Development and Characterization of Antiviral Drugs That

抗病毒药物的开发和表征

• 批准号: 7338640
• 负责人: VINAY K. PATHAK
• 金额: --
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7338640
• 关键词: Development; Characterization; Antiviral; Drugs

Through collaborative studies, we have identified azido-group containing beta-diketo acid derivatives (DKAs) as potent inhibitors of HIV-1 replication. We have also analyzed the effects of the DKAs on the frequency of 2-LTR circles as well as the sequences at the 2-LTR-circle junctions. These results suggest that the structure of the DKAs can influence the extent of degradation of viral DNA ends by host nucleases and the frequency of deletions at the 2-LTR-circle junctions. A suitable animal model system for testing of HIV-1 IN inhibitors is currently not available. We are constructing simian immunodeficiency virus (SIV)-HIV-1 hybrid viruses (SHIVs) that are dependent on the HIV-1 RT and IN for replication (SHIV-RT-IN). The SHIV-RT-IN viruses should provide a suitable animal model system to evaluate combination antiviral therapies containing anti-IN inhibitors alone or in combination with RT inhibitors and to characterize mutations in HIV-1 IN that confer resistance to the IN inhibitors. In collaboration with Dr. Kvaratskhelia's laboratory, we have performed mass spectrometric analysis to identify the HIV-1 IN binding site for a nucleotide analog inhibitor of HIV-1 IN, pyridoxal 5'-phosphate. These results indicate for the first time that a small inhibitor molecule can inhibit IN activity by binding to the protein C-terminus. In collaborative studies, we are exploring novel methods to identify lead compounds that inhibit HIV-1 IN. The purpose of the project is to discover candidate inhibitors of HIV-1 IN on the basis of latest technologies in bioinformatics and computer-aided drug design. The capability of all candidate compounds to inhibit HIV-1 integrase will be tested in vitro, and the most potent substances will be prepared in sufficient amounts. The promising lead compounds will be tested in the cell-based assays that we have developed to determine their ability to inhibit HIV-1 replication and to determine the molecular targets of antiviral activity. In conjunction with these studies, we are developing in vivo assays to define the intracellular mechanisms of action of HIV-1 IN inhibitors and are developing quantitative methods to determine the effects of potential HIV-1 IN inhibitors of 2-LTR circle formation. Finally, we are developing novel in vivo assays to identify new lead compounds that inhibit HIV-1 IN.

通过协作研究，我们已经确定了含有β-二甲酸衍生物（DKAS）的氮杂组是HIV-1复制的有效抑制剂。我们还分析了DKA对2-LTR圆圈频率以及2-LTR圆连接处的序列的影响。这些结果表明，DKA的结构可以影响病毒DNA终止的降解程度，并在2-Ltr-Circle连接处的宿主核酸酶和缺失频率。目前尚不可用用于抑制剂中HIV-1测试的合适动物模型系统。我们正在构建依赖于HIV-1 RT并进行复制的邻居免疫缺陷病毒（SIV）-HIV-1混合病毒（SHIVS）（SHIV-RT-IN）。 SHIV-RT-IN病毒应提供合适的动物模型系统，以评估单独或与RT抑制剂组合的组合抗病毒疗法，并在HIV-1中表征HIV-1中的突变，以赋予对IN抑制剂的抗性。通过与Kvaratskhelia博士的实验室合作，我们进行了质谱分析，以鉴定HIV-1中HIV-1的HIV-1 IN，HIV-1 IN，吡啶多毒素5'-磷酸的HIV-1。这些结果首次表明，小抑制剂分子可以通过与蛋白C末端结合而抑制活性。在协作研究中，我们正在探索新的方法，以识别抑制HIV-1 IN的铅化合物。该项目的目的是根据生物信息学和计算机辅助药物设计的最新技术发现HIV-1的候选抑制剂。所有候选化合物抑制HIV-1整合酶的能力将在体外进行测试，并且最有效的物质将以足够的量制备。有希望的铅化合物将在基于细胞的测定中进行测试，即我们已经开发出来确定它们抑制HIV-1复制的能力并确定抗病毒活性的分子靶标。结合这些研究，我们正在开发体内测定，以定义HIV-1在抑制剂中的细胞内作用机理，并正在开发定量方法，以确定潜在的HIV-1在2-LTR圆形抑制剂中的影响。最后，我们正在开发新型的体内测定法，以识别抑制HIV-1 IN的新铅化合物。