Discovery of SARS-CoV-2 antivirals using a replicon assay

使用复制子测定发现 SARS-CoV-2 抗病毒药物

• 批准号: 10673119
• 负责人: Stefan G Sarafianos
• 金额: $ 64.35万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10673119
• 关键词: 2019-nCoV; Active Sites; Address; Affect; Air; Antiviral Agents; Binding; Biochemical; Biological Assay; Biophysics; COVID-19; COVID-19 mortality; Caco-2 Cells; Cell Line; Cells; Cessation of life; Collection; Combined Modality Therapy; Coronavirus; Data; Development; Disclosure; Disease; Disease Outbreaks; Drug Kinetics; Drug resistance; Effectiveness; Enzyme Kinetics; Fluorescence Microscopy; Generations; Genes; Genetic Engineering; Genomics; Grant; HIV-1; HIV-2; Hepatitis B Virus; Hepatitis C virus; Human; In Vitro; Individual; Influenza; International; Kinetics; Lead; Legal patent; Libraries; Liquid substance; Measurement; Measures; Metals; Molecular; Mus; Mutation; Nonstructural Protein; Pharmaceutical Chemistry; Plasmids; Preparation; Proteins; Public Health; Published Comment; Publishing; RNA; Replicon; Reporting; Resistance; SARS coronavirus; SARS-CoV-2 B.1.1.7; SARS-CoV-2 B.1.351; SARS-CoV-2 CAL.20C; SARS-CoV-2 P.1; SARS-CoV-2 antiviral; SARS-CoV-2 inhibitor; Severe Acute Respiratory Syndrome; System; Technology; Testing; Time; Transfection; Untranslated Regions; Vaccines; Variant; Viral; Viral Physiology; Virus; Virus Diseases; Visualization; Washington; Work; ZIKA; airway epithelium; analog; cell preparation; combat; cytotoxicity; drug candidate; drug discovery; efficacy study; experimental study; expression cloning; fitness; helicase; high throughput screening; improved; in vivo; inhibitor; innovation; invention; lead optimization; meter; miniaturize; monolayer; mortality; mouse model; nanomolar; novel; pandemic disease; remdesivir; resistance mechanism; resistance mutation; screening; small molecule inhibitor; small molecule libraries; socioeconomics; stable cell line; success; synergism; tool; viral entry inhibitor

PROJECT SUMMARY Severe acute respiratory syndrome Coronavirus 2 (SARS-CoV-2) cause severe diseases in humans (COVID- 19) that presents a major threat for global public health. Since it was first reported in 12/2019, COVID-19 has become a pandemic that continues to spread, with >246 million confirmed cases and >5 million deaths as of 11/02/2021. In addition to the human tragedy, the magnitude of the pandemic-driven implosion of global economies is enormous. Although vaccines are now available, their efficacy appears to be reduced with spreading viral strains. Remdesivir is the only approved antiviral targeting SARS-CoV-2, but it has little effect on COVID-19 mortality. Therefore, it is critical to identify and develop additional antivirals to combat viral infection. The following strong preliminary data enable targeting of SARS-CoV-2: 1) Construction of an extensive collection of SARS-CoV-2 replicon systems that enable cutting-edge, rapid, and economical high-throughput screening. 2) Preparation of cell lines that stably express SARS-CoV-2 replicon. 3) Preliminary screening of chemical libraries led to the discovery of a novel SARS-CoV-2 antiviral, which has already been improved with one round of optimization through medicinal chemistry efforts. 4) Multiplex visualization of single-genomic or subgenomic (+) or (-) SARS-CoV-2 RNA and simultaneously viral and/or host proteins in individual infected cells. 5) Cutting-edge rapid high-throughput infectious virus BSL3 assays that enable kinetic, mechanistic, drug resistance studies. 6) In-house cloning expression and purification of 10 SARS-CoV-2 non-structural proteins (nsps). 7) Biochemical and biophysical assays to measure the enzymatic activities of several SARS-CoV and SARS-CoV-2 nsp proteins and to measure compound binding to nsps. We hypothesize that our recently developed replicon systems can be used for the discovery of anti-SARS- CoV-2 hits, which upon hit-to-lead optimization can become COVID-19 drug candidates. To address this hypothesis, we propose the following specific aims: 1. Use of SARS-CoV-2 replicon plasmid systems and SARS-CoV-2 replicon-expressing cell lines to screen chemical libraries for antiviral hits. 2. Inhibition and resistance studies with prioritized inhibitors. 3. Hit-to-lead optimization These studies will lead to antivirals with strong potency and pharmacokinetic profiles, setting the stage for development of SARS-CoV-2 antivirals and combination therapies.

