Project 6 - Development of Antivirals against Alphaviruses

项目 6 - 开发抗甲病毒的抗病毒药物

基本信息

批准号：
10513947
负责人：
MARGARET KIELIAN
金额：
$ 293.23万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
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/10513947
关键词：
Active Sites Acute Alphavirus Alphavirus Infections Antiviral Agents Antiviral Therapy Arthritogenic Automobile Driving Biochemical Biological Assay Cell Culture System Cell Culture Techniques Cell Line Cells Chemicals Chemistry Chikungunya virus Chronic Chronic Phase Collaborations Complex Data Development Disease Dose Encephalitis Viruses Equine Encephalomyelitis Genomics Goals Health Homology Modeling Human Infection Libraries Mayaro virus Mediating Modeling Mus Musculoskeletal Diseases Noise Nonstructural Protein Oral Pathogenicity Peptide Hydrolases Performance Pharmaceutical Chemistry Primary Infection Prodrugs Production Prophylactic treatment Protease Inhibitor Proteins Public Health Publishing RNA Viruses RNA replication Replicon Reporter Resistance Resistance profile Ribonucleosides Ross river virus Sensitivity and Specificity Signal Transduction Specificity Structure System Testing Therapeutic Time Tissues Vaccine Therapy Venezuelan Venezuelan Equine Encephalitis Virus Viral Viremia Virus Virus Diseases Virus Replication Western Equine Encephalitis Virus Work analog antiviral drug development arthropathies base biodefense chikungunya infection chronic infection counterscreen cytotoxicity design drug structure efficacy evaluation fitness high throughput screening human pathogen in silico in vivo inhibitor innovation joint infection mouse model nanoluciferase novel nucleoside inhibitor preclinical development prevent protein structure resistance mutation response small molecule inhibitor small molecule libraries tool

项目摘要

Alphaviruses are enveloped plus-sense RNA viruses that include a number of important human pathogens such as the arthritogenic alphaviruses chikungunya virus (CHIKV), Mayaro virus, and Ross River virus, and encephalitic alphaviruses such as Eastern and Venezuelan equine encephalitis viruses. These viruses have emerged as world-wide public health and/or biodefense threats, but to date there are no licensed vaccines or antiviral therapies. Project 6 in the AC/DC seeks to develop orally available direct-acting antivirals against alphavirus infection. We focus on targeting the RNA replication complex, which is formed by the coordinated activities of the four alphavirus non-structural proteins (nsP1-4). Promising preliminary data with our existing chemical assets demonstrate inhibition of CHIKV RNA replication by nucleosides and by inhibitors of the essential protease activity of nsP2. Moreover, prophylactic treatment of mice with our ribonucleoside EIDD-2749 prevented CHIKV viremia and disease. We will build on these findings through the following Aims, in close collaboration with the AC/DC Cores: 1. Optimize and characterize inhibition by the previously identified nucleoside inhibitors EIDD-2749, EIDD- 2997, and additional prodrugs/analogs developed from AC/DC SAR studies. We will define their mechanism of action using our panel of available protein, cell-based, and virus infection assays. We will determine the breadth of inhibition across other alphaviruses and determine resistance profiles and effects on virus replication in cell culture. 2. Perform high throughput screening for inhibitors of CHIKV RNA replication using a cell-based replicon reporter system. Hits will be progressed to preclinical development in collaboration with Cores B and C, and mechanisms defined as in Aim 1. 3. Optimize and characterize inhibitors of the nsP2 protease. We will optimize and further develop our initial inhibitors of nsP2 protease, and use a cell-based screen to identify additional nsP2 protease inhibitors. We will define their mechanisms by using cell-free nsP2 enzymatic assays and virus infection, as well as by applying the strategies described in Aims 1 and 2. 4. Characterize in vivo efficacy. We will use established mouse models to determine the in vivo efficacy of EIDD-2749 and early leads from Aims 1-3 against acute and chronic CHIKV infection and disease. We will also develop a novel reporter mouse line with an integrated alphavirus minigenome template that can detect alphavirus-mediated RNA replication with high specificity and sensitivity. This strategy will be used to follow infection by unmodified alphaviruses, to identify target cells during the acute and chronic phases of CHIKV infection, and to evaluate antiviral therapies developed in this proposal.

α病毒是包裹的正义RNA病毒，其中包括许多重要的人类病原体作为关节炎α病毒Chikungunya病毒（CHIKV），玛雅罗病毒和罗斯河病毒，以及脑甲状腺病毒，例如东部和委内瑞拉马脑炎病毒。这些病毒具有出现是全球公共卫生和/或生物消除威胁，但迄今为止没有许可疫苗或抗病毒疗法。 AC/DC中的项目6旨在开发针对口服的直接作用抗病毒药 α病毒感染。我们专注于靶向RNA复制复合络合物，该复制复合络合物由协调形成四种非结构蛋白（NSP1-4）的活性。我们现有的有希望的初步数据化学资产表明核苷抑制了CHIKV RNA复制和抑制剂。 NSP2的必需蛋白酶活性。此外，用核糖核苷EIDD-2749对小鼠进行预防治疗预防CHIKV病毒血症和疾病。我们将通过以下目标建立这些发现，并在与AC/DC核心的合作： 1。通过先前鉴定的核苷抑制剂EIDD-2749，EIDD-优化和表征抑制 2997，以及来自AC/DC SAR研究开发的其他前药/类似物。我们将定义它们的机制使用我们的可用蛋白质，基于细胞和病毒感染测定法的面板的作用。我们将确定广度在其他α病毒之间的抑制作用，并确定细胞中病毒复制的抗性谱和影响文化。 2。使用基于细胞的复制子进行CHIKV RNA复制抑制剂的高吞吐量筛选记者系统。通过与核心B和C合作，将以临床前开发进行启动，并将机制定义为AIM 1。 3。优化和表征NSP2蛋白酶的抑制剂。我们将优化并进一步发展我们的初始 NSP2蛋白酶的抑制剂，并使用基于细胞的筛选来识别其他NSP2蛋白酶抑制剂。我们将通过使用无细胞NSP2酶试验和病毒感染来定义其机制，并应用目标1和2中描述的策略。 4。表征体内功效。我们将使用已建立的鼠标模型来确定体内功效 EIDD-2749和AIMS 1-3的早期导线针对急性和慢性CHIKV感染和疾病。我们也会使用一个可以检测 α病毒介导的RNA复制具有高特异性和灵敏度。该策略将用于遵循通过未修饰的α病毒感染，以鉴定CHIKV急性和慢性阶段的靶细胞感染，并评估本提案中开发的抗病毒疗法。