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病毒感染性因子(VIF)是HIV-1在自然目标细胞如CD4+T细胞和巨噬细胞中复制所必需的调节蛋白。我们团队和其他人最近的研究表明，Vif是一种关键的病毒对抗防御，它特异性地中和由人APOBEC3G(A3G)和相关胞苷脱氨酶介导的固有抗病毒防御机制。这些抗病毒蛋白在新合成的负链病毒DNA中诱导胞嘧啶到尿嘧啶的致死性修饰，导致在没有逆转录病毒Vif蛋白的情况下产生无功能的病毒。VIF劫持细胞Cullin5(Cul5)、ElonginB和ElonginC，形成针对A3G的病毒E3泛素连接酶，以实现多泛素化和降解。我们已确定N，N，N‘，N’-四-(2-吡啶甲基)-乙二胺(TPEN)为VIF的小分子抑制剂。该项目的总体目标是进一步表征新型小分子Vif抑制剂及其衍生物，以恢复强大的天然抗病毒防御能力。特别是，我们提出了1.检测TPEN对多种HIV-1亚型和HIV-2的Vif功能的影响。我们观察到，TPEN是HIV-1 Vif(NL4-3)功能的有效抑制剂，使抗病毒胞苷脱氨酶A3G甚至可以抑制野生型HIV-1的感染性。目前尚不清楚TPEN是否可以抑制多种HIV-1亚型的Vif分子。我们将构建HIV-1和HIV-2 Vif分子的所有主要亚型的表达载体，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。这一目的中提出的研究旨在进一步证实TPEN的抗艾滋病毒活性。2.检测TPEN衍生物对Vif功能的影响及A3G的抗病毒活性。我们观察到TPEN是一种有效的HIV-1 Vif功能的抑制剂，它可以使抗病毒的哺乳动物胞苷脱氨酶APOBEC3G(A3G)甚至抑制野生型HIV-1的感染性。目前尚不清楚TPEN是否仅通过锌螯合来抑制VIF，因为另一种膜透性锌螯合剂氯喹酚无效。为了解决这个问题，我们将评估不能与锌结合的TPEN衍生物的抗Vif活性。改变了疏水性、亲水性或带电性质的TPEN衍生物也将被评估其抗VIF功能。在这一目标中提出的研究旨在进一步表征TPEN的抗HIV活性的机制，并寻找更有效的抗Vif抑制剂，具有广阔的治疗窗口。公共卫生相关性：HIV-1病毒粒子传染性因子(Vif)是HIV-1在自然靶细胞(如CD4+T细胞和巨噬细胞)中复制所必需的必要调节蛋白。我们团队和其他人最近的研究表明，Vif是一种关键的病毒对抗防御，它特异性地中和由人APOBEC3G(A3G)和相关胞苷脱氨酶介导的固有抗病毒防御机制。我们已经确定了一种VIF的小分子抑制剂。该项目的总体目标是进一步表征新型小分子Vif抑制剂及其衍生物，以恢复强大的天然抗病毒防御能力。