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

• 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
• 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

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.

项目摘要 登革病毒（DENV）和其他黄病毒是引起重大疾病的主要人类病原体。 这些病毒由广泛分布的蚊子传播，其中许多病毒传播迅速， 在没有事先免疫的情况下，对公共卫生造成毁灭性影响。因此，非常需要 应对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 目前的主要问题是，经过验证的抗病毒靶点数量相对较少，其中大部分是病毒酶 (e.g.,聚合酶，蛋白酶）;当直接作用的抗病毒药物用作 单一疗法;以及大多数这些药剂的窄谱活性（“一种虫子，一种药物”）。新类 这些靶点可以介导针对相关病毒的广谱活性，并且对 特别需要抵抗力来对抗新出现的病毒，因为我们通常缺乏足够的时间， 一旦这些病毒构成重大威胁，就需要在有效的时间范围内开发新药的资源。 靶向黄病毒包膜蛋白E的小分子具有模拟体液免疫的潜力。 通过在细胞外接合它们的靶点并在复制周期的早期阻断病毒进入来增强免疫应答。 我们已经鉴定了多种小分子抑制剂系列，其结合DENV包膜蛋白E， 在病毒进入过程中抑制E介导的膜融合，即使在颗粒上只有少数E拷贝时 都在走廊里这些化合物结合在结构域I和II之间的口袋中，并抑制西尼罗病毒、寨卡病毒、 和日本脑炎病毒，因为这个网站至少部分的保护。我们最近建立了一个 靶向测定并验证其在鉴定DENV和Zika E蛋白的新抑制剂中的用途 结合在保守的口袋里，比我们原来的抑制剂有更多的药物样特性。建筑 在这项工作中，我们现在提出了一个全面的计划，以合理优化小分子抑制剂， DENV E蛋白作为潜在的抗病毒策略。为此，我们将联合收割机建模和结构- 使用互补的基于靶点的生物化学的有效筛选级联的指导药物设计， 细胞和机制分析，以实现两个化学上不同的铅系列的有效优化。我们 这项工作的主要目标是证明在DENV感染的小鼠模型中的抗病毒功效，从而奠定 该公司是一流的直接作用抗病毒药物的基础，用于治疗DENV构成的日益增长的全球威胁。