Screening and development of anti-Ebola entry inhibitors

埃博拉和马尔堡病毒属于丝状病毒科，可导致致命的出血热 以广泛的组织破坏为特征，潜伏期为4-14天。因为 出于安全考虑，这些病毒被指定为生物安全4级制剂。黄曲霉毒素的研究进展 部分出于安全考虑，丝状病毒进入的机制和致病机制受到了阻碍。 目前还没有针对丝状病毒感染和发病机制的有效疫苗或治疗方法。 在人类身上。此外，为大部分人接种疫苗可能永远不是完全实际或可取的。 由于接种疫苗所需的最短时间至少是 一个月，丝状病毒的暴发是零星的。因此，必须确定和 开发有效的抑制丝状病毒感染的药物。在这个应用程序中，我们演示了埃博拉病毒 糖蛋白(GP)可以有效地结合到HIV病毒颗粒上，而假型病毒粒子可以 介导有效的病毒进入。更重要的是，在初步的实验中，我们已经证明了这一点 系统可用作一种强大的分析方法来筛选和识别可以阻止埃博拉病毒进入 目标单元格。这项应用的总体目标是筛选和开发能够 特别是抑制埃博拉病毒进入。本申请将侧重于以下三个具体目标：(1) 迭代合成化学优化先导化合物的研究。(2)小鼠的筛选和鉴定 使用高通量筛查(HTS)抑制埃博拉病毒进入的化合物。和(3)验证和 埃博拉进入抑制剂的开发。这些研究应该识别和开发有效的进入抑制剂。 对抗埃博拉病毒感染和疾病。 摘要 在原提案《抗埃博拉病毒进入抑制剂的筛选与开发》(NIH UO1 AI77767- 01)，提出了三个具体目标：(1)。迭代合成法优化先导化合物 化学，(2)。筛选和鉴定抑制埃博拉病毒侵入的小分子化合物，以及(3)。验证 以及埃博拉进入抑制剂的开发。因为我们在原计划中提出了五年的研究计划 应用程序，我们现在希望修改原始应用程序的范围，以反映 申请(两年而不是五年)。在此，我们将重点介绍 以下各节将详细介绍当前的应用程序和这些更改： 1.当前申请将具有与原始申请相同的三个具体目标 (见上)，但拟议的实验将更集中于反映较短的持续时间 研究计划。 2.在最初的申请中，我们提出了用动物来测试一系列埃博拉病毒进入抑制剂 模特们。然而，我们认为，动物实验不太可能像最初那样 拟议的活动将在两年内进行。我们要求删除该部分的 所有拟议的实验，以及Viktor Volchkov博士每年75,000美元的分包合同 集团在法国。因此，当前的应用不需要动物协议。 3.在最初的应用中，我们提出了测试和验证最有希望的进入抑制剂 在BSL-4设施中使用传染性埃博拉病毒(目标3)。这些实验在 目前的申请，他们将与丽莎·亨斯利博士和吉恩博士合作进行 奥林格在USAMRIID。我们已经与这些埃博拉研究人员达成了合作协议 专家(见所附协议)。然而，我们不要求这一部分的任何预算 由于汉斯利博士和奥林格博士有其他财政支持来实施拟议的 实验。 需要强调的是，由于其对人类的高致病性，埃博拉病毒被认为是一种 严重的公共卫生威胁，是美国HHS公共卫生紧急医疗体系的组成部分 确保企业战略(HHS PHEMCE战略，《联邦公报》，2007年，72(53)，13109- 13114)。因此，目前的应用对国家安全具有重大影响。

Ebola and Marburg viruses belong to the family Filoviridae and can cause fatal hemorrhagic fevers characterized by widespread tissue destruction with an incubation period of 4-14 days. Because of the safety concerns, these viruses are designated as the biosafety level 4 agents. Progress in elucidation of the mechanisms of filoviral entry and pathogenesis has been hampered partly due to the safety concern. Currently there is no effective vaccine or therapeutic treatment against filoviral infection and pathogenesis in human. Further, it may never be entirely practical or desirable to vaccinate large portions of the population against this disease due to the facts that the minimal time required for vaccination is at least one month and that the filoviral outbreaks are sporadic in nature. Thus it is imperative to identify and develop potent inhibitors against filoviral infection. In this application, we demonstrate that Ebola glycoproteins (GP) can be efficiently incorporated onto HIV viral particles, and the pseudotyped virions can mediate efficent viral entry. More importantly, in the preliminary experiments, we have shown that this system can be used as a powerful assay to screen and identify molecules that can block Ebola entry to the target cells. The overall objective of this application is to screen and develop molecules which can specifically inhibit Ebola viral entry. This application will focus on the following three specific aims: (1) Optimization of lead compounds by iterative synthetic chemistry. (2)Screening and identification of small compounds which inhibit Ebola entry using high throughput screenings (HTS). And (3) Validation and development of Ebola entry inhibitors. These studies should identify and develop potent entry inhibitors against Ebola viral infection and disease. Summary In the original proposal "Screening and development of anti-Ebola entry inhibitors" (NIH UO1 AI77767- 01), we proposed three specific aims: (1). Optimization of the lead compounds by iterative synthetic chemistry, (2). Screening and identification of small compounds that inhibit Ebola entry, and (3). Validation and development of Ebola entry inhibitors. Since we proposed the five years of research plan in the original application, we now wish to modify the scope of the original application to reflect the shorter duration of the application (two years instead of five years). Here we will highlight the proposed major modifications in the current application and these changes will be elaborated in the following sections: 1. The current application will have the same three specific aims as stated in the original application (see above), but the proposed experiments will be more focused to reflect the shorter duration of the research plan. 2. In the original application, we proposed to test a series of Ebola entry inhibitors using animal models. However, we believe that it is highly unlikely that animal experiments as originally proposed will be performed during a two-year period. We request to remove that part of the proposed experiments altogether, and the subcontract of $75,000/per year for Dr. Viktor Volchkov's group in France. Therefore no animal protocol is needed for the current application. 3. In the original application, we proposed to test and validate the most promising entry inhibitors using infectious Ebola virus in a BSL-4 facility (Aim 3). These experiments are important in the current application, and they will be performed in collaboration with Drs. Lisa Hensley and Gene Olinger at USAMRIID. We have set up a collaboration agreement with these Ebola research experts (see the attached agreement). However, we do not request any budget for this part of work since Drs. Hansley and Olinger have other financial supports to carry out the proposed experiments. It is important to emphasize that due to its highly pathogenic nature to humans, Ebola virus is considered a serious public health threat, and it is an integral part in the US HHS Public Health Emergency Medical Contermeansures Enterprise Strategy (HHS PHEMCE Strategy, Federal Register, 2007, 72(53), 13109- 13114). Therefore, the current application has great implications for the national security.