RNA As A Therapeutic Target

RNA作为治疗靶点

• 批准号: 8325074
• 负责人: Webster L Santos
• 金额: $ 29.83万
• 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/8325074
• 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; 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被确定为艾滋病的病原体以来已经过去了26年，但随着每年新感染的发生和HIV治疗延长生命，HIV感染者的比例稳步上升。2007年，估计有3300万人感染艾滋病毒。目前用于治疗艾滋病的大多数药物是通过联合靶向HIV逆转录酶和/或蛋白酶（HAART，高活性抗逆转录病毒治疗）和/或gp41的酶活性来起作用的。这些治疗仍然很昂贵，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传统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，开发具有新型作用方式的新药迫在眉睫。我们建议开发对HIV生命周期至关重要的蛋白质- rna相互作用抑制剂，这是医学界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，具有明确的靶向结构。靶向这些RNA结构是有吸引力的，因为它影响病毒特异性相互作用，可能导致病毒复制的特异性抑制，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，这种策略可以降低耐药性的发生率。我们最近报道了支链肽是HIV-1 TAR的良好选择性配体。我们建议进行短支肽组合文库的筛选。这是完成了特定的目的1，其中一个更大的文库的多样化的支链肽和硼链支链肽被合成。由于靶向RNA的主要挑战是赋予特定序列/结构的选择性，因此需要高度严格的筛选试验。Aim 1也讨论了这一点。我们认识到，在高通量筛选试验中，假阳性是可能的，RNA结合剂不一定表明所需的蛋白质/RNA相互作用的扰动。在目标2中，我们使用一系列基于细胞的实验来测试hit支链肽对HIV-1 Tat/TAR和Rev/RRE相互作用的抑制活性。此外，我们还研究了支链肽抑制剂的选择性和可能的细胞毒性。