Preparation And Properties Research Of Docetaxel-loaded Phosphatidylcholine/Bile Salt Mixed Micelles

ObjectiveThis research chose docetaxel?DTX?as a model drug to prepare docetaxel-loaded phosphatidylcholine/bilesalt-mixedmicelles?DTXPC/BS-MMs?,inwhich PC/BS-MMs was used to improve the solubility of hydrophobic drugs DTX in water and then discussed its related properties.Solubilizer polysorbate 80 was not been used in preparation,PC/BS-MMs provided a new direction for the clinical use of DTX.Methods1.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 usedto investigate dissolving capacityof 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 safety 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.Results1.Micelles 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?was 40%,the concentration of total material was 5%,the hydration media was pure water and hydration volume was 10ml in the optimal prescription.The average size and average zeta potential of DTX PC/BS-MMs were 18.56±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 about 10s and the solution after dissolution was clear.In 120 minutes the dilution of the micelle solution was not precipitated.The hemolysis rate was less than 5%in hemolytic test.And all mouse survived more than 48h in the abnormal toxicity experiment.The IC₅₀ values in Cytotoxic experiment were respectively 0.84±0.03?M?BT474?and 0.88±0.05?M?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.Conclusion1.We used PC and SDS as materials to prepared DTX PC/BS-MMs by coprecipitation method in this experiment,the mixed micelles were clear and pale yellow.2.Hemolytic experiment and abnormal toxicity experiment found that the preparation had safety in vitro.3.The cytotoxic experiment and the pharmacodynamic experiment in vivo showed that DTX PC/BS-MMs had a considerable inhibitory effect for tumor cell.The preparation of DTX PC/BS-MMs was simple and the solution was stable.Therefore the preparation have hope to be developed into a new drug delivery system.