Preparation And Reversal Of Multug Resistance Mechanisms Mediadiated Epidermal Growth Factor Trioxide Albumin Nanoparticles

[Object]:Preparation of arsenic trioxide(As2O3)BSA nanoparticles,which needs its morphology,particle size distribution,drug loading,a final coating rate for quality inspection;Coupling epidermal growth factor(EGF)and nanoparticles process research and identification;Inspecting its behavior of in vitro release;Study of pharmacokinetics behavior which after the rat caudal vein injection in vivo,and calculate the related parameters;A preliminary study of multi-drug resistance which for epidermal growth factor coupling with arsenic trioxide nanoparticles.[METHOD]:By Hydride Generation-Atomic Fluorescence Spectrometry to establish analytical method of As2O3;In the means of Secondary phacoemulsification method and Solvent evaporation technique for preparation of arsenic trioxide(As2O3)BSA nanoparticles,according to the L9(34)orthogonal test counsels for prescription optimization,ultimately,with a final coating rate as examining index to inspect the best process;With appearance and dispersion as examining index to investigate the best technology of freeze-dried,Using transmission electron microscope to observe nanoparticles morphology,Using Zeta potential analyzer and nanometer particle to measure potential,particle size and distribution of nanoparticles;Ascertain the freeze-dried process of the nanoparticles and the stability of its product;By dynamic dialysis method to investigate drug release characteristics for As2O3-NP in vitro;According 3're pharmacokinetic software to calculate pharmacokinetic parameters of after dosing rats;Applied to The MTT methods,nanoparticles of coupling with epidermal growth factor be influence to reversal fold of MCF 7/ADR cells[Results]:By using the optimum prescription to preparate of three sample groups,Stability is excellent,the encapsulation efficiency is of 81.73%,drug-loading rate is of 8.97%;Frozen avoiding powder of As2O3-NP be with a better redispersibility in injecting water.Under the electron microscope be observed kind of spherical form but irregularity,transparent solution are light blue and with milk light shape,by using particle size analyzer to measure size distribution which is found more uniform,the average particle size is about 165.81 nm,Zeta potential is of 1.384 mV;PDI value is of 0.221,there be no hemolysis and flocculation phenomenon for injection producing,abnormal toxicity test is qualified;As to meet the requirements of the quality of the injection,Freeze-dried for 6 month and encapsulation efficiency fell by 3.26%,research shows that its stability is favorable;Establish the method of coupling EGF and nanoparticles;Inspect the testing method of As2O3-NP releasing in vitro,preliminary inspect for appropriate release medium in vitro release and to which makes a preliminary study on the in vitro drug release,As2O3-NP dialyzes to release medium of PBS buffer solution(pH=7),which cumulatives release is of 48.46%within 24 h,and was of up to 50.66%Within 48 h,but it was in a slow speed in vitro release,As2O3API releases faster than As2O3-NP,As2O3-NP has a certain extent of sustained-release effect;The result of pharmacokinetic parameters of the rat in vivo shows that:medicine-time curve of As2O3-NP accords with weight of 1/C2 2 chamber model,with the same dosage EGF-NPs would be in a slow metabolism,physiological saline group's is 1.61 times of EGF-NPs group's;AUC of EGF-NPs is 1.65 times of saline group's,the area under medicine-time curve enlarger significantly;Reduce plasma clearance,CL of EGF-NPs group is 0.63 times of saline group's.To choice multi-drug resistance reversal agents,As2O3 and EGF-NP,IC50 of ADM to MCF-7/ADR is 8.68 and 6.15,fold reversal is 1.58 and 3.09.[CONCLUSION]:Applicate of secondary ultrasonic emulsification volatilization method and solvent evaporation technique to preparate of As2O3-NP successfully,which method is simple and easy,the average particle size is of small and uniform,form is of a excellent,the encapsulation rate is of higher;As2O3-NP releases more slowly than As2O3API in vitro,which has a slow-release effect but no sudden release effect;And As2O3-NP can significantly extend the elimination half-life of rats;MCF-7/ADR reverse multi-drug resistant effect on human breast cancer cell,which effect on EGF-NP is better than As2O3's.