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    • 17. 发明授权
    • Enhanced adenovirus-assisted transfection composition and method
    • 增强腺病毒辅助转染组合物和方法
    • US5830730A
    • 1998-11-03
    • US852934
    • 1997-05-08
    • Michael S. GermanFrancis C. Szoka, Jr.
    • Michael S. GermanFrancis C. Szoka, Jr.
    • A61K48/00C12N15/861C12N15/87C12N15/63C12N7/00C12N15/11
    • C12N15/86A61K48/00C12N15/87C12N2710/10343
    • A composition for transfecting eukaryotic cells comprising a cationic polymer which has protonatable groups which serve to buffer the acidic endosome, protecting the endocytosed polynucleotide from degradation and a viral agent is used to target uptake into and/or lysis from endosomes in the desired eukaryotic cell. By co-infecting the eukaryotic cells with cationic polymer, polynucleotide, and the viral agent, the polynucleotide is brought into the cell and then released. Preferably, polyamidoamine dendrimers are used as the cationic polymer and adenovirus is used as the viral agent. The dendrimers help associate plasmid DNA with the adenovirus, which then provokes receptor-mediated endocytosis. Within the endosome, the tertiary amine groups of the dendrimer buffer the pH change in the endosome. Then, the endosomalytic activity of the adenovirus releases the plasmid DNA into the cell. Also preferably, the adenovirus and the dendrimers are mixed before addition to the polynucleotide and the cells are washed after about 1.5 hours of incubation.
    • 用于转染真核细胞的组合物包含阳离子聚合物,其具有用于缓冲酸性内体的可质子基团,保护内吞多核苷酸免于降解和病毒剂的阳离子聚合物用于靶向所需真核细胞内内体的摄取和/或裂解。 通过用阳离子聚合物,多核苷酸和病毒剂共同感染真核细胞,将多核苷酸引入细胞中,然后释放。 优选使用聚酰胺胺树枝状大分子作为阳离子聚合物,并使用腺病毒作为病毒剂。 树枝状大分子帮助质粒DNA与腺病毒相关联,其然后引发受体介导的内吞作用。 在内体内,树状聚合物的叔胺基团缓冲了内体内的pH变化。 然后,腺病毒的内分泌活性将质粒DNA释放到细胞中。 还优选地,在加入多核苷酸之前将腺病毒和树枝状大分子混合,并在孵育约1.5小时后洗涤细胞。