| Project/Area Number |
12557201
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Chemical pharmacy
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| Research Institution | Osaka University |
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Principal Investigator |
IMANISHI Takeshi Osaka University, Graduate School of Pharmaceutical Sciences, Professor, 薬学研究科, 教授 (40028866)
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| Co-Investigator(Kenkyū-buntansha) |
OBIKA Satoshi Osaka University, Graduate School of Pharmaceutical Sciences, Research Associate, 薬学研究科, 助手 (80243252)
DOI Takefumi Osaka University, Graduate School of Pharmaceutical Sciences, Professor, 薬学研究科, 教授 (00211409)
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| Project Period (FY) |
2000 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥12,200,000 (Direct Cost: ¥12,200,000)
Fiscal Year 2002: ¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 2001: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2000: ¥5,900,000 (Direct Cost: ¥5,900,000)
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| Keywords | 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 |
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| Research Abstract |
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.
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