紫杉醇聚乳酸微球的制备及其抗胃癌活性评价  

Preparation of PTX-PLLA-MPS and evaluation of its anti-gastric cancer activity

作　　者：孙彬成 刘静 李君 SUN Bincheng;LIU Jing;LI Jun(Department of Gastroenterology,Bayannur Hospital,Bayannur 015000,China;不详)

机构地区：[1]巴彦淖尔市医院消化内科,内蒙古巴彦淖尔015000 [2]内蒙古医科大学药学院

出　　处：《山东医药》2023年第22期31-35,共5页Shandong Medical Journal

基　　金：内蒙古自治区高等学校科学研究项目(NJZY23135);内蒙古医科大学校级青年项目(YKD2022QN004)。

摘　　要：目的制备载紫杉醇聚乳酸微球(PTX-PLLA-MPS),建立含量测定方法,优化制备工艺,并对其体外释放行为及抗胃癌活性进行初步评价。方法采用溶剂挥发法制备PTX-PLLA-MPS;采用高效液相色谱法测定PTX-PLLA-MPS中紫杉醇含量并计算载药量及包封率;以载药量、包封率为评价指标,通过正交试验优化制备工艺;采用光学显微镜观察其形态,采用粒径分析仪测定其粒径及Zeta电位,采用高效液相色谱法考察体外释放行为;胃癌SGC7901细胞调整细胞浓度至4×10^(6)/mL,将细胞接种96孔板。过夜培养后,随机设置空白微球组、PTX组、PTXPLLA-MPS组,分别加入30μg/mL空白微球、30μg/mL PTX、30μg/mL PTX-PLLA-MPS处理24、48、72 h。加入CCK-8溶液计算各组细胞活力;使用胰蛋白酶消化人胃癌细胞SGC7901,调整细胞浓度后,皮下注射到裸鼠前肢腋下,注射体积为0.2 mL/只,建立裸鼠荷瘤模型。选取成瘤裸鼠15只,按体质量随机分为3组:模型组、PTX组和PTX-PLLA-MPS组,尾静脉注射给药,2 d一次,给药剂量15 mg/kg,绘制肿瘤生长曲线。结果PTX-PLLA-MPS最佳制备工艺:PVA水溶液为1.0%,油/水体积比1∶10,PLLA浓度为6%,PLLA/PTX比为6∶1,所得微球形态圆整,其平均粒径为(1.23±0.21)μm、Zeta电位为(3.67±1.42)mV、平均载药量为(18.23±1.40)%、包封率为(74.00±1.49)%,体外21天累计释放率为(76.20±3.46)%,空白微球组、PTX组、PTX-PLLA-MPS组细胞活力随着时间的延长依次降低(P均<0.05)。模型组、PTX组和PTX-PLLA-MPS组肿瘤体积随着时间延长呈增大趋势,28 d后PTXPLLA-MPS组肿瘤体积大于PTX组(P均<0.05),PTX组肿瘤体积小于模型组(P均<0.05)。结论成功制备PTX-PLLA-MPS,其形态良好,包封率载药量高,具有缓释特性和显著抗胃癌活性。Objective To prepare the paclitaxel-loaded polylactic acid microspheres(PTX-PLLA-MPS),to establish the content determination method,to optimize the preparation process,and to preliminarily evaluate its in vitro release behavior and anti-gastric cancer activity.Methods PTX-PLLA-MPS was prepared by solvent evaporation method.The content of paclitaxel in PTX-PLLA-MPS was determined by high-performance liquid chromatography and the drug loading and encapsulation efficiency were calculated.The preparation process was optimized through orthogonal experiments based on the evaluation indicators of drug loading and encapsulation efficiency.The morphology was observed with an optical microscope,the particle size and Zeta potential were measured by a particle size analyzer,and the release behavior in vitro was investigated by high performance liquid chromatography.We adjusted the cell concentration of gastric cancer SGC7901 cells to 4×10^(6)/mL,and inoculated the cells in 96-well plates.After overnight culture,the blank microsphere group,PTX group,and PTX-PLLA-MPS group were randomly set up,and 30μg/mL blank microspheres,30μg/mL PTX,and 30μg/mL PTX-PLLA-MPS were added for treatment of 24,48,72 h.We added CCK-8 solution to calculate cell viability in each group,used trypsin to digest human gastric cancer cells SGC7901,adjusted the cell concentration,and injected subcutaneously into the axilla of the forelimb of nude mice with an injection volume of 0.2 mL/mouse to establish the nude mouse tumor-bearing models.Fifteen nude mice with tumors were selected and randomly divided into three groups according to their body weight:model group,PTX group,and PTX-PLLA-MPS group.They were injected through the tail vein,once every 2 days,at a dose of 15 mg/kg,and the tumor growth curve was plotted.Results It was showed that PTX-PLLA-MPS was optimally prepared with an aqueous PVA solution of 1.0%,an oil/water volume ratio of 1:10,a PLLA concentration of 6%,and a PLLA/PTX ratio of 6:1,and the resulting microspheres were round with a mean

关 键 词：胃癌 紫杉醇聚乳酸微球 含量测定 制备工艺优化 紫杉醇新剂型 抗胃癌活性 

分 类 号：R735.2[医药卫生—肿瘤]