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找到方法，利用对疾病转化和进展机制的不断了解来开发艾滋病的新治疗策略。针对参与 HIV-1 增殖和存活的特定 RNA-蛋白质相互作用，例如 Tat-TAR 或 Rev-RRE，是一种有前景的方法。我们的初步结果表明，新型配体能够稳定 TAR RNA、在纳摩尔浓度下抑制 Tat-TAR 相互作用以及抑制 MT-2 细胞中的 HIV-1。现在将在这些初步结果的基础上开发一个缀合物库，以靶向 Tat-TAR 相互作用，以高亲和力和特异性与 TAR 结合。拟议的研究将进一步帮助确定这种方法的有效性。这里提出的工作是一项涵盖有机合成、生物物理化学和 HIV 发病机制的多学科努力，描述了小分子介导的 Tat-TAR 抑制剂抑制作为 HIV-1 疗法的发展。这项工作的成功将是对目前艾滋病治疗和 RNA 靶向中可用的蛋白质特异性方法的重大补充。我们建议使用 31 nt TAR 靶序列来设计可用于抑制 Tat-TAR 相互作用的缀合物；为开发小分子 RNA 靶向 HIV-1 疗法开辟了可能性。 公共健康相关性：几十年来对 RNA 结构的研究表明它是一个既定的药物靶点，众所周知是小分子抗生素的受体。尽管自 20 世纪 40 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，但仍迫切需要新的抗菌和抗病毒方法来对抗耐药性，这严重限制了现有抗生素的有效性。 为了研究基于小分子的特异性与电荷/形状互补性相结合的优势，我们启动了一项计划，开发一种使用多聚配体（由具有独立结合位点的配体组成）的方法，该方法可用于靶向特定的 RNA。该提案重点关注小分子氨基糖（新胺）缀合物的开发，作为该方法的一个例子。识别多种病原体中基本药物靶点的综合方法可以与我们开发小分子的补充方法相结合，这些小分子以特定的方式与先前已知的以及快速识别的新 RNA 靶点以高亲和力结合。 抑制 Tat/TAR 相互作用可促进 HIV RNA 转录，随后阻止 HIV 复制。本申请的中心假设是，具有独立结合位点的两个配体的缀合可以与适当的接头缀合，以提供高亲和力的TAR特异性配体，能够在纳摩尔浓度下抑制Tat/TAR相互作用。此外，该测定适用于基于 RNA 的药物发现，其中可以组合两个具有独立结合位点的药效团来选择高亲和力配体。最终，与现有分子相比，TAR 结合配体的亲和力和特异性得到改善，这将为人们更好地了解新靶点在抗击 HIV 中的潜在用途提供帮助。 NUBAD 具备良好的能力来合成分子并进行抑制的生物物理测定。从测定中鉴定出的在纳摩尔 Kd 下抑制 tat-TAR 相互作用的精选化合物将被测试对 HIV 的抑制作用。