RNA As A Therapeutic Target

RNA作为治疗靶点

• 批准号: 8537941
• 负责人: Webster L Santos
• 金额: $ 30.21万
• 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/8537941
• 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; Series; Specificity; Structure; Structure-Activity Relationship; 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; screening; 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(申请人提供)：虽然自艾滋病毒-1被确定为艾滋病的致病因素以来已经26年了，但随着每年发生新的感染病例和艾滋病毒治疗延长生命，艾滋病毒携带者的比例稳步上升。据估计，2007年有3300万人携带艾滋病毒。目前用于治疗艾滋病的大多数药物都是通过联合靶向HIV逆转录酶和/或蛋白酶(HAART，高效抗逆转录病毒疗法)和/或gp41的酶活性发挥作用的。这些治疗方法仍然昂贵，而且患者往往不能很好地耐受。由于耐药病毒的出现通常是经典艾滋病毒治疗的结果，因此仍然非常需要继续寻找针对其他基本病毒活动的替代疗法。因此，开发具有新作用模式的新药迫在眉睫。我们建议开发对艾滋病毒生命周期绝对必要的蛋白质-RNA相互作用的抑制剂--这是社区广泛认可的优秀目标。TAR和RRE RNAs是病毒基因组中保守的非编码序列，具有明确的靶向结构。以这些RNA结构为靶点是很有吸引力的，因为它们会影响病毒特异性的相互作用，从而可能导致对病毒复制的特定抑制，而对其他细胞功能的副作用可能最小。此外，由于HIV-1转录的调节需要病毒和细胞成分的相互作用，因此这一策略可以导致较低的耐药性发生率。我们最近报道了支链多肽是HIV-1TAR的良好选择性配体。我们建议开展短支链多肽组合文库的筛选。这是通过特定目标1实现的，其中合成了更大的多样化支化肽和硼支化肽的文库。由于靶向RNA的主要挑战是赋予特定序列/结构的选择性，因此需要一种非常严格的筛选方法。目标1中也提到了这一点。我们认识到，在高通量筛选试验中，假阳性是可能的，RNA结合子并不一定表明所需的蛋白质/RNA相互作用受到干扰。在目标2中，我们使用一系列基于细胞的方法来测试HIT支链多肽对HIV-1 TAT/TAR和REV/RRE相互作用的抑制活性。此外，我们还研究了支链多肽抑制剂的选择性和可能的细胞毒性。 与公共卫生相关：拟议的研究旨在发现和开发中等大小的分子，这些分子选择性地抑制蛋白质-RNA相互作用，而蛋白质-RNA相互作用对HIV-1病毒复制绝对必要。如果成功，这些分子将成为下一代具有抗艾滋病毒治疗作用模式的药物。