Novel Small Molecule Inhibitors of HIV

新型 HIV 小分子抑制剂

• 批准号: 7895567
• 负责人: Xiao-Fang Yu
• 金额: $ 20.5万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-17 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7895567
• 关键词: Address; Antiviral Agents; Binding; Biological Models; Boxing; CD4 Positive T Lymphocytes; Cells; Charge; Chelating Agents; Clioquinol; Complex; Cytidine Deaminase; Cytosine; Defense Mechanisms; Enzymes; Ethylenediamines; Goals; HIV; HIV-1; HIV-2; Individual; Integration Host Factors; Mediating; Membrane; Modification; Polyubiquitination; Production; Property; Proteins; Research; Techniques; Testing; Therapeutic; Uracil; Viral; Viral Physiology; Viral Proteins; Virion; Virus; Zinc; chelation; design; drug development; expression vector; genetic regulatory protein; human CEM15 protein; inhibitor/antagonist; insight; macrophage; novel; public health relevance; receptor; receptor binding; small molecule; ubiquitin-protein ligase; vif Gene Products; viral DNA

DESCRIPTION (provided by applicant): 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. Recent studies from our group and others have demonstrated that Vif is a critical viral counter defense that specifically neutralizes an innate antiviral defense mechanism, mediated by human APOBEC3G (A3G) and related cytidine deaminases. These antiviral proteins induce lethal modification of cytosines to uracils in newly synthesized minus-strand viral DNA, resulting in the production of non-functional viruses in the absence of the retroviral Vif protein. Vif hijacks cellular Cullin5 (Cul5), ElonginB, and ElonginC to form a viral E3 ubiquitin ligase that targets A3G for polyubiquitination and degradation. We have identified N,N,N',N'-Tetrakis-(2-pyridylmethyl)-Ethylenediamine (TPEN) as a small molecule inhibitor of Vif. The overall goal of this project is to further characterize the novel small molecule Vif inhibitor and its derivatives that restore the potent innate antiviral defense. In particular we propose 1. To examine the effect of TPEN on Vif functions from multiple HIV-1 subtypes as well as HIV-2. We have observed that TPEN is a potent inhibitor of HIV-1 Vif (NL4-3) function which allows the anti-viral cytidine deaminase A3G to suppress even the wild-type HIV-1 infectivity. It is not clear whether TPEN could inhibit Vif molecules from a wide range of HIV-1 subtypes. We will construct expression vectors for all the major subtypes of HIV-1 and HIV-2 Vif molecules and test the effect of TPEN on these Vif molecules. We will also determine whether TPEN could inhibit HIV-1 replication in primary CD4+ T cells which express endogenous A3G. Studies proposed in this aim are designed to further substantiate the effect of TPEN anti-HIV activity. 2. To examine the effect of derivatives of TPEN on Vif function and the anti-viral activity of A3G. We have observed that TPEN is a potent inhibitor of HIV-1 Vif function which allows the anti-viral mammalian cytidine deaminase APOBEC3G (A3G) to suppress even the wild-type HIV-1 infectivity. It is not clear whether TPEN inhibits Vif solely through zinc chelation, since another membrane permeable zinc chelator, chloroquinol, was not effective. To address this issue, we will evaluate TPEN derivatives that do not have the ability to bind zinc for anti-Vif activity. TPEN derivatives that have altered hydrophobic, hydrophilic, or charged properties will also be evaluated for their anti-Vif function. Studies proposed in this aim are designed to further characterize the mechanism of TPEN anti-HIV activity and to identify more potent anti-Vif inhibitors with a broad therapeutic window. 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. Recent studies from our group and others have demonstrated that Vif is a critical viral counter defense that specifically neutralizes an innate antiviral defense mechanism, mediated by human APOBEC3G (A3G) and related cytidine deaminases. We have identified a small molecule inhibitor of Vif. The overall goal of this project is to further characterize the novel small molecule Vif inhibitor and its derivatives that restore the potent innate antiviral defense.

描述（由申请人提供）：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。我们小组和其他人的最新研究表明，VIF是一项关键的病毒反式防御，该防御专门中和由人Apobec3g（A3G）和相关的胞苷脱氨酶介导的先天抗病毒防御机制。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，从而导致在没有逆转录病毒VIF蛋白的情况下产生非功能性病毒。 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。我们已经将N，N，N'，N'-Tetrakis-（2-吡啶基甲基） - 乙二胺（TPEN）视为VIF的小分子抑制剂。该项目的总体目标是进一步描述新型的小分子VIF抑制剂及其衍生物，以恢复有效的先天抗病毒防御。特别是我们建议1。检查TPEN对多种HIV-1亚型和HIV-2的VIF功能的影响。我们已经观察到TPEN是HIV-1 VIF（NL4-3）功能的有效抑制剂，它允许抗病毒胞苷脱氨酶A3G抑制甚至抑制野生型HIV-1感染性。目前尚不清楚TPEN是否可以抑制来自多种HIV-1亚型的VIF分子。我们将为HIV-1和HIV-2 VIF分子的所有主要亚型构建表达矢量，并测试TPEN对这些VIF分子的影响。我们还将确定TPEN是否可以抑制表达内源性A3G的原代CD4+ T细胞中的HIV-1复制。该目标中提出的研究旨在进一步证实TPEN抗HIV活性的影响。 2。检查TPEN衍生物对VIF功能和A3G病毒活性的影响。我们已经观察到TPEN是HIV-1 VIF功能的有效抑制剂，它允许抗病毒哺乳动物胞苷脱氨酸酶Apobec3g（A3G）抑制甚至抑制野生型HIV-1感染性。目前尚不清楚TPEN是否仅通过锌螯合抑制VIF，因为另一个膜可渗透的锌螯合剂氯喹酚无效。为了解决这个问题，我们将评估没有结合抗VIF活性的锌的TPEN衍生物。还将评估改变疏水性，疏水性或带电性能的TPEN衍生物的抗VIF功能。该目标中提出的研究旨在进一步表征TPEN抗HIV活性的机制，并使用广泛的治疗窗口鉴定出更有效的抗VIF抑制剂。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。我们小组和其他人的最新研究表明，VIF是一项关键的病毒反式防御，该防御专门中和由人Apobec3g（A3G）和相关的胞苷脱氨酶介导的先天抗病毒防御机制。我们已经确定了VIF的小分子抑制剂。该项目的总体目标是进一步描述新型的小分子VIF抑制剂及其衍生物，以恢复有效的先天抗病毒防御。