Development of intracellular controlled release system for optimization of gene therapy

开发细胞内控释系统以优化基因治疗

• 批准号: 13470515
• 负责人: MAYUMI Tadanori
• 金额: $ 8.77万
• 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: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470515/
• 关键词: intracellular controlled release; nanoparticles; fusogenic liposomes; drug delivery system; oligonucleotide; confocal laser microscope; gene therapy; ナノパーティクル; 細胞内徐放; 細胞内導入; FACS; ナノスフェアー; アンチセンス核酸; 薬物動態; ポリビニルアミン

In recent years, sustained or controlled drug-release system using nanospheres or microspheres is noticed in systemic pharmacokinetics. However, in the near future, not only "systemic pharmacokinetics" but also "intracellular pharmacokinetics seems to be important in Drug Delivery System research. Technology for delivering sustained release particles such as nanospheres into cytoplasm is indispensable to control the intracellular pharmacokinetics. Although some systems have been already achieved to deliver soluble drugs into cytoplasm, no system has been successful in delivering nanoparticles into cytoplasm for further sustained regulation of drug release in intracellular compartments. In this study, we established a protocol to encapsulate nanoparticles into liposomes. Additionally, the, liposomes were fur.ther fused with ultraviolet-inactivated Sendai virus in order to make fusogenic liposomes. We demonstrated that fusogenic liposomes could deliver the encapsulated nanoparticless into cytoplasm via fusion-dependent manner rather thanrendocytosis. Also nanoparticles were introduced in 94% of the 105 cells treated with fusogenic liposomes encapsulating nanoparticles. The mean number of nanoparticles introduced into cytoplasm was about 10 particles / ce11. Additionally, to evaluate intracellular slow release of drugs, nanoparticles that had adsorbed FITC-labeled oligonucleotide was introduced into the cytoplasm by fusogenic liposome. FITC-labeled oligonucleotide was confirmed to be released gradually into the cytoplasm for more than 7 days. We conclude that the technology, which can be used for the encapsulation of any functional nanoparticles, is valuable for regulation of intracellular pharmacokinetics.

近年来，使用纳米球或微球的持续或控制药物释放系统在全身药代动力学中受到关注。然而，在不久的将来，在药物递送系统研究中，不仅“全身药代动力学”而且“细胞内药代动力学”似乎也很重要。将纳米球等缓释颗粒递送到细胞质中的技术对于控制细胞内药代动力学是必不可少的。已经实现了将可溶性药物递送到细胞质中，但还没有系统成功地将纳米颗粒递送到细胞质中以进一步持续调节细胞内的药物释放。在这项研究中，我们建立了一个方案。此外，脂质体进一步与紫外线灭活的仙台病毒融合，以制成融合脂质体，并且我们证明融合脂质体可以通过融合依赖性方式将封装的纳米颗粒递送到细胞质中。在用包裹纳米颗粒的融合脂质体处理的 105 个细胞中，94% 的细胞中引入了纳米颗粒。引入细胞质的纳米颗粒的平均数量约为 10 个颗粒/ ce11。此外，为了评估药物的细胞内缓慢释放，通过融合脂质体将吸附有 FITC 标记寡核苷酸的纳米颗粒引入细胞质中。 FITC标记的寡核苷酸被证实在7天以上的时间内逐渐释放到细胞质中。我们的结论是，该技术可用于封装任何功能性纳米颗粒，对于细胞内药代动力学的调节具有重要价值。

项目成果

S.Nakagawa: "Development of novel technology of DDS for gene therapy"Drug Metabol.Pharmacokin.. 18. 223-229 (2003)

S.Nakakawa：“用于基因治疗的 DDS 新技术的开发”Drug Metabol.Pharmacokin.. 18. 223-229 (2003)

中川晋作: "遺伝子医薬品のDDS"医薬ジャーナル. 37. 1559-1565 (2001)

Shinsaku Nakakawa：“基因药物的 DDS”医药杂志 37. 1559-1565 (2001)。

真弓忠範: "DDSとナノ粒子 特集によせて"生物工学会誌. 81. 177-177 (2003)

Tadanori Mayumi：“DDS 和纳米粒子的专题”生物工程学会杂志 81. 177-177 (2003)。

J.Kunisawa, I.Takahashi, A.Okudaira, T.Hiroi, K.Katayama, T.Ariyama, Y.Tsutsumi, S.Nakagawa, H.Kiyono, T.Mayumi: "Lack of antigen-specific_immune responses in anti-IL-77 receptor α chain antibody-treated Peyer's path-null mice following intestinal immuniz

J.Kunisawa、I.Takahashi、A.Okudaira、T.Hiroi、K.Katayama、T.Ariyama、Y.Tsutsumi、S.Nakakawa、H.Kiyono、T.Mayumi：“抗-免疫反应中缺乏抗原特异性免疫反应” IL-77受体α链抗体治疗的派尔氏通路无效小鼠肠道免疫后

S.Nakagawa, T.Mayumi: "Development of novel technology of DDS for gene therapy."Drug Metabol. Pharmacokin.. 18. 223-229 (2003)

S.Nakakawa、T.Mayumi：“用于基因治疗的 DDS 新技术的开发。”药物代谢。