目的 本实验以聚乙二醇 (polyethylene glycol,PEG) 和多柔比星(doxorubicin, DOX)合成的刺激敏感型前药聚合物(PEG-DOX)包载美法仑(melphalan,MEL),制备MEL/PEG-DOX纳米胶束,考察其体外协同抗肿瘤作用。方法 通过希夫碱反应将PEG化的二硫代二丙酸二酰肼(TPH)与DOX结合,形成pH/还原敏感型PEG-DOX前药纳米载体,通过其自组装性能制备MEL/PEG-DOX载药胶束。用透射电镜观察其形态,粒径仪测定其粒径和电位,用超滤法测定载药量和包封率,用透析法评价胶束的药物释放,采用MTT法评价细胞毒性。结果 通过核磁共振氢谱验证了所制备的刺激响应型PEG-DOX前药聚合物;其PEG-DOX载体平均粒径为(188±2.4)nm,多分散系数(PDI)为(0.255±0.008);MEL/PEG-DOX载药胶束的平均粒径为(299.7±2.4)nm,多分散系数(PDI)为(0.301±0.03),Zeta电位为(-0.385±0.02) mV;DOX载药量为(14.85±0.24)%,包封率是(85.78±0.37)%,MEL载药量为(7.36±0.36)%,包封率为(38.79±0.42)%;体外药物释放实验结果表明,MEL/PEG-DOX胶束具有还原敏感和pH敏感性,且还原敏感性大于pH敏感性;细胞毒性实验分析,DOX和MEL在细胞内共同释放,实现了对肿瘤细胞的联合杀伤。结论 MEL/PEG-DOX可以在肿瘤微环境特异性释放,对肿瘤细胞具有协调治疗的作用,具有良好的应用前景。
Abstract
OBJECTIVE To synthesize a stimulus-sensitive precursor polymer (PEG-DOX) from polyethylene glycol and doxorubicin, encapsulate melphalan(MEL) to prepare MEL/PEG-DOX nanometer micelles, and investigate its synergistic anti-tumor effect in vitro. METHODS Through Schiff base reaction, the pegylated dithionyl hydrazide (TPH) was combined with DOX to form a pH/reduction sensitive PEG-DOX prodrug nanocarrier, and MEL/PEG-DOX loaded micelle was prepared by its self-assembly property.The morphology was observed by transmission electron microscopy, the particle size and potential were measured by particle size analyzer, drug loading and encapsulation rate were measured by ultrafiltration, drug release of micelles was evaluated by dialysis, and cytotoxicity was evaluated by MTT. RESULTS The stimulation response PEG-DOX precursor polymer was verified by H-NMR.The mean particle size of peg-dox carrier was (188±2.4) nm, and the polydispersion coefficient (PDI) was (0.255±0.008). The mean particle size of MEL/PEG-DOX loaded micelles was (299.7±2.4) nm, the polydispersion coefficient (PDI) was 0.301±0.03, and the Zeta potential was (-0.385±0.02) mV. DOX drug-polymer interactions (14.85±0.24)%, the encapsulation rate was (85.78±0.37)%, MEL drug-polymer interactions (7.36±0.36)%, the encapsulation rate was (38.79±0.42)%.The results of drug release experiments in vitro showed that MEL/PEG-DOX micelles were sensitive to reduction and pH, and the reduction sensitivity was greater than that of pH. Cytotoxicity analysis showed that DOX and MEL were released together in cells, achieving the joint killing of tumor cells. CONCLUSION MEL/PEG-DOX can be specifically released in the tumor microenvironment, which has a coordinating effect on tumor cells and a good application prospect.
关键词
聚乙二醇 /
纳米前药 /
多柔比星 /
美法仑
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Key words
polyethylene glycol /
nanoprodrug /
doxorubicin /
melphalan
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中图分类号：
R944
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脚注
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基金
国家自然科学基金项目资助(81503007,21574059)
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