酶响应纳米粒子共输送地西他滨阿霉素逆转乳腺癌多药耐药  

Enzyme Responsive Nanoparticles Co-Deliver Doxorubicin and Decitabine to Reverse Breast Cancer Multi-Drug Resistance

作　　者：刘耀阳 王荣梅[1] LIU Yao-yang;WANG Rong-mei(The No.2 Hospital of Shandong University,Department of Pharmacy,Jinan 250000,China)

机构地区：[1]山东大学第二医院药剂科,山东济南250000

出　　处：《药物生物技术》2018年第4期288-293,共6页Pharmaceutical Biotechnology

基　　金：北京医卫健康公益基金会(No.YWJKJJKYJJ-B16211)

摘　　要：合成两亲性基质金属蛋白酶(Matrix metalloproteinases,MMPs)响应的m PEG-CPLGLAGG(PDDC)聚合物,与地西他滨、阿霉素组装形成纳米粒子,并初步评价其逆转乳腺癌多药耐药的效果。以碳二亚胺/N-羟基琥珀酰亚胺为缩合剂,将MMPs响应的多肽(CPLGLAGG)通过酰胺化反应接支到m PEG上,采用核磁共振和红外光谱对其结构进行确证。将PDDC与阿霉素、地西他滨超分子组装形成纳米粒子,HPLC和荧光光谱测定载药量、包封率和体外释放。在MCF-7/ADR细胞中,采用CCK-8试剂盒检测细胞毒性,蛋白印迹检测p-糖蛋白的表达,荧光光谱检测细胞内阿霉素含量,流式检测细胞凋亡率,并通过荷瘤小鼠模型初步评价其体内药效。实验结果显示,PDDC聚合物与药物共组装形成粒径130 nm左右的纳米粒子,具有很好的稳定性和MMP-2响应性,可通过增强细胞毒性、诱导细胞凋亡、提高细胞内阿霉素浓度、抑制p-糖蛋白的表达等逆转MCF-7/ADR细胞多药耐药,并可显著降低荷瘤小鼠肿瘤体积,对体重无明显影响。该MMPs响应性纳米粒子表现出良好的生物安全性和明显的逆转阿霉素耐药的效果,可作为潜在的药物载体用于肿瘤多药耐药的治疗。To design and synthesize a matrix metalloproteinases（ MMPs） responsive amphiphilic m PEG-CPLGLAGG（ PDDC） polymer,which can be assembled with decitabine and doxorubicin to form nanoparticles,and preliminarily evaluate its effects of reversal of multidrug resistance in vitro and in vivo. PDDC was synthesized via amide bond,with EDC/NHS as condensing agent,and its chemical structure was characterized by1 H-NMR and FT-IR. PDDC was then assembled with decitabine and doxorubicin to form nanoparticles. HPLC and fluorescence spectra were used to assess drug loading capacity,entrapment efficiency and in vitro drug release. Then,cytotoxicity,expression of P glycoprotein,intracellular doxorubicin concentration and apoptosis were detected respectively by CCK-8 Kit,western-blotting,fluorescence spectra and flow cytometry,to evaluate the activity of drug-loaded nanoparticles in MCF-7/ADR cells. Tumor-bearing mice were prepared to preliminarily evaluate the curative effect of nanoparticles in vivo. The results showed the drug-loaded nanoparticles were stable and had good MMP-2 responsiveness,with average particle size being about 130 nm. In MCF-7/ADR cells,the nanoparticles showed obvious anti-multidrug resistance effects by increasing cytotoxicity and intracellular doxorubicin concentration,inhibiting expression of P glycoprotein and inducing cell apoptosis. The nanoparticles could also reduce the tumor volume in mice,with no obvious influence on body weight. In conclusion,the MMPs responsive nanoparticles exhibited reversal ability of multidrug resistance,indicating that PDDC could be used as a potential drug carrier for cancer therapy in the future.

关 键 词：多药耐药 纳米药物 基质金属蛋白酶 乳腺癌 阿霉素 地西他滨 

分 类 号：R979.1[医药卫生—药品]