RNA As A Therapeutic Target

RNA作为治疗靶点

基本信息

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
项目类别：
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

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

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项目摘要

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）：尽管自HIV-1被确定为艾滋病的病原体以来已经过去了26年，但随着每年新的感染发生，艾滋病毒感染者的百分比稳步增加，并且随着艾滋病毒治疗延长寿命。估计有3,300万人在2007年患有艾滋病毒。目前用于治疗艾滋病工作的大多数药物通过靶向HIV逆转录酶和/或蛋白酶的酶活性（HAART，高度活跃的抗逆转录病毒疗法）和/或GP41的组合。这些治疗仍然很昂贵，并且常常无法容忍患者。由于通常由于经典HIV治疗而出现的耐药病毒出现，因此仍然需要继续寻找针对其他必要病毒活性的替代疗法。因此，以新颖的作用方式开发新药是最紧迫的。我们建议开发蛋白实 - RNA相互作用的抑制剂，这些抑制剂对于社区广泛认可的HIV生命周期 - 细胞性目标绝对必不可少。 TAR和RRE RNA是具有定义可靶向结构的病毒基因组中保存良好的非编码序列。靶向这些RNA结构是有吸引力的，因为影响病毒特异性相互作用可能会导致特异性抑制病毒复制，并且可能对其他细胞功能的副作用最小。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，因此该策略可能导致耐药性的发生率较低。我们最近报道说，分支肽是HIV-1 TAR的良好选择配体。我们建议对短分支肽的组合文库进行筛选。这是通过特定的目标1来完成的，其中合成了较大的多元化支枝肽和硼链肽的图书馆。由于靶向RNA的主要挑战是对特定序列/结构赋予选择性，因此必须进行高度严格的筛选测定。 AIM 1也解决了这一点。我们认识到，在高吞吐量筛选测定中，假阳性是可能的，RNA粘合剂不一定表明所需的蛋白质/RNA相互作用的扰动。在AIM 2中，我们使用一系列基于细胞的测定方法来测试对HIV-1 TAT/TAT和REV/RRE相互作用的命中分支肽的抑制作用。此外，我们研究了分支肽抑制剂的选择性以及可能的细胞毒性。公共卫生相关性：拟议的研究旨在发现和开发中型分子，这些分子有选择地抑制蛋白RNA相互作用的相互作用，这对于HIV-1病毒复制绝对必不可少。如果成功，这些分子将是具有新型作用方式作为抗HIV治疗的下一代药物。