肿瘤酸度响应性聚(L-赖氨酸)-阿霉素键合药的制备与表征  

Synthesis and Characterization of Tumor-acidity-sensitive Poly(L-lysine)-doxorubicin Conjugates

作　　者：张建成[1,2] 丁建勋[2,3] 肖春生[2] 贺超良[2] 庄秀丽[2] 杨亚楠[1] 陈学思[2] 

机构地区：[1]长春工业大学化学工程学院,长春130012 [2]中国科学院长春应用化学研究所,生态环境高分子材料重点实验室,长春130022 [3]中国科学院研究生院,北京100049

出　　处：《高等学校化学学报》2012年第12期2809-2815,共7页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金(批准号:51103015;50733003;20904053;51003103;21174142和50973108)资助

摘　　要：用丁二酸酐(SA)和顺式乌头酸酐(CA)分别对阿霉素(DOX)进行修饰,得到非酸响应的SA-DOX(SAD)和酸响应的CA-DOX(CAD).通过SAD或CAD、端羧基化的聚乙二醇单甲醚(mPEG-COOH)与聚(L-赖氨酸)(PLL)的缩合反应,制得非酸响应的PLL-g-mPEG/SAD和酸响应的PLL-g-mPEG/CAD键合药.通过核磁共振氢谱和红外光谱表征键合药的化学结构,并通过紫外-可见分光光度计测定药物键合量.动态激光光散射研究结果表明,两亲性的PLL-DOX键合药可以在pH=7.4的磷酸缓冲溶液中自组装形成稳定的纳米微粒.体外释放实验及噻唑蓝检测结果表明,PLL-g-mPEG/SAD在实验pH范围和时间段内只释放出少量DOX,不具有酸响应特性,且对HeLa细胞增殖抑制作用较小.而PLL-g-mPEG/CAD在生理条件(pH=7.4)下相对稳定,在弱酸性条件(pH=5.3,6.8)下,CAD中酸响应的酰胺键能快速水解并释放出DOX,表现出较强的HeLa细胞增殖抑制效果.Succinic anhydride （SA） and cis-aconitic anhydride （CA） were used to modify doxorubicin （ DOX）, obtaining acid-insensitive SA-DOX （SAD） and acid-sensitive CA-DOX （ CAD）, respectively. SAD or CAD, and carboxyl group terminated monomethoxyl poly（ ethylene glycol） （mPEG-COOH） were conjugated onto poly （L-lysine） （PLL）, yielding acid-insensitive PLL-g-mPEG/SAD and acid-sensitive PLL-g-mPEG/ CAD, respectively. The chemical structures of PLL-DOX conjugates were characterized by ^1 H NMR and FTIR. The drug conjugating content was determined with UV-Vis spectrophotometer. Dynamic laser scattering （DLS） measurements revealed that the amphiphilic PLL-DOX conjugates could self-assemble into nanoparticles in phosphate buffer（PB） at pH = 7.4. In vitro release profiles revealed that the DOX release from PLL-g mPEG/CAD could be accelerated at acid conditions（ pH=5.3 and 6. 8 ）, while that from PLL-g-mPEG/SAD was slow at all test pH （5.3, 6. 8 and 7.4）. The acid-sensitive DOX release from PLL-g-mPEG/CAD conjugates ensured higher concentration of free DOX in tumor and more pronounced antitumor efficacy. In vitro methyl thiazolyl tetrazolium assay demonstrated that PLL-g-mPEG/CAD had enhanced tumor proliferation inhibition activity comparing with acid-insensitive PLL-g-mPEG/SAD. Therefore, PLL-g-mPEG/CAD conjugates might be further developed as potential smart antitumor drugs with controlled DOX release.

关 键 词：阿霉素 聚(L-赖氨酸) 肿瘤酸度响应性 肿瘤细胞增殖抑制 

分 类 号：O632[理学—高分子化学] O629.7[理学—化学]