TRIPLEX FORMING OLIGONUCLEOTIDES TARGETED AGAINST HIV 1

描述：（改编自申请人的摘要）这是一个修订的 11月研究组先前审查的申请。先前 该实验室的实验提供了证据， 三螺旋形成寡核苷酸（TFO），用于聚集在 病毒LTR可以抑制HIV-1的转录， 感染的单核细胞系。本提案的总体目标是 制定合理的药物设计计划，通过以下方式改善TFO结合 采用先进的分子建模技术，核磁共振和最近的 核酸化学的进展。提案的具体目标 （1）深化对三链体结构的认识， 特别是水和离子结合的作用，采用分子 建模和2DNMR方法。(2)使用建模、NMR和相关物理 评价咪唑-核苷同系物在 TFO。(3)设计可以附着在 TFO的末端，以增强结合复合物的稳定性。（四） 合成含有这种碱取代基的HIV 33类TFO， 稳定末端修饰，并评估其结构、稳定性 和与LTR结合的选择性。(5)利用表面上庞大的 可能伴随RRY三螺旋形成的螺旋扭曲变化 使用“定相”质粒测定。本试验将探索 TFO与HIV-1启动子区结合后可能发生的变化， 还将探索聚合物连接元件的作用， 合成到TFO的中心。(6)采用亲脂性3 '-末端 胆固醇类型的修饰和新的阳离子脂质， HIV 33的TFO的细胞摄取和细胞内分配 课(7)测量急性和慢性感染者的抗艾滋病毒活性 显示增强的LTR的HIV 33类的修饰的TFO的细胞 结合、增强的摄取或结合位点的扭转变形。

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.