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

本工作的主要目的是制备新的脂质囊泡，其具有特异性靶向癌细胞的功能以应用于癌症治疗。选择麒麟菜凝集素（Eucheuma serra，ESA）和抗肿瘤细胞抗原的单时性抗体（monochronal antibody，mAb）作为癌细胞“导弹装置”的靶向配体，ESA对高甘露糖具有特异性亲和力。发现ESA固定的囊泡与癌细胞如结肠腺癌（COLO 201）特异性联合收割机结合。囊泡在家兔上动脉内流动顺畅。此外，将固定了ESA的囊泡注射到大鼠的体内实验表明，由于大鼠在5周内生活而体重没有下降，因此该囊泡具有生命安全性。通过两步乳化技术将N+ C5 Ala 2C 16（缩写为CPL）的阳离子肽脂质加入到Span 80（脱水山梨糖醇单油酸酯）中制备阳离子囊泡。然后，将作为模型配体的抗IgM抗体固定到囊泡（免疫囊泡）。抗体的抗原是IgM。IgM在人-人杂交瘤（HB 4C 5细胞）的表面产生。选择HB 4C 5细胞作为模型靶向细胞，其是来自肺癌患者的人淋巴细胞和人淋巴瘤细胞系NAT-30的融合产物。荧光显微镜观察证实了免疫囊泡与HB 4C 5细胞的特异性结合和融合。将CPL添加到Span 80中增强了免疫囊泡与HB 4C 5细胞之间的融合。还证实了包埋在免疫囊泡中的DNA对HB 4C 5的转染。因此，脂质囊泡有望应用于DDS或基因转染。

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.