| Project/Area Number |
14572158
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
応用薬理学・医療系薬学
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| Research Institution | Kumamoto University |
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Principal Investigator |
ARIMA Hidetoshi Kumamoto University, Graduate School of Medical and Pharmaceutical Sciences, Associate Professor, 大学院・医学薬学研究部, 助教授 (50260964)
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| Co-Investigator(Kenkyū-buntansha) |
HIRAYAMA Fumitoshi Kumamoto University, Graduate School of Medical and Pharmaceutical Sciences, Professor, 大学院・医学薬学研究部, 助教授 (90094036)
UEKAMA Kaneto Kumamoto University, Graduate School of Medical and Pharmaceutical Sciences, Professor, 大学院・医学薬学研究部, 教授 (90040328)
KAI Hirofumi Kumamoto University, Graduate School of Medical and Pharmaceutical Sciences, Professor, 大学院・医学薬学研究部, 教授 (30194658)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2003: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 2002: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | cyclodextrin / dendrimer / conjugate / gene transfer / cytotoxicity / 遺伝子キャリアー / ガラクトース / マンノース / 核移行シグナル |
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| Research Abstract |
To design a novel gene transfer carrier, we prepared dendrimer conjugates with α-cyclodextrin (α-CDE conjugates) with its various degree of substitution (DS) and investigated the chemical structure, physicochemical properties, in vitro and in vivo gene transfer activity and cytotoxicity. α-CDE conjugates formed the complexes with plasmid DNA (pDNA), resulting in a change of the particle sizes of pDNA complexes, but the distinction of physicochemical properties among their vector/pDNA complexes was only very slight. The membrane-disruptive ability of α-CDE conjugates on liposomes encapsulating calcein and their cytotoxicity to NIH3T3 and HepG2 increased with an increase in the DS value of α-cyclodextrin. In vitro gene transfer activity of α-CDE conjugates in both NIH3T3 and HepG2 cells augmented as the charge ratio (vector/pDNA) increased, and the activity of α-CDE conjugate (G3, DS 2.4) was the highest at higher charge ratios among dendrimer (G3), the three α-CDE conjugates, and commercial transfection reagents such as Lipofectin and TransFast. After intravenous administration of pDNA complexes in mice, α-CDE conjugate (DS 2.4) delivered pDNA more efficiently in spleen, liver, and kidney, compared with dendrimer and other α-CDE conjugates. The potential use of α-CDE conjugate (G3, DS 2.4) could be expected as a nonviral vector in vitro and in vivo, and these data may be useful for design of α-CyD conjugates with other nonviral vectors. Furthermore, α-CDE conjugates attached with galactose (GalCDE conjugates) improved gene transfer activity more than parent α-CDE conjugates in HepG2 and NIH3T3 cells, suggesting that GalCDE conjugates are superior gene transfer carrier to the other commercially-available transfection agents.
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