低分子肝素-阿霉素聚合物胶束的制备及体外细胞增殖抑制作用研究  

Preparation of low molecular weight heparin-doxorubicin(LMWH-DOX)polymer micelles and inhibition of cell proliferation in vitro

作　　者：蒋燕平 牛丽娜[1] 梁桂贤[1] 谢茵[1] 范博[1] JIANG Yan-ping;NIU Li-na;LIANG Gui-xian;XIE Yin;FAN Bo(Pharmaceutical Sciences School of Shanxi Medical University,Taiyuan 030000,China)

机构地区：[1]山西医科大学药学院,太原030000

出　　处：《中国新药杂志》2019年第2期195-201,共7页Chinese Journal of New Drugs

基　　金：山西医科大学博士启动基金资助项目(03201620)

摘　　要：目的:利用低分子肝素(low molecular weight heparin,LMWH)的亲水性和阿霉素(doxorubicin,DOX)疏水性的特点,合成并制备同时递送2种作用机制不同的药物的LMWH-DOX聚合物胶束,用于抗肿瘤治疗。方法:以乙二胺为连接臂,通过酰胺化和希夫氏碱2步反应合成LMWH-DOX,采用透析法使其在水溶液中自组装形成"核-壳"结构的聚合物胶束。通过1H NMR对合成的LMWH-DOX聚合物进行结构确证。透射电镜观察胶束形态,粒度仪测定聚合物胶束的粒径及Zeta电位,UV测定其载药量。采用芘荧光探针法测定LMWH-DOX胶束的临界胶束浓度(critical micelle concentration,CMC);动态透析法考察LMWH-DOX胶束的体外释药特征;噻唑蓝法考察LMWH-DOX聚合物胶束对4T1细胞及EA. hy926细胞存活率的影响。结果:LMWH-DOX胶束呈类球形;粒径为144. 4 nm; PDI为0. 121; Zeta电位为-36. 5 m V;临界胶束浓度为6. 92×10-6μg·m L-1,载药量为(12. 09±0. 69)%;其体外释药行为表现出酸敏感性特征,对2种细胞株也表现出良好的增殖抑制作用。结论:LMWH-DOX聚合物胶束具有良好的理化性质,作为抗肿瘤药物递送系统具有良好的研究前景。Objective:To prepare low molecular weight heparin-doxorubicin (LMWH-DOX)polymer micelles by conjugating the two kinds of drugs with different water-solubilities and biological effects together. Methods:The DOX grafted LMWH was first synthesized via imine linkage and characterized by ^1H NMR.Then the amphiphilic polymer LMWH-DOX was self-assembled into micelles with core-shell structure in aqueous solution. The morphology of LMWH-DOX micelles was observed by transmission electron microscopy (TEM).The particle size and Zeta potential were measured using a Malvern particle size analyzer.The critical micelle concentration (CMC)of LMWH-DOX micelles was detected by fluorescent probe method.The loading capacity and in vitro release of DOX from LMWH-DOX were also investigated.Furthermore,MTT assay was used to study the viability of 4T1 and EA.hy 926 cells after LMWH-DOX micelles treatment.Results :The LMWH-DOX micelles were spherical with the size of 144.4 nm and PDI of 0.121.The Zeta potential was -36.5 mV,and CMC was 6.92×10^-6 μg·mL^-1.The average drug loading capacity was (12.09±0.69)%.The in vitro drug release behavior of LMWH-DOX micelles showed an acid-sensitive characteristic.More importantly,LMWH-DOX micelles displayed good inhibitory effects on cell proliferation of both 4T1 and EA.hy 926 cells.Conclusion:The prepared LMWH-DOX micelles have good physicochemical properties and are a promising drug delivery system for antitumor treatment.

关 键 词：低分子肝素 阿霉素 聚合物胶束 细胞增殖抑制 

分 类 号：R944.9[医药卫生—药剂学]