HER2靶向聚酰胺-胺树枝状聚合物递药系统的构建及评价  

Construction and evaluation of HER2-targeted PAMAM dendrimer drug delivery system

作　　者：马鹏凯[1] 张雪梅 李金明 倪玲 王正[1] 张峰溥 孙考祥[1] 

机构地区：[1]烟台大学药学院,烟台264000 [2]山东绿叶制药有限公司,烟台264000

出　　处：《中国新药杂志》2015年第10期1171-1177,1186,共8页Chinese Journal of New Drugs

基　　金：国家重点基础研究发展计划(973计划)前期研究专项基金(2012CB724003)

摘　　要：目的：构建曲妥珠单抗-聚乙二醇-PAMAM（trastuzumab-PEG-PAMAM）靶向递药系统,对其体内和体外靶向性进行评价。方法：用FITC或CY7-NHS标记PAMAM后,通过NHS-PEG-MAL将巯基化抗体与PAMAM连接。采用HNMR对共轭物进行化学结构表征;粒度仪和透射电镜观察共轭物粒径大小和形态特征;荧光显微镜和流式细胞仪考察细胞摄取;小动物活体成像考察在裸鼠体内分布。结果：HNMR结果显示PEG,FITC和CY7成功偶联到PAMAM;SDS-PAGE结果显示曲妥珠单抗与PEG-PAMAM偶联成功。trastuzumab-PEG-PAMAM-FITC/CY7共轭物粒径在10~40 nm。trastuzumab-PEG-PAMAM较PAMAM更多被BT474细胞摄取,而且更多分布在肿瘤组织中,经细胞摄取后主要存在于细胞质中。结论：细胞摄取与小动物成像实验证明其具有很好的靶向效果,作为抗肿瘤药的靶向递药载体具有广阔的前景。Objective： To synthesize trastuzumab-PEG-PAMAM HER2-targeted drug delivery system and evaluate its targeting ability in vitro and in vivo. Methods： FITC or CY7-NHS was conjugated with PAMAM to syn- thesize PAMAM-FITC/CY7. Thiolated trastuzumab was conjugated with PAMAM dendrimer through bi-functional polyethylene glycol NHS-PEG-MAL to synthesize different conjugates of trastuzumab-PEG-PAMAM-FITC/CY7. The structure was characterized using HNMR. The size, zeta potential and morphology were characterized using particle size analyzer and transmission electron microscope. Fluorescent microscopy and flow cytometer were used to evalu- ate cellular uptake. In vivo fx pro was used to evaluate its distribution in nude mice. Results： The results of HNMR indicated that PEG, FITC, and CY7 had been conjugated with PAMAM successfully. The results of SDS-PAGE in- dicated that trastuzumab had been conjugated with PEG-PAMAM successfully. The particle sizes were 10 -40 nm. Cellular uptake of trastuzumab-PEG-PAMAM was more than that of PAMAM alone, and it was located in the cyto- plasm. Its distribution in tumor tissue was more than that of PAMAM alone. Conclusion： Targeting ability in vitro and in vivo is satisfactorv. It will he a promising targeted drug delivery system for anti-cancer drugs.

关 键 词：曲妥珠单抗 PAMAM树枝状聚合物 主动靶向 

分 类 号：R979.1[医药卫生—药品]