肝靶向肽修饰的负载阿霉素纳米粒的制备及其体外作用评价  

Preparation and in vitro evaluation of liver-targeting peptide-modified doxorubicin-loaded nanoparticles

作　　者：邝启健 詹壹斐 高焯巧 王晓曼 朱凯琳 张颖妍 马艳[3] KUANG Qijian;ZHAN Yifei;GAO Zhuoqiao;WANG Xiaoman;ZHU Kailin;ZHAGN Yingyan;MA Yan(School of Public Health,Guangdong Pharmaceutical University,Guangzhou 510006,China;School of Life Science and Biopharmaceutics,Guangdong Pharmaceutical University,Guangzhou 510006,China;School of Bisic Medical Sciences,Guangdong Pharmaceutical University,Guangzhou 510006,China)

机构地区：[1]广东药科大学公共卫生学院,广东广州510006 [2]广东药科大学生命科学与生物制药学院,广东广州510006 [3]广东药科大学基础医学院,广东广州510006

出　　处：《广东药科大学学报》2025年第2期16-23,共8页Journal of Guangdong Pharmaceutical University

基　　金：广东省级大学生创新创业训练计划项目(202210573036);国家自然科学基金项目(81502520)。

摘　　要：目的以马来酰亚胺-聚乙二醇(Mal-PEG,PP)为纳米载体,制备肝靶向肽修饰的负载阿霉素(doxorubicin,DOX)纳米粒并体外评价其活性。方法人工合成多肽与不同来源的细胞孵育,筛选与肝癌细胞特异性结合的多肽;通过优化PP与DOX比例,确定制备粒径较小纳米粒的投料比;制备肝靶向肽修饰的负载DOX纳米粒,对其形貌、粒径、包封率进行表征,CCK8法、流式细胞仪和荧光显微镜对其细胞毒性、抑癌作用和细胞摄取进行初步评价。结果多肽A54和CSP I-plus与肝癌细胞有特异性结合,制备了球形且粒径均一的偶联肝靶向肽-阿霉素的纳米粒A54-PP-DOX和CSP-PP-DOX,阿霉素包封率分别为53.2%和54.8%。纳米粒的细胞毒性呈浓度依赖性,肝靶向肽修饰的纳米粒对肝癌细胞的抑制作用明显优于无靶向肽修饰的纳米粒,且更容易被肝癌细胞Bel7402和HepG2摄取,提高药物进入肝癌细胞的浓度,降低毒副作用。结论成功制备了主动靶向肝癌细胞负载DOX的纳米粒,为靶向治疗肝细胞癌及有效降低化疗药物不良反应提供了实验依据。Objective To prepare liver-targeting peptide-modified doxorubicin(DOX)-loaded nanoparticles with maleimide polyethylene glycol(Mal-PEG,PP)and evaluate their activity in vitro.Methods Firstly,fluorescein isothiocyanate(FITC)modified peptides were synthesized and incubated with different cells to screen the peptides,which specifically bound to hepatoma cells.By optimizing the size of nanoparticles,the ratio of PP to DOX was determined.Then the liver-targeting peptide-modified DOX-loaded nanoparticles were prepared,and their morphology,particle size and encapsulation rate were characterized.Finally,their cytotoxicity and cell uptake activity in vitro were preliminarily evaluated by CCK8,flow cytometry and fluorescence microscopy.Results CSP I-plus peptide and A54 peptide specifically bound to hepatocellular carcinoma cells.Coupled A54 maleimidepolyethylene glycol-doxorubicin nanoparticles A54-PP-DOX and coupled CSP I-plus maleimide-polyethylene glycol-doxorubicin nanoparticles CSP-PP-DOX were successfully prepared with the encapsulation rates of 53.2% and 62.8%,respectively.The nanoparticles had dose-dependent cytotoxicity.The inhibition effect of liver-targeting peptide modified nanoparticles on hepatocellular carcinoma cells was significantly better than that of nanoparticles without peptide modification.Liver-targeting peptide modified nanoparticles were more easily ingestion by hepatocellular carcinoma cells Bel7402 and HepG2,which increased the concentration of drugs within hepatocellular carcinoma cells and reduced the toxic side effects of drugs.Conclusion DOX-loaded nanoparticles that actively target hepatocellular carcinoma have been prepared successfully,which provides an experimental basis for the targeted treatment of hepatocellular carcinoma and effectively reduces the adverse reactions of chemotherapy drugs.

关 键 词：肝细胞癌 阿霉素 纳米粒 肝靶向肽 靶向给药 

分 类 号：R944.2[医药卫生—药剂学]