Study to develop novel genomic drugs by using superfunctional oligonucleotide analogues

利用超功能寡核苷酸类似物开发新型基因组药物的研究

• 批准号: 12557201
• 负责人: IMANISHI Takeshi
• 金额: $ 7.81万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12557201/
• 关键词: Sugar conformation; Nucleoside analogue; Oligonucleotide analogue; Artificial nucleic acid; Synthesis; Antisense; Antigene; BNA; 酵素耐性; ホスホロアミダート結合

Antisense and antigene strategies are new methodology for genomic drugs discovery. An antisense oligonucleotide hybridizes with a target mRNA and inhibits gene expression sequence-selectively in a living cell. On the other hand, an antigene oligonucleotide (triplex-forming oligonucleotides, TFOs) binds with genomic dsDNA to result in gene expression control. We previously developed various kinds of conformationally locked nucleic acids (Bridged Nucleic Aclds, BNAs). Ologonucleotides modified with BNA monomers were widely used to evaluate physical and biological properties. These results are summarized below.1) TFOs containing the 2',4'-BNA monomers were found to stabilize significantly triplex formation with the homopurine-homopyrimidine dsDNA target under near physiological conditions.2) Efficient synthetic routes for the 2',4'-BNA monomers bearing unnatural nucleobases wore developed.3) There is no natural nucleoside to recognize a CG base pair, and this psoes a serious problem in antigene strategy. A novel nucleoside analogue, 2',4'-BNA with a 2-pyridone base-unit was designed, synthesized and proved to be excellent for recognition of a CG base pair.4) Another modification was made on a phosphodiester linkage or the sugar moiety of the 2',4'-BNA oligonucleotide. The newly modified oligonucleotides showed high binding affinity with dsDNA or ssRNA target. Excellent enzymatic stability of these oligonucleotide analogues was also observed.5) Antisense 2',4'-BNA oligonucleotides inhibited expression of the target gene in living cell with high sequence-selectivity and non-cytotxicity.

反义和反基因策略是基因组药物发现的新方法。反义寡核苷酸与靶mRNA杂交并在活细胞中序列选择性地抑制基因表达。另一方面，反基因寡核苷酸（三链体形成寡核苷酸，TFO）与基因组dsDNA结合以导致基因表达控制。我们以前开发了各种构象锁定核酸（桥核酸，BNAs）。用BNA单体修饰的寡核苷酸被广泛用于评价其物理和生物学性质。这些结果总结如下：1）发现含有2 '，4'-BNA单体的TFO在接近生理条件下显著稳定了与高嘌呤-高嘧啶dsDNA靶的三链体形成。2）开发了携带非天然核碱基的2 '，4'-BNA单体的有效合成路线。3）没有天然核苷来识别CG碱基对，这是抗基因策略中的一个严重问题。设计、合成了一种新的核苷类似物2 '，4'-BNA，它具有2-吡啶酮碱基单元，并被证明对CG碱基对具有良好的识别能力。4）对2 '，4'-BNA寡核苷酸的磷酸二酯键或糖部分进行了另一种修饰。新修饰的寡核苷酸显示出与dsDNA或ssRNA靶标的高结合亲和力。5）反义2 '，4'-BNA寡核苷酸抑制目的基因在活细胞中的表达，具有高度的序列选择性和非细胞毒性。

项目成果

Obika, S., Hari, Y., Sugimoto, T., Sekiguchi, M., Imanishi, T.: "Triplex-forming enhancement with high sequence selectivity by single 2'-O,4'-C-methylene bridged nucleic acid (2',4'-BNA) modification"Tetrahedron Lett.. 41. 8923-8927 (2000)

Obika, S.、Hari, Y.、Sugimoto, T.、Sekiguchi, M.、Imanishi, T.：“单 2-O,4-C-亚甲基桥核酸具有高序列选择性的三链体形成增强作用

Uchida, M., Hino, N., Yamanaka, T., Hukushima, H., Imanishi, T., uchiyama, Y., Kodama, T., Doi, T.: "Hepatitis C virus core protein binds to a C-terminal region of NS5B RNA polymerase"Hepatology Res.. 22. 297-306 (2002)

Uchida, M.、Hino, N.、Yamanaka, T.、Hukushima, H.、Imanishi, T.、uchiyama, Y.、Kodama, T.、Doi, T.：“丙型肝炎病毒核心蛋白与 C

Yamanaka, T., Uchida, M., Doi, T.: "Innate form of HCV core protein plays an important role in the localization and the function of HCV core protein"Biochem.Biophys.Res.Commun.. 294. 521-527 (2002)

Yamanaka, T.、Uchida, M.、Doi, T.：“HCV 核心蛋白的天然形式在 HCV 核心蛋白的定位和功能中发挥着重要作用”Biochem.Biophys.Res.Commun. 294. 521-

Yamanaka, T., Kodama, T., Doi, T.: "Subcellular localization of HCV core protein regulates its ability for p53 activation and p21 suppression"Biochem.Biophys.Res.Commun.. 294. 528-534 (2002)

Yamanaka, T.、Kodama, T.、Doi, T.：“HCV 核心蛋白的亚细胞定位调节其 p53 激活和 p21 抑制的能力”Biochem.Biophys.Res.Commun.. 294. 528-534 (2002)

Satoshi OBIKA, Mayumi ONODA, Koji MORITA, Jun-ichi ANDOH, Makoto KOIZUMI and Takeshi IMANISHI: "3'-Amino-2',4'-BNA: Novel bridged nucleic acids having an N'3→P5' phosphoramidate linkage"Chem. Commun.. 1992-1993 (2001)

Satoshi OBIKA、Mayumi ONODA、Koji MORITA、Jun-ichi ANDOH、Makoto KOIZUMI 和 Takeshi IMANISHI：“3-Amino-2,4-BNA：具有 N3→P5 氨基磷酸酯键的新型桥接核酸”Chem通讯.. 1992-1993 (2001)