Preparation Of Galactosylated Liposomes Loaded With Gemcitabine Hydrochloride And Preliminary Study On Anti-tumor Activity

Objective In this experiment, gemcitabine hydrochloride as the model drug, stearyl alcohol galactosidase as the guide molecule to prepare galactosed gemcitabine hydrochloride liver targeting liposomes, expecting to improve gemcitabine hydrochloride liver targe ting, so that side effects decreased to obtain the most good therapeutic effect.Methods Before gemcitabine hydrochloride prescription studies, methods of determination and validation of gemcitabine hydrochloride was established and studied its physico chemical properties. Dextran gel microcolumn centrifugation method-UV and cation exchange resin microcolumn centrifugation-HPCE were used to detect entrapment efficiency of galactose gemcitabine hydrochloride liposomes. The entrapment efficiency as the evaluation index, Box-Behnken response surface design method was used to optimize the formulation, Galactosylated proliposomes loaded with gemcitabine hydrochloride was evaluated by encapsulation efficiency, drug loading, morphology, particle size, stability, and in vitro release. The inhibitory effect of lipo somes on SMMC-7721 and A549 cells was tested by MTT assay. Apoptotic rate of SMMC-7721 cells was detected by Annexin V-FITC/PI flow cytometry. Investigated the targeting property of liposomes initially.Results This experiment has established the best method for the determination of content of gemcitabine hydrochloride. Two methods can be used to encapsulate liposomes rate determination. The optimized formulation was as follows:the concentration of phospholipid was 3.10:1, ammonium sulfate concentration was 0.19mol·L-1, lipid ratio was 20.99:1, the average of liposome entrapment efficiency was(70.8±2.8)%, the average particle size was(78.8±8.9) nm, the particle size distribution is narrow, good stability and slow release characteristics. MTT showed that for the SMMC-7721 cells, Galactosylated proliposomes loaded with gemcitabine hydrochloride were the highest inhibition rate.For A549 cells, the inhibition rate of alactosylated gemcitabine hydrochloride proliposomes and gemcitabine hydrochloride proliposomes were not significant difference, but were higher than the gemcitabine hydrochloride. The results showed apoptotic effect of the alactosylated gemcitabine hydrochloride proliposomes were significantly higher than the original drug and gemcitabine hydrochloride proliposomes. Initially identified that Galactosylated proliposomes loaded with gemcitabine hydrochloride were capable of liver targeting.Conclusions In this experiment, Galactosylated proliposomes loaded with gemcitabine hydrochloride were prepared with high encapsulation efficiency, uniform particle size distribution, with certain liver targeting property.