Evaluation and development of E1-TopoI as a target for anti-HPV therapeutics

E1-TopoI 作为抗 HPV 治疗靶点的评估和开发

• 批准号: 8297142
• 负责人: THOMAS MELENDY
• 金额: $ 43.8万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8297142
• 关键词: AIDS/HIV problem; Amino Acids; Anogenital venereal warts; Antiviral Agents; Binding; Bioinformatics; Biological Assay; C-terminal; Cell Culture Techniques; Cells; Cervical; Chemicals; Computer software; DNA Primase; DNA biosynthesis; Data; Development; Docking; Enzyme-Linked Immunosorbent Assay; Enzymes; Evaluation; Excision; Generations; Genome; Goals; Health Professional; Herpesviridae; Highly Active Antiretroviral Therapy; Human; Human Activities; Human Papilloma Virus Vaccine; Human Papillomavirus; Human papilloma virus infection; Human papillomavirus 16 E1 protein; Imiquimod; Immune; Immune system; In Vitro; Individual; Infection prevention; Lesion; Malignant Neoplasms; Maps; Modeling; Molecular; Mutagenesis; Mutation; Mutation Analysis; Papillomavirus; Patients; Peptides; Prevention; Protein Databases; Proteins; Reliance; Research; Screening procedure; Sexually Transmitted Diseases; Structure; Testing; Therapeutic; Topoisomerase; Topoisomerase Interaction; Type I DNA Topoisomerases; Vaccines; Viral; Virus; Visit; X-Ray Crystallography; base; expression vector; helicase; immunological intervention; immunoregulation; inhibitor/antagonist; malignant mouth neoplasm; mutant; novel strategies; nucleotide analog; prevent; prophylactic; small molecule; small molecule libraries; success; weapons

DESCRIPTION (provided by applicant): Human papillomavirus (HPV) is the most common sexually-transmitted infection, and the cause of nearly all cervical and anogenital, and over half of oral cancers. Current HPV treatment is by lesion removal or through immunological intervention (imiquimod as an immune stimulant, or the HPV vaccines to prevent infection of the most common HPVs). While antiviral agents have been developed against many types of viruses, to date no true antivirals are available against HPV. With the heavy reliance on the immune system for HPV treatment/prevention, HPV infections and cancers remain a major problem for HIV/AIDS patients, even after HAART treatment. A true HPV antiviral that acts directly against HPV would be an important weapon against HPV infections and cancers. Recent successes of small molecule inhibitors that interfere with the herpesvirus primase- helicase interaction justify using such an approach against HPV. We have identified an interaction between the HPV DNA replication helicase, E1, and human Topoisomerase I that is vital for HPV genome duplication. Aim 1 will evaluate a panel of E1 mutations predicted to disrupt the interaction with Topoisomerase to more fully define this interaction. In Aim 2 peptide binding will be used to further define the interaction domain, and our E1-Topoisomerase interaction assays will be used to screen panels of peptides and small molecules to identify first-generation inhibitors of this interaction. In Aim 3 the structure of this interaction will be evaluted using predictive software, NMR, X-ray crystallography, and SAXS. Each Aim can be achieved independently, yet information from each synergizes and enhances the others. Multiple achieveable approaches in each Aim allows for attaining the goals without requiring success of all approaches in each Aim. Results will provide chemical and structural information that will be used in developing second-generation inhibitors that will be investigated as potential antiviral therapeutics against papillomaviruses. PUBLIC HEALTH RELEVANCE: We have identified an interaction between the human papillomavirus (HPV) DNA replication protein, E1, and a cellular enzyme, Topoisomerase I, which appears essential for papillomavirus genome duplication. In the proposed research we will define the E1-Topoisomerase I interaction through mutational analysis, identification of peptide and small molecule inhibitors of this interaction, and through structural studies, with the long-term goal of developing highly specific peptide or small molecule inhibitors of the interaction that can act as HPV antiviral agents.

描述（由申请人提供）：人乳头瘤病毒 (HPV) 是最常见的性传播感染，也是几乎所有宫颈癌和肛门生殖器癌以及一半以上口腔癌的病因。目前的 HPV 治疗是通过病灶去除或通过免疫干预（咪喹莫特作为免疫刺激剂，或 HPV 疫苗来预防最常见的 HPV 感染）。虽然已经开发出针对多种病毒的抗病毒药物，但迄今为止还没有真正针对 HPV 的抗病毒药物。由于HPV治疗/预防严重依赖免疫系统，因此即使在HAART治疗之后，HPV感染和癌症仍然是HIV/AIDS患者的主要问题。直接对抗 HPV 的真正 HPV 抗病毒药物将是对抗 HPV 感染和癌症的重要武器。最近干扰疱疹病毒引物酶-解旋酶相互作用的小分子抑制剂的成功证明了使用这种方法对抗 HPV 的合理性。我们已经确定了 HPV DNA 复制解旋酶 E1 和人类拓扑异构酶 I 之间的相互作用，这对于 HPV 基因组复制至关重要。目标 1 将评估一组预计会破坏与拓扑异构酶相互作用的 E1 突变，以更全面地定义这种相互作用。在 Aim 2 中，肽结合将用于进一步定义相互作用域，我们的 E1-拓扑异构酶相互作用测定将用于筛选肽和小分子组，以鉴定这种相互作用的第一代抑制剂。在目标 3 中，将使用预测软件、NMR、X 射线晶体学和 SAXS 评估这种相互作用的结构。每个目标都可以独立实现，但每个目标的信息可以协同并增强其他目标。每个目标中的多种可实现的方法允许实现目标，而不需要每个目标中的所有方法都成功。结果将提供化学和结构信息，用于开发第二代抑制剂，这些抑制剂将作为针对乳头瘤病毒的潜在抗病毒疗法进行研究。 公共健康相关性：我们已经确定了人乳头瘤病毒 (HPV) DNA 复制蛋白 E1 与细胞酶拓扑异构酶 I 之间的相互作用，该酶对于乳头瘤病毒基因组复制至关重要。在拟议的研究中，我们将通过突变分析、鉴定这种相互作用的肽和小分子抑制剂以及通过结构研究来定义 E1-拓扑异构酶 I 相互作用。 长期目标是开发高度特异性的肽或小分子相互作用抑制剂，作为 HPV 抗病毒剂。