MECHANISM OF SYNTHETIC RIBOZYME ACTION

合成核酶的作用机制

• 批准号: 2518965
• 负责人: PETER T GILHAM
• 金额: $ 18.16万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-07-09 至 1999-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2518965
• 关键词: chemical substitution; enzyme mechanism; nucleic acid chemical synthesis; nucleotide analog; oligonucleotides; ribozymes

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).

描述：（改编自申请人的摘要）本提案旨在 继续研究具有不可切割核苷酸间键的核酶 并利用核苷酸间的特殊化学修饰 在研究和开发新的治疗药物， 针对核酸中的特定靶序列， 疾病的控制。这些修改是“内联”主干 其中磷酸二酯被转化为 膦酰基（5 ′）亚甲基或膦酰基（3 ′）亚甲基连接， 5 ′-氧或3 ′-氧分别为亚甲基。这 （1）特设工作组和 5 ′-氧被亚甲基取代的ATP类似物 作为多核苷酸磷酸化酶和噬菌体T3 RNA的底物 聚合酶。(2)含有所有的寡核苷酸 膦酰基（5 ′）亚甲基键似乎与它们的天然配体是等排的， 因为它们能够参与三链 阵(3)由于P-C键的存在， 含有这种键的化合物完全抵抗 化学和核酸酶切割，涉及断裂之间的 磷和5 '-氧在它们的天然对应物中。深入 将进行化学、物理、酶和生物学研究 寡核苷酸和核酸类似物 含有膦酰基亚甲基键。此外，替换a 核酶中切割点上的膦酰基（5 '）亚甲基键 底物应允许核酶活性构象的形成 在分析过程中没有自毁的风险。第二特殊 本申请涉及蒙帕肽类似物的潜在用途 它们本身作为治疗药物，似乎可能是一种或多种 所述核糖-和脱氧核糖-核苷三磷酸类似物含有 将发现亚甲基取代5 ′-羟基起到 一种或多种人聚合酶（包括反向聚合酶）的底物 转录酶）。