阿霉素前药纳米粒/姜黄素联合递送系统的构建及其抗肿瘤研究  

Construction of a co-delivery system based on doxorubicin prodrug nanoparticle and curcumin and its application in anti-tumor study

作　　者：褚丽萍[1] 刘金剑[1] 杨翠红[1] 黄帆[1] 刘鉴峰[1] 张玉民[1] 

机构地区：[1]北京协和医学院&中国医学科学院放射医学研究所,天津300192

出　　处：《中国生化药物杂志》2016年第1期12-15,19,共5页Chinese Journal of Biochemical Pharmaceutics

基　　金：国家自然科学基金(81471727);"协和青年基金资助";"中央高校基本科研业务费专项资金资助"(3332015100);北京协和医学院2015年度"协和新星"项目(1579);中国医学科学院放射医学研究所发展基金(1609)

摘　　要：目的以酸敏感阿霉素前药纳米粒为基础,通过负载姜黄素构建联合递送系统,从而在提高抗肿瘤细胞增殖效果的同时降低由阿霉素引起的心肌细胞毒性。方法利用阿霉素(doxorubicin,DOX)的氨基与醛基化聚乙二醇(PEG-CHO)的醛基进行席夫碱反应得到PEG-DOX前药聚合物,通过疏水作用将疏水性抗肿瘤药物姜黄素(curcumin,Cur)负载到PEG-DOX的疏水内核中,最后通过纳米沉淀技术得到同时负载两种药物的前药纳米粒PEG-DOX/Cur NPs。通过核磁(1H-NMR)对PEG-DOX前药进行结构表征,利用动态光散射(DLS)和透射电镜(TEM)对PEG-DOX/Cur NPs的粒径和形貌进行表征;利用反相高效液相色谱法(RP-HPLC)研究PEG-DOX/Cur NPs在酸性条件下的药物释放行为;利用MTT法研究PEG-DOX/Cur NPs的肿瘤细胞(H1975)增殖抑制效果以及评价PEG-DOX/Cur NPs对心肌细胞(H2C9)的毒性;通过H2C9细胞内的氧化自由基(ROS)水平检测(2',7'-二氯荧光素二乙酸盐法,DCFH-DA法)揭示PEG-DOX/Cur NPs对DOX心肌细胞毒性改善的机理。结果 PEG-DOX/Cur NPs为直径约90 nm的均一的球形结构,在酸性条件下能够同步释放DOX和Cur;MTT结果表明PEG-DOX/Cur NPs的H1975细胞增殖抑制效果优于DOX和PEG-DOX NPs(P<0.05),且PEG-DOX/Cur NPs对H2C9细胞的毒性明显低于DOX和PEG-DOX NPs(P<0.05);H2C9细胞ROS含量检测结果表明PEG-DOX/Cur NPs处理的心肌细胞具有最低的ROS水平(P<0.05),由此说明PEG-DOX/Cur NPs较低的心肌细胞毒性可能源于Cur引起的胞内ROS的降低所致。结论基于DOX前药纳米粒负载Cur构建联合递送系统在实现了理想的肿瘤细胞增殖抑制效果的同时,明显的降低了由DOX引起的心肌细胞毒性,实现了联合治疗的效益最大化。Objective To construct a joint delivery system by loading curcumin,acid sensitive adriamycin nanoparticles as the basis,as to improve the anti-tumor cell proliferation activity and simultaneously decrease the cardiotoxicity induced by doxorubicin. Methods The PEG-DOX conjugate was synthesized by schiff base reaction between the amino of doxorubicin and the aldehyde group of aldehyde functionalized polyethylene glycol,the hydrophobic curcumin was loaded by PEG-DOX conjugate through the hydrophobic interaction between curcumin and the core of PEG-DOX conjugate,and then the prodrug nanoparticle PEG-DOX / Cur NPs was obtained by nanoprecipitation technique. The structure of PEG-DOX conjugate was confirmed by1 H NMR. The diameter and morphology of PEG-DOX / Cur NPs were detected by dynamic light scattering（ DLS） and transmission electron microscope（ TEM）,and the DOX and Cur release kinetics under physiological and acidic condition were detected by reversed-phase high performance liquid chromatography（ RP-HPLC） method. The in vitro combined antitumor effects and the cardiotoxicity of PEG-DOX / Cur NPs were evaluated by MTT method. The reactive oxygen species（ ROS） within the myocardial cells was estimated by 2＇,7＇-dichlorofluorescin diacetate（ DCFH-DA） method.Results The PEG-DOX / Cur NPs showed spherical structure with diameter of about 100 nm with a narrow PDI. The PEG-DOX / Cur NPs could release DOX and Cur,simultaneously. The MTT results showed that PEG-DOX / Cur NPs had the best cytotoxicity on H197 cells and the lowest cardiotoxicity on H2C9 cells. Compared with DOX and PEG-DOX / Cur NPs,H2C9 cells administrated by PEG-DOX / Cur NPs showed the lowest ROS level,which indicated that the lowest cardiac myocyte toxicity of PEG-DOX / Cur NPs may result from the ROS clearance function of curcumin. Conclusion The codelivery system of PEG-DOX / Cur NPs based on PEG-DOX conjugate and curcumin achieved the maximum of effectiveness,with desirable anti-tumor proliferation effect and significantly impr

关 键 词：纳米粒前药 阿霉素 姜黄素 联合治疗 心肌毒性 

分 类 号：R914.5[医药卫生—药物化学]