Small molecule inhibitors targeting filoviral infections

针对丝状病毒感染的小分子抑制剂

• 批准号: 7670045
• 负责人: Paul Bates
• 金额: $ 28.85万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-01 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7670045
• 关键词: Antiviral Agents; Binding; Biochemical; Biological Assay; Cathepsin L; Cathepsins; Cathepsins B; Cell Culture Techniques; Cell physiology; Cell surface; Cells; Cleaved cell; Coronavirus Infections; Country; Diamond; Drug resistance; Ebola virus; Endocytosis; Endosomes; Enzyme Inhibitor Drugs; Enzyme Inhibitors; Enzymes; Escape Mutant; Generations; Genetic; Glycoproteins; Human; Infection; Inhibitory Concentration 50; Knock-out; Lead; Libraries; Life; Mediating; Membrane Fusion; Methodology; Methods; Molecular; Molecular Bank; Mus; Nipah Virus; Play; Process; Protease Inhibitor; Role; SARS coronavirus; Screening procedure; Severe Acute Respiratory Syndrome; Specificity; Structure-Activity Relationship; System; Testing; Therapeutic; Time; United States National Institutes of Health; Vesicular stomatitis Indiana virus; Viral; Virus; Virus Diseases; Virus-like particle; base; biodefense; drug development; efficacy testing; genetic inhibitor; high throughput screening; improved; in vivo; inhibitor/antagonist; novel; novel virus; pathogen; resistance mechanism; small molecule; small molecule libraries; therapeutic development; vector

Recent evidence demonstrates an essential role for cellular cathepsins in viral glycoprotein processing and cellular entry for the highly pathogenic viruses Ebola, SARS coronavirus and Nipah/Hendra. Cathepsin L appears to be important for Nipah and SARS coronavirus virus glycoprotein activation while Ebola requires both cathepsins L and B for viral entry. Inhibitors of these enzymes effectively block viral entry and replication in cell culture. Studies in mice with broad spectrum cathepsin inhibitors as well as specific inhibitors and genetic knockouts all suggest that cathepsin activity can be impaired in vivo without significant detrimental effects. In preliminary studies we have used a novel methodology to screen chemical libraries on microarrays and have identified two previously unrecognized cathepsin L inhibitors. One of these inhibitors, PC-185, is highly specific for cathepsin L and blocks Ebola entry with an IC50 of ~193nM. This proposal will build upon these significant preliminary findings and will develop cathepsin inhibitors as therapeutics for Ebola. For this application, we propose the following aims: Specific Aim 1) Utilize high throughput screening of diverse libraries to identify additional new cathepsin inhibitors. Analyze structureactivity relationships (SAR), optimize lead compounds, and develop second-generation agents and screening libraries. Specific Aim 2) Test the candidate compounds for inhibition of viral entry using a rapid, quantitative and safe assay employing viral pseudotypes carrying the glycoproteins of Ebola virus. A novel virus-like particle system that we developed to study Ebola entry will be employed to confirm the ability of lead compounds to block filamentous Ebola infection. Additionally, we will send promising candidates to collaborators at USAMRIID for efficacy testing against Ebola infection. Effective candidate compounds will be cross screened against SARS-CoV and Hendra infections. Specific Aim 3) Determine the mechanism(s) by which viruses can escape cathepsin inhibition by selection of escape mutants using a novel VSV vector that relies upon Ebola GP for replication (VSV-GP).

最近的证据表明，细胞组织蛋白酶在病毒糖蛋白加工和 高度致病性病毒埃博拉病毒，SARS冠状病毒和Nipah/Hendra的细胞进入。组织蛋白酶l 对于Nipah和Sars冠状病毒病毒糖蛋白激活似乎很重要，而埃博拉则需要 两个组织蛋白酶L和B用于病毒。这些酶的抑制剂有效地阻断了病毒的进入和 细胞培养中的复制。在具有广谱组织蛋白酶抑制剂以及特定的小鼠中进行的研究 抑制剂和遗传基因敲除所有都表明组织蛋白酶活性在体内可能受到损害 有害影响。在初步研究中，我们使用了一种新颖的方法来筛选化学文库 在微阵列上，已经确定了两个先前未识别的组织蛋白酶L抑制剂。其中之一 PC-185抑制剂对组织蛋白酶L高度特异，并以〜193nm的IC50阻断埃博拉病毒的进入。这 提案将以这些重要的初步发现为基础 埃博拉病毒的治疗剂。对于此应用，我们提出以下目的：特定目的1） 吞吐量筛选不同的文库，以识别其他新的组织蛋白酶抑制剂。分析结构性 关系（SAR），优化铅化合物，并开发第二代药物和 筛选库。特定目标2）测试候选化合物，使用快速的， 使用含有埃博拉病毒糖蛋白的病毒型型的定量和安全测定。小说 我们开发用于研究埃博拉病毒进入的病毒样颗粒系统将被用来确认 铅化合物可阻断丝状埃博拉病毒感染。此外，我们将向有希望的候选人发送 USAMRIID的合作者针对埃博拉病毒感染有效测试。有效的候选化合物将 在SARS-COV和HENDRA感染上进行交叉筛选。特定目标3）确定机制 通过使用新型VSV载体选择逃生突变体，病毒可以通过选择逃避组织蛋白酶抑制 这依赖于埃博拉病毒的复制（VSV-GP）。