双重响应靶向双药协同递送纳米系统的制备及体外评价

Preparation and in vitro evaluation of dual-response targeting dual drug-loaded synergistic nanosystem

目的:制备双重响应靶向性双载药纳米粒(DOX/CMCS-ss-MTX NPs),并进行体外抗肿瘤考察。方法:甲氨蝶呤(MTX)通过二硫键接枝到羧甲基壳聚糖(CMCS)上,核磁共振氢谱(1H-NMR)、红外光谱(IR)确证结构。离子交联法制备纳米粒,对粒径、Zeta电位进行表征;测定载阿霉素(DOX)和甲氨蝶呤(MTX)的DOX/CMCS-ss-MTX NPs包封率和载药量;MTT实验考察DOX/CMCS-ss-MTX NPs体外抗肿瘤活性。结果:DOX/CMCS-ss-MTX NPs粒径为(148.3±2.5)nm,阿霉素载药量为(17.10±0.28)%,包封率为(71.32±3.54)%;甲氨蝶呤载药量为(19.35±0.33)%,包封率为(86.9±2.35)%。实验证明其具有pH和还原响应,肿瘤微环境下可快速释放。MTT实验表明DOX/CMCS-ss-MTX NPs对正常肝细胞没有明显影响,对肝癌细胞有明显抑制作用。结论:DOX/CMCS-ss-MTX NPs粒径均一、载药量较高,具有良好的pH响应性和还原响应性,共同递送甲氨蝶呤和阿霉素,具有协同抗肿瘤效果。

OBJECTIVE Toprepare dual-drug loaded nanoparticles with dual response targeting(DOX/CMCS-ss-MTX NPs)and its anti-tumor effect in vitro.METHODS Methotrexate(MTX)was grafted onto carboxymethyl chitosan(CMCS)backbone through disulfide bonds,and the structure of the co-polymer was confirmed by hydrogen nuclear magnetic resonance(1H-NMR)and infrared spectroscopy(IR).Then nanoparticles were prepared by ion crosslinking method,and their particle size and Zeta potential were characterized.The encapsulation efficiency and drug loading of DOX/CMCS-ss-MTX NPs were determined.The in vitro antitumor activity of DOX/CMCS-ss-MTX NPs was investigated by MTT assay.RESULTS The size of DOX/CMCS-ss-MTX nanoparticles was(148.3±2.5)nm.The drug loading and encapsulation efficiency of DOX were(17.1±0.28)%and(71.3±3.54)%.The drug loading and encapsulation efficiency of MTX were(19.3±0.33)%and(86.9±2.35)%.The experiments have shown that it has pH and reduction responses and can be rapidly released in the tumor microenvironment.The results of MTT experiments showed that DOX/CMCS-ss-MTX NPs had no significant inhibitory effect on normal hepatocytes,but had a significant inhibitory effect on hepatoma cells.CONCLUSION The prepared DOX/CMCS-ss-MTX NPs had uniform particle size,with high drug loading,and good pH and reduction responsiveness.The co-delivery of MTX and DOX improved the anti-tumor effect.