Identification of Small Molecular Inhibitors of Rift Valley Fever Virus Replication

裂谷热病毒复制小分子抑制剂的鉴定

• 批准号: 10250003
• 负责人: Wenjun Ma
• 金额: $ 18.24万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-08 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10250003
• 关键词: Acute; Affect; Africa; African; Animals; Antiviral Agents; Biological Assay; Blood - brain barrier anatomy; Brain; Bypass; Cattle; Cells; Chemicals; Collection; Country; Development; Disease; Disease Outbreaks; Dose; Encephalitis; Evaluation; Fever; Goals; Goat; Health; Hemorrhage; Hepatitis; Human; In Vitro; Inbred BALB C Mice; Infection; Kansas; Knockout Mice; Knowledge; Laboratories; Libraries; Mitoxantrone; Mus; Pharmaceutical Preparations; Polymerase; Public Health; Renilla; Renilla Luciferases; Research; Research Activity; Retinitis; Ribavirin; Rift Valley Fever; Rift Valley fever virus; STAT1 gene; Sheep; Structure; Syndrome; Testing; Therapeutic; United States National Institutes of Health; Universities; Vaccines; Virulent; Virus; Virus Diseases; Virus Inhibitors; Virus Replication; Work; Zoonoses; abortion; base; biocontainment facility; biosafety level 2 facility; cytotoxicity; design; detection assay; drug candidate; high throughput screening; in vivo; inhibitor/antagonist; insight; juvenile animal; mortality; mosquito-borne; mouse model; novel; novel therapeutics; screening; small molecular inhibitor; small molecule; small molecule libraries

Project Summary Rift Valley fever caused by Rift Valley fever virus (RVFV) is an acute, mosquito-borne, fever-causing zoonotic disease that affects both humans and animals. Large Rift Valley fever outbreaks have occurred throughout Africa and more recently in the Arabian Peninsula. Because RVFV is no longer restricted to African countries, it has raised concerns that the disease could spread worldwide. The RVFV is a select agent that requires high biocontainment facilities. This limitation has hampered the development of RVFV antivirals and vaccines. Despite the significant impact of the disease to the economy and public health, there are no fully licensed vaccine and antivirals available in the US for human and animal use. It is urgent to identify and develop effective inhibitors against RVFV to treat exposed and infected humans and animals. Dr. Ma’s group has developed a cell-based screening assay based on the RVFV MP12 vaccine strain that expresses Renilla Luciferase using Renilla as readout to identify RVFV inhibitors, and established a STAT1-KO mouse model susceptible to infection with MP12 vaccine strain that can be used in a BSL-2 facility. Furthermore, they have screened 727 compounds from the NIH collections of which two candidates including 6-azauridine and mitoxantrone inhibited replication of MP12. They hypothesize that effective inhibitors against RVFV can be identified by screening large compound collections and by further optimization of their structures and activities, and the mechanisms of inhibitory effects of identified candidates can be determined. Thus, they plan to use the developed high-throughput assay to identify inhibitors against RVFV, evaluate their efficacy in vitro and in mice, and understand the underlying mechanisms of inhibitory effects of identified candidates through two specific aims in this R21 proposal. In specific aim 1, the libraries assembled by the University of Kansas High Throughput Screening Laboratory that contain approximately 26,000 chemical compounds will be screened. The 26,000 compounds are predicted to cross the blood-brain barrier. Hits from the library will help overcome the challenge for RVFV antivirals to reach the brain through the blood-brain barrier to protect encephalitis. Primary hits will be confirmed by in vitro and in vivo assays. In specific aim 2, 6-azauridine and mitoxantrone as well as others identified in Aim 1 will be evaluated in vitro and in the BALB/c mice using virulent RVFV. To understand the underlying mechanisms of their inhibitory effects, whether the compounds block virus entry, inhibit virus replication and polymerase activity will be investigated using different designed assays. The results of this study could identify novel effective inhibitors against RVFV and understand the underlying mechanisms of their inhibitory effects, and offer novel insights toward the design of novel antiviral drugs against this zoonotic disease that will benefit both human and animal health.

项目摘要 裂谷谷热病毒（RVFV）引起的裂谷谷热是一种急性，蚊子引起的发烧 动物和动物都会影响人的动物疾病。发生了大裂谷热暴发 通过非洲，最近在阿拉伯半岛。因为RVFV不再限于非洲 国家，人们一直担心这种疾病可能在全球范围内传播。 RVFV是一个选择代理 需要高生物内在设施。这种限制阻碍了RVFV抗病毒药的发展 疫苗。尽管该疾病对经济和公共卫生产生了重大影响，但没有完全 美国和动物使用的有执照的疫苗和抗病毒药可用于人类和动物。迫切要识别和发展 对RVFV的有效抑制剂治疗暴露和感染的人和动物。 MA博士的小组基于RVFV MP12疫苗菌株开发了基于细胞的筛查测定法 使用Renilla作为读数来识别RVFV抑制剂，并建立了STAT1-KO 小鼠模型可在BSL-2设施中使用MP12疫苗菌株感染。此外， 他们从NIH系列中筛选了727种化合物，其中两个候选人在内 Mitoxantrone抑制MP12的复制。他们假设有效针对RVFV的抑制剂可能是 通过筛选大型化合物收集并进一步优化其结构和活动， 并且可以确定已确定的候选物的抑制作用机制。那就是他们计划使用 开发了高通量测定法以鉴定针对RVFV的抑制剂，评估其体外和小鼠的效​​率， 并了解通过两个特定的识别候选物的抑制作用的潜在机制 目的是该R21提案。在特定目标1中，图书馆由堪萨斯大学高通量组装 将筛选大约26,000种化合物的筛选实验室。 26,000 预计化合物可以越过血脑屏障。图书馆的命中将有助于克服挑战 为了使RVFV抗病毒药通过血脑屏障到达大脑以保护脑炎。主要热门单曲将是 通过体外和体内屁股确认。在特定的目标2中，6-唑啉和mitoxantrone以及其他 在AIM 1中鉴定的，将在体外和BALB/C小鼠中使用毒力RVFV进行评估。理解 它们的抑制作用的基本机制，这些化合物是否会阻断病毒的进入，抑制病毒 将使用不同的设计测定法研究复制和聚合酶活性。这项研究的结果 可以识别出针对RVFV的新型有效抑制剂，并了解其基本机制 抑制作用，并提供针对这种动物疾病的新型抗病毒药物设计的新见解 这将使人类和动物健康受益。