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).

合成了一种2‘-O-甲基尿酸衍生物，其环状结构连接在尿嘧啶残基的5位和5’-磷酸基团之间。核磁共振谱分析表明该环丙烯酸衍生物具有独特的C3‘-Endo构象，有利于与RNA形成双链构象。利用完全保护的环丙烯酸衍生物在固相合成中合成了两个在5’-末端含有该环丙烯酸衍生物的寡核苷酸[pc3Um(PT)9和pc3Um(Pu)9]。为了考察这种环结构对修饰寡核苷酸及其互补寡核苷酸之间双链热稳定性的实际影响，还合成了两个具有非环结构的寡核苷酸[PUM(PT)_9和PUM(PU)_9]。以da(Pda)_9和A(Pa)_9为互补寡核苷酸，对这些5‘端修饰的寡核苷酸进行TM实验。测量了所有可能的双链的Tm值。结果表明，A(Pa)_9/Pc_3Um(PT)_9的Tm比A(Pa)_9/T(PT)_9的Tm高5.5ﾟC。此外，A(Pa)_9/Pc3Um(PT)_9的Tm值比A(Pa)_9/PUM(PT)_9高4.5ﾟC。这一结果表明，只有在修饰齐聚物和十腺苷的情况下，环状结构才能显著地稳定双链体。

项目成果

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）。

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。

Mitsuo Sekine et al.: "Studies on Steric and Electronic Control of 2'-3' Phosphoryl Migration in 2'-Phosphorylated Oligouridylates" Journal of Organic Chemistry. 61・12. 4087-4100 (1996)

Mitsuo Sekine 等：“2-磷酸化寡尿苷酸中 2-3 磷酰基迁移的空间和电子控制研究”有机化学杂志 61・12（1996）。

Hiroyuki Tsuruoka et al.: "Synthesis and Structural and Thermodynamic Properties of Oligonucleotides Containing 2'-O-Phosphorylated Ribonucleotides" Tetrahedron Letters. 37・37. 6741-6744 (1996)

Hiroyuki Tsuruoka 等人：“含有 2-O-磷酸化核糖核苷酸的寡核苷酸的合成以及结构和热力学性质”Tetrahedron Letters 37・37 (1996)。

K.Seio, O.Kurasawa, T.Wada, M.Sekine, et al: "Synthesis and Properties of Conformationally Rigid Cyclouridylic Acids Having Covalent Bond Between the Uracil 5-Position and the 5'-Phosphate Group" Nucleoside Nucleotides. 16・7-9. 1023-1032 (1997)

K.Seio、O.Kurasawa、T.Wada、M.Sekine 等人：“尿嘧啶 5 位和 5-磷酸基团之间具有共价键的构象刚性环尿苷酸的合成和性质”核苷核苷酸 16・。 7-9。1023-1032（1997）