TRIPLEX FORMING OLIGONUCLEOTIDES TARGETED AGAINST HIV 1

描述：(改编自申请者摘要)这是一个修订后的 11月份研究科以前审查过的申请。上一首 这个实验室的实验已经提供了证据，证明了 形成聚集性Sp1位点的三螺旋寡核苷酸(TFO) 病毒LTR可长期抑制HIV-1的转录 感染的单核细胞系。这项提案的总体目标是 制定合理的药物设计方案，通过以下方式改善TFO的结合 利用先进的分子建模技术，核磁共振和最新的 核酸化学研究进展。该提案的具体目的 是：(1)提炼对三链结构的总体认识， 尤其是水和离子结合的作用，采用分子 建模方法和二维核磁共振方法。(2)使用建模、核磁共振和相关物理方法 评价咪唑-核苷同系物在临床中的应用 TFOS。(3)设计主要的沟槽粘结剂，这些粘合剂可粘贴到 TFOS的末端，从而增强结合络合物的稳定性。(4) 合成含有这种碱基取代基的HIV33类TFOS和 稳定末端改性并评价其结构、稳定性 以及与LTR结合的选择性。(5)利用看似庞大的资源 可能伴随RRY三螺旋形成的螺旋扭度的变化 使用“阶段性”的质粒法。这项分析将探索这一扭曲 当TFO与HIV-1启动子区域结合时可能发生的变化 还将探索聚合物连接元件的影响，这些元件是 合成到TFO的中心。(6)使用亲脂性的3‘端 修饰胆固醇类型和新型阳离子脂类以增强 HIV-33对TFOS的细胞摄取和细胞内分配 班级。(7)检测急性和慢性感染者的抗艾滋病毒活性 HIV 33类修饰的TFOS细胞显示增强的LTR 结合、增强摄取或结合部位的扭曲变形。

DESCRIPTION:(Adapted from Applicant's Abstract) This is a revised application previously reviewed by the November Study Section. Previous experiments form this laboratory have provided evidence that binding of a triple helix forming oligonucleotide (TFO) to clustered Sp1 sites in the viral LTR could repress transcription of HIV-1 in a chronically infected monocyte cell line. The overall goal of this proposal is to develop a program of rational drug design to improve TFO binding by employing advanced molecular modelling techniques, NMR and recent advances in nucleic acids chemistry. The specific aims of the proposal are: (1) Refine the general understanding of triplex structure, especially the role of water and ion binding, employing molecular modeling and 2DNMR methods. (2) Use modeling, NMR and related physical methods to evaluate the utility of imidazole-nucleoside homologues in TFOs. (3) Design major groove binding agents which may be affixed to the ends of TFOs so as to enhance the stability of the bound complex. (4) Synthesize HIV33-class TFOs which contain such base substituents and stabilizing end modifications and evaluate their structure, stability and selectivity of binding to the LTR. (5) Exploit the apparently large change of helix twist which may accompany RRY triple helix formation using a "phasing" plasmid assay. This assay will explore the twist change which may occur upon TFO binding to the HIV-1 promoter region and will also explore effect of polymeric linker elements which are synthesized into the center of the TFO. (6) Employ lipophilic 3'- end modification of the cholesterol type and novel cationic lipids to enhance the cellular uptake and intracellular partitioning of TFOs of the HIV 33 class. (7) Measure anti-HIV activity in acutely and chronically infected cells of modified TFOs of the HIV 33 class which display enhanced LTR binding, enhanced uptake, or torsional distortion of the binding site.