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.

项目总结