白芨多糖衍生物的制备及其作为抗癌药物多西他赛载体的研究

发布时间：2018-02-24 06:40

  本文关键词： 白芨多糖 两亲性聚合物 聚合物胶束 载药量 包封率　出处：《吉林大学》2017年硕士论文　论文类型：学位论文

【摘要】：为增加白芨多糖的疏水性,以硬脂酸对白芨多糖进行疏水修饰,制备了白芨多糖两亲性聚合物(SA-BSPs)药物载体。采用红外光谱及氢核磁共振光谱对其进行表征,其取代度利用氢核磁共振光谱中峰面积进行计算。并以多西他赛为模型药物,采用溶剂挥发法制备多西他赛硬脂酸修饰白芨多糖(DTX-SA-BSPs)胶束,采用激光粒度测定仪对胶束粒径、粒度分布及zeta电位进行测定。以高效液相色谱法(HPLC)测定其载药量及包封率。采用透析法对其体外释放度进行考察。小鼠单剂量尾静脉注射给予DTX-SA-BSPs聚合物胶束及多西他赛注射液,采用高效液相色谱法测定不同时间点血药浓度,绘制药-时曲线,使用DAS2.1药动学软件进行相关药动学参数的计算,考察其在小鼠体内药动学。测定各组织脏器中药物浓度,评价多西他赛制成胶束后其在小鼠体内的分布。通过MTT法和红细胞溶血实验对硬脂酸修饰的白芨多糖衍生物进行初步生物安全性评价。结果表明,硬脂酸已成功接枝到白芨多糖的羟基上,取代度为12.94%。DTX-SA-BSPs聚合物胶束平均粒径为(97.01±3.17)nm,zeta电位为(-19.56±0.22)mV、载药量为(9.13±0.17)%、包封率达(81.11±0.18)%。结果表明,在pH7.4磷酸盐缓冲液中多西他赛从胶束释放量随时间延长,体外累积释药量逐渐增加,在48 h时体外累积释药量达到(66.93±1.79)%,且呈现出缓慢释放的特性。MTT实验表明,0.5μg/mL DTX-SA-BSPs聚合物胶束孵育72 h时,肝癌细胞抑制率为(83.7±1.0)%,对肝癌细胞具有一定的抑制作用。小鼠体内药代动力学研究结果表明,DTX-SA-BSPs聚合物胶束相较于多西他赛注射液在小鼠体内具有更高的血药浓度、更长的维持时间(MRT 1.42±0.02h vs 1.82±0.11 h,p0.05),绝对生物利用度是多西他赛注射液的1.39倍(AUC0-∞65.39±5.21μg/mL·h vs 47.73±0.49μg/mL·h,p0.05)。组织分布实验表明,DTX-SA-BSPs聚合物胶束在心、肝、肺组织具有较高的药物分布,而在脾、肾药物分布较低,表明DTX-SA-BSPs聚合物胶束可改变药物的组织分布,是一种治疗癌症的可用方法。
[Abstract]:In order to increase the hydrophobicity of Bletilla striata polysaccharide, stearic acid was used to modify Bletilla striata polysaccharide hydrophobically. The amphiphilic polymer of Bletilla striata polysaccharide SA-BSPswas prepared and characterized by IR and HNMR. The degree of substitution was calculated by means of peak area in hydrogen nuclear magnetic resonance spectroscopy, and docetaxel was used as model drug to prepare DTX-SA-BSPs micelles modified with docetaxel stearic acid by solvent volatilization. The micelle size was measured by laser particle size analyzer. The particle size distribution and zeta potential were measured. The drug loading and entrapment efficiency were determined by HPLC. The release in vitro was investigated by dialysis. Single dose tail vein injection of DTX-SA-BSPs polymer micelle and docetaxel injection was used in mice. High performance liquid chromatography (HPLC) was used to determine the drug concentration at different time points and draw the drug-time curve. The pharmacokinetic parameters were calculated by DAS2.1 software, and the pharmacokinetics in mice was investigated. To evaluate the distribution of docetaxel micelles in mice. The biological safety of stearic acid modified Bletilla striata polysaccharide derivatives was evaluated by MTT method and erythrocyte hemolysis test. Stearic acid was successfully grafted onto the hydroxyl groups of Bletilla striata polysaccharides. The degree of substitution was 12.94. The average particle size of DTX-SA-BSPs polymer micelles was 97.01 卤3.17nmmZeta potential was -19.56 卤0.22mV, the drug loading capacity was 9.13 卤0.17mV, the entrapment efficiency was 81.11 卤0.180.The results showed that. In pH7.4 phosphate buffer, the amount of docetaxel released from micelles increased with time, and the cumulative release of docetaxel gradually increased with time. At 48 h, the cumulative drug release reached 66.93 卤1.79%, and showed slow release. The results showed that 0.5 渭 g / mL DTX-SA-BSPs polymer micelles were incubated for 72 h. The inhibition rate of hepatoma cells was 83.7 卤1.0, which had a certain inhibitory effect on hepatoma cells. The results of pharmacokinetic study in mice showed that DTX-SA-BSPs polymer micelles had higher blood drug concentration than docetaxel injection in mice. The longer maintenance time was 1.42 卤0.02 h vs 1.82 卤0.11 h (p 0.05), and the absolute bioavailability was 1.39 times higher than that of docetaxel injection. AUC0- 鈭，

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