Inhibitors of Coronavirus Fidelity and Cap Methylation as Broadly Applicable

冠状病毒保真度和帽甲基化抑制剂广泛适用

基本信息

批准号：
8995621
负责人：
Mark R Denison
金额：
$ 88.38万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8995621
关键词：
5&apos -exoribonuclease Ablation Acute Animals Antiviral Agents Area Attenuated Binding Biological Biological Assay Biological Availability Chiroptera Coronavirus Data Set Detection Development Disease Drug Targeting Drug resistance Enzymes Europe Exons Family Family member Frequencies Genes Genetic Genome Goals Histology Human Immune Immune response Impairment In Vitro Infection Interferons Lead Lower Respiratory Tract Infection Mediating Methylation Methyltransferase Middle East Modeling Monitor Morbidity - disease rate Movement Mus Mutagenesis Mutagens Mutation Nonstructural Protein Pathogenesis Pathway interactions Pharmaceutical Chemistry Phenotype Process RNA RNA Caps RNA-Directed RNA Polymerase Replication-Associated Process Respiratory physiology Ribonucleosides SARS coronavirus Severity of illness Solubility Technology Testing Therapeutic Viral Viral Proteins Virulence Virulent Virus Virus Replication Work analog attenuation drug development drug discovery efficacy testing experience fitness high throughput screening human disease in vivo inhibitor/antagonist mortality mouse model novel programs protein function public health priorities replicase reverse genetics screening small molecule inhibitor small molecule libraries therapeutic target transmission process viral RNA viral fitness viral resistance

项目摘要

Both the emergence and subsequent human-to-human transmission of SARS-CoV in 2002-2003, and ofthe highly virulent human coronavirus HCoV-EMC in the Middle East and Europe in 2012-2013 exemplifies CoV movement potential and transmissibility, and underscores the urgent and critical need for a broadly efficacious therapeutics. The overall goal of Project 2 is to identify inhibitors of two highly conserved CoV processes, replication fidelity and RNA capping, that are essential for SARS-CoV virulence and survival in vivo. Multiple viral proteins and enzymatic activities are critical for these processes, including CoV 3'-to-5' exoribonuclease (fidelity; nsp14-ExoN) and 2'-0-methyltransferase (capping; nsp16-0MTase) activities. Consistent with the importance of these processes, we have shown that decreased replication fidelity and ablation of RNA capping through genetic inactivation of either ExoN or OMTase, respectively, results in replication competent viruses that are profoundly attenuated in vivo. In Aims 1 and 2, we will work with the Screening Core (Core B) and the Medicinal Chemistry lead Development Core (Core C) to identify, characterize, and optimize small molecule inhibitors of SARS-CoV fidelity and RNA capping. Once active compounds are identified, we will define their mechanism of action, test for the development of virus resistance, and determine their activity across the CoV family. In Aim S, we will work with Core C to chemically optimize and test the in vivo efficacy of lead compounds in progressively tiered models of SARSCoV disease severity, and assess the development of drug resistance in vivo. The complementary expertise ofthe Denison and Baric Labs, extensive preliminary datasets, state-of-the-art technologies, and the expertise of SR in the areas of medicinal chemistry, high-throughput screening, and drug development will contribute significantly to the successful identification, confirmation, and in vivo testing of lead compounds. Ultimately, inhibiting these two conserved and distinct pathways required for in vivo pathogenesis will allow for the treatment of endemic and emerging CoVs and potentially reduce the emergence of viral resistance.

2002-2003 年 SARS-CoV 的出现和随后的人际传播，以及 2012-2013 年中东和欧洲高毒力人类冠状病毒 HCoV-EMC 的出现和随后的人际传播都说明了 CoV 的运动潜力和传播性，并强调了紧迫性迫切需要一种广泛有效的治疗方法。项目 2 的总体目标是确定两个高度保守的 CoV 过程（复制保真度和 RNA 封端）的抑制剂，这对于 SARS-CoV 的毒力和体内存活至关重要。多种病毒蛋白和酶活性对于这些过程至关重要，包括 CoV 3'-to 5' 核糖核酸外切酶（保真度；nsp14-ExoN）和 2'-0-甲基转移酶（加帽；nsp16-0MTase）活性。与这些过程的重要性一致，我们已经表明，分别通过 ExoN 或 OMTase 的基因失活来降低复制保真度和消除 RNA 帽，导致具有复制能力的病毒在体内深度减毒。在目标 1 和 2 中，我们将与筛选核心（核心 B）和药物化学先导开发核心（核心 C）合作，识别、表征和优化 SARS-CoV 保真度和 RNA 封端的小分子抑制剂。一旦确定了活性化合物，我们将定义它们的作用机制，测试病毒抗性的发展，并确定它们在冠状病毒家族中的活性。在 Aim S 中，我们将与 Core C 合作，在 SARSCoV 疾病严重程度的分级模型中进行化学优化和测试先导化合物的体内功效，并评估体内耐药性的发展。 Denison 和 Baric 实验室的互补专业知识、广泛的初步数据集、最先进的技术以及 SR 在药物化学、高通量筛选和药物开发领域的专业知识将为成功识别、先导化合物的确认和体内测试。最终，抑制体内发病机制所需的这两种保守且不同的途径将有助于治疗地方性和新出现的冠状病毒，并有可能减少病毒耐药性的出现。