thus laying the foundation for first-in-class direct acting antivirals to treat the growing global threat that DENV poses.

项目概要登革热病毒（DENV）和其他黄病毒是导致严重疾病的主要人类病原体。这些病毒由分布广泛的蚊子物种传播，许多病毒传播迅速，并且可以产生在不存在先前免疫力的情况下，对公众健康造成毁灭性影响，因此非常需要这种疫苗。对抗当前和未来黄病毒威胁的对策当前抗病毒药物的主要局限性。开发中经过验证的抗病毒靶标数量相对较少，其中大部分是病毒酶（例如，聚合酶、蛋白酶）；使用直接作用抗病毒药物时，耐药性屏障较低单一疗法；以及大多数这些药物的窄谱活性（“一种细菌，一种药物”）。可以介导针对相关病毒的广谱活性并且具有高屏障的目标由于我们通常缺乏足够的时间和能力，因此特别需要抵抗新出现的病毒一旦这些病毒构成重大威胁，就可以在有用的时间范围内开发新药的资源。靶向黄病毒包膜蛋白 E 的小分子有可能模拟体液通过在细胞外接合其靶标并在复制周期的早期阻止病毒进入来产生免疫反应。我们已经鉴定出多种小分子抑制剂包膜系列，可与 DENV 蛋白、E 和即使颗粒上只有少数 E 拷贝，也能在病毒进入过程中抑制 E 介导的膜融合这些化合物结合在结构域 I 和 II 之间的口袋中，抑制西尼罗河病毒、寨卡病毒、和日本脑炎病毒，由于至少部分保护了该网站，我们最近建立了一个。基于目标的测定并验证其在识别 DENV 和 Zika E 蛋白新抑制剂中的应用结合在保守的口袋中，并且比我们最初的抑制剂具有更多的药物特性。在这项工作中，我们现在提出了一个综合方案，合理优化小分子抑制剂 DENV E蛋白作为一种潜在的抗病毒策略，为此，我们将结合建模和结构。使用互补的基于靶标的生化物质，通过有效的筛选级联来指导药物设计，细胞和机械分析，可有效优化两个化学上不同的先导系列。这项工作的主要目标是在 DENV 感染的小鼠模型中证明抗病毒功效，从而奠定为一流的直接作用抗病毒药物治疗 DENV 造成的日益严重的全球威胁奠定了基础。