Targeting RNA conformation for drug development

药物开发中的靶向 RNA 构象

• 批准号: 8252970
• 负责人: DEV PRIYA ARYA
• 金额: $ 29.46万
• 依托单位: NUBAD, LLC
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-15 至 2014-07-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8252970
• 关键词: AIDS therapy; Acquired Immunodeficiency Syndrome; Affinity; Amino Sugars; Aminoglycoside Antibiotics; Aminoglycosides; Anti-Bacterial Agents; Antibiotics; Antiviral Agents; Benzimidazoles; Binding; Binding Sites; Biological; Biological Assay; Cells; Charge; Collaborations; Communicable Diseases; Country; Coupled; Cytostatics; Development; Disease; Drug Delivery Systems; Drug resistance; Effectiveness; Epidemic; Essential Drugs; Genetic Transcription; Goals; HIV; HIV-1; In Vitro; Knowledge; Length; Libraries; Ligand Binding; Ligands; Measures; Mediating; Molecular; Molecular Conformation; Nucleic Acids; Organic Synthesis; Outcomes Research; Phase; Phenols; Procedures; Protein Binding; Protein Biosynthesis; Proteins; RNA; RNA Conformation; RNA-Protein Interaction; Relative (related person); Research; Response Elements; Ribosomes; Screening procedure; Series; Shapes; Site; Specificity; Streptomycin; Structure; Techniques; Testing; Therapeutic; Transactivation; Variant; Viral Proteins; Virus Inhibitors; Virus Replication; Work; base; benzimidazole; biophysical chemistry; combat; design; drug development; drug discovery; fight against; high throughput screening; improved; inhibitor/antagonist; innovation; multidisciplinary; novel; novel therapeutics; pathogen; pharmacophore; programs; receptor; response; small molecule; success; tat Protein; tool; viral RNA; virus pathogenesis

DESCRIPTION (provided by applicant): One of the challenges of research in infectious diseases is to find ways to use the increasing knowledge of the mechanisms underlying disease transformation and progression to develop novel therapeutic strategies for AIDS. Targeting specific RNA- protein interactions, such as Tat-TAR or Rev-RRE, which are involved in proliferation and survival of HIV-1 is a promising approach. Our preliminary results show the ability of novel ligands to stabilize TAR RNA, inhibit Tat-TAR interaction at nanomolar concentrations and inhibit HIV-1 in MT-2 cells. These preliminary results will now be built upon to develop a library of conjugates to target Tat-TAR interaction that bind with high affinity and specificity to TAR. Proposed studies will further help establish the efficacy of this approach. The work proposed here, a multidisciplinary effort encompassing organic synthesis, biophysical chemistry and HIV pathogenesis describes the development of small molecule mediated inhibition of Tat-TAR inhibitors as HIV-1 therapeutics. The success of the proposed work would be a significant addition to currently available protein- specific approaches in AIDS therapy and RNA targeting. We propose using a 31 nt TAR target sequences to design conjugates that can be employed to inhibit Tat-TAR interaction; opening possibilities for developing small molecule RNA targeted HIV-1 therapeutics. PUBLIC HEALTH RELEVANCE: Several decades of research on the RNA structure has shown it to be an established drug target, well known as a receptor for small molecule antibiotics. Though the bacterial ribosome has been a well known receptor for antibiotics blocking protein synthesis since the discovery of streptomycin in the 1940s, new antibacterial and antiviral approaches are urgently needed to combat drug resistance, which severely limits the effectiveness of current antibiotics. To investigate the advantage of small molecule-based specificity coupled with charge/shape complementarity, we have initiated a program in the development of a approaches using multimeric ligands (consisting of ligands with independent binding sites) that can be used to target a specific RNA. This proposal focuses on the development of small molecule aminosugars (neamine) conjugates as an example of this approach. A comprehensive approach to identifying essential drug targets in multiple pathogens can be combined with our complementary approach of developing small molecules that bind with high affinity in a specific fashion to previously known as well as rapidly identified, new RNA targets. The inhibition of the Tat/TAR interaction, which facilitates HIV RNA transcription subsequently arrests HIV replication. The central hypothesis of this application is that conjugation of two ligands with an independent binding sites can be conjugated with an appropriate linker to provide a high affinity TAR specific ligand, capable of inhibiting the Tat/TAR interaction at nanomolar concentrations. Furthermore, the assay is applicable to RNA based drug discovery where two pharmacophores with independent binding sites can be combined to select a high affinity ligand. Ultimately, the discovery of a TAR binding ligand with improved affinity and specificity over currently available molecules will provide a better understanding for the potential use of a novel target for implementation in the fight against HIV. NUBAD is well equipped to synthesize the molecules and carry out the biophysical assays for inhibition. Select compounds identified from the assay that inhibit tat-TAR interaction at nanomolar Kd will be tested for inhibition of HIV.

描述（由申请人提供）：传染病研究的挑战之一是寻找方法来利用对疾病转化和进步的机制的越来越多的了解，以开发新的艾滋病治疗策略。针对特定的RNA蛋白相互作用，例如TAT-TAR或REV-RRE，这些相互作用与HIV-1的增殖和存活有关是一种有前途的方法。我们的初步结果表明，新型配体能够稳定焦油RNA，抑制纳摩尔浓度下的TAT-TAR相互作用，并抑制MT-2细胞中的HIV-1。现在将建立这些初步结果，以开发一个结合物库，以靶向与焦油高亲和力和特异性结合的TAT-TAR相互作用。拟议的研究将进一步帮助确定这种方法的功效。这里提出的工作是一种多学科的努力，其中包括有机合成，生物物理化学和HIV发病机理，描述了小分子介导的TAT-TAR抑制剂作为HIV-1治疗剂的抑制作用的发展。拟议工作的成功将是当前可用的蛋白质治疗和RNA靶向方法的重要补充。我们建议使用31 NT TAR目标序列设计可用于抑制TAT-TAR相互作用的共轭物。开发针对小分子RNA靶向HIV-1疗法的可能性。 公共卫生相关性：关于RNA结构的数十年研究表明它是一个已建立的药物靶标，众所周知是小分子抗生素的受体。尽管自1940年代发现链霉素以来，细菌核糖体一直是抗生素阻断蛋白质合成的众所周知的受体，但迫切需要采用新的抗菌和抗病毒方法来打击耐药性，这严重限制了当前抗生素的有效性。 为了研究基于小分子的特异性以及电荷/形状互补性的优势，我们在使用多聚体配体（由具有独立结合位点的配体组成的配体）​​开发方法中启动了一个程序，可用于靶向特定的RNA。该提案的重点是小分子氨基糖（Neamine）偶联的开发，以这种方法的一个例子。可以将多种病原体中基本药物靶标的综合方法与我们的补充方法结合在一起，即开发小分子以特定方式与先前已知的以及迅速鉴定出的新RNA靶标结合高亲和力。 TAT/TAR相互作用的抑制作用，促进HIV RNA转录随后逮捕HIV复制。该应用的中心假设是，可以将两个配体与独立结合位点的结合与适当的接头偶联以提供高亲和力焦油特异性配体，能够抑制纳摩尔浓度下的TAT/TAR相互作用。此外，该测定法适用于基于RNA的药物发现，其中两个具有独立结合位点的药体可以合并以选择高亲和力配体。最终，发现与当前可用分子具有改善亲和力和特异性的焦油结合配体的发现将为潜在使用新的目标在与HIV斗争中实施。 Nubad具有良好的配合，可以合成分子并进行抑制作用。从测定中鉴定出的选择化合物将测试纳摩尔KD处的TAT-TAR相互作用以抑制HIV。