The RCMI Program in Health Disparities Research at Meharry Medical College (Supplement)

梅哈里医学院的 RCMI 健康差异研究项目（补充）

• 批准号: 10874884
• 负责人: Samuel Evans Adunyah
• 金额: $ 21.82万
• 依托单位: MEHARRY MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-30 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10874884
• 关键词: APOCEC3G gene; Acquired Immunodeficiency Syndrome; Address; African American population; Antiviral Agents; Applications Grants; Award; Biological Assay; CD4 Positive T Lymphocytes; Cell Line; Cell Separation; Cells; Cessation of life; Chemicals; Clinical Trials; Collaborations; Collection; Complementary DNA; Cytidine Deaminase; Data; Development; Drug Targeting; Exclusion; Exhibits; Failure; Fostering; Foundations; Frequencies; Goals; Grant; HIV; HIV antiretroviral; HIV-1; HIV-1 drug resistance; Health Disparities Research; Health Services Accessibility; Incidence; Infection; Laboratories; Libraries; Mediating; Mentors; Mentorship; Michigan; Minority Groups; Morphologic artifacts; Multi-Drug Resistance; Mutation; Natural Products; Persons; Pharmaceutical Preparations; Pilot Projects; Reaction; Reproducibility; Research; Resources; Science; Solid; Tennessee; Testing; Universities; Viral; Viral Drug Resistance; Viral Physiology; Viral Proteins; Virus Inhibitors; Virus Replication; antiretroviral therapy; caucasian American; compound 30; cytotoxicity; drug development; experimental study; health disparity; high throughput screening; infection rate; inhibitor; medical schools; multicatalytic endopeptidase complex; novel; novel therapeutics; pandemic disease; pharmacologic; programs; screening; small molecule; small molecule libraries

AIDS disproportionately targets several minority groups, including the African American population. The burden is due to the high frequency of HIV-1 infection rates and the high incidence of virologic failure of HIV antiretroviral therapy, owing to the multidrug resistance in the African American population. To control the emergence of multidrug-resistant HIV-1 and solve the AIDS Health Disparities issue, it is important to develop drugs against novel viral targets. HIV-1 viral infectivity factor (Vif) is essential for virus replication. The primary function of Vif is to counteract APOBEC3G (A3G), a potent host restriction factor for HIV. Vif-A3G interaction has been the target for drug development. Accordingly, several groups have developed HTS assays to screen for small molecules to inhibit Vif-mediated A3G degradation. However, none have advanced to clinical trials due to their moderate potency in rescuing A3G antiviral function. Notably, there is evidence that Vif directly inhibits A3G cytidine deaminase activity (CDA). Given that the efforts to identify effective inhibitors of Vif-mediated A3G degradation have not been successful and A3G CDA is critical for its antiviral function, we will target Vif-mediated A3G CDA inhibition function for identifying novel Vif inhibitors. To develop a chemical probe and lay the foundation for discovering a new class of HIV drugs, we have established a novel and robust assay to screen small molecules that target this function of Vif with excellent reproducibility and a calculated Z-score of 0.83. In preliminary studies, we screened ~5500 compounds from a combination of several small compound libraries. We obtained one hit, Quinobene, which showed potent antiviral activity (IC50:0.75-1.25 μM) by restoring A3G function. The data validate our assay and support the hypothesis that screening a pharmacologically diverse small molecule library can lead to the identification of Vif-specific inhibitors that target A3G CDA activity. In this proposal, we propose to screen a collection of Natural Product Extracts (3K extracts, >30 compounds per extract, totaling ~10K chemicals) in collaboration with Dr. Ashu Tripathi, Director of the Natural Products Discovery Core at the University of Michigan (Aim 1). Natural Product Extracts (NPEs) with a high potency of inhibiting Vif function will be selected for further validating the potency of Vif inhibitors and perform secondary assays to verify their A3G-dependent anti-HIV activity in CD4+T cell lines (Aim 2). Finally, we will utilize the data-intensive platform of Tripathi's lab to rapidly de-convolute identified hits to characterize active leads and biologically characterize the promising leads in primary CD4+ T cells isolated from Caucasian and African American (AA) populations (Aim 3). The proposed experiments will (i) develop chemical probes that can be used to elucidate the function of Vif in counteracting A3G and (ii) lead to the discovery of potent anti-HIV inhibitors for the development of new class antiviral drugs to address multidrug resistance issue disproportionately targeting AA population.

艾滋病主要针对几个少数群体，包括非裔美国人。负担 是由于艾滋病毒-1感染率高和艾滋病毒抗逆转录病毒治疗病毒学失败的发生率高 由于非裔美国人人口中存在多药耐药性，治疗受到了限制。控制……的出现 多药耐药HIV-1和解决艾滋病健康差距问题，开发针对艾滋病的药物非常重要 新的病毒靶标。HIV-1病毒感染性因子(VIF)是病毒复制所必需的。VIF的主要功能 是为了对抗APOBEC3G(A3G)，A3G是HIV的一个强有力的宿主限制因素。VIF-A3G互动一直是 药物开发的目标。因此，几个组织已经开发了HTS检测方法来筛查小型 抑制Vif介导的A3G降解的分子。然而，由于他们的原因，没有一种进入临床试验 挽救A3G抗病毒功能的中等效力。值得注意的是，有证据表明，VIF直接抑制了A3G 胞苷脱氨酶活性(CDA)。鉴于寻找Vif介导的A3G有效抑制剂的努力 降解并不成功，A3G CDA对于其抗病毒功能至关重要，我们将针对Vif介导的 A3G CDA抑制功能用于识别新的Vif抑制剂。开发一种化学探针，并铺设 为发现一类新的艾滋病毒药物奠定了基础，我们建立了一种新的和强大的检测方法来筛选 靶向VIF这一功能的小分子具有极好的重复性和计算的Z-Score为0.83。在……里面 初步研究，我们从几个小化合物文库的组合中筛选出~5500个化合物。 我们获得了一种Hit，Quinobene，它通过恢复A3G显示出强大的抗病毒活性(IC50：0.75-1.25μM) 功能。这些数据验证了我们的分析，并支持了这样的假设，即筛选出一种药理上不同的 小分子文库可以帮助鉴定针对A3G CDA活性的Vif特异性抑制物。在这 建议，我们建议筛选一组天然产品提取物(3K提取物，每种提取物30种化合物， 总计约10K种化学品)与天然产品发现核心主任阿苏·特里帕蒂博士合作 在密歇根大学(目标1)。高效抑制血管生成的天然产物提取物(NPE) 将选择功能以进一步验证Vif抑制剂的效力，并进行二次检测以验证 它们在CD4+T细胞系中依赖A3G的抗HIV活性(目标2)。最后，我们将利用数据密集型 Tripari实验室的平台，用于快速分解已识别的HIT，以表征活性线索和生物学特征 从高加索人和非裔美国人(AA)分离的原代CD4+T细胞中有希望的线索的特征 人口(目标3)。拟议中的实验将(I)开发出可用于阐明 Vif在对抗A3G中的作用和(Ii)导致发现有效的抗HIV抑制剂 开发新型抗病毒药物以解决不成比例地针对再障的多药耐药问题 人口。