Inhibitors of virus glycoprotein-LAMP1 receptor binding for Lassa virus therapy

用于拉沙病毒治疗的病毒糖蛋白-LAMP1受体结合抑制剂

基本信息

批准号：
9465714
负责人：
Gai Liu
金额：
$ 29.94万
依托单位：
MICROBIOTIX, INC
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-01-15 至 2019-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9465714
关键词：
Address Adverse effects Aerosols Affect Affinity Africa Animals Antiviral Agents Antiviral Therapy Arenavirus Binding Biochemical Biological Assay Biotin Case Fatality Rates Cells Cellular Assay Cellular biology Cessation of life Chemicals Chemistry Complex DNA Viruses Disease Distal Ebola virus Environment Exhibits FDA approved Family GP2 gene Glycoproteins Goals Guanosine Health Human In Vitro Infection Institutes Interferometry Junin virus Lassa virus Lujo virus Lymphocytic choriomeningitis virus Measures Medical Membrane Membrane Proteins Methods Modeling Mus NPC1 gene Pharmaceutical Preparations Phase Property Proteolytic Processing RNA Viruses Recombinants Ribavirin Series Specificity Streptavidin Structure Surface Technology Therapeutic Translating Travel Vaccines Vesicular stomatitis Indiana virus Viral Virus Virus Diseases Virus Inhibitors analog base cellular targeting cytotoxicity experience extracellular global health high throughput screening in vivo inhibitor/antagonist medical schools novel pathogen prevent prophylactic receptor receptor binding scaffold screening small molecule small molecule inhibitor small molecule libraries success virus host interaction

项目摘要

Summary Since its discovery in 1969, Lassa virus (LASV), a bi-segmented RNA virus from the family Arenaviridae, has been recognized as the cause of disease affecting a quarter million people per year, resulting in ~5,000 deaths annually in West Africa. Due to global travel, there have been numerous confirmed cases of LASV infection outside West Africa including the US. Its ability to spread as an aerosol and a case fatality rate of ~15% make LASV a major threat to human health and a BSL 4 pathogen. Unfortunately, no FDA-approved drugs or vaccines are available for treatment of LASV. The overall goal of this project is to address this medical need by identifying and validating small molecule inhibitors of LASV infection as prophylactics and/or therapeutics. The strategy of this project is to identify small molecule inhibitors that target the cellular entry of LASV. Being the first step in viral infection and occurring in the extracellular/endosomal environment, viral entry is a susceptible and accessible target for antiviral therapy. The approach is to leverage our experience with a homogeneous, biochemical, high-throughput screening (HTS) method, AlphaLISA, to identify small molecules that prevent interactions between the glycoprotein of LASV and its host receptor LAMP1. Previously, we developed and applied a biochemical HTS based on AlphaLISA technology to identify compounds that block the Ebola glycoprotein (GP) binding to its host receptor NPC1. Two distinct scaffolds were identified and one exhibited potency against infectious Ebola virus in a murine in vivo study. In Phase I, for Aim 1, an AlphaLISA HTS will be developed and optimized for the identification of small molecules that inhibit the interaction between LASV GP1 and its receptor LAMP1. In Aim 2, a biolayer interferometry (BLI) biochemical assay and cell-based secondary assays utilizing recombinant vesicular stomatitis virus (VSV) carrying arenavirus glcyoproteins (GP) in place of VSV-GP will be built and optimized to confirm initial hits from the primary screen, to determine which interacting partner they bind, and to approximate the affinity of that interaction. In Aim 3, the HTS will be applied to diverse chemical libraries, and hits will be confirmed in the secondary assays. In Aim 4, hits will be validated in infectious Lassa virus assays and prioritized by drug-like structural features and in vitro ADME properties. Together, these assays will identify and validate compounds that suppress LASV infection by inhibiting viral entry and will provide valuable information for prioritizing those inhibitors. In Phase II, we will chemically optimize priority inhibitors for potency and selectivity and evaluate them in animal infection models.

概括自 1969 年发现以来，拉沙病毒 (LASV) 是一种来自沙粒病毒科的双节段 RNA 病毒，被认为是每年影响 25 万人的疾病原因，导致约 5,000 人死亡每年在西非。由于全球旅行，已出现多起 LASV 感染确诊病例西非以外地区，包括美国。它以气溶胶形式传播的能力和约 15% 的病死率使得 LASV 是对人类健康的主要威胁，也是 BSL 4 级病原体。不幸的是，没有 FDA 批准的药物或疫苗可用于治疗 LASV。该项目的总体目标是解决这一医疗需求通过鉴定和验证 LASV 感染的小分子抑制剂作为预防剂和/或治疗剂。该项目的策略是鉴定针对 LASV 细胞进入的小分子抑制剂。存在病毒感染的第一步发生在细胞外/内体环境中，病毒进入是抗病毒治疗的易感且可及的靶点。该方法是利用我们的经验均质、生化、高通量筛选 (HTS) 方法 AlphaLISA，用于鉴定小分子阻止 LASV 糖蛋白与其宿主受体 LAMP1 之间的相互作用。此前，我们开发并应用基于 AlphaLISA 技术的生化 HTS 来识别阻断化合物埃博拉糖蛋白（GP）与其宿主受体NPC1结合。确定了两种不同的支架，一种在小鼠体内研究中表现出对抗传染性埃博拉病毒的效力。在第一阶段，针对目标 1，AlphaLISA HTS 将被开发和优化，用于识别抑制相互作用的小分子 LASV GP1 及其受体 LAMP1 之间。在目标 2 中，生物层干涉测量法 (BLI) 生化检测和利用携带沙粒病毒的重组水泡性口炎病毒 (VSV) 进行基于细胞的二次测定将构建和优化代替 VSV-GP 的糖蛋白 (GP)，以确认主屏幕的初始命中，确定它们结合的相互作用伙伴，并估计该相互作用的亲和力。在目标 3 中， HTS 将应用于不同的化学文库，命中结果将在二次测定中得到确认。瞄准 4、命中将在传染性拉沙病毒检测中进行验证，并根据药物样结构特征和在体外 ADME 特性。这些检测方法将共同鉴定和验证抑制 LASV 的化合物通过抑制病毒进入来抑制感染，并将为优先考虑这些抑制剂提供有价值的信息。同相 II，我们将对优先抑制剂的效力和选择性进行化学优化，并在动物中对其进行评估感染模型。