ENHANCED EFFICACY OF TRIPLEX OLIGONUCLEOTIDES

该项目的长期目标是开发一类新的 基于三螺旋形成寡核苷酸(TFOS)的治疗。在……里面 这项建议，将亲脂基团附着在抗癌和抗癌药物上 将对HSV-2 TFOS进行检测，以增强其细胞效力。 具体地说，胆固醇修饰的TFOS旨在结合到 普适孕酮反应元件与HSV-2的LE 175启动子 将被详细审查。初步数据显示，细胞 序列特异性TFOS的有效性显著提高，因为 附着性胆固醇。在L阶段的研究中，重点将是 确认这些结果，并进行系统的调查 这种增强的机制。此外，摄取、稳定性、细胞 TFOS在小鼠体内的分布、毒性和聚集特性 将对水溶液进行研究。附加胆固醇的影响 通过不同长度的新型连接物以及可逆的二硫化物 此外，还将研究它们之间的联系。 在第二阶段，使用我们目前正在开发的方法 实验室大规模合成，克量的这些改性 将合成TFOS。用脂质体包裹TFOS将是 已执行。胆固醇TFOS和封装的TFOS将用于 小鼠体内药代动力学和细胞分布研究。

The long term objective of this project is to develop a novel class of therapeutics based on Triple-Helix Forming Oligonucleotides (TFOs). In this proposal, attachment of lipophilic groups to anti-cancer and anti- HSV-2 TFOs will be examined to enhance their cellular efficacy. Specifically, cholesteryl modified TFOs that are designed to bind to the consensus progesterone response element and the lE 175 promoter of HSV-2 will be examined in detail. Preliminary data indicate that the cellular efficacy of sequence specific TFOs are increased considerably due to the attachment of cholesterol. In the Phase l study, the emphasis will be to confirm these results and undertake a systematic investigation on the mechanism of this enhancement. In addition, uptake, stability, cellular distribution, toxicity and the aggregation properties of the TFOs in aqueous solutions will be studied. The effects of attaching cholesterol through novel linkers of various lengths as well as reversible disulfide linkages will also be examined. In Phase II, using methods that are currently being developed in our laboratory for large scale synthesis, gram quantities of these modified TFOs will be synthesized. Encapsulation of the TFOs with liposomes will be performed. Cholesteryl TFOs and the encapsulated TFOs will be used for pharmacokinetic and cellular distribution studies in mice.