Identification and characterization of novel anti-HIV inhibitors

新型抗 HIV 抑制剂的鉴定和表征

• 批准号: 8012537
• 负责人: Xiao-Fang Yu
• 金额: $ 24.6万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8012537
• 关键词: 26S proteasome; Acquired Immunodeficiency Syndrome; Antiviral Agents; Biological Assay; Biological Models; CD4 Positive T Lymphocytes; Cell Line; Cells; Cytidine Deaminase; Cytosine; Development; Goals; HIV; HIV-1; Host Defense; Human; Intervention; Knowledge; Libraries; Luminescent Measurements; Mediating; Modification; Molecular; Polyubiquitination; Proteins; Research; Reverse Transcription; Screening procedure; System; Testing; Toxic effect; Uracil; Viral; Virion; Virus; Virus Diseases; Virus Inhibitors; antiretroviral therapy; base; counterscreen; design; effective intervention; genetic regulatory protein; high throughput screening; human CEM15 protein; inhibitor/antagonist; macrophage; novel; novel strategies; public health relevance; small molecule; ubiquitin-protein ligase; vif Gene Products; viral DNA

DESCRIPTION (provided by applicant): Human cytidine deaminases APOBEC3 (A3) proteins are potent host defenses against HIV. These antiviral proteins induce lethal modification of cytosines to uracils in newly synthesized minus-strand viral DNA, resulting in abortive viral infection. HIV must overcome these host cellular defenses for successful viral replication. HIV-1 encodes a protein, Vif, which suppresses the antiviral effects of A3 proteins by targeting them for degradation through the 26S proteasome. Vif hijacks cellular Cullin5 (Cul5), ElonginB, and ElonginC to form a viral E3 ubiquitin ligase that targets A3G for polyubiquitination and degradation. Thus, identification of novel strategies to preserve the antiviral functions of A3 is an exciting new target for antiretroviral therapy. In this application, we propose to capitalize our expertise in HIV-1 Vif/A3 system and our new understanding of the viral evasion mechanism to develop a rapid cell-based assay for the identification of small molecule inhibitors of Vif and to further optimize and adapt the system for application to high throughput molecular screening of large compound libraries to identify molecules that inhibit HIV-1 replication. We will also characterize several small molecule compounds that have been identified to inhibit Vif mediated A3G degradation in our initial screening. PUBLIC HEALTH RELEVANCE: The HIV-1 virion infectivity factor (Vif) is an essential regulatory protein required for HIV- 1 replication in natural target cells such as CD4+ T-cells and macrophages which express innate antiviral human APOBEC3G (A3G) and related cytidine deaminases. We have identified a small molecule inhibitor of Vif. The overall goal of this project is to develop a rapid cell-based assay for the identification of small molecule inhibitors of Vif and to further optimize and adapt the system for application to high throughput molecular screening of large compound libraries to identify molecules that inhibit HIV-1 replication.

描述（由申请人提供）：人胞苷脱氨酶 APOBEC3 (A3) 蛋白是宿主针对 HIV 的有效防御。这些抗病毒蛋白诱导新合成的负链病毒DNA中胞嘧啶对尿嘧啶的致命修饰，导致病毒感染失败。 HIV 必须克服这些宿主细胞防御才能成功复制病毒。 HIV-1 编码一种蛋白质 Vif，它通过 26S 蛋白酶体降解 A3 蛋白来抑制 A3 蛋白的抗病毒作用。 Vif 劫持细胞 Cullin5 (Cul5)、ElonginB 和 ElonginC，形成病毒 E3 泛素连接酶，以 A3G 为目标进行多泛素化和降解。因此，确定保留 A3 抗病毒功能的新策略是抗逆转录病毒治疗的一个令人兴奋的新目标。在此应用中，我们建议利用我们在 HIV-1 Vif/A3 系统方面的专业知识以及我们对病毒逃避机制的新理解，开发一种基于细胞的快速检测方法来识别 Vif 小分子抑制剂，并进一步优化和调整该系统，以应用于大型化合物库的高通量分子筛选，以识别抑制 HIV-1 复制的分子。我们还将表征几种小分子化合物，这些化合物在我们的初步筛选中已被鉴定可抑制 Vif 介导的 A3G 降解。 公共健康相关性：HIV-1 病毒粒子感染因子 (Vif) 是 HIV-1 在天然靶细胞（例如 CD4+ T 细胞和表达先天抗病毒人 APOBEC3G (A3G) 和相关胞苷脱氨酶的巨噬细胞）中复制所需的重要调节蛋白。我们已经鉴定出 Vif 的小分子抑制剂。该项目的总体目标是开发一种基于细胞的快速检测方法，用于鉴定 Vif 小分子抑制剂，并进一步优化和调整该系统，以应用于大型化合物库的高通量分子筛选，以鉴定抑制 HIV-1 复制的分子。