MOLECULAR DESIGN OF NEW EFFECTIVE ANTISENSE NUCLEIC ACIDS CAPABLE OF HYBRIDIZATION TO THE TARGET NENES

能够与目标九烯杂交的新型有效反义核酸的分子设计

• 批准号: 07408014
• 负责人: SEKINE Mitsuo
• 金额: $ 19.65万
• 依托单位: TOKYO INSTITUTE OF TECHNOLOGY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07408014/
• 关键词: ANTISENCE NUCLEIC ACID; CONFORMATION; 3'-ENDO; CYCLOURIDYLIC ACID; INTRAMOLECULAR CYCLIZATION; MOLECULAR MECHANICS; SOLID PHASE SYNTHESIS; DUPLEX FORMATION; 3′-endo; シクロヌクレオチド; 遺伝子治療; 分子内水素結合; H-ホスホネートDNA

A 2'-O-methyluridylic acid derivative having a cyclic structure linked between the 5-position of the uracil residue and the 5'-phosphate group was synthesized. The NMR analysis of suggests that this cyclouridylic acid derivative has exclusively the C3'-endo conformation which is in favor of duplex formation with RNA.Two oligonucleotides [pc3Um(pT)_9 and pc3Um(pU)_9]incorporating this cyclouridylic acid unit at the 5'-terminal site were synthesized by using the fully protected cyclouridylic acid 3'-phosphoramidite derivative in the solid phase synthsis. To examine the actual effect of this cyclic structure on the thermal stability of duplexes between the modified oligonucleotides and their complementary oligonucleotides, two oligonucleotides [pUm(pT)_9 and pUm(pU)_9] having an acyclic structure were also synthesized. As the complementary oligonucleotides, dA(pdA)_9 and A(pA)_9 were used for Tm experiments with these 5'-terminal modified oligonucleotides. The Tm values of all possible duplexes were measured. These results clearly show that there the duplex of A(pA)_9/pc3Um(pT)_9 has a higher Tm value by 5.5ﾟC than that of A(pA)_9/T(pT)_9. This is rather significant compared with all other cases. Moreover, the Tm value of A(pA)_9/pc3Um(pT)_9 is 4.5ﾟC higher than that of A(pA)_9/pUm(pT)_9. This result suggests that the cyclic structure can considerably contribute to stabilization of the duplex only in the case of the modified oligomer (DNA) and decaadenylate (RNA).

合成了具有连接在尿嘧啶残基的5位和5'-磷酸基团之间的环状结构的2'-O-甲基尿苷酸衍生物。NMR分析表明该环尿苷酸衍生物仅具有C3'-内基。有利于与 RNA 形成双链体的构象。两个寡核苷酸 [pc3Um(pT)_9 和 pc3Um(pU)_9] 包含此构象通过在固相合成中使用完全保护的环尿苷酸3'-亚磷酰胺衍生物合成了5'端位点的环尿苷酸单元，以检查该环状结构对修饰的寡核苷酸和双链体之间的热稳定性的实际影响。还合成了具有非环状结构的两种寡核苷酸[pUm(pT)_9和pUm(pU)_9]作为互补寡核苷酸。 dA(pdA)_9 和 A(pA)_9 用于这些 5' 末端修饰寡核苷酸的 Tm 实验，测量了所有可能的双链体的 Tm 值。这些结果清楚地表明存在 A( 的双链体。 pA)_9/pc3Um(pT)_9 的 Tm 值比 A(pA)_9/T(pT)_9 高 5.5°C。与所有其他情况相比，A(pA)_9/pc3Um(pT)_9 的 Tm 值比 A(pA)_9/pUm(pT)_9 高 4.5°C。仅在修饰寡聚物 (DNA) 和十腺苷酸 (RNA) 的情况下，环状结构才能显着促进双链体的稳定。

项目成果

T.Wada, F.Honda, Y.Seto, S.Kawahara, M.Sekine: "Chemical Synthesis of H-Phosphonate DNA without Using N-Protecting Groups" Nucleic Acids Symposium Series. 37. 19-20 (1997)

T.Wada、F.Honda、Y.Seto、S.Kawahara、M.Sekine：“不使用 N 保护基团的 H-磷酸 DNA 的化学合成”核酸研讨会系列。

Kohji Seio et al.: "Chemical Synthesis and Conformational Properties of a New Cyclouridylic acid Having an Ethylene Bridge between the Uracil 5-Position and 5′-Phosphate Group" Journal of Organic Chemistry. 61・4. 1500-1504 (1996)

Kohji Seio 等人：“尿嘧啶 5 位和 5-磷酸基团之间具有乙烯桥的新型环尿苷酸的化学合成和构象性质”有机化学杂志 61・4（1996 年）。

Shun-ichi Kawahar et al.: "1:1 and 1:2 Complexes of Bu_4 NF and BF_4 Et_2 O:New Reagents for Rapid Cleavage of Silyl Ethers" Tetrahedron Lett.37. 509-512 (1996)

Shun-ichi Kawahar 等人：“Bu_4 NF 和 BF_4 Et_2 O 的 1:1 和 1:2 复合物：用于快速裂解硅醚的新试剂”Tetrahedron Lett.37。

K.Seio, T.Wada, K.Sakamoto, S.Yokoyama, M.Sekine: "Chemical Synthesis and Properties of Conformationally Flxed Diuridine Monophosphates as Building Blocks of RNA Tum Motif" Journal of Organic Chemistry. 印刷中

K.Seio、T.Wada、K.Sakamoto、S.Yokoyama、M.Sekine：“作为 RNA Tum 基序构建模块的构象固定二尿苷单磷酸的化学合成和特性”有机化学杂志正在出版。

T.Wada, Y.Sato, F.Honda, S.Kawahara, M.Sekine: "Chemical Synthesiss of Oligodeoxyribonucleotides Using N-Unprotected H-Phosphonate Monomers and Carbonium and Phosphoniun Condensing Reagents" Journal of American Chemical Society. 119・52. 12710-12721 (1997)

T.Wada、Y.Sato、F.Honda、S.Kawahara、M.Sekine：“使用 N-未保护的 H-磷酸酯单体和碳和膦缩合试剂化学合成寡脱氧核糖核苷酸”美国化学会杂志 119・52。 12710-12721 (1997)