Multiplex Small Molecule Discovery to Identify Broad-Acting Viral Protease Inhibitors

多重小分子发现来鉴定广泛作用的病毒蛋白酶抑制剂

• 批准号: 10513925
• 负责人: DAVID D HO
• 金额: $ 354.56万
• 依托单位: HACKENSACK UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-16 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10513925
• 关键词: 2019-nCoV; Address; Amino Acids; Animal Model; Animals; Antiviral Agents; Attention; Bar Codes; Cells; Chemicals; Chymase; Collaborations; Computational Biology; Coronavirus; Coupled; DNA; Data; Development; Disease Outbreaks; Dose; Drug Screening; Drug Targeting; Drug resistance; Economic Burden; Endoribonucleases; Exhibits; Family; Flaviviridae; Flavivirus; Goals; Human; Lead; Libraries; Metagenomics; Methods; Methyltransferase; Middle East Respiratory Syndrome; Mutagenesis; Mutation; Patients; Peptide Hydrolases; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacology; Population; Process; Property; Protease Inhibitor; Proteins; Resistance; SARS-CoV-2 infection; Severe Acute Respiratory Syndrome; Societies; Stream; Structural Chemistry; Surveys; Technology; Testing; Therapeutic; Time; Triage; Variant; Viral; Viral Proteins; Virus; Virus Diseases; Virus Replication; Work; ZIKA; base; cost; drug development; drug discovery; global health; health economics; human coronavirus; improved; in vivo; inhibitor; innovation; lead optimization; metropolitan; mutant; mutation screening; novel; novel strategies; pandemic disease; pandemic preparedness; pathogen; pathogenic virus; pre-clinical; preclinical development; process optimization; programs; resistance mechanism; response; screening; small molecule; structural biology; viral resistance; virology

ABSTRACT Viral pathogens present a serious health and economic burden to society, yet for the majority of viruses, there are no approved antiviral compounds. Worse still, this treatment gap continues to widen due to a continuous stream of emerging viral pathogens (e.g. MERS, Zika, and SARS-CoV-2). The objective of this proposal is to utilize a high-throughput, multiplexed approach for drug screening in combination with a novel approach to surveying drug resistant variants to guide the development of broad-acting antivirals with the long-term goal of bridging the existing antiviral therapeutic gap. Our proposal is based on the central hypothesis that DNA- barcoding technology coupled with deep mutational scans (DMS) of essential viral proteins can be used to rapidly search through chemical space and guide the hit-to-lead small molecule discovery process. The rationale underlying this proposal is that, if successful, we will be able to develop optimized leads that are active against multiple viruses and robust to viral escape at a fraction of the time, cost, and effort of traditional approaches. Given the ongoing pandemic and their proven pandemic potential, during the initial stages of our proposal, we will focus on generating broadly active inhibitors against coronavirus proteases. In later years, we will target other essential viral proteins (e.g. methyltransferase) and viral families (e.g. Flaviviridae). Our preliminary data support the feasibility of our approach for screening for inhibitors to dozens of viral proteases at the same time, along with our ability to characterize the effects of hundreds of mutations on the response of a viral protease to chemical inhibitors. To achieve our project’s goals, we will pursue the following three aims: 1) Increase the number of viral targets to be simultaneously screened to ≥100 and perform small molecule screens against them; 2) Test our screening hits against live virus, evolve their potency and drug-like properties, and demonstrate their in vivo efficacy; and 3) Use comprehensive mutagenesis to understand drug-target interactions and guide our drug development efforts. This proposal is innovative because it presents a multiplex method of small molecule screening that increases the quantity and richness of the data obtained. It also develops a method of studying the response of thousands of mutant variants of essential viral proteins to chemical inhibition, and uses this information to guide the hit-to-lead optimization process. This work is significant and is expected to have a positive impact by identifying a set of promising broad-acting protease inhibitors against human and animal viral pathogens, developing a highly-scalable approach to drug screening, and providing a framework for merging resistance profiling with structural and medicinal chemistry throughout the drug discovery process.

摘要 病毒病原体给社会带来了严重的健康和经济负担，但对于大多数病毒， 没有被批准的抗病毒化合物。更糟糕的是，这种治疗差距继续扩大，由于持续的 新出现的病毒病原体（如MERS、Zika和SARS-CoV-2）。这项建议的目的是 利用高通量、多重方法进行药物筛选，并结合新方法， 调查耐药变异，以指导广泛作用的抗病毒药物的开发，其长期目标是 弥合现有的抗病毒治疗差距。我们的建议是基于核心假设，即DNA- 条形码技术与基本病毒蛋白的深度突变扫描（DMS）相结合，可用于快速 通过化学空间搜索和引导命中领先的小分子发现过程。的理由 该提案的基础是，如果成功，我们将能够开发积极针对以下目标的优化潜在客户 多个病毒和强大的病毒逃逸在一小部分的时间，成本和努力的传统方法。 鉴于目前的大流行及其已证实的大流行潜力，在我们建议的最初阶段，我们 将专注于产生广泛活性的冠状病毒蛋白酶抑制剂。在以后的几年里，我们将针对 其他必需的病毒蛋白（例如甲基转移酶）和病毒家族（例如黄病毒科）。我们的初步数据 支持我们同时筛选几十种病毒蛋白酶抑制剂的方法的可行性， 沿着我们的能力，以表征数百种突变对病毒蛋白酶反应的影响， 化学抑制剂为了实现我们的项目目标，我们将追求以下三个目标：1）增加 同时筛选的病毒靶点数量≥100个，并对其进行小分子筛选; 2)测试我们针对活病毒的筛选命中，进化其效力和药物样特性，并证明其 体内疗效; 3）使用综合诱变来了解药物-靶点相互作用，并指导我们的 药物开发工作。该方案具有创新性，因为它提出了一种小分子的多重方法， 增加所获得数据的数量和丰富性的筛选。它还发展了一种研究 基本病毒蛋白质的数千种突变变体对化学抑制的反应，并利用这种 信息，以指导命中到领先的优化过程。这项工作意义重大，预计将有一个 通过鉴定一组有前途的针对人类和动物病毒的广泛作用的蛋白酶抑制剂 病原体，开发一种高度可扩展的药物筛选方法，并提供一个框架， 在整个药物发现过程中，利用结构和药物化学进行耐药性分析。