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Development of liposomes for clinical research and application

临床研究及应用脂质体的开发

基本信息

批准号：
08557082
负责人：
YOSHIDA Jun
金额：
$ 6.27万
依托单位：
Nagoya university
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08557082/
关键词：
LIPOSOME CLINICAL RESEARCH BRAIN TUMOR GENE-REGULATION DRUG 遺伝子安全性純度検定

项目摘要

We have promoted the development of cationic liposomes for human gene therapy against malignant tumors. A lot of kinds of cationic liposomes were made and performed the evaluation of the transduction efficiency. As a result, we confirmed TMAG-liposomes and DDAB-liposomes were very useful to transfer the foreign genes to. The former is a cationic liposome composed in a molar ratio of 1 : 2 : 2 as N- (alpha-trimethylammonioacetyl)- didodecyl-D-glutamate chloride (TMAG) : dilauroyl phosphatidylcholine (DLPC) : diolenyl phosphatidyl ethanolamine (DOPE) . The latter is a cationic liposome composed in a molar ratio of 1 : 2 : 2 or 1 : 1 : 1 as dimethyl-dioctadecylammonium bromide (DDAB) : dilauroyl phosphatidylcholine (DLPC) : dioleoyl phosphatidylethanolamine (DOPE) . We also found to be able to give these liposomes cell - or tissue-specificity by conjugating with a monoclonal antibody or asialoglycoprotein. In contrast, we also confinned that adeno-associated virus (AAV) vector-associated liposomes were more effective for in vitro gene transfer to human glioma cells than are liposomes containing plasmid DNA.Using vector-associated liposomes increased transduction efficiency more than 1O-fold compared to liposomes containing plasmid DNA and more than 6-fold compared to AAV alone. From these results, AAV vector-associated liposomes appear to be good candidates for in vivo gene delivery to malignant tumors. Furthermore, we investigated the effectiveness of the combination of cationic liposomes containing the plasmids driven by heat shock protein promoter and small iron ball magnetite, and induced remarkable antitumor effect.On the other hand, we established human gene therapy vector producing facility in Nagoya University Hospital and confirmed that liposomes for clinical research could prepared in the facility.

我们推动了用于人类恶性肿瘤基因治疗的阳离子脂质体的开发。制备了多种阳离子脂质体并对其转导效率进行了评价。结果，我们证实TMAG-脂质体和DDAB-脂质体对于转移外源基因非常有用。前者是由N-(α-三甲基氨乙酰基)-双十二烷基-D-谷氨酸氯化物(TMAG):二月桂酰磷脂酰胆碱(DLPC):二烯基磷脂酰乙醇胺(DOPE)以1:2:2的摩尔比组成的阳离子脂质体。后者是由二甲基双十八烷基溴化铵(DDAB):二月桂酰磷脂酰胆碱(DLPC):二油酰磷脂酰乙醇胺(DOPE)以1:2:2或1:1:1的摩尔比组成的阳离子脂质体。我们还发现，通过与单克隆抗体或脱唾液酸糖蛋白缀合，能够赋予这些脂质体细胞或组织特异性。相比之下，我们还发现，与含有质粒DNA的脂质体相比，腺相关病毒（AAV）载体相关脂质体对于体外基因转移至人胶质瘤细胞更有效。与含有质粒DNA的脂质体相比，使用载体相关脂质体的转导效率提高了1O倍以上，与单独的AAV相比提高了6倍以上。从这些结果来看，AAV 载体相关脂质体似乎是体内基因递送至恶性肿瘤的良好候选者。此外，我们还研究了含有热休克蛋白启动子驱动的质粒和小铁球磁铁矿的阳离子脂质体组合的有效性，并诱导了显着的抗肿瘤效果。另一方面，我们在名古屋大学医院建立了人类基因治疗载体生产设施，并证实该设施可以制备用于临床研究的脂质体。