天津医药

天津医药 ›› 2017, Vol. 45 ›› Issue (4): 349-354.doi: 10.11958/20170288

• 细胞与分子生物学 • 上一篇    下一篇

黄帆#， 高阳#， 杨丽军， 任春华， 褚丽萍， 张玉民△   

  1. 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 （邮编 300192） #共同第一作者
  • 收稿日期:2017-03-08 修回日期:2017-03-16 出版日期:2017-04-15 发布日期:2017-04-15
  • 通讯作者: △通讯作者 E-mail: zhangyumin21@163.com E-mail:zhangyumin21@163.com
  • 作者简介:共同第一作者 黄帆 （1987）， 男， 助理研究员， 博士， 主要从事高分子自组装方面的研究； 高阳 （1993）， 女， 硕士在读， 主要从事纳米药物载体方 面的研究
  • 基金资助:
    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）

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

HUANG Fan#, GAO Yang#, YANG Li-jun, REN Chun-hua, CHU Li-ping, ZHANG Yu-min△   

  1. 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
  • Received:2017-03-08 Revised:2017-03-16 Published:2017-04-15 Online:2017-04-15
  • Contact: △Corresponding Author E-mail：zhangyumin21@163.com E-mail:zhangyumin21@163.com

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摘要： 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法研究比较纳米粒对肿瘤细胞（MCF-7）的增殖抑制效果。结果 PEG-2S-PTX、PEG-SA-PTX能够分别自组装成直径92、113 nm左右的纳米粒；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细胞增殖抑制均呈现浓度依赖性，当浓度为0.05、0.1、5、10、50、100 mg/L时PEG-2S-PTX NPs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。

关键词: 活性氧, 纳米粒子, ROS 响应, 紫杉醇前药, 自组装

Abstract: 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 by 1H-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-7 cells 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-7 cells compared with PEG-SA-PTX NPs. They both showed concentration dependent anti-tumor effects, but the cytotoxicity of PEG-2S-PTX NPs was stronger than that of PEG-SA-PTX NPs in the concentrations of 0.05, 0.1, 5, 10, 50 and 100 mg/L (P＜0.05). Conclusion As paclitaxel prodrug nanoparticles with ROS responsive ability, PEG-2S-PTX NPs can rapidly release PTX in response to ROS in tumor cells, and exhibit great anti-tumor activity in vitro.

Key words: reactive oxygen species, nanoparticles, ROS-responsive, paclitaxel prodrug, self-assembly