RNA As A Therapeutic Target

RNA作为治疗靶点

• 批准号: 7930377
• 负责人: Webster L Santos
• 金额: $ 33.08万
• 依托单位: VIRGINIA POLYTECHNIC INST AND ST UNIV
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-13 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7930377
• 关键词: AIDS/HIV problem; Acquired Immunodeficiency Syndrome; Address; Adverse effects; Affect; Affinity; Alternative Therapies; Amino Acids; Anti-HIV Agents; Anti-HIV Therapy; Binding; Binding Sites; Biological; Biological Assay; Boron; Cell physiology; Cells; Clinic; Combined Modality Therapy; Communities; Development; Disease; Drug resistance; Elements; Genetic Transcription; Goals; HIV; HIV-1; Highly Active Antiretroviral Therapy; In Vitro; Incidence; Infection; Lead; Length; Libraries; Life; Life Cycle Stages; Ligands; Measurement; Measures; Methods; Molecular; Nucleic Acids; Pathway interactions; Patients; Peptide Hydrolases; Peptide Library; Peptides; Pharmaceutical Preparations; Property; Protein Binding; Proteins; RNA; RNA Binding; RNA Interference; RNA-Directed DNA Polymerase; RNA-Protein Interaction; Regulation; Reporting; Research; Resistance; Screening procedure; Series; Specificity; Structure; Structure-Activity Relationship; Tars; Testing; Viral; Viral Genome; Viral Physiology; Virus; Work; analog; base; combinatorial; cytotoxicity; drug resistant virus; high throughput screening; in vitro Assay; inhibitor/antagonist; innovation; next generation; novel; novel strategies; public health relevance; small molecule; therapeutic target

DESCRIPTION (provided by applicant): Although 26 years has passed since HIV-1 was identified as the causative agent for AIDS, the percentage of people living with HIV has steadily increased as new infections occur each year and as HIV treatments extend life. An estimated 33 million people were living with HIV in 2007. Most of the current drugs in use for the treatment of AIDS work by combination targeting of the enzymatic activities of the HIV reverse transcriptase and/or protease (HAART, highly active anti-retroviral therapy) and/or gp41. These treatments remain expensive and are often not well- tolerated by patients. Because of the emergence of drug-resistant virus that commonly occurs as the result of classical HIV treatment, there remains a great need to continue the search for alternative therapies that target other essential viral activities. Thus, the development of new drugs with novel mode of action is of utmost urgency. We propose to develop inhibitors of protein-RNA interactions that are absolutely essential for the HIV life cycle-excellent targets widely recognized by the community. TAR and RRE RNAs are well-conserved noncoding sequences in the viral genome with defined targetable structures. Targeting these RNA structures is attractive because the affect virus-specific interactions that could potentially lead to specific inhibition of viral replication, with possibly minimal side effects on other cellular functions. Additionally, this strategy can result in a lower incidence of drug resistance since the regulation of HIV-1 transcription requires the interplay of both viral and cellular components. We have recently reported that branched peptides are good selective ligands for HIV-1 TAR. We propose to carry-out the screening of combinatorial libraries of short branched peptides. This is accomplished with specific aim 1, wherein a larger library of diversified branched peptides and borono-branched peptides is synthesized. Because the major challenge in targeting RNA is imparting selectivity to a particular sequence/structure, a highly stringent screening assay is necessary. This is also addressed in Aim 1. We recognize that during a high throughput screening assay, false positives are possible and that an RNA binder does not necessarily indicate perturbation of the desired protein/RNA interaction. In aim 2, we use a series of cell based assays to test the inhibitory activities of hit branched peptides against HIV-1 Tat/TAR and Rev/RRE interactions. Additionally, we investigate the selectivity as well as possible cytotoxicity of branched peptide inhibitors. PUBLIC HEALTH RELEVANCE: The proposed research aims to discover and develop medium-sized molecules that selectively inhibit the interaction of protein-RNA interactions that are absolutely essential for HIV-1 viral replication. If successful, these molecules will be the next generation drugs with novel mode of action as anti-HIV therapy.

描述（由申请人提供）：虽然自 HIV-1 被确定为艾滋病病原体以来已经过去了 26 年，但随着每年新感染的出现以及艾滋病毒治疗延长寿命，艾滋病毒感染者的比例稳步上升。据估计，2007 年有 3300 万人感染了艾滋病毒。目前用于治疗艾滋病的大多数药物都是通过联合靶向艾滋病毒逆转录酶和/或蛋白酶的酶活性（HAART，高效抗逆转录病毒疗法）来发挥作用。 ）和/或 gp41。这些治疗方法仍然昂贵并且通常不能被患者很好地耐受。由于经典艾滋病毒治疗通常会出现耐药病毒，因此仍然非常需要继续寻找针对其他基本病毒活性的替代疗法。因此，开发具有新颖作用方式的新药刻不容缓。我们建议开发蛋白质-RNA 相互作用的抑制剂，这对于 HIV 生命周期绝对必要，是得到社区广泛认可的优秀靶点。 TAR 和 RRE RNA 是病毒基因组中高度保守的非编码序列，具有明确的可靶向结构。针对这些 RNA 结构很有吸引力，因为它们会影响病毒特异性相互作用，从而可能导致病毒复制的特异性抑制，同时对其他细胞功能的副作用可能最小。此外，这种策略可以降低耐药性的发生率，因为 HIV-1 转录的调节需要病毒和细胞成分的相互作用。我们最近报道支链肽是 HIV-1 TAR 良好的选择性配体。我们建议进行短支化肽组合文库的筛选。这是通过具体目标 1 实现的，其中合成了更大的多样化支链肽和硼代支链肽库。由于靶向 RNA 的主要挑战是赋予特定序列/结构选择性，因此需要高度严格的筛选测定。目标 1 中也解决了这个问题。我们认识到，在高通量筛选测定中，可能会出现假阳性，并且 RNA 结合物并不一定表明对所需蛋白质/RNA 相互作用的干扰。在目标 2 中，我们使用一系列基于细胞的测定来测试命中分支肽对 HIV-1 Tat/TAR 和 Rev/RRE 相互作用的抑制活性。此外，我们研究了支链肽抑制剂的选择性以及可能的细胞毒性。 公共健康相关性：拟议的研究旨在发现和开发中等大小的分子，选择性抑制蛋白质-RNA 相互作用，而蛋白质-RNA 相互作用对于 HIV-1 病毒复制至关重要。如果成功，这些分子将成为具有新颖作用方式的下一代抗艾滋病毒治疗药物。