TRIPLEX FORMING OLIGONUCLEOTIDES TARGETED AGAINST HIV-1

描述：（改编自申请人摘要）HIV感染模型 为指导三相成形的化学细化提供了理论依据 寡核苷酸（TFO）。 该项目的主要目标是提高 TFO-双链DNA与 具体重点是提高HIV 38 p类TFO的疗效。 的 方法是集中使用核苷同系物， 在TA或CG倒位位点的稳定碱基三联体， 氢键势（黄嘌呤核苷和福霉素类）或通过 改变的主链构型（拉伸的主链类）。 实现 这个TFO增强计划，调查人员利用了一个正在进行的 具有分子建模、自然科学和生物技术专业知识的团队之间的合作 产品和核酸化学和核酸生物物理学。 的 第二个目标是尝试开发有用的终端修改。 还 作者将试图提高TFO稳定性方面的细胞 核酸酶，以最大化TFO摄入细胞核的速率， 探索使用可共价结合的TFO-烷基化剂缀合物， 与它们的双链体DNA靶位点交联。 研究者 特别感兴趣的是这些烷化剂共轭物作为测量 TFO在体内的结合以及作为增强功效的机制 TF 0介导的转录停滞。 双链体DNA结合潜力 建议的TFO同系物将通过结构和 热力学分析采用紫外光谱，核磁共振，带位移和 足迹法 显示出增强的“前导”HIV 38 p同系物 结合亲和力、稳定性或细胞吸收特征，或具有 已被证明具有有效的TFO介导的能力， 交联，将评估U937中增强的抗病毒活性， MT4测定。

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.