Studies On Self-assembly Nanoparticles Of Curcumin-Vitamin-E Derivative

Objective:Curcumin-Vitamin E(CUR-VE),a derivative with the same pharmacological activity as Curcumin(CUR),was prepared by the best synthetic process screened in the laboratory.Using the self-assembly characteristics of CUR-VE,the self-assembly nanoparticles of Curcumin-Vitamin E derivative(CUR-VE-NPs)with high drug loading were designed and prepared by nanometer precipitation method.With stability,oral pharmacokinetic behavior as evaluation index,comparing with CUR,develop drug loadings,high stability and high pharmacokinetic behavior good,has potential clinical application of CUR-VE-NPs,with its as replacement drugs of CUR to make up for the deficiency of nanomaterials of CUR.Investigating the mechanism of CUR-VE-NPs’ self-assembly characteristic,thus supporting the scientific researches and clinical applications of derivatives and nanomaterials of CUR with data and evidence.Methods:1.Establishing the analysis methods of HPLC and UPLC-MS/MS for the analysis and measurement of CUR and CUR-VE,thus providing basic detection methods for subsequent experiments2.Preparing CUR-VE-NPs by nanoprecipitation method and determining the optimal prescription and preparation processes of CUR-VE-NPs,while taking particle sizes,PDI,Zeta potential,encapsulation efficiency and drug loading capacity as assessment indexes.3.Using Differential Scanning Calorimetry(DSC),X-ray Powder Diffraction(PXRD),Fourier Transform Infrared Spectrometer(FT-IR)and infrared difference spectrum analysis method to explore the mechanism of the self-assembly process.4.MTT assay was used to investigate the cytotoxic effects of CUR and CUR-VE on human breast cancer MCF-7 cells,and to investigate whether the introduction of D-α-tocopherol in the lead compound of CUR caused changes in drug activity.5.Establishing a simulated environment model of gastric fluids,intestinal fluids,plasma,and liver homogenate,to investigate the stability of CUR-VE-NPs,CUR-VE and CUR.6.Taking the Area Under the Curve(AUC0-∞),Half-life(T1/2),Tmax and Cmax as evaluation indexes,evaluating the pharmacokinetic behavior differences between CUR-VE-NPS and CUR through pharmacokinetic experiments on SD rats.Results:1.Established the analysis method of HPLC and UPLC-MS/MS for the determination of CUR and CUR-VE,which were suitable for the testing needs of in vitro formulation evaluation,formulation stability experiments and in vivo pharmacokinetic experiments.2.Screened the optimal prescription and preparation process through single factor inspection:dosing amount was 4 mg/mL,stirring adjustable was 400 rpm,the reaction temperature was 80 ℃,reaction time was 0.5 h,the stabilizer was 5%(w/w)DSPE-PEG2000.CUR-VE-NPs prepared by the nano-precipitation method and the average particle size was(172.9 ± 3.29)nm,the PDI was 0.133± 0.009,the zeta potential was(-46.5 ± 7.88)mV,the encapsulation efficiency was(81.66 ± 0.54)%,and the drug loading was(93.63 ± 0.84)%.In the 72 h release experiment,the cumulative release of CUR-VE-NPS was(44.97 ± 1.26)%.3.There was no crystal form change during CUR-VE’s self-assembly process,and no crystalline substances such as nanocrystals were formed.By exploring the mechanism showed that CUR-VE’s self-assembly process mainly relies on driving forces including van der Waals forces,hydrogen bonding forces,and π-π stacking.4.MTT results showed that CUR and CUR-VE had significant concentration dependence on the proliferation of MCF-7 cells,and there was no significant difference in the effect of CUR and CUR-VE on the viability of MCF-7 cells(P>0.05).The two drugs had similar cytotoxic effects on MCF-7 cancer cells.5.In the stability experiment with the simulated environmental model of gastric and intestinal fluids,plasma and liver homogenate,T1/2 of CUR-VE and CUR-VE-NPs were significantly different from that of CUR(P<0.05).All the orders of stability were:CUR-VE>CUR-VE-NPs>CUR.This indicated that CUR’s weak stability can be improved,either with structure modification(CUR-VE)or with further dosage form transformation(CUR-VE-NPs).6.The pharmacokinetic experiments on SD rats showed that pharmacokinetic parameters including the AUC0-∞,T1/2,Tmax and Cmax of CUR-VE-NPS were significantly higher than those of CUR,which means that the results were statistically significant(P<0.05).Comparing with CUR,CUR-VE-NPS can be better absorbed and released into the blood environment after oral administration.Conclusion:In this study,the optimal prescription and preparation processes of CUR-VE-NPs were screened out through single factor test,and the CUR-VE-NPs drug delivery system was established.Relying on intermolecular forces such as van der Waals forces and electrostatic repulsion,hydrogen bonding forces,and π-π stacking,CUR-VE-NPs can be formed and aggregate without other carrier materials or stabilizers.For the CUR-VE,in the structure of which the D-α-tocopherol replaces one molecule of the phenolic hydroxyl active group of CUR,its stability was better than CUR in simulated in vitro environments of gastric and intestinal fluids,plasma,and liver homogenate,however,the modification of CUR hydroxyl sites did not affect its anti-tumor activity.As CUR-VE-NPs can be dispersed as fine particles suspended in an aqueous dispersion medium without precipitation,the drug’s water dispersibility was increased significantly.The in vivo pharmacokinetic experiments indicated that comparing with CUR suspension,the T1/2 of CUR-VE-NPs was significantly prolonged(P<0.01,P<0.001),which indicated that the elimination rate of CUR-VE in vivo has slowed down.Then the AUC0-∞ and Cmax of CUR-VE were significantly different(P<0.001,P<0.01),indicating that the degree of oral absorption was significantly improved and pharmacokinetic behavior was significantly improved.