MECHANISM OF SYNTHETIC RIBOZYME ACTION

描述：(改编自申请者摘要)本提案旨在 具有不可切割核苷酸间连接的核酶的继续研究 并利用核苷酸间的特殊化学修饰 在研究和开发新的治疗剂方面的联系 针对核酸中的特定靶序列作为一种机制 疾病的控制。这些修改是“串联”主干 将磷酸二酯转化为 亚膦(5‘)亚甲基或亚膦(3’)亚甲基取代 分别用于5‘-氧或3’-氧的亚甲基基团。这 该提案基于以下最近的观察结果：(1)ADP和 5‘-氧基被亚甲基取代的ATP类似物 作为多核苷酸磷酸化酶和噬菌体T3RNA的底物 分别为聚合酶。(2)含All的寡核苷酸 膦(5‘)亚甲基键似乎与它们的天然键是等位异构键。 因为它们能够参与三股链的作用 队形。(3)由于P-C键的存在，齐聚物 包含这样的连接完全抵抗类型的 化学和核酸酶的裂解，涉及到 磷和5‘-氧在它们的天然对应物中。深入探讨 将进行化学、物理、酶和生物学研究 关于单核苷酸、寡核苷酸和核酸类似物 含有亚磷酸亚甲基键。此外，还替换了 核酶中裂解点上的膦(5‘)亚甲基连接 底物应允许形成核酶的活性构象 在分析过程中不会有自我毁灭的风险。第二个特别节目 应用涉及单核苷酸类似物的潜在用途 作为治疗药物，而且看起来很可能是一个或多个 所述核糖核苷三磷酸类似物含有 取代5‘-羟基的亚甲基将被发现用于 一种或多种人聚合酶的底物(包括逆转录 转录酶)。

DESCRIPTION: (Adapted from Applicant's Abstract) This proposal seeks to continue studies on ribozymes with non-cleavable internucleotide linkages and to exploit particular chemical modifications in internucleotide linkages in the study and development of new therapeutic agents that are aimed at specific target sequences in nucleic acids as a mechanism for the control of diseases. The modifications are "in-line" backbone changes in which the phosphodiester is converted to either a phosphono(5')methylene or phosphono(3')methylene linkage by substituting a methylene group for the 5'-oxygen or 3'-oxygen, respectively. This proposal is based on the following recent observations (1) The ADP and ATP analogues in which the 5'-oxygens are replaced with methylene groups serve as substrates for polynucleotide phosphorylase and phage T3 RNA polymerase, respectively. (2) Oligonucleotides containing all phosphono(5')methylene linkages appear to be isosteric with their natural counterparts in that they are able to participate in triple strand formation. (3) Because of the presence of the P-C bond, oligomers containing such linkages are completely resistant to the types of chemical and nuclease cleavage that involve breakage between the phosphorous and the 5'-oxygen in their natural counterparts. In-depth chemical, physical, enzymatic and biological studies will be performed on mononucleotides, oligonucleotide and nucleic acid analogues containing phosphonomethylene linkages. In addition substitution of a phosphono(5')methylene linkage at the cleavage point in a ribozyme substrate should permit formation of the ribozyme's active conformation without risk of its self-destruction during analysis. A second special application concerns the potential use of the mononucleotide analogues themselves as therapeutic drugs and it seems likely that one or more of the ribo- and deoxyribo-nucleoside triphosphate analogues containing a methylene group in place of the 5'-hydroxyl will be found to serve a substrates for one or more of the human polymerases (including reverse transcriptase).