Preparation And Study Of Magnetic Fe₃O₄@NH₂-?-CD Nanoparticles Loading Gambogic Acid

Objective:Based on the design principle of magnetic nanodrug-loading,Gambogic acid?GA?,magnetic iron tetraoxide nanoparticles?Fe₃O₄ MNPs?and aminated beta cyclodextrin??-CD?were used as raw materials to prepare GA loaded or loading magnetic Fe₃O₄@NH₂-?-CD nanoparticles?Fe₃O₄@NH₂-?-CD@GA MNPs?to improve the low bioavailability and the great vascular irritation of gambogic acid.Then,its physical properties and the biological activity was studied.And hope to obtain a novel magnetic nanomedicine with strong targeting and biological activity.Methods:1.Fe₃O₄ MNPs were prepared by co-precipitation method and modified with citric acid?CA?and aminated?-cyclodextrin?6-NH₂-?-CD?to prepare Fe₃O₄@NH₂-?-CD@GA MNPs.Indicated by the yield,a single factor investigation was conducted on the synthesis of 6-NH₂-?-CD,and the optimal synthesis process was discussed.The physical and chemical characteristics of the magnetic nanoparticles were characterized by means of FTIR,XRD,Zeta,VSM and HRTEM.2.Dialysis method was used to evaluate the drug-loading rate and drug release performance of Fe₃O₄@NH₂-?-CD@GA MNPs,based on in vivo environment of human was simulated.3.The hepatoma cell line HepG2 and leukemia cell line HL-60 were used as research objects,and the MTT assay was used to investigate the anti-tumor effects of the nano drug delivery system on solid tumors and non-solid tumor cells,and the IC₅₀was calculated.In addition,a method for the determination of GA in rat plasma by HPLC was established.The GA and Fe₃O₄@NH₂-?-CD@GA MNPs as medicines were administered by tail vein injection.Plasm was taken and determined at different time points.DAS2.0 software was used to process plasma concentration-time data,fit its pharmacokinetic model,and calculate the main pharmacokinetic parameters.4.Rabbits were used as experiment objects,the vascular irritation of Fe₃O₄@NH₂-?-CD@GA MNPs was observed by HE staining in optical microscope.Results:FTIR results showed that Fe₃O₄@NH₂-?-CD MNPs andFe₃O₄@NH₂-?-CD@GA MNPs have been synthesized and the particle sizes was119.4 nm and 147.4 nm,PDI was 0.201 and 0.189,potentials was-30.8 mV and-29.3mV,respectively,so the magnetic nano-drug delivery system was stable.The XRD and HRTEM spectra showed that the crystal lattice of Fe₃O₄@NH₂-?-CD MNPs and Fe₃O₄@NH₂-?-CD@GA MNPs were spherical or spheroidal.The crystal sizes of Fe₃O₄@NH₂-?-CD MNPs and Fe₃O₄@NH₂-?-CD@GA MNPs reaches 13.07 nm and15.60 nm,respectively.In addition,the H-B curves show that the saturation magnetizationofFe₃O₄MNPs,Fe₃O₄@NH₂-?-CDMNPsand Fe₃O₄@NH₂-?-CD@GA MNPs were significantly different,reaching 56.85,44.87and 41.76 emu/g.Moreover,the cumulative release curves was obtained from simulated human condition,which were divided into two stages of rapid release stage and slow release stage with a cumulative release time of 1080 min..The results of MTT assay demonstrated that Fe₃O₄@NH₂-?-CD@GA MNPs could inhibit cell lines HepG2 and cell lines HL-60,the IC₅₀ was 1.18?g/mL and 0.41?g/mL separately,lower than GA,significantly.Pharmacokinetic test found that the half-life(t_1/2)of Fe₃O₄@NH₂-?-CD@GA MNPs was significantly increase.Besides,the MRT has been greatly extended,AUC_0-t,AUC_0-?were also increased by 1.48 and 1.42 times.HE staining of the pathological section indicated that the vascular irritation was decreased of Fe₃O₄@NH₂-?-CD@GA MNPs.Conclusion:Fe₃O₄@NH₂-?-CD@GA MNPs were successfully prepared and their physical properties were studied.In vitro cell assays showed that Fe₃O₄@NH₂-?-CD@GA MNPs had a significant inhibitory effect on both solid tumor HepG2 and non-solid tumor HL-60 cells.Compared with GA,studies have shown that Fe₃O₄@NH₂-?-CD@GA MNPs have the advantages of small vascular irritation,long biological half-life,et al.And its could significantly improve the shortcomings of GA elimination and serious vascular irritation.A kind of new nano anti-tumor magnetic targeting agents was deserved further research and development.