针对K-Ras(G12C)突变肿瘤的还原响应型纳米胶束的制备及评价  

Preparation and evaluation of reduction-responsive nanomicelles for K-Ras (G12C) mutant tumors

作　　者：卢贵茹 刘阳[1] 刘晶阳 冯友成 马孟城 张壮壮 付志远 LU Gui-ru;LIU Yang;LIU Jing-yang;FENG You-cheng;MA Meng-cheng;ZHANG Zhuang-zhuang;FU Zhi-yuan(Department of Pharmacy,the Affiliated Hospital of Xinyang Vocational and Technical College,Xinyang Henan 464000;Department of Pharmacy,Xinyang Central Hospital,Xinyang Henan 464000;Ningbo Baoting Biotechnology Co.,Ltd.,Ningbo Zhejiang 315000;State Key Laboratory of Biotherapy,Chengdu 610000)

机构地区：[1]信阳职业技术学院附属医院药学部,河南信阳464000 [2]信阳市中心医院药学部,河南信阳464000 [3]宁波宝亭生物科技有限公司,浙江宁波315000 [4]生物治疗全国重点实验室,成都610000

出　　处：《中南药学》2024年第12期3156-3161,共6页Central South Pharmacy

基　　金：信阳市软科学研究计划项目(No.20230068)。

摘　　要：目的 以不同分子量的巯基化聚乙二醇和K-Ras(G12C)抑制剂6合成一系列还原敏感型高分子前药(PERI),制备并筛选出最优纳米胶束(PERIs胶束),考察其理化性质和体外抗肿瘤作用。方法 通过二硫键将巯基化聚乙二醇与小分子抑制剂结合,得到还原敏感型PERIs高分子前药,通过其自组装制备PERIs胶束。透射电镜观察其形态;粒度仪测定其粒径和表面电位;芘荧光探针法测定其临界胶束浓度;间接法测定其载药量及包封率;透析法测定其在不同还原条件下的体外释药行为;采用CCK-8 法评价其细胞增殖毒性。结果 核磁共振氢谱表明PERIs成功合成;PERIs胶束(Mw:2000)的平均粒径为(95.76±1.8)nm,多分散系数为(0.189±0.0084),Zeta电位为-(7.07±0.14)mV,临界胶束浓度为 0.875 μg·mL^(-1),载药量为(11.5±0.16)%,包封率为(71.6±0.83)%;体外释放度实验表明PERIs胶束具有还原敏感性;细胞毒性实验表明PERIs胶束对肿瘤细胞有较强的增殖抑制作用。结论 制备的PERIs胶束(Mw:2000)粒径均一,分散性及稳定性良好,具有控释能力,并对肿瘤细胞有较强的增殖抑制作用。Objective To synthesize a series of reduction-sensitive macromolecular predrugs from thiolated polyethylene glycol of different molecular weights and K-Ras (G12C) inhibitor 6 to prepare nanomicelles,and to screened the optimal nanomicelle (PERIs micelles) to evaluate its physicochemical property and anti-tumor effect in vitro.Methods Through disulfide bonds,the thiolated polyethylene glycol was combined with K-Ras (G12C) inhibitor 6 to form reduction sensitive PERIs predrug nanocarriers,and PERIs micelles were prepared by its self-assembly property.The morphology was observed by transmission electron microscopy.The particle size and Zeta potential were measured by particle size analyzer.The critical micelle concentration was determined by pyrene-based fluorescence probe.Drug loading and encapsulation rate were measured indirectly.Drug release of the micelles was evaluated by dialysis,and cytotoxicity was evaluated by CCK-8.Results A series of reduction-sensitive macromolecular predrugs (PERI) were verified by ~1H-NMR.Based on the size,surface potential and drug loading,PERIs micelles (Mw:2000) was screened for subsequent experiments.The mean particle size was (95.76±1.8) nm with a polydispersity index of (0.189±0.0084),Zeta potential was -(7.07±0.14) mV,critical micelle concentration was 0.875 μg·mL^(-1),the drug loading was (11.5±0.16)% and the encapsulation rate was (71.6±0.83)%.The drug release experiments in vitro showed that PERIs were sensitive to reduction.Cytotoxicity analysis showed that PERIs effectively inhibited the proliferation of tumor cells.Conclusion The prepared PERIs micelles (Mw:2000) exhibits uniform particle size,good dispersion and stability,sustained drug release,and strong proliferation inhibition of tumor cells.

关 键 词：巯基化聚乙二醇 K-Ras(G12C)抑制剂6 还原敏感 前药纳米胶束 

分 类 号：R943[医药卫生—药剂学]