Structure-based Antiviral Design against HTLV-1 Protease

基于结构的 HTLV-1 蛋白酶抗病毒设计

• 批准号: 10750889
• 负责人: Sarah N Zvornicanin
• 金额: $ 3.5万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10750889
• 关键词: 2019-nCoV; Active Sites; Adult; Adult T-Cell Leukemia/Lymphoma; Advisory Committees; Advocate; Affect; Affinity; Amino Acid Sequence; Antiviral Agents; Aspartic Endopeptidases; Binding; Biochemical; Biological Assay; Carcinogens; Communities; Crystallography; Disease; Environment; Evolution; Family; HIV; HIV Protease; HIV-1; HIV-1 protease; HIV/HCV; Health; Hepatitis C virus; Human; Human T-lymphotropic virus 1; Individual; Infection; Inflammatory; Investigation; Laboratories; Lead; Life Cycle Stages; Molecular; Mutation; Oncogenic; Patients; Peptide Hydrolases; Persons; Polyproteins; Population; Positioning Attribute; Prevalence; Protease Inhibitor; Proteins; Relapse; Reporting; Research; Resistance; Retroviridae; SARS coronavirus; Series; Shapes; Site; Specificity; Structure; Substrate Interaction; Substrate Specificity; T-Cell Lymphoma; Techniques; Testing; Training; Translating; Vaccines; Viral; Viral Proteins; Virus; X-Ray Crystallography; analog; chronic infection; design; experience; infection rate; inhibitor; insight; laboratory experience; meter; molecular dynamics; novel; pressure; prevent; protease So; protein purification; rational design; skills

Project Summary Human T-cell leukemia virus type-1 (HTLV-1) is an oncogenic human retrovirus affecting over 20 million people worldwide. HTLV-1 infection can cause adult T-cell lymphoma (ATL) and other serious inflammatory diseases. Estimates report that 5-10% of HTLV-1 infected patients will develop a serious condition such as ATL, which has poor 4-year survival and high relapse rates. HTLV-1 has persistent infection rates across the globe and reaches up to 45% prevalence in certain communities. Despite this impact on human health, there are no direct-acting antivirals (DAAs) or vaccines against HTLV-1. HIV-1 and HTLV-1 are from the same viral family and encode for a homodimeric aspartyl protease crucial for cleavage of functional proteins from viral polyproteins. The activity of HIV-1 and HTLV-1 protease is essential to their viral life cycles. The Schiffer laboratory has extensive experience with viral protease crystallography and inhibition, especially with viral proteases for HIV-1, HCV NS3/4A, and SARS-CoV-2 main protease. This expertise uniquely positions me to design, synthesize, and characterize potent, resistance-thwarting protease inhibitors against HTLV-1 protease. Resistance-preventing DAA design is essential because of the selective pressure applied during DAA treatment. An ideal and proven strategy for developing a highly potent and resistance-preventing viral protease inhibitor is to target the active site through rational design using the substrate envelope. The substrate envelope for HTLV-1 protease has not been characterized and we lack a detailed understanding of the protease substrate specificity. I hypothesize that by translating strategies from our design of HIV-1 protease inhibitors, namely characterizing HTLV-1 protease’s substrate specificity, I can design potent and resistance-preventing DAAs for HTLV-1 protease. Aim 1: Characterize the structural basis for HTLV-1 protease substrate specificity. HTLV-1 protease cleaves six substrates by recognizing cleavage sites between individual proteins of the viral polyprotein. I will investigate the molecular basis of this recognition underlying protease specificity by determining cocrystal structures of the protease with bound substrates. The conserved volume inhabited by the substrates will define the substrate envelope and inform inhibitor design for HTLV-1 protease. Aim 2: Rationally design, synthesize, and characterize inhibitors of HTLV-1 protease to optimize potency. HIV-1 and HTLV-1 proteases share an active site amino acid sequence identity of 45% and high structural similarity. Therefore, I will begin inhibitor design by testing a selection of our in-house HIV-1 protease inhibitors, which have already shown low (1 µM) to moderate (30 nM) potency against HTLV-1 protease. I will combine experimental inhibition assays with cocrystal structure analysis to identify lead compounds for inhibitor design. I will leverage substrate specificity of the protease by moving inhibitor design towards compounds that mimic the shape of substrates, leveraging the substrate envelope (Aim 1), and the interactions between protease and substrate. I aim to produce novel, highly potent (sub-nM) inhibitors that will be promising DAAs for further investigation against HTLV-1.

项目摘要 人类T细胞白血病病毒类型1（HTLV-1）是一种影响超过2000万人的致癌人逆转录病毒 全世界。 HTLV-1感染会引起成人T细胞淋巴瘤（ATL）和其他严重的炎症性疾病。 估计报告说，有5-10％的HTLV-1感染患者将发展出严重的状况，例如ATL 4年生存率和高继电器率较差。 HTLV-1在全球范围内具有持续的感染率 在某些社区中，多达45％的患病率。尽管对人类健康的影响，但没有直接行动 针对HTLV-1的抗病毒药（DAA）或疫苗。 HIV-1和HTLV-1来自同一病毒家族，并编码 同二聚体蛋白酶对于从病毒多蛋白裂解功能蛋白的裂解至关重要。活动 HIV-1和HTLV-1蛋白酶的病毒生命周期至关重要。 Schiffer实验室有广泛的 具有病毒蛋白酶晶体学和抑制作用的经验，尤其是用于HIV-1的病毒蛋白酶，HCV NS3/4A和SARS-COV-2主要蛋白酶。这种专业知识独特地定位了我设计，合成和 表征了针对HTLV-1蛋白酶的有效抗抗性蛋白酶抑制剂。预防性 DAA设计是必不可少的，因为在DAA处理过程中施加了选择性压力。理想和经过验证的 开发高潜力和耐药性的病毒蛋白酶抑制剂的策略是针对活动 通过使用底物信封的理性设计站点。 HTLV-1蛋白酶的底物信封尚未 我们的特征是对蛋白酶底物特异性的详细理解。我假设这一点 通过翻译我们的HIV-1蛋白酶抑制剂设计的策略，即表征HTLV-1蛋白酶的策略 底物特异性，我可以针对HTLV-1蛋白酶设计潜在的和抗性的DAA。 目标1：表征HTLV-1蛋白酶底物特异性的结构基础。 HTLV-1蛋白酶 通过识别病毒多蛋白的各个蛋白质之间的切割位点来切割六个底物。我会 通过确定共晶的蛋白酶特异性来研究这种识别基础蛋白酶特异性的分子基础 蛋白酶的结构与结合的底物。由基板感染的配置卷将定义 底物包络并告知HTLV-1蛋白酶的抑制剂设计。目标2：合理设计，合成， 并表征HTLV-1蛋白酶的抑制剂以优化效力。 HIV-1和HTLV-1蛋白酶共享 活性位点氨基酸序列身份为45％，结构相似性高。因此，我将开始抑制剂 通过测试我们的内部HIV-1蛋白酶抑制剂的选择，这些抑制剂已经显示出低（1 µm） 针对HTLV-1蛋白酶的中度（30 nm）效力。我将将实验抑制测定法与共晶法相结合 结构分析以识别抑制剂设计的铅化合物。我将利用 蛋白酶通过将抑制剂设计转向模仿底物形状的化合物，利用 底物包膜（AIM 1）以及保护酶和底物之间的相互作用。我的目标是制作小说 潜在的（子-NM）抑制剂将有助于DAA，以进一步研究HTLV-1。