HIV-1 Inhibition using Tat Peptide Derivatives

使用 Tat 肽衍生物抑制 HIV-1

• 批准号: 7367875
• 负责人: Ajit Kumar
• 金额: $ 31.76万
• 依托单位: GEORGE WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-07-01 至 2011-04-19
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7367875
• 关键词: Acquired Immunodeficiency Syndrome; Affect; Algorithms; Animals; Binding; Biochemical; Biological; Biological Assay; Biological Availability; Cell Cycle; Cells; Chemistry; Complex; Computer Simulation; Cultured Cells; Cyclization; DNA; DNA-dependent protein kinase; Data; Data Analyses; Data Set; Disease; Drug resistance; Endopeptidases; Environment; Gene Expression Profile; Gene Expression Profiling; Genes; Goals; Graph; HIV; HIV-1; In Vitro; Infection; Maps; Methods; Microarray Analysis; Modeling; Mus; Pathogenesis; Peptide Hydrolases; Peptides; Peripheral Blood Mononuclear Cell; Phase; Phosphotransferases; Procedures; Proteins; RNA-Directed DNA Polymerase; Range; Resistance; Resistance development; Simian virus 40; Standards of Weights and Measures; T-Lymphocyte; Tars; Testing; Toxic effect; Toxicity Tests; Trans-Activators; Transfection; Tropism; Validation; Viral; Virus; cytotoxicity; in vivo; inhibitor/antagonist; mouse model; peptide I; success

DESCRIPTION (provided by applicant): Human Immunodeficiency virus (HIV) is the etiologic agent of AIDS. The pathogenesis of HIV -induced disease is complex and multifactoral. Several key HIV and cellular proteins have been assigned to be necessary for the course of infection including the transactivator Tat. Viral clones deficient in Tat will not replicate in vitro or in vivo to high titers. The current proposal aims to extend our previous results, which were obtained during the past four years on the effect of Tat peptide derivatives in inhibition of HIV-1. Previous specific aims were successfully met in a timely manner, and they included determination of the target(s) of the Tat peptide inhibitor, defining the minimum structural requirement for the inhibitoryactivity of the Tat core peptide derivative, and determining the range of inhibition on a few HIV-1 isolates. More preliminary data is included to show the target of these Tat peptide inhibitors using in vitro and in vivo assays. Tat peptides were also cyclized at small scale and used in non-specific toxicity assays as well as bioavailability assays in animals with great success. Therefore, we have proposed to continue our initial goal, which was to develop HIV-1 Tat inhibitors that are specific to the Tat/TAR complex. We will optimize peptide cyclization of the short Tat peptide by using new cyclization strategies includingthe use of Cys 4 and Cys 2 compounds. Tat peptides will also contain either Tat or SV40 NLS sequences for efficient delivery. Biological inhibitory activities in addition various biochemical and virological methods will be utilized. To define the breath of Tat peptide inhibition, we will determine the inhibitoryactivity of the Tat peptide on various HIV- 1 isolates including drug-resistant strains. We will assess the cytotoxicity of Tat peptides on replicating T-cells and PBMCs as well as determining the inhibitory activity of these compounds on various HIV-1 wild type, reverse transcriptase and protease resistant viruses with different tropisms. Also, we will utilize the SCID-Hu Thy/Liv mice model to test toxicity and viral inhibition in vivo. Finally, to define whether these inhibitors have any non-specific cellular toxicity, we will utilize gene expression profiling of Tat peptides in treated cells. This will be accomplished through the use of clustering and data analysis methods in PBMC infected cells. Validation of the data will initially include in silico prediction algorithms, independent array data sets, and functional assays using probabilistic rational models, GenMAPP, and Pubgene.

描述（由申请人提供）：人类免疫缺陷病毒（HIV）是艾滋病的病原体。艾滋病病毒引起的疾病的发病机制是复杂和多因素的。几种关键的HIV和细胞蛋白质已被指定为感染过程所必需的，包括反式激活因子达特。缺乏达特的病毒克隆在体外或体内不会复制到高滴度。目前的提议旨在扩展我们以前的结果，这些结果是在过去四年中获得的关于达特肽衍生物抑制HIV-1的效果。先前的特定目标被及时成功地满足，它们包括确定达特肽抑制剂的靶标，定义达特核心肽衍生物的药理活性的最低结构要求，以及确定对一些HIV-1分离株的抑制范围。包括更多的初步数据，以显示这些达特肽抑制剂的目标，在体外和体内测定。达特肽也在小规模环化，并用于非特异性毒性测定以及动物中的生物利用度测定，取得了巨大成功。因此，我们建议继续我们最初的目标，即开发对达特/TAR复合物具有特异性的HIV-1达特抑制剂。我们将通过使用新的环化策略优化短达特肽的环化，包括使用Cys 4和Cys 2化合物。达特肽还将含有达特或SV 40 NLS序列以用于有效递送。除生物学抑制活性外，还将使用各种生物化学和病毒学方法。为了确定达特肽抑制的范围，我们将测定达特肽对各种HIV- 1分离株包括耐药株的药理活性。我们将评估达特肽对复制T细胞和PBMC的细胞毒性，以及确定这些化合物对具有不同向性的各种HIV-1野生型、逆转录酶和蛋白酶抗性病毒的抑制活性。此外，我们将利用SCID-Hu Thy/Liv小鼠模型来测试体内毒性和病毒抑制。最后，为了确定这些抑制剂是否具有任何非特异性细胞毒性，我们将利用达特肽在处理的细胞中的基因表达谱。这将通过在PBMC感染的细胞中使用聚类和数据分析方法来实现。数据验证最初将包括计算机模拟预测算法、独立阵列数据集和使用概率理性模型、GenMAPP和Pubgene的功能测定。