Identification of Ebola Virus Entry Inhibitors

埃博拉病毒进入抑制剂的鉴定

• 批准号: 7155577
• 负责人: Arnab Basu
• 金额: $ 35.64万
• 依托单位: MICROBIOTIX, INC
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-01 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7155577
• 关键词: cytotoxicity; infection; lead; library; life cycle; model; small molecule; virus infection mechanism

DESCRIPTION (provided by applicant): Ebola virus (EBOV) causes periodic outbreaks of severe viral hemorrhagic fevers in Africa with high mortality rates in infected patients. EBOV can be used in acts of bio-terrorism because of its highly infectious nature and stability in aerosolized form. It is classified as a Category A bioweapons agent by the Centers for Disease Control and Prevention (CDC). Currently, there is no FDA approved vaccine or antiviral drug that is effective against EBOV infections in humans. However, vaccines will only be partially effective in controlling an EBOV outbreak or bioterrorist attack since rapid progression of EBOV infection offers little opportunity for developing acquired immunity. Therefore, there is a critical need for to respond to postexposure or bioterrorist attack. EBOV infection is initiated by the fusion between viral and host cell membranes, which is mediated by the viral membrane glycoprotein (GP). This selective interaction between EBOV GP and host cell surface receptor molecules is essential for the initiation and establishment of the infection. Therefore, blocking of EBOV entry into its target cell will be advantageous since this will lead to suppression of viral infectivity early in its life cycle. Our objective is to discover and develop small molecule entry inhibitors against EBOV infection. Considering the aggressive nature of EBOV infection, and its extreme virulence, biosafety level 4 containment facilities are required to study the virus. To circumvent these stringent biohazard conditions, we will generate EBOV pseudotype virus as surrogate model to study EBOV attachment/entry. This model system allows the study of viral entry into the cell without replicating other aspects of the viral life cycle and therefore, can be performed in a biosafety level 2 facility. In Phase I, we will develop a high-throughput screening (HTS) assay utilizing the EBOV pseudotype virus to measure virus infection. This assay will be used to screen libraries of structurally diverse small molecules in "Microbiotix Inc's" repository, to identify potent inhibitors of EBOV entry. Inhibitors will be further confirmed for antiviral activity in a secondary EBOV pseudotype virus plaque assay, and infectious EBOV plaque assay. They will also be assessed for mammalian cytotoxicity. In Phase II, we will progress the most promising scaffolds through a rational drug design program, and lead compounds will be tested for efficacy and toxicity in animal models. The most active compound, with the least toxicity, will advance to IND enabling studies (Phase III). Therefore the specific aims of this Phase I application are to: 1. Develop a high-throughput screening (HTS) assay for selecting inhibitors of EBOV attachment/entry; 2. Screen a diverse compound library "Microbiotix Inc."s repository to identify and confirm selective inhibitors of EBOV; 3. EBOV inhibitor confirmation; and 4. Prioritize screening hits based on spectrum, cytotoxicity and mechanism. The end result of Phase 1 work will be identification of hits specific for EBOV entry inhibition.

描述（由申请人提供）：埃博拉病毒（EBOV）导致非洲严重病毒出血发作的周期性暴发，感染患者死亡率很高。 EBOV可以用于生物恐怖主义的行为，因为它具有高度感染性和稳定性。它被归类为疾病控制与预防中心（CDC）的生物武器代理类别。当前，尚无FDA批准的疫苗或抗病毒药，可有效抵抗人类EBOV感染。但是，疫苗只能在控制EBOV爆发或生物恐怖主义攻击方面有效地有效，因为EBOV感染的快速发展几乎没有机会开发获得的免疫力。因此，迫切需要应对暴露后或生物恐怖主义攻击。 EBOV感染是由病毒和宿主细胞膜之间的融合引发的，该病毒细胞膜是由病毒膜糖蛋白（GP）介导的。 EBOV GP与宿主细胞表面受体分子之间的这种选择性相互作用对于开始和建立感染至关重要。因此，阻止EBOV进入其目标细胞将是有利的，因为这将导致其生命周期早期抑制病毒感染性。我们的目标是发现和开发针对EBOV感染的小分子进入抑制剂。考虑到EBOV感染的侵略性及其极端的毒力，生物安全4级遏制设施需要研究病毒。为了避免这些严格的生物危害条件，我们将生成EBOV伪型病毒作为替代模型，以研究EBOV附着/进入。该模型系统允许研究病毒进入细胞，而无需复制病毒生命周期的其他方面，因此可以在生物安全2级设施中进行。在第一阶段，我们将利用EBOV伪型病毒来测量病毒感染，开发高通量筛选（HTS）测定。该测定法将用于筛选“ Microbiotix Inc”存储库中结构多样的小分子的库，以鉴定EBOV进入的有效抑制剂。将进一步证实抑制剂在继发性EBOV伪型病毒斑块测定和感染性EBOV斑块测定中的抗病毒活性。还将评估它们的哺乳动物细胞毒性。在第二阶段，我们将通过合理的药物设计程序来促进最有希望的脚手架，并将测试铅化合物在动物模型中的疗效和毒性。最活跃的化合物，最少的毒性将促进IND促进研究（第三阶段）。因此，该阶段应用程序的具体目的是：1。开发用于选择EBOV附件/进入的抑制剂的高通量筛选（HTS）测定； 2.屏幕一个多样化的复合库“ Microbiotix Inc.”的存储库来识别和确认EBOV的选择性抑制剂； 3。EBOV抑制剂确认；和4。根据频谱，细胞毒性和机制确定筛选的优先次序。第1阶段工作的最终结果将是识别特定于EBOV进入抑制的命中。