CHEMICAL SYNTHESIS OF A NEW MATERIAL OF ANTISENSE NUCLEIC ACID "2'-PHOSPHORYLATED RNAS" -DIRECTED TOWARD ITS BASIC STRUCTURAL STUDIES AND REGULATION OF EXPRESSION OF HIV VIRUS-

反义核酸新材料“2-磷酸化RNAS”的化学合成-针对其基础结构研究和HIV病毒表达调控-

• 批准号: 05558090
• 负责人: SEKINE Mitsuo
• 金额: $ 9.92万
• 依托单位: TOKYO INSTITUTE OF TECHNOLOGY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-05558090/
• 关键词: ANTISENCE NUCLEIC ACID; 2'-PHOSPHATE; RNA; CHEMICAL SYNTHESIS; HIV; SOLID PHASE SYNTHESIS; ENZYME RESISTANCE; AMIDITE

This year, we have tried to study the development of new organic reactions required for the synthesis of 2'-phosphorylated RNAs. As the result, we have achieved the following results : First, we found that bis (2-cyano-1,1-dimethylethyl) phosphoryl can be used not only as a 2'-hydroxyl protecting groups but also as a precursor of the 2'-phosphate group. This protected phosphoryl group was found to be quite stable upon removal of a 3', 5-cyclic silyl protecting group with HF/Py in dichloromethane and not to migrate from the 3'-2'hydroxyl functions. This finding led us to realize the construction of key intermediates, 2'-phosphorylated 3'-phosphoramide units which were required throughout this study. The building units were utilized for the solid phase synthesis of 2'-phosphorylated uridylate derivatives. We could synthesize 2,4,6, and 10mers of 2'-phosphorylated uridylates in good yields. They were found to be resistant to acid and alkaline conditions as well as enzymatic digestion with spleen phosphodiesterase. In particular, the 10 mer proved to have an unique structure as evidenced by NMR.At the present time, we are studying the binding ability of these new nucleic acid derivatives for target DNA or RNA fragmets.

今年，我们试图研究合成2 '-磷酸化RNA所需的新有机反应的发展。首先，我们发现双（2-氰基-1，1-二甲基乙基）磷酰基不仅可以作为2 ′-羟基的保护基，而且可以作为2 ′-磷酸酯基的前体。发现这种保护的磷酰基在用HF/Py在二氯甲烷中除去3 ′，5-环甲硅烷基保护基时非常稳定，并且不从3 ′-2 ′羟基官能团迁移。这一发现使我们实现了关键中间体的构建，即整个研究所需的2 '-磷酸化3'-磷酰胺单元。构建单元用于固相合成2 '-磷酸化尿苷酸衍生物。我们可以合成2，4，6和10聚体的2 '-磷酸化尿苷酸在良好的产率。他们被发现是耐酸性和碱性条件，以及酶消化脾磷酸二酯酶。特别是10聚体的NMR证实了其独特的结构。目前，我们正在研究这些新的核酸衍生物与靶DNA或RNA片段的结合能力。

项目成果

T. Wada et al.: "2-Trimethylsilylethyl as a phosphate protecting groups in oligonucleotide synthesis" Tetrahedron Letters. 35. 757-760 (1994)

T. Wada 等人：“2-三甲基甲硅烷基乙基作为寡核苷酸合成中的磷酸盐保护基团”四面体快报。

T.Wada et al.: "Synthesis and Propertise of N-Phosphorylated Nucleosides" J.Am.Chem.Soc.116. 9901-9911 (1994)

T.Wada 等人：“N-磷酸化核苷的合成和特性”J.Am.Chem.Soc.116。

T.Wada et al.: "2-Trimethylsilylethyl as a phosphate protecting group in oligonucleotide synthesis" Tetrahedron Letters. 35. 757-760 (1994)

T.Wada 等人：“2-三甲基甲硅烷基乙基作为寡核苷酸合成中的磷酸盐保护基团”四面体快报。

M.Katahira et.al: "A Raman spectroscopic study on conformations of DNA oligomers" Nucleosides & Nucleotides. 6&7. 1467-1487 (1994)

M.Katahira 等人：“DNA 寡聚物构象的拉曼光谱研究”

M. Sekine et al.: "New method for removal of modified trityl and pixyl groups by use of an acid species generated" Nucleosides & Nucleotides. 6&7. 1397-1414 (1994)

M. Sekine 等人：“通过使用生成的酸物质去除修饰的三苯甲基和像素基团的新方法”