Small molecule-induced degradation of dengue proteins as an antiviral strategy

小分子诱导的登革热蛋白降解作为抗病毒策略

• 批准号: 10472071
• 负责人: NATHANAEL Schiander GRAY
• 金额: $ 81.09万
• 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: 2020-07-23 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10472071
• 关键词: Ablation; Affect; Affinity; Antiviral Agents; Antiviral resistance; Binding; Biological Assay; Cancer Biology; Cell Culture Techniques; Cells; Core Protein; Dengue; Dengue Infection; Dengue Virus; Development; Dose; Drug Kinetics; Event; Excision; Flavivirus; Foundations; Genetic Variation; Genotype; Goals; HIV; Hepatitis C; Hepatitis C virus; Human; In Vitro; Length; Ligands; Link; Mediating; Membrane; Methods; Monitor; Mutation; Peptide Hydrolases; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacology; Polymerase; Predisposition; Process; Proteins; RNA Viruses; RNA replication; RNA-Directed RNA Polymerase; Resistance; Resistance profile; Serial Passage; Serotyping; Specificity; Suppressor Mutations; Testing; Time; Ubiquitination; Vaccines; Viral; Viral Genome; Viral Proteins; Virus; Virus Replication; Work; anti-viral efficacy; antiviral drug development; chemotherapy; combat; cytotoxicity; design; efficacy testing; experimental study; improved; in vivo; inhibitor; mouse model; mutant; pathogen; pathogenic virus; protein degradation; protein function; recruit; residence; resistance mechanism; small molecule; tool; ubiquitin-protein ligase; vaccine development; virus core

PROJECT SUMMARY / ABSTRACT Dengue virus (DENV) is a pathogen of high biomedical significance against which we lack effective countermeasures. Although targeted chemotherapy using combinations of direct-acting antivirals (DAAs) has proven highly successful against hepatitis C virus infection and HIV, efforts to develop analogous drugs against DENV have not been successful. The genetic diversity of DENV due to replication by an RNA-dependent RNA polymerase that lacks proofreading function presents additional challenges by making it difficult to develop vaccines and antivirals with broad-spectrum coverage of all genotypes within one viral species and facilitating the rapid development of antiviral resistance when DAAs are used as monotherapies. Recently developed methods for small molecule-induced degradation of specific proteins rely on chimeric molecules (“PROTACs,” “degronimids,” “degraders”) that have a target-specific ligand linked to a moiety that binds an E3 ubiquitin ligase (e.g., cereblon, VHL). Small molecule-binding leads to ubiquitination and proteasomal degradation of the target. This results in event-driven rather than occupancy-driven pharmacology leading to efficient removal of the target from the cell and functional ablation of all of the protein's functions. Since pharmacological activity does not require constant, stoichiometric engagement of the target, even modest affinity ligands can be effective degraders. In addition, this mechanism of action can have higher natural barriers to resistance than conventional inhibitors, as has been demonstrated in the cancer biology field. While these potential advantages are attractive for antivirals development, it remains unclear the extent to which they can be leveraged to attain significant antiviral effects. In particular, strong viral expression and localization of viral processes (and their effectors) on or near specialized membranes may limit the susceptibility of DENV and other viruses to this pharmacological strategy. Here we propose to explore whether we can successfully deploy targeted protein degradation against three essential DENV proteins: core, NS4B, and NS5. As there are currently no approved anti-DENV drugs, there is an urgent need to find new pharmacological strategies to target this virus. Starting with known inhibitors as targeting ligands for degrader development, we will develop and validate antiviral degraders. We will then use these as tools to systematically explore potential points of differentiation between degraders and conventional inhibitors in terms of affinity, potency, selectivity, duration of action and susceptibility to resistance. We will also optimize validated antiviral degraders to test the efficacy of this antiviral approach in vivo. The overall goal is to validate degradation of one or more of these targets as an antiviral strategy with high natural barrier to resistance and to advance first-in-class degraders as leads for the development of antivirals. In pursuit of this goal, we will also establish important proof of concept and the foundation for more broadly developing antiviral degraders against other viral pathogens.

项目概要/摘要 登革热病毒（DENV）是一种具有高度生物医学意义的病原体，我们缺乏有效的对抗方法 对策。尽管使用直接作用抗病毒药物（DAA）组合的靶向化疗已 事实证明，在对抗丙型肝炎病毒感染和艾滋病毒方面非常成功，因此正在努力开发类似的药物来对抗 DENV 尚未成功。由于依赖RNA的RNA复制而导致的DENV遗传多样性 缺乏校对功能的聚合酶因其难以开发而带来额外的挑战 广谱覆盖一种病毒物种内所有基因型的疫苗和抗病毒药物，并促进 当 DAA 用作单一疗法时，抗病毒耐药性迅速发展。 最近开发的小分子诱导特定蛋白质降解的方法依赖于嵌合体 分子（“PROTAC”、“degronimids”、“降解剂”）具有与某个部分相连的目标特异性配体 结合 E3 泛素连接酶（例如 cereblon、VHL）。小分子结合导致泛素化 靶标的蛋白酶体降解。这导致了事件驱动而非占用驱动的药理学 导致从细胞中有效去除靶标并功能性消除所有蛋白质的功能。 由于药理活性不需要靶标持续的、化学计量的参与，即使是适度的参与 亲和配体可以是有效的降解剂。另外，这种作用机制可以具有较高的天然屏障 正如癌症生物学领域所证明的那样，它比传统抑制剂具有更高的耐药性。虽然这些 潜在的优势对于抗病毒药物的开发很有吸引力，但目前尚不清楚它们能在多大程度上发挥作用 以获得显着的抗病毒效果。特别是，病毒的强烈表达和病毒定位 特殊膜上或附近的过程（及其效应器）可能会限制 DENV 和其他病毒的敏感性 病毒对这种药理学策略的影响。这里我们建议探讨一下能否成功部署 针对三种基本 DENV 蛋白的靶向蛋白质降解：核心、NS4B 和 NS5。由于有 目前尚无批准的抗DENV药物，迫切需要寻找新的药理学策略来靶向 该病毒。从已知的抑制剂作为降解剂开发的靶向配体开始，我们将开发和 验证抗病毒降解剂。然后我们将使用这些作为工具来系统地探索潜在的点 降解剂和传统抑制剂在亲和力、效力、选择性、持续时间方面的区别 行动和抵抗力的敏感性。我们还将优化经过验证的抗病毒降解剂，以测试其功效 这种体内抗病毒方法。总体目标是验证这些目标中的一个或多个目标的退化情况 具有高天然耐药屏障的抗病毒策略，并推进一流的降解剂作为领先 抗病毒药物的开发。为了实现这一目标，我们还将建立重要的概念验证和 为更广泛地开发针对其他病毒病原体的抗病毒降解剂奠定了基础。