Preparation,Characterization And Preliminary Pharmacokinetics Research Of Folate Modified Non-ionic Surfactant Vesicles Loaded Gambogenic Acid

Objective:Gambogenic acid is one of the main anti-tumor active ingredients of Gamboge.Previous reports indicated that GNA has broad anti-tumor spectrum and inhibited several tumor cells proliferation.However,it was limited due to its low aqueous solubility,short elimination half-life,poor bioavailability and excessive irritation to blood vessel by intravenous administration.The folic acid modified non-ionic surfactant vesicles loaded gambogenic acid was prepared,using non-ionic surfactant vesicles as a drug carrier,GNA as the model drug.It was modified with targeting ligand folate-polyethyleneglycol₂₀₀₀-phosphatidylethanolamine(FA-PEG₂₀₀₀-DSPE)in niosomes carrie surface.The purpose of this study was to improve the poor water solubility,short half-life,low bioavailability and vascular irritation of GNA.It will support for theoretical of GNA basic research.Methods:?1?It were prepared from the methodnon-ionic surfactant of Spans60,membrane stabilizer of cholesterol,polyethylene glycol monostearate?PEG-SA?by improvingethanolinjection anditwasmodifiedwithtargetingligand FA-PEG₂₀₀₀-DSPE,which was grafted onto the surface of niosomes carrie.The optimum formulation was optimized through single factor and orthogonal experiment.?2?Characterization of FA-GNA-NSVs:to investigate the morphology of the folic acid modified non-ionic surfactant vesicles loaded gambogenic acid,it could be seen from the transmission electron microscopy?TEM?observation of morphology.The average particle size,Zeta potential and dispersion coefficient?PDI?were determined by dynamic light scattering with zetasizer.HPLC was calculated encapsulation efficiency?EE%?and drug loading?DL?.The DSC analysis by Differential scanning calorimetry.The folic acid modified non-ionic surfactant vesicles loaded gambogenic acid were investigated physical placement the stability of 30 d in 4?refrigeration and 25?room temperature.The drug release characteristics was evaluated using dialysis technique by simulation of normal cell environment in vitro.?3?It was evaluated that the pharmacokinetic behavior in vivo using SPF SD rats as animal model.?4?The safety of FA-GNA-NISVs was evaluated by of vascular irritation experiment and hemolytic experiment in rabbits.Results:?1?The optimal prescription of the folic acid modified non-ionic surfactant vesicles loaded gambogenic acid was Spans60:25.0 mg,Chol:20.0 mg,PEG-SA:7.0mg,FA-PEG₂₀₀₀-DSPE:3.0 mg,GNA:7.0mg,ethanol:3.0 mL,DMSO:150.0?L,pure water 25.0 mL by orthogonal design.?2?The folic acid modified non-ionic surfactant vesicles loaded gambogenic acid was observed smooth surface and no adhesion by the transmission electron microscopy?TEM?.Average particle size?161.85±1.19?nm,Zeta potential?-27.33±0.33?mV,the polydispersity index 0.065±0.026 and encapsulation efficiency?87.84±0.0064?%.DSC analysis showed the GNA had been loaded by FA-GNA-NISVs,and the GNA was not a crystalline state but a new phase.Furthermore,The folic acid modified non-ionic surfactant vesicles loaded gambogenic acid was stabilized within 30 d at 4?refrigeration and 25?room temperature.The drug release characteristics of the folic acid modified non-ionic surfactant vesicles loaded gambogenic acid showed sustained release in 12 h,the cumulative release rate?67.49±0.0015?%,with a slow and sustained release drug characteristics,in accordance with First-order kinetics equation.?3?Pharmacokinetic studies in rats after a single i.v.injection showed that the FA-GNA-NISVs AUC?up to 5.4-fold?,C_max?up to4.4-fold?,t_1/2?up to 2.4-fold?,MRT?up to 1.2-fold?compared to GNA solution.?4?The results of vascular stimulation test showed that the stimulation of FA-GNA-NISVs on blood vessels was significantly lower than that of GNA,and the hemolysis test showed that FA-GNA-NISVs had no hemolysis in 240 min.Conclusion:The results confirmed that the folic acid modified non-ionic surfactant vesicles loaded gambogenic acid has small particle size,high encapsulation efficiency and better stability.It can effectively improve the GNA of the poor water solubility,short of half-life,vascular stimulation and bioavailability.The folic acid modified non-ionic surfactant vesicles loaded gambogenic acid can significantly enhance the killing effect on tumor cells.It will be expected to a new anti-tumor drug delivery system.