摘要
目的合成两亲性聚合物聚乙二醇-聚L-天冬氨酸[methoxy poly(ethylene glycol)-b-poly(β-benzyl L-aspartate),PEGb-PBLA]和pH敏感性聚合物[octadecylamine-p(API-Asp)10,OAPI],并利用PEG-b-PBLA包载OAPI和抗肿瘤药物姜黄素,构建pH敏感性载药胶束,考察其理化性质,并评价其体内外抗非小细胞肺癌(non-small cell lung cancer,NSCLC)效果。方法利用甲氧基聚乙二醇胺[methoxy poly(ethylene glycol)amine,mPEG2000-NH2]引发L-天冬氨酸-4-苄酯-N-羧基环内酸酐[benzyl(S)-2,5-dioxooxazolidine-4-acetate,BLA-NCA]开环聚合制备两亲性聚合物PEG-b-PBLA。利用硬脂胺引发BLA-NCA开环聚合和氨解反应合成具有pH敏感特性的聚合物OAPI。采用透析法制备载姜黄素的具pH敏感特性胶束(CNP),并对其粒径、稳定性和pH响应性体外药物释放等性质进行了考察。最后,利用NSCLC A549细胞、三维肿瘤球模型以及A549细胞建立的荷瘤小鼠模型考察了C-NP胶束的体内外抗肿瘤效果。结果透射电子显微镜结果显示,所制备的CNP呈类球形,具有明显的核壳结构,且分布均匀,其粒径为(165.32±1.53)nm,ζ电位为(−5.24±0.42)mV,载药量为(5.27±0.14)%,包封率为(88.46±3.28)%,临界胶束浓度为4.82μg/mL。体外稳定性实验显示在生理条件下,C-NP具有较好的稳定性,敏感性实验显示在酸性条件下,C-NP不稳定,具pH敏感特性。体外药物释放结果表明,在pH 5.0的介质中孵育72 h后,C-NP的累积释放量高达(81.44±5.90)%,表明C-NP胶束具有良好的pH敏感性。细胞毒性实验、肿瘤球生长抑制实验证明C-NP能增强姜黄素体外抗肿瘤活性。细胞共定位实验显示,相比较pH不敏感组C-B-NP,C-NP可以在酸性溶酶体中快速分解,从而逃逸发挥抗肿瘤效应。体内药效学实验结果表明,C-NP组显著抑制了荷瘤裸鼠的肿瘤生长。结论成功构建了载有姜黄素的pH敏感性胶束C-NP,其具有良好的理化性质和pH敏感性,且通过多种模型初步证实其对NSCLC非小细胞肺癌具有较强的抑制作用。
Objective To synthesize the amphiphilic polymer methoxy poly(ethylene glycol)-b-poly(β-benzyl L-aspartate)(PEG-bPBLA)and pH-sensitive polypeptide(OAPI),then,PEG-b-PBLA was used to construct pH-sensitive drug-carrying micelles containing OAPI and curcumin,finally,the physicochemical properties and anti-non-small cell lung cancer(NSCLC)effects of the micelles were evaluated in vivo and in vitro.Methods The methoxy poly(ethylene glycol)amine(mPEG2000-NH2)was utilized to initiate benzyl(S)-2,5-dioxooxazolidine-4-acetate(BLA-NCA)to synthesize the amphiphilic polymer PEG-b-PBLA by the ring-opening polymerization,and the pH-sensitive polypeptide OAPI was synthesized by ring-opening polymerization of BLA-NCA triggered by stearamine and ammonolysis reaction.Then,the pH sensitive micelle(C-NP)containing curcumin was prepared by dialysis method.The particle size,stability and pH-sensitive drug release properties of C-NP were investigated.Finally,the in vitro and in vivo antitumor effects of C-NP micelle were evaluated by using A549 cells,three-dimensional tumor ball models and tumor bearing mice model of NSCLC.Results The prepared C-NP showed a uniform spherical shape with obvious core-shell structure,uniform distribution under transmission electron microscopy.Its particle size was(165.32±1.53)nm,ζpotential was(−5.24±0.42)mV,drug loading was(5.27±0.14)%,and encapsulation effenciency was(88.46±3.28)%.The critical micellar concentration was 4.82μg/mL.In vitro stability experiments showed that under physiological conditions,C-NP had good stability,sensitivity experiments showed that under acidic conditions,C-NP was unstable,with pH sensitive characteristics.In vitro release results showed that the cumulative release of C-NP reached(81.44±5.90)%after 72 h incubation in pH 5.0 medium,indicating that C-NP micelles had good pH sensitivity.The cytotoxicity,tumor spheroid growth inhibition experiments revealed C-NP can enhance the anti-tumor activity in vitro.The cell colocalization experiment showed that,compared with the pH-insensitive group C-B-NP,C-NP could be decomposed rapidly in acid lysosomes,thus escaping to play an anti-tumor effect.In vivo pharmacodynamic experiment results showed that C-NP group significantly inhibited tumor growth in nude mice.Conclusion In this experiment,the pH sensitive C-NP micelle was successfully constructed,which has good physicochemical properties,pH sensitivity,and strong inhibitory effect on NSCLC preliminarily confirmed by a variety of models.
作者
侍慧慧
周美玲
柳晔
张可娇
李飞飞
SHI Hui-hui;ZHOU Mei-ling;LIU Ye;ZHANG Ke-jiao;LI Fei-fei(Kangda College of Nanjing Medical University,Lianyungang 222000,China;School of Medicine,Southeast University,Nanjing 210009,China;The People’s Hospital of Sheyang County,Yancheng 224300,China;Harbin Medical University,Harbin 150081,China)
出处
《中草药》
CAS
CSCD
北大核心
2023年第19期6264-6275,共12页
Chinese Traditional and Herbal Drugs
基金
2021年度江苏省高校基础科学(自然科学)面上项目(21KJD350004)
2020年度江苏省高校基础科学(自然科学)面上项目(20KJD350007)
江苏省大学生创新创业训练计划项目(202113980003Y)
南京医科大学康达学院2021年度科研发展基金(KD2021KYJJZD010)
南京医科大学康达学院第二期“科研创新团队项目”(KD2023KYCXTD003)。
关键词
PH敏感
非小细胞肺癌
姜黄素
纳米递药系统
聚合物
胶束
pH sensitive
non-small cell lung cancer
curcumin
nano delivery systems
polymer
micelle
