Identification and characterization of novel anti-HIV inhibitors

新型抗 HIV 抑制剂的鉴定和表征

• 批准号: 8132453
• 负责人: Xiao-Fang Yu
• 金额: $ 20.3万
• 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/8132453
• 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; 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）：人类细胞丁胺脱氨酶APOBEC3（A3）蛋白是对HIV的有效宿主防御能力。这些抗病毒蛋白会在新合成的负链病毒DNA中诱导细胞儿的致命修饰，从而导致流产的病毒感染。艾滋病毒必须克服这些宿主细胞防御，以成功的病毒复制。 HIV-1编码一种蛋白质VIF，该蛋白通过靶向通过26S蛋白酶体降解来抑制A3蛋白的抗病毒作用。 VIF劫持细胞Cullin5（Cul5），Elonginb和Elonginc形成一种病毒E3泛素连接酶，该连接酶靶向A3G用于多泛素化和降解。因此，确定保留A3抗病毒功能的新型策略是抗逆转录病毒疗法的令人兴奋的新目标。在此应用程序中，我们建议将我们在HIV-1 VIF/A3系统方面的专业知识以及对病毒逃避机制的新了解，以开发一种基于细胞的快速测定法，以鉴定VIF的小分子抑制剂，并进一步优化和适应该系统，以应用和适用于对大型化合物库的高吞吐量分子筛选，以识别构倍分子的高素质hebiv-1。我们还将表征几种已鉴定的小分子化合物，这些化合物在初始筛选中抑制VIF介导的A3G降解。 公共卫生相关性：HIV-1病毒感染因子（VIF）是自然靶细胞中HIV-1复制所需的必不可少的调节蛋白，例如CD4+ T细胞和巨噬细胞，表达先天性抗病毒抗病毒人Apobec3g（A3G）和相关的胞苷甲激酶。我们已经确定了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测定，以鉴定VIF的小分子抑制剂，并进一步优化和适应系统以应用于大型化合物库的高吞吐量分子筛选，以识别抑制HIV-1复制的分子。