奥沙利铂肝靶向脂质体的制备及其体外药效学的初步研究

发布时间：2018-04-15 22:22

本文选题：奥沙利铂 + 脂质体　；参考：《辽宁医学院》2014年硕士论文

【摘要】：目的本实验以实验室自制的肝靶向导向分子-硬脂醇半乳糖苷作为修饰材料，，制备硬脂醇半乳糖苷修饰的奥沙利铂肝靶向脂质体，提高奥沙利铂的肝靶向性，降低其不良反应，以获得最佳的治疗效果。方法对奥沙利铂进行处方前研究，并进行方法学验证，建立奥沙利铂含量测定的方法，并考察其理化性质。采用葡聚糖凝胶微柱离心-HPLC法测定奥沙利铂肝靶向脂质体的包封率。以包封率作为评价指标，选择合适的脂质体制备方法，并以单因素考察为基础，采用Box-Benhnken试验设计法优化奥沙利铂肝靶向脂质体的制备处方。按照筛选出的最优处方制备肝靶向脂质体，并考察该靶向脂质体的形态、粒径及包封率等理化性质。利用体外培养的人肝癌细胞株SMMC-7721作为研究对象，采用细胞毒性试验及流式细胞仪检测细胞凋亡实验，在细胞水平评价硬脂醇半乳糖苷修饰的奥沙利铂肝靶向脂质体对人肝癌细胞株SMMC-7721的抑制率及凋亡程度，初步考察奥沙利铂肝靶向脂质体的靶向性。结果本实验建立了奥沙利铂的含量测定方法。葡聚糖凝胶微柱离心-HPLC法适用于测定本实验制备的奥沙利铂肝靶向脂质体的包封率。经Box-Benhnken试验设计法优化得到的奥沙利铂肝靶向脂质体的最优处方如下：磷脂质量浓度为10.34g·L-1，脂质与Ox质量比为30.9∶1，脂质与18-Gal之比为21.5∶1。按照此处方制备的奥沙利铂肝靶向脂质体的平均包封率为76.7%，其平均粒径为198.4nm，电镜下观察该靶向脂质体的形态近似球形。MTT细胞毒性试验结果表明，奥沙利铂肝靶向脂质体对SMMC-7721细胞的抑制率明显优于普通脂质体及游离奥沙利铂的抑制率。流式细胞术实验结果表明，在相同药物浓度作用下，奥沙利铂肝靶向脂质体能诱导更多的SMMC-7721细胞凋亡。初步判定奥沙利铂肝靶向脂质体具有一定的肝靶向性。结论本实验制备的奥沙利铂肝靶向脂质体，具有较高的包封率，粒径分布均匀，可抑制肝癌细胞生长，诱导肝癌细胞凋亡，具有一定肝靶向性。
[Abstract]:PurposeIn this experiment, the liposome of oxaliplatin liposome modified by stearic galactoside was prepared by using the self-made liver targeting molecule, stearic galactoside, in order to improve the liver targeting of oxaliplatin and reduce the adverse reaction of oxaliplatin.In order to obtain the best therapeutic effect.MethodA method for the determination of oxaliplatin was established and its physical and chemical properties were investigated.The encapsulation efficiency of oxaliplatin liposomes was determined by dextran gel microcolumn centrifugation-HPLC.Taking the encapsulation efficiency as the evaluation index, the suitable preparation method of liposome was selected, and the preparation prescription of oxaliplatin liposome targeting liver was optimized by Box-Benhnken method based on single factor investigation.Liver targeting liposome was prepared according to the optimized prescription, and its physical and chemical properties such as morphology, particle size and encapsulation efficiency were investigated.Using human hepatoma cell line SMMC-7721 in vitro as the research object, cell toxicity test and flow cytometry were used to detect the apoptosis of human hepatoma cell line.To evaluate the inhibition rate and apoptosis degree of oxaliplatin liposomes modified with stearic galactoside on human hepatoma cell line SMMC-7721 and to investigate the targeting of oxaliplatin liposomes.ResultA method for the determination of oxaliplatin was established.The method of dextran gel microcolumn centrifugation and HPLC is suitable for the determination of encapsulation efficiency of oxaliplatin liposomes.The optimal formulation of oxaliplatin liposome for liver targeting was obtained by Box-Benhnken design method. The optimal formulation was as follows: the mass concentration of phospholipid was 10.34 g / L ~ (-1), the mass ratio of lipid to ox was 30.9: 1, and the ratio of lipid to 18-Gal was 21.5: 1.The average encapsulation efficiency of oxaliplatin liposome was 76.7nm and the average diameter was 198.4nm. the morphology of the liposome was approximately spherical. MTT cytotoxicity test showed that the liposome was similar in shape.The inhibition rate of oxaliplatin liposomes on SMMC-7721 cells was significantly higher than that of common liposomes and free oxaliplatin.Flow cytometry showed that oxaliplatin liposomes could induce more apoptosis of SMMC-7721 cells under the same concentration.It was preliminarily determined that oxaliplatin liposomes were liver targeting.ConclusionThe liposomes prepared in this study have high encapsulation efficiency and uniform particle size distribution, which can inhibit the growth of hepatoma cells and induce apoptosis of hepatoma cells.
【学位授予单位】：辽宁医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R943;R96

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