DNA桥接的Hela细胞靶向组装型树状大分子的制备  

Synthesis and research of assembled PAMAM dendrimers targeting to cancer cells

作　　者：王秀萍[1] 范伟[2] 费瀚雨[2] 张奇[2] 

机构地区：[1]华中科技大学同济医学院梨园医院,湖北武汉430077 [2]华中科技大学同济药学院天然药物化学与资源评价湖北省重点实验室,湖北武汉430030

出　　处：《中国医院药学杂志》2012年第11期833-837,共5页Chinese Journal of Hospital Pharmacy

基　　金：国家自然基金青年基金项目(编号:30801455);湖北省自然科学基金项目(编号:2008CDB245)

摘　　要：目的:设计一种叶酸受体介导的经DNA桥接的靶向组装型聚酰胺-胺树状大分子,探讨靶向树状大分子库及功能树状大分子库经DNA组合可行性。方法:将第五代聚酰胺-胺树状大分子(G5 PAMAM)部分酰化后,分2组分别连接叶酸(FA)以及异硫氰酸荧光素(FITC),再分别与经EDC活化的互补单链DNA相连,最后完成DNA杂交合成终产物,荧光显微镜观察其对HELA细胞的靶向性。结果:(1)G5 PAMAM的酰化按投料比获得末端酰化率约为70%的PAMAM。(2)氢谱显示FA和FITC与部分酰化G5 PAMAM连接后,产物出现相应的苯环氢,通过积分计算出每个部分酰化的G5 PAMAM连接约5个FA或2.5个FITC,且经TLC检测,产物经葡聚糖凝胶层析和透析后可达色谱纯。(3)经DNA桥接的PAMAM-FA及PAMAM-FITC加入HELA细胞后相对于没有连接叶酸的桥接产物荧光显微镜在细胞内可观察到明显的荧光,并且FA可竞争性阻断。结论:由DNA桥接的肿瘤靶向组装型树状大分子具一定的可行性。OBJECTIVE To design a kind of assembled PAMAM dendrimers linked together by using complementary DNA oligonucleotides targeted to cancer cells, and discuss the possiblity and effect of coupled functional PAMAM dendrimers. METHODS G5 PAMAM dendrimers was first partially acetylated and then conjugated with FITC or FA, followed by the covalent attachment of complementary, 5-phosphate-modified oligonueleotides. Hybridization of these oligonucleo-bindtide conjugates led to the self-assembly of the FITC and FA-conjugated dendrimers. The targeting to Hela cells was observed by fluorescence microscope. RESULTS （1）Acetylation ratio of terminal primary amine of dendrimer was 70%. （2）^1 H-WMR showed that the corresponding unsaturated hydrogens of FA and FITC was found. The number of FA molecules was calculated to he 5. 0 while the FITC was 2.5. The TLC results showed that the products were chromatography pure after dialysis and chromatography. （3）Obvious fluorescence was observed from assembled PAMAM dendrimers, while the fluorescence of reference sample which FA was not connected was unobvious. CONCLUSION Assembled PAMAM dendrimers which are linked together using complementary DNA oligonucleotides to target to cancer cells is possible.

关 键 词：叶酸靶向 聚酰胺-胺树状大分子 DNA 组装型 FITC FA 

分 类 号：R943[医药卫生—药剂学]