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

基本信息

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

项目摘要

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.

反义和抗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，并在活细胞中抑制基因表达序列。另一方面，抗基因寡核苷酸（三核苷酸，TFOS）与基因组DSDNA结合，从而导致基因表达控制。我们以前开发了各种构象锁定的核酸（桥接核ACLD，BNA）。用BNA单体修饰的遗传核苷酸被广泛用于评估物理和生物学特性。在下面总结了这些结果。1）含有2'BNA单体的TFO，发现与同孕胺甲酰胺丁烷dsDNA靶标显着稳定，在接近生理条件下均具有三酰胺的合成途径。 PSOE在抗固国策略中是一个严重的问题。设计，合成并证明具有2-吡啶酮基单元的2'，4'-BNA，具有2-吡啶酮碱基单位。新修饰的寡核苷酸与dsDNA或ssRNA靶标的高结合亲和力。还观察到了这些寡核苷酸类似物的出色酶促稳定性。5）反义2'，4'-BNA寡核苷酸抑制了靶基因在活细胞中具有较高序列序列和非胞毒性的活细胞中的表达。