项目概要 严重急性呼吸综合征冠状病毒 2 (SARS-CoV-2) 会导致人类严重疾病 (COVID-19) 19）这对全球公共卫生构成重大威胁。自 12/2019 年首次报告以来，COVID-19 已 成为一种持续蔓延的大流行病，截至目前确诊病例超过 2.46 亿，死亡人数超过 500 万人 2021 年 11 月 2 日。除了人类悲剧之外，大流行引发的全球经济崩溃的严重程度 经济规模巨大。尽管现在已有疫苗，但其功效似乎会随着时间的推移而降低 传播病毒株。瑞德西韦是唯一被批准的针对 SARS-CoV-2 的抗病毒药物，但对治疗效果甚微 COVID-19 死亡率。因此，识别和开发额外的抗病毒药物来对抗病毒感染至关重要。 以下强有力的初步数据能够针对 SARS-CoV-2：1) 构建广泛的 SARS-CoV-2 复制子系统的集合，可实现尖端、快速且经济的高通量 筛选。 2）稳定表达SARS-CoV-2复制子的细胞系的制备。 3) 初步筛选 化学库导致了一种新型 SARS-CoV-2 抗病毒药物的发现，该药物已通过以下方法进行了改进： 通过药物化学努力进行一轮优化。 4) 单基因组或基因组的多重可视化 单个感染细胞中的亚基因组 (+) 或 (-) SARS-CoV-2 RNA 以及同时的病毒和/或宿主蛋白。 5) 尖端的快速高通量感染性病毒 BSL3 检测，可实现动力学、机械、药物分析 耐药性研究。 6) 10种SARS-CoV-2非结构蛋白的内部克隆表达和纯化 （国家统计局）。 7) 生化和生物物理测定，测量几种 SARS-CoV 的酶活性和 SARS-CoV-2 nsp 蛋白并测量化合物与 nsp 的结合。 我们假设我们最近开发的复制子系统可用于发现抗 SARS- CoV-2 命中，经过命中到先导优化后可以成为 COVID-19 候选药物。为了解决这个问题 假设，我们提出以下具体目标： 1.利用SARS-CoV-2复制子质粒系统和SARS-CoV-2复制子表达细胞系进行筛选 抗病毒药物的化学库。 2.优先抑制剂的抑制和耐药性研究。 3. 命中引导优化 这些研究将产生具有强大效力和药代动力学特征的抗病毒药物，为 SARS-CoV-2 抗病毒药物和联合疗法的开发。

项目成果

Mechanisms by which the cystic fibrosis transmembrane conductance regulator may influence SARS-CoV-2 infection and COVID-19 disease severity.

囊性纤维化跨膜电导调节剂可能影响 SARS-CoV-2 感染和 COVID-19 疾病严重程度的机制。

• 发表时间: 2023-11
• 作者: Tedbury, Philip R;Manfredi, Candela;Degenhardt, Frauke;Conway, Joseph;Horwath, Michael C;McCracken, Courtney;Sorscher, Adam J;Moreau, Sandy;Wright, Christine;Edwards, Carolina;Brewer, Jo;Guarner, Jeannette;de Wit, Emmie;Williamson, Brandi N
• 通讯作者: Williamson, Brandi N