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 登革热病毒（DENV）和其他黄素病毒是引起严重疾病的主要人类病原体。 通过宽度蚊子物种传播，其中许多病毒迅速扩散，可以具有 对事先免疫力不存在的对公共卫生的毁灭性影响。因此，有很大的需求 对抗当前和未来的黄病毒威胁的对策。当前抗病毒药的主要局限性 开发是相对少量的经过验证的抗病毒靶标，其中大多数是病毒酶 （例如，聚合酶，蛋白酶）；当直接作用抗病毒药用作电阻的低障碍物 单层；以及大多数这些药物的狭窄活性（“一种虫子，一种药物”）。新课程 可以针对相关病毒介导广泛活性并具有很高障碍的靶标 尤其需要抵抗来打击新兴病毒，因为我们通常缺乏足够的时间，并且 一旦这些病毒构成了重大威胁，可以在有用的时间范围内开发新药的资源。 针对黄病毒包膜蛋白E的小分子E具有模仿体液的潜力 免疫反应通过细胞外接合并在复制周期早期阻止病毒进入。 我们已经确定了与DENV包膜蛋白，E和 即使在粒子上只有少数E的E副本，在病毒进入过程中抑制电子介导的膜融合 是抑制剂结合的。这些化合物在域I和II之间的袋中结合，并抑制西尼罗河，寨卡 和日本脑炎病毒至少部分保护该地点。我们最近建立了一个 基于目标的测定并验证了其在识别DENV和Zika E蛋白的新抑制剂中的使用 与我们的原始抑制剂相比，在配置的口袋中结合并且具有更多的药物样性能。建筑 在这项工作中，我们现在提出了一个全面的计划，以合理地优化小分子抑制剂 DENV E蛋白是一种潜在的抗病毒策略。为此，我们将结合建模和结构 - 使用完整的基于目标的生化，具有有效筛选级联的指导药物设计， 细胞和机械测定能够有效地优化两个化学上不同的铅系列。我们的 这项工作的主要目标是在DENV感染的鼠模型中证明抗病毒效率，从而铺设 第一类直接表演抗病毒药的基础，以治疗DENV所拥有的日益增长的全球威胁。