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的潜在机制的日益增长的知识来开发艾滋病的新治疗策略。靶向特定的RNA-蛋白质相互作用，如TAT-TAR或REV-RRE，参与HIV-1的增殖和生存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TAR RNA，在纳摩尔浓度下抑制TAT-TAR相互作用，并抑制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靶向TAT-TAR相互作用的连接物库，这些连接物与TAR具有高亲和力和特异性。拟议的研究将进一步有助于确定这一方法的有效性。这项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，描述了作为HIV-1治疗药物的小分子介导的TAT-TAR抑制剂的发展。拟议工作的成功将是对目前在艾滋病治疗和RNA靶向方面可用的蛋白质特异性方法的重大补充。我们建议使用31个核苷酸的TAR靶序列来设计可用于抑制TAT-TAR相互作用的结合物；为开发针对HIV-1的小分子RNA靶向疗法打开了可能性。 与公共健康相关：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，它是小分子抗生素的受体。虽然自20世纪40年代发现链霉素以来，细菌核糖体一直是众所周知的抗生素受体，阻止蛋白质合成，但迫切需要新的抗菌和抗病毒方法来对抗耐药性，这严重限制了当前抗生素的有效性。为了研究基于小分子的特异性结合电荷/形状互补的优势，我们启动了一项计划，开发一种使用多聚体配体(由具有独立结合位点的配体组成)的方法来靶向特定的RNA。本提案侧重于开发小分子氨基糖(新胺)共轭化合物，作为这一方法的一个例子。一种在多种病原体中识别基本药物靶点的综合方法可以与我们的互补方法相结合，即开发以特定方式与先前已知的以及快速识别的新RNA靶点具有高亲和力的小分子。TAT/TAR相互作用的抑制促进了艾滋病毒RNA的转录，从而阻止了艾滋病毒的复制。这一应用的中心假设是，两个具有独立结合位点的配体可以与适当的连接物结合，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的相互作用。此外，该方法适用于基于RNA的药物发现，其中具有独立结合位点的两个药效团可以结合以选择高亲和力的配体。最终，与现有分子相比，具有更高亲和力和特异性的焦油结合配体的发现将为在抗击艾滋病毒方面实现新靶点的潜在用途提供更好的理解。NUBAD公司装备精良，可以合成分子并进行生物物理抑制试验。从检测中确定的在纳米分子KD处抑制TAT-TAR相互作用的选定化合物将进行HIV抑制试验。