ROS响应性纳米前药的制备及其体外抗肿瘤研究  

Preparation of ROS responsive nano prodrug and its anti-tumor activity in vitro

作　　者：黄帆[1] 高阳[1] 杨丽军[1] 任春华[1] 褚丽萍[1] 张玉民[1] HUANG Fan;GAO Yang;YANG Li-jun;REN Chun-hua;CHU Li-ping;ZHANG Yu-min(Tianjin Key Laboratory of Radiation Medicine and Molecular Nuclear Medicine, Institute of Radiation Medicine,Chinese Academy of Medical Science and Peking Union Medical College, Tianjin 300192, China)

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,天津市放射医学与分子核医学重点实验室,300192

出　　处：《天津医药》2017年第4期349-354,共6页Tianjin Medical Journal

基　　金：国家自然科学基金资助项目(51603231);中国医学科学院医学与健康科技创新工程项目(2016-I2M-3-022);天津市自然科学基金项目(16JCQNJC02500);"协和青年基金资助"和"中央高校基本科研业务费专项资金资助"(3332016098);中国医学科学院;北京协和医学院"中央级公益性科研院所基本科研业务费(2016ZX310079)

摘　　要：目的设计合成一类新的具有活性氧自由基(ROS)响应性的紫杉醇前药纳米粒,对其结构进行表征,并研究其稳定性、体外响应性释药行为、细胞摄取情况和体外抗肿瘤作用。方法通过硫醚间隔基(2S)连接亲水性的聚乙二醇(PEG)与疏水性的紫杉醇(PTX),得到前药聚合物PEG-2S-PTX单体,通过自组装制备前药纳米粒PEG-2S-PTX NPs。同时合成以丁二酸酐(SA)为间隔基的PEG-SA-PTX单体,并制备前药纳米粒PEG-SA-PTX NPs作为对照。利用核磁(1H-NMR)对前药进行结构表征,利用动态光散射(DLS)对纳米粒的粒径进行表征并考察其稳定性。采用高效液相色谱法(HPLC)研究纳米粒在氧化条件下的释放行为,通过荧光显微镜观察人乳腺癌MCF-7细胞对纳米粒的摄取行为,利用MTT法比较纳米粒对MCF-7的增殖抑制效果。结果 PEG-2S-PTX、PEG-SA-PTX能够分别自组装成粒径为(92.15±12.42)nm、(113.20±12.16)nm的纳米粒;在氧化条件下PEG-2S-PTX NPs能够快速响应性释放PTX,而PEG-SA-PTX NPs只产生微弱的响应性;PEG-2S-PTX NPs较PEG-SA-PTX NPs能被MCF-7细胞更快速地摄取,对MCF-7细胞增殖抑制均呈浓度依赖性,当浓度为0.05、0.1、5、10、50、100 mg/L时PEG-2S-PTXNPs体外细胞毒性强于PEG-SA-PTX NPs(P<0.05)。结论 PEG-2S-PTX NPs作为具有ROS响应性的紫杉醇前药纳米粒,能在被摄取进入细胞后以ROS响应的方式在肿瘤细胞内快速释放PTX,发挥良好的体外抗肿瘤作用。Objective To design and synthesize a novel paclitaxel loaded nanoparticle with reactive oxygen species(ROS)response,and characterize its structure,and investigate its stability,in vitro drug responsive release,cellular uptake and in vitro antitumor activity.Methods The PEG-2S-PTX monomer was synthesized by coupling the hydrophilic polyethylene glycol(PEG)with hydrophobic paclitaxel(PTX)via a thioether chain(2S),and the prodrug nanoparticles(PEG-2S-PTX NPs)were prepared by self-assembly.Meanwhile,using succinic anhydride(SA)as the linking group to synthesize the PEG-SA-PTX monomer and prepare the other prodrug nanoparticles(PEG-SA-PTX NPs)as control.The structures of PEG-2S-PTX and PEG-SA-PTX monomer were confirmed by1H-NMR.The diameter and stability of the nanoparticles were detected by dynamic light scattering(DLS).The PTX release kinetics under oxidizing condition was detected by high performance liquid chromatography(HPLC)method.And the cellular uptake efficiency of nanoparticles by MCF-7cells was observed by fluorescence microscope.The in vitro antitumor effects of nanoparticles were compared by MTT assay.Results PEG-2S-PTX and PEG-SA-PTX could both be self-assemble into nanoparticles with the diameter of(92.15±12.42)nm and(113.20±12.16)nm.PEG-2S-PTX NPs could rapidly release PTX under oxidative condition while PEG-SA-PTX NPs only showed weak responsiveness.PEG-2S-PTX NPs could be more rapidly taken up by MCF-7cells compared with PEGSA-PTX NPs.They both showed concentration dependent anti-tumor effects,but the cytotoxicity of PEG-2S-PTX NPs wasstronger than that of PEG-SA-PTX NPs in the concentrations of0.05,0.1,5,10,50and100mg/L(P<0.05).ConclusionAs paclitaxel prodrug nanoparticles with ROS responsive ability,PEG-2S-PTX NPs can rapidly release PTX in response toROS in tumor cells,and exhibit great anti-tumor activity in vitro.

关 键 词：活性氧 纳米粒子 ROS响应 紫杉醇前药 自组装 

分 类 号：R318.08[医药卫生—生物医学工程]