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

基本信息

批准号：
10661608
负责人：
NATHANAEL Schiander GRAY
金额：
$ 83.59万
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：
2020-07-23 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10661608
关键词：
Ablation Affect Affinity Antiviral Agents Antiviral resistance Binding Biological Assay Cancer Biology Cell Culture Techniques Cells Dengue Dengue Infection Dengue Virus Development Dose Drug Kinetics Event Excision Experimental Designs 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 Ubiquitination Vaccines Viral Viral Genome Viral Physiology Viral Proteins Virus Virus Replication Work anti-viral efficacy antiviral drug development chemotherapy combat cytotoxicity efficacy testing improved in vivo inhibitor mouse model mutant pathogen pathogenic virus pharmacologic protein degradation recruit residence resilience resistance mechanism small molecule tool ubiquitin-protein ligase vaccine development virus core

项目摘要

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.

项目摘要 /摘要登革热病毒（DENV）是一种具有高生物医学意义的病原体，我们缺乏有效对策。尽管使用直接作用抗病毒药（DAA）的组合有针对性化疗的具有被证明对丙型肝炎病毒感染和HIV非常成功，努力开发针对的类似药物 DENV没有成功。由RNA依赖性RNA复制而引起的DENV的遗传多样性缺乏校对功能的聚合酶通过使开发难以开发带来其他挑战疫苗和抗病毒药具有广谱覆盖在一种病毒物种中的所有基因型的范围，并促进当将DAA用作单一疗法时，抗病毒抗性的快速发展。最近开发的用于小分子诱导的特定蛋白降解的方法依赖于嵌合分子（“ protacs”，“脱基酰亚胺”，“降解器”），其具有特定于靶的配体与一个与部分相关的配体结合E3泛素连接酶（例如Cereblon，VHL）。小分子结合导致泛素化和靶标的蛋白酶体降解。这导致事件驱动而不是占用驱动的药理学导致从细胞中有效去除靶标，并使所有蛋白质功能的功能消融。由于药物活动不需要恒定，因此靶标的化学计量参与甚至适度亲和力配体可以有效降级。此外，这种作用机理可以具有更高的自然障碍抗药性比常规抑制剂，如癌症生物学领域所证明的那样。而这些潜在的优势对于抗病毒药的开发很有吸引力，目前尚不清楚它们在多大程度上可以杠杆以获得明显的抗病毒作用。特别是，强烈的病毒表达和病毒的定位过程（及其影响）对或附近的专业机制可能会限制DENV和其他的敏感性该药物策略的病毒。在这里，我们建议探索我们是否可以成功部署针对三种必需DENV蛋白的靶向蛋白质降解：核，NS4B和NS5。如有目前尚无批准的抗Denv药物，迫切需要寻找新的药物来针对目标该病毒。从已知的抑制剂作为靶向降解器开发的配体开始，我们将开发和验证抗病毒降解器。然后，我们将使用这些作为系统探索的工具从亲和力，效能，选择性，持续时间方面，降解器和常规抑制剂之间的差异动作和抵抗的敏感性。我们还将优化经过验证的抗病毒降解器，以测试这种抗病毒在体内。总体目标是验证其中一个或多个目标的降解作为一个具有高自然障碍的抗病毒策略，阻力并推进第一类降级器作为铅抗病毒药的发展。为了实现这一目标，我们还将建立重要的概念证明和针对其他病毒病原体的更广泛发展抗病毒药降解者的基础。