TRIPLEX FORMING OLIGONUCLEOTIDES TARGETED AGAINST HIV-1

描述：(改编自申请者摘要)HIV感染模型 用来指导三相成形的化学细化。 寡核苷酸(TFOS)。该项目的主要目标是增强 TFO-双链DNA与TFO结合的结合常数和结合位点特异性 特别侧重于提高HIV38P类TFO的疗效。这个 方法是重点使用核苷同系物，这可以形成 蚀变法研究TA或CG反转部位的稳定碱基三联体 氢键潜力(黄嘌呤和甲霉素类)或通过一个 更改了主干配置(扩展主干类)。要实现 在这个TFO增强计划中，调查者利用了正在进行的 拥有分子建模专业知识的团队之间的协作，自然 产品与核酸化学和核酸生物物理学。这个 第二个目标是尝试开发有用的终端修改。另外， 作者将尝试提高TFO在细胞方面的稳定性 核酸酶，以最大限度地提高TFO摄取到细胞核的速度并 探索TFO-烷基化偶联物的用途 与他们的双链DNA目标位点相关联。调查员是 对作为测量工具的这些烷基化合物特别感兴趣 TFO在体内的结合并作为一种增强疗效的机制 TFO介导的转录抑制。双链DNA结合势 建议的TFO同源化合物将通过结构和 用紫外可见光谱、核磁共振、谱带位移和红外光谱进行热力学分析 足迹方法。HIV38P的“先导”同系物表现出增强 结合亲和力、稳定性或细胞摄取特性，或具有 已被证明具有高效的TFO介导的能力 将评估U937和U937中增强的抗病毒活性 MT4检测。

DESCRIPTION: (Adapted from applicant's abstract) An HIV infection model is presented to direct the chemical refinement of triplex forming oligonucleotides (TFOs). The primary goal of this project is to enhance the binding constant and site specificity of TFO-duplex DNA binding with specific focus on enhancing the efficacy of the HIV38p class of TFO. The approach is to focus of the use of nucleoside homologues which can form stable base triplets at sites of TA or CG inversion by means of altered H-bonding potential (the xanthosine and formycin class) or by means of an altered backbone configuration (the stretched backbone class). To achieve this program of TFO enhancement, the investigator exploits an on-going collaboration between groups with expertise in molecular modeling, natural products and nucleic acid chemistry and nucleic acid biophysics. The second goal is to try to develop useful terminal modifications. Also, the author will try to enhance TFO stability with respect to cellular nucleases, to maximize the rate of TFO uptake into the nucleus and to explore the use of TFO-alkylator conjugates which can be covalently crosslinked to their duplex DNA target site. The investigator is particularly interested in these alkylator conjugates as tools to measure TFO binding in vivo and as a mechanism to enhance the efficacy of TFO-mediated transcription arrest. The duplex DNA binding potential of the proposed TFO homologues will be explored by structural and thermodynamic analyses employing UV spectroscopy, NMR, band shift and footprinting methods. The "lead" HIV38p homologues which exhibit enhanced binding affinity, stability or cell uptake characteristics, or which have been shown to possess the capacity for efficient TFO-mediated crosslinking, will be assessed for enhanced antiviral activity in U937 and MT4 assays.