ENHANCED EFFICACY OF TRIPLEX OLIGONUCLEOTIDES

该项目的长期目标是开发一种新型的 基于三阻碍形成寡核苷酸（TFO）的治疗剂。在 该提议，将亲脂性基团连接到抗癌和抗- 将检查HSV-2 TFO以增强其细胞功效。 具体地，胆固醇修饰的TFO被设计为结合到 HSV-2的共有孕酮反应元件和lE 175启动子 将详细审查。初步数据显示， 序列特异性TFO的功效显著增加， 胆固醇的附着。在I期研究中，重点是 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 分布，毒性和聚集特性的TFOs在 将研究水溶液。附着胆固醇的影响 通过各种长度的新型连接体以及可逆二硫键 还将审查各种联系。 在第二阶段，使用我们目前正在开发的方法， 大规模合成的实验室，克量的这些改性 将合成TFO。将用脂质体包封TFO。 执行。胆固醇基TFO和包封的TFO将用于 在小鼠中的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.