Identification and characterization of novel anti-HIV inhibitors

描述（由申请人提供）：人胞苷脱氨酶APOBEC 3（A3）蛋白是宿主对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。HIV-1编码一种名为Vif的蛋白质，通过26 S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Cullin 5（Cul 5）、ElonginB和ElonginC以形成病毒E3泛素连接酶，其靶向A3 G进行聚泛素化和降解。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靶点。在本申请中，我们建议利用我们在HIV-1 Vif/A3系统中的专业知识和我们对病毒逃逸机制的新理解，开发用于鉴定Vif小分子抑制剂的快速基于细胞的检测方法，并进一步优化和调整该系统，以应用于大化合物库的高通量分子筛选，以鉴定抑制HIV-1复制的分子。我们还将表征在我们的初步筛选中已鉴定为抑制Vif介导的A3 G降解的几种小分子化合物。 公共卫生相关性：HIV-1病毒体感染因子（Vif）是HIV- 1在天然靶细胞如CD 4 + T细胞和巨噬细胞中复制所需的必需调节蛋白，其表达先天性抗病毒人APOBEC 3G（A3 G）和相关胞苷脱氨酶。我们已经鉴定了Vif的小分子抑制剂。该项目的总体目标是开发一种快速的基于细胞的检测方法，用于鉴定Vif的小分子抑制剂，并进一步优化和调整该系统，以应用于大化合物文库的高通量分子筛选，以鉴定抑制HIV-1复制的分子。

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.