Preparation Of Evodiamine Liposome And The Evaluation In Vitro And In Vivo

Objective: To establish method for the determination of Evodiamine liposomes and their biological samples. Prepare Evodiamine liposomes with high encapsulation efficiency and stable quality. Investigate the release characteristics and antitumor effects of liposomes in vitro.Study on the pharmacokinetics of evodiamine liposome in rat blood,and distribution of tissues in mice.Methods:Determinate the encapsulation efficiency of Evodiamine liposome with high performance liquid chromatography. Combined single factor experiment and response surface to optimizate the formulation and process. Evaluate the appearance, particle size,drug loading, encapsulation efficiency, stability and other indicators of Evodiamine liposome. Investigate the drug release in vitro of Evodiamine liposome and solution by dialysis. MTT assay was used to investigate the antitumor activity in vitro of Evodiamine liposomes and Evodiamine solution on hepatoma cells SMMC-7721. Rats were intravenously injected Evodiamine liposomes and solution, timing orbital blood to determine the content of the Evodiamine pharmacokinetic characteristics of the two formulations were compared. Mice were intravenously injected Evodiamine liposomes and solution, timing sacrificed to determine the Evodiamine content of heart, liver, spleen, lung,kidney tissue, study the tissue distribution of two formulations in vivo,conduct the targeting preliminary preliminary evaluation.Results: The best process for the preparation of liposomes was film dispersion method,the organic solvent was ether,forming temperature was 35℃,p H of PBS buffer was7.4,microjet pressure was 120 psig,cycles for 12 times, mass ratio of phospholipid and V Ewas 15:1,mass ratio of phospholipid to drug was 30.58:1,mass ratio of phospholipid to cholesterol was 15.22:1, the concentration of phospholipid was 42.26 mg·m L-1.Evodiamine liposome was spherical or oval pellets, the average particle size was 126 nm, Zeta potential-48.1 m V, p H was 6.94, the average encapsulation efficiency was 92.99%, the average drug loading was 2.54%, liposomes was stable at 4 ℃. The release in vitro of evodiamine liposomes comply with Weibull model, only 29.79% was released in 6 h. SMMC-7721 cells was strongly inhibited by evodiamine liposome and solution. The IC50 of evodiamine liposome in 24 h was 7.394 μg·m L-1.The IC50 of evodiamine liposome in 48 h was 4.217μg·m L-1.The IC50 of evodiamine solution in 24 h was 0.628 μg·m L-1. The IC50 of evodiamine solution in 48 h was 0.936 μg·m L-1.The pharmacokinetic of evodiamine solution in line with a three-compartment model,and liposomes in line with two-compartment model. CL of evodiamine solution group was 5.395 L/h/kg, AUC(0-t)was 2.996 mg/L*h, CL of liposome group was 3.512 L/h/kg, AUC(0-t) was 4.891 mg/L*h.After the mice were intravenously injected liposomes, liver had the largest distribution,the relative uptake(Re) was 3.21, peak concentration ratio(Ce) was 1.21.Conclusions:Preparation of evodiamine liposomes was stable and feasible, quality of liposomes was quite stable. evodiamine liposomes had significant sustained-release,anti-tumor activity and liver targeting.