Development of Novel Antivirals Targeting Viral RNA Methylation

针对病毒 RNA 甲基化的新型抗病毒药物的开发

• 批准号: 10512630
• 负责人: Danica Galonic Fujimori
• 金额: $ 404.9万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 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/10512630
• 关键词: 2019-nCoV; Animal Model; Antigen Presentation; Antiviral Agents; Binding; Binding Sites; Biochemistry; Biological Assay; C-terminal; Cell model; Cells; Communicable Diseases; Complex; Coronavirus; Coronavirus Infections; Development; Docking; Drug Targeting; Enzymes; Exonuclease; Exoribonucleases; Family; Genetic Transcription; Guanidines; Guanosine Triphosphate; Head; Human; Immune Evasion; Immune response; In Vitro; Industry Collaboration; Innate Immune System; Interferons; Letters; Libraries; Ligand Binding; Link; MHC Class I Genes; Messenger RNA; Methylation; Methyltransferase; Middle East Respiratory Syndrome Coronavirus; N-terminal; Natural Immunity; Nonstructural Protein; Nucleotides; Open Reading Frames; Pharmaceutical Chemistry; Pharmaceutical Preparations; Proteins; Proteomics; RNA; RNA Caps; RNA methylation; Role; S-Adenosylmethionine; SARS coronavirus; SARS-CoV-2 genome; SARS-CoV-2 infection; Series; Structure; Testing; Therapeutic; Translations; Viral; Viral Proteins; Virulence; Virus; Virus Diseases; Virus Replication; Work; adaptive immunity; antiviral drug development; base; cofactor; high throughput screening; in vivo; industry partner; inhibitor; lead optimization; loss of function mutation; mRNA capping; mutant; novel; pandemic disease; poly A specific exoribonuclease; programs; response; screening; small molecule; small molecule inhibitor; small molecule libraries; structural biology; targeted agent; viral RNA; virology

PROJECT 4: DEVELOPMENT OF NOVEL ANTIVIRALS TARGETING VIRAL RNA METHYLATION SUMMARY Coronaviruses cap their RNA by coordinated action of two methyltransferases (MTase): Nsp14, which catalyzes N7-guanidine methylation of GTP at the 5′ terminus of viral RNAs, and Nsp16, which forms C2′-O-methyl- ribosyladenine at the subsequent nucleotide. By mimicking mRNA of the host cell, the resulting cap structure is critical for immune evasion, stabilization of viral RNA and efficient translation. While loss of function mutations in the MTase domain of Nsp14 are lethal to SARS-CoV-2, SARS-CoV strains that carry mutant Nsp16 have low virulence, suggesting that targeting of these enzymes, either alone or in combination with other viral proteins, has strong therapeutic potential. In this application, we propose to develop antiviral agents that target MTase activities of Nsp14 and Nsp16. Our approach will assess the potential of RNA capping MTases as a novel target family for development of antiviral agents. Importantly, since both enzymes are conserved across coronaviruses known to infect humans, this approach could provide a footprint for development of pan-coronaviral acting agents. Inhibitors for each of the MTases will be identified using a combination of small molecule discovery approaches: computational docking, fragment linking and merging, and high throughput screening (HTS) to identify novel chemotypes. Enabled by the recent developments in availability of make-on-demand libraries, we propose to use ultra-large library docking to identify candidate inhibitors. Availability of drug-like compounds in in-house small molecule libraries will facilitate inhibitor identification through HTS. Hit compounds will be tested in a series of activity assays and validated using direct binding strategies. Experimentally determined structures of MTases in complex with small molecule inhibitors will be used to guide optimization, aided by access to make-on-demand libraries. The identified inhibitors will be prioritized based on their selectivity against a comprehensive panel of human MTases and antiviral activity in cellular models of SARS-CoV-2 infection. The subsequent medicinal chemistry optimization and assessment of antiviral activity in animal models is expected to result in Optimized Lead compounds, which will be further elaborated by our industry partners (Roche).

项目 4：开发针对病毒 RNA 甲基化的新型抗病毒药物 概括 冠状病毒通过两种甲基转移酶 (MTase) 的协调作用来限制其 RNA：Nsp14，其催化 病毒 RNA 5' 末端 GTP 的 N7-胍甲基化，以及 Nsp16，形成 C2'-O-甲基- 随后的核苷酸处的核糖腺嘌呤。通过模仿宿主细胞的 mRNA，得到的帽结构是 对于免疫逃避、病毒 RNA 的稳定和有效翻译至关重要。当功能丧失突变时 Nsp14 的 MTase 结构域中的 Nsp14 对 SARS-CoV-2 具有致死性，携带突变 Nsp16 的 SARS-CoV 毒株具有低 毒力，表明这些酶的靶向，无论是单独的还是与其他病毒蛋白组合， 具有很强的治疗潜力。在此应用中，我们建议开发针对 MTase 的抗病毒药物 Nsp14 和 Nsp16 的活性。我们的方法将评估 RNA 加帽 MTase 作为新靶点的潜力 家族致力于开发抗病毒药物。重要的是，由于这两种酶在冠状病毒中都是保守的 已知会感染人类，这种方法可以为泛冠状病毒作用的发展提供足迹 代理。 将使用小分子发现方法的组合来鉴定每种 MTase 的抑制剂： 计算对接、片段连接和合并以及高通量筛选 (HTS) 来识别新的 化学型。由于按需制作库的可用性的最新发展，我们建议 使用超大型库对接来识别候选抑制剂。内部类药化合物的可用性 小分子文库将有助于通过 HTS 识别抑制剂。热门化合物将进行一系列测试 活性测定并使用直接结合策略进行验证。实验确定的 MTase 结构 与小分子抑制剂的复合物将用于指导优化，并通过获取按需定制的帮助 图书馆。已确定的抑制剂将根据其对综合组的选择性进行优先排序 SARS-CoV-2 感染细胞模型中的人类 MTase 和抗​​病毒活性。随后的药用 化学优化和动物模型抗病毒活性评估预计将导致优化 铅化合物，将由我们的行业合作伙伴（罗氏）进一步阐述。