Preparation of the vesicle as surfactant assemblies having a functional molecular -recognition -toward to a cancer therapy in DDS-

制备具有功能性分子识别功能的表面活性剂组装体囊泡，用于 DDS 中的癌症治疗

• 批准号: 10450297
• 负责人: KATO Keiichi
• 金额: $ 7.1万
• 依托单位: Ehime University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10450297/
• 关键词: DDS; Cancer Therapy; Gene transfection; Lipid vesicle; Liposome; Immunovesicle; Alga lectin; Hybridoma

The main purpose of this work is the new preparation of the lipid vesicle, which has a function of specific targeting to a cancer cell to apply to a cancer therapy. As a targeting ligand of "missile device" to a cancer cell, two species of ligands were chosen : one is Eucheuma serra (alga lectin) and another is a monochronal antibody to an antigen of a cancer cell.The ESA showed the specific affinity to a high-mannose. The ESA-immobilized vesicle was found to specifically combine to cancer cells, such asolon adenocarcinoma (COLO201). The vesicle flew smoothly in the supenor artery of rabbit. Moreover, the in-vivo experiments on the injection of the ESA-immobilized vesicle to a rat suggested a life safety of this vesicle because of the rat's living during 5 weeks without decrease in the weight.Another was an investigation on a cationic immnovesicle. The cationic vesicles were prepared by adding the cationic peptide lipids of N+C5Ala2C16 (abbreviated as CPL) to Span80 (sorbitan monooleate), by means of a two-step emulsification technique. Then, the anti-IgM antibody as a model ligand was immobilized to the vesicle (immunovesicle). The antigen of the antibody was the IgM. The IgM was produced at the surface of the human-human hybridoma (HB4C5 cell). The HB4C5 cell was chosen as a model-targeting cell, which was a fusion product of a human lymphocyte from lung cancer patient and a human lymphoma line, NAT-30. The specific binding and the fusion between the immnovesicles and the HB4C5 cells were confirmed by the fluorescence microscopic observation. The addition of the CPL to Span80 enhanced the fusion between the immunovesicles and HB4C5 cells. The transfection of the DNA entrapped in the immunovesicles to HB4C5 was also confirmed. Thus, the lipid vesicles were expected to be applied in a DDS or a gene transfection.

本工作的主要目的是制备具有特异性靶向癌细胞功能的脂质囊泡的新制剂，并将其应用于癌症治疗。作为针对癌细胞的“导弹装置”的靶向配体，选择了两种配体：一种是Eucheuma serra（藻类凝集素），另一种是针对癌细胞抗原的单时抗体。ESA 显示了对癌细胞抗原的特异性亲和力。高甘露糖。发现 ESA 固定的囊泡与癌细胞（如索隆腺癌 (COLO201)）特异性结合。囊泡在兔上动脉中平稳飞行。此外，向大鼠注射固定有ESA的囊泡的体内实验表明该囊泡具有生命安全性，因为大鼠存活5周而体重没有下降。另一个是对阳离子免疫囊泡的研究。将N+C5Ala2C16（简称CPL）的阳离子肽脂质添加到Span80（失水山梨糖醇单油酸酯）中，通过两步乳化技术制备阳离子囊泡。然后，将作为模型配体的抗IgM抗体固定化至囊泡（免疫囊泡）。抗体的抗原是IgM。 IgM 是在人-人杂交瘤（HB4C5 细胞）表面产生的。 HB4C5细胞被选为模型靶向细胞，它是来自肺癌患者的人淋巴细胞与人淋巴瘤细胞系NAT-30的融合产物。通过荧光显微镜观察证实了免疫囊泡与HB4C5细胞之间的特异性结合和融合。在 Span80 中添加 CPL 增强了免疫囊泡和 HB4C5 细胞之间的融合。还证实了包埋在免疫囊泡中的DNA转染至HB4C5。因此，脂质囊泡有望应用于DDS或基因转染。