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目的:制备共轭亚油酸-吉西他滨共轭物脂质体(CGLP)并考察其相关性质。方法:通过正交设计筛选出CGLP的最优制备方法。采用激光纳米粒度仪和透射电子显微镜测定粒径和形态;差式扫描量热法(DSC)对脂质体各组分进行检测;动态透析法测定药物释放和稳定性;采用SRB法考察CGLP在不同时间点对MCF-7乳腺癌细胞的抑制作用。结果:单因素考察法确立采用薄膜分散法制备CGLP。采用正交设计实验,获得CGLP的最佳处方为磷脂与胆固醇的比例15∶1、药脂比1∶8、超声时间15 min。在最佳处方条件下,平均包封率为94.35%。平均粒径在(69.73±2.09)nm,PDI为(0.235±0.042)。脂质体电镜照片表明粒子分散状态良好。DSC表明脂质体中由于CLA-GEM的加入,磷脂双分子层结构发生变化,利于载药。体外释放试验表明原料药溶液在6 h后体外累积释药量达到96.39%,而CGLP 24 h后仅释放24.07%,具有明显的缓释效应。稳定性实验表明4℃条件更适合脂质体贮藏。细胞毒实验表明CGLP比共轭物溶液的抑制能力更强。二者相较于未修饰的吉西他滨有显著性差异(P<0.001)。结论:本研究制备了一种新的CGLP,制备方法简单、包封率高,具有较好的抗肿瘤活性。
Objective: To prepare conjugated linoleic acid-gemcitabine conjugate liposomes (CGLP) and investigate its related properties. Methods: The optimum preparation method of CGLP was screened by orthogonal design. Particle size and morphology were determined by laser nanoparticle size analyzer and transmission electron microscopy. Differential scanning calorimetry (DSC) was used to detect the components of liposomes. The drug release and stability were determined by dynamic dialysis method. Inhibition of MCF-7 breast cancer cells at different time points. Results: Single factor method was used to establish CGLP by thin film dispersion method. Orthogonal design experiments using CGLP the best prescription for the phospholipid and cholesterol ratio of 15: 1, lipid ratio of 1: 8, ultrasonic time of 15 min. The best prescription conditions, the average entrapment efficiency of 94.35%. The mean diameter was (69.73 ± 2.09) nm, PDI was (0.235 ± 0.042). Electron microscopy of liposomes showed that the particles were well dispersed. DSC shows that due to the addition of CLA-GEM in liposomes, the structure of the phospholipid bilayer changes, which facilitates the drug loading. In vitro release tests showed that the cumulative drug release reached 96.39% after 6 h in vitro and 24.07% after 24 h in CGLP, which showed a sustained release effect. Stability experiments showed that the 4 ℃ conditions are more suitable for liposome storage. Cytotoxicity assays show that CGLP is more potent than the conjugate solution. There was a significant difference between the two groups (P <0.001) compared with unmodified gemcitabine. Conclusion: This study prepared a new CGLP, preparation method is simple, high encapsulation efficiency, has good anti-tumor activity.