英文缩略词
英文缩写英文全名中文全名BS Bile salt胆盐
CMC Critical micelle concentration临界胶束浓度DTX Docetaxel多西他赛 西华大学图书馆
DTX PC/BS-MMs Docetaxel Phosphatidylcholine－
bile salt-mixed micelles 多西他赛磷脂胆盐复合胶束 HPLC High performance liquid
chromatography
高效液相谱MTT3-(4,5-dimethylthiazol-2-yl)-2,5-
diphenyltetrazolium bromide
噻唑蓝
劳合乔治
PC Phoshatidylcholine磷脂（磷脂酰胆碱）PC/BS-MMs Phosphatidylcholine－bile
salt-mixed micelles
磷脂胆盐复合胶束PBS Phosphate buffered solution磷酸盐缓冲液SDC Sodium deoxycholate去氧胆酸钠
SGC Sodium glycocholate甘氨胆酸钠
TEM Transmission electron microscope透射电子显微镜UV-Vis Ultra-violet and visible紫外-可见光
多西他赛磷脂胆盐复合胶束的制备及其性质研究安徽新长江投资集团
聚四氟乙烯烧结炉
摘要
目的：
太中银铁路本研究选用多西他赛（docetaxel，DTX）作为模型药物，制备多西他赛磷脂胆盐复合胶束，利用复合胶束对疏水药物多西他赛增溶，并对其相关性质进行探讨。制备中不使用增溶剂吐温-80，以期为临床
使用多西他赛提供一个新的方向。方法：
1.磷脂胆盐复合胶束的制备与理化性质表征
分别采用直接溶解法和共沉淀法制备复合胶束，考察制备中的实验条件（反应溶剂、时间和温度等）对胶束制备的影响，初步确立制备方法及条件。采用高效液相谱法测定复合胶束中多西他赛的含量。以单因素法和星点设计法筛选制备多西他赛复合胶束的最优处方，利用紫外可见-分光光度法考察胆盐对磷脂的溶解能力，高效液相谱法考察不同影响因素（辅料比例及种类、辅料总质量、水化液的pH及离子强度等）下多西他赛在溶液中的浓度。使用透射电镜鉴定胶束的粒径和形态，激光粒度仪测定胶束的Zeta电位和粒径。 2.磷脂胆盐复合胶束的体内外安全性与有效性初步评价
制备复合胶束冻干制剂，并测定其包封率、粒径；稀释实验考察胶束的稳定性。采用体外溶血实验和异常毒性实验初步考察制剂体外安全性。使用细胞毒性实验（MTT法）考察制剂对BT474和A549细胞的生长抑制作用。构建荷瘤小鼠模型，对制剂的安全性和有效性进行初步评价。
结果：
1.使用共沉淀法制备了胶束溶液（反应溶剂：无水乙醇，反应温度：40℃，反应时间：1h，实验用胆
盐：去氧胆酸钠）。最佳处方中磷脂/总辅料质量比为40%，辅料总浓度为5%，水化介质为纯水，水化体积为10ml。载药胶束平均粒径为18.56±
2.21nm，Zeta电位为-28.7±2.3mV，透射电镜图显示胶束为类圆形，大小分布均匀。
2.复合胶束的冻干制剂为白疏松状固体，无需添加赋形剂。冻干制剂复溶时间在10s左右，复溶后为澄清溶液。复合胶束溶液稀释后放置120分钟内未出现析出现象。溶血试验中溶血率小于5%，异常毒性试验中给药小鼠生存时间均超过48h。细胞毒性试验测定的IC50值分别为0.84±0.03μM（BT474）及0.88±0.05μM（A549）。载药复合胶束组小鼠最终平均肿瘤体积(1500mm3)显著小于生理盐水组(2500mm3)，且载药复合胶束组小鼠体重未出现明显下降。
结论：
1.本实验以磷脂（主要为磷脂酰胆碱）和去氧胆酸钠作为辅料，通过共沉淀法制备了多西他赛磷脂胆盐复合胶束（DTX PC/BS-MMs），载药胶束为澄清淡黄溶液。
2.溶血实验和异常毒性实验提示制剂有一定的体外安全性。
道门秘术3.细胞毒性实验和体内药效学实验发现，多西他赛磷脂胆盐复合胶束对实验采用的肿瘤细胞的生长有一定的抑制作用。且DTX PC/BS-MMs制备工艺简单，稳定性好，有望开发成新的药物传递系统。
关键词：多西他赛；磷脂酰胆碱；胆盐；复合胶束
Preparation and Properties Research of Docetaxel-loaded Phosphatidylcholine/Bile Salt Mixed Micelles
Abstract
Objective
This research chose docetaxel(DTX)as a model drug to prepare docetaxel-loaded phosphatidylcholine/bile salt-mixed micelles(DTX PC/BS-MMs),in which PC/BS-MMs was used to improve the solubility of hydrophobic drugs DTX in water and then discussed its related properties.Solubilizer polysorbate80was not been used in preparation,PC/BS-MMs provided a new direction for the clinical use of DTX. Methods
1.Preparation and physicochemical properties evaluation of PC/BS-MMs.
Direct dissolution and coprecipitation method were respectively carried out for preparation of PC/BS-MMs,the influence of the experimental conditions(reaction solvents,time,temperature,etc.)were studied,preparation method and conditions were established preliminary.High performance liquid chromatography(HPLC)was established for content determination of DTX in DTX PC/BS-MMs.The single factor and star design method were conducted to screen the optimal prescription,ultraviolet visible-spectrophotometry was used to investigate dissolving capacity of phospholipids in bile salt,the concentration of DTX in solution was studied by HPLC method in different influence factors(material proportion and types,total mass of material,the pH and ionic strength of hydrated solution etc.).The particle size and appearance of micelles were determined by transmission electron microscopy(TEM), the Zeta potential and particle size of the micelle were detected by laser granulometer.
2.The evaluation of safety and effectiveness for DTX PC/BS-MMs in vitro and vivo
The DTX PC/BS-MMs were prepared by the method of freeze-drying,then its encapsulation rate and particle diameter were detected;the stability of micelle was investigated by dilution experiment.The s
afety of the preparation was preliminarily examined by hemolytic test in vitro and abnormal toxicity experiment.The anti-tumor effect,safety and effectiveness of DTX PC/BS-MMs were studied by cytotoxic experiment(MTT method)and building the tumor-bearing model in mice respectively.
Results
1.M icelles solution was prepared by coprecipitation method(reaction solvent: anhydrous ethanol,reaction temperature:40℃,reaction time:1h,experimental bile salts:sodium deoxycholate（SDC）).The mass ratio of PC/total（PC+SDC）was40%, the concentration of total material was5%,the hydration media was pure water and hydration volume was10ml in the optimal prescription.The average size and average zeta potential of DTX PC/BS-MMs were18.56±
2.21nm and-28.7±2.3mV respectively.A round like shape was observed by TEM,the size and distribution were pretty uniform.
2.The freeze-drying preparations of PC/BS-MMs were white loose solid and prepared without adding any excipient.The dissolution time was about10s and the solution after dissolution was clear.In120minutes the dilution of the micelle solution was not precipitated.The hemolysis rate was less than5%in hemolytic test.And all mouse survived more than48h in the abnormal toxicity experim
ent.The IC50values in Cytotoxic experiment were respectively0.84±0.03μM（BT474）and0.88±0.05μM （A549）.The final mean tumor volume(1200mm3)of DTX PC/BS-MMs group was significantly smaller than the saline group(3200mm3),and the weight of mice in the DTX PC/BS-MMs group did not decrease significantly.
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