共载阿霉素和siRNA的还原敏感型前药纳米粒的制备及体外评价  

Preparation and in vitro evaluation of reduction-sensitive prodrug nanoparticles for coloaded DOX and siRNA

作　　者：马菊 吕灵灵 胡青 沈龙华 MA Ju;LYU Lingling;HU Qing;SHEN Longhua(Department of Pharmaceutics,School of Pharmacy,Fujian Medical University,Fujian Fuzhou 350122,China)

机构地区：[1]福建医科大学药学院药剂学系,福建福州350122

出　　处：《中国医院药学杂志》2024年第5期550-556,共7页Chinese Journal of Hospital Pharmacy

基　　金：国家自然科学基金项目(编号:82003663);福建省科技创新联合资金项目(编号:2019Y9006)。

摘　　要：目的:构建共载阿霉素(DOX)和siRNA的还原敏感型前药纳米粒(PSCSD NPs),并对其理化性质、细胞摄取和体外细胞毒性进行考察。方法:采用核磁共振氢谱(^(1)H NMR)和傅里叶红外光谱(FT-IR)对羧甲基壳聚糖-二硫键-DOX(CMCSS-DOX)进行结构表征;超声法制备PSCSD NPs,考察其粒径、载药量、包封率、血清稳定性、溶血率和体外释药特性等;采用荧光显微镜和流式细胞术考察4T1细胞对PSCSD NPs的摄取;通过MTT实验测定PSCSD NPs的体外细胞毒性。结果:^(1)H NMR和FT-IR结果表明CMC-SS-DOX成功合成;PSCSD NPs的粒径为(155.1±4.0)nm(PDI=0.144±0.028),Zeta电位为(-29.9±1.0)mV,载药量为(8.25±0.47)%,包封率为(78.41±4.52)%;透射电镜观察PSCSD NPs为球形,且具有良好的血液相容性和血清稳定性。PSCSD NPs具有还原响应释药特性,可在肿瘤部位快速释药。细胞摄取和MTT实验结果表明PSCSD NPs可以有效共载DOX和siRNA进入肿瘤细胞内以发挥抗肿瘤作用。结论:该研究成功制备了PSCSD NPs,共载DOX和siRNA以实现化疗和免疫治疗协同增效。OBJECTIVE To construct a kind of reduction-sensitive prodrug nanoparticles(PSCSD NPs)for coloaded doxorubicin(DOX)and siRNA,and to evaluate its physicochemical properties,cellular uptake and in vitro cytotoxicity.METHODS Carboxymethyl chitosan-disulfide bond-DOX(CMC-SS-DOX)was characterized by ^(1)H nuclear magnetic resonance spectroscopy(^(1)H NMR)and Fourier transform infrared spectroscopy(FT-IR).PSCSD NPs were prepared by ultrasonic method,and the properties,such as particle size,drug loading,encapsulation efficiency,serum stability,hemolysis rate and in vitro release behavior,were investigated.Both fluorescence microscopy and flow cytometry were used to determine the uptake of PSCSD NPs by 4T1 cells.MTT assay was used to determine the cytotoxicity of PSCSD NPs in vitro.RESULTS CMC-SS-DOX was successfully synthesized.The particle size of PSCSD NPs was(155.1±4.0)nm(PDI=0.144±0.028).The Zeta potential was(-29.9±1.0)mV,the drug loading was(8.25±0.47)%,and the encapsulation efficiency was(78.41±4.52)%.The prepared PSCSD NPs were spherical with good blood compatibility and serum stability.PSCSD NPs had reductive response drug release properties and were expected to release drugs quickly at the tumor site.The results of cell uptake and MTT assay showed that PSCSD could efficiently co-delivery DOX and siRNA into tumor cells to exert anti-tumor effect.CONCLUSION The PSCSD NPs were successfully prepared and co-loaded with DOX and siRNA to achieve synergistic efficiency between chemotherapy and immunotherapy.

关 键 词：阿霉素 SIRNA 联合治疗 前药纳米粒 还原响应 

分 类 号：R944[医药卫生—药剂学]