Construction Of Docetaxel Targeted Nanoparticles And The Studies On The Anti-tumor Activities

At present,cancer has become one of the the main causes of death in China due to the increasing pressure of life and the deterioration of the environment and other factors.Surgical excision,radiotherapy and chemotherapy are the main methods for cancer treatment,but they all have some limitations.In order to improve the problem of chemotherapeutic drugs,the research of new drug delivery system is a hotspot of anti-cancer drugs.Nano-drug delivery system can be built by biodegradable materials to improve drug solubility,enhance targeting and reduce damage to other tissues.Objective: Docetaxel,taxanes,semisynthetic antineoplastic drugs,tubulin inhibitors.Docetaxel was used as the main drug,lecithin was used as carrier material,O/W emulsified solvent evaporation method was used to prepare nanoparticles.Stearamine was used as positive charge regulator.DSPE-PEG2000 was used to modify the nanoparticles.The beta-Ganoderma lucidum dextran was coated on the surface of positive charge nanoparticles by positive and negative charge adsorption.Zeta potential measurements showed that Ganoderma lucidum polysaccharide adsorbed docetaxel nanoparticles with negative charge.The encapsulation efficiency,drug loading,particle size and Zeta potential of nanoparticles were characterized.The release and stability of nanoparticles in vitro were investigated,and the cytotoxicity of nanoparticles on tumor cells and the anti-tumor effect in vivo in tumor-bearing mice were evaluated.The results provide theoretical guidance for the further study of nanoparticles.Methods: Docetaxel nanoparticles were prepared by O/W emulsifying solvent evaporation method.A method for the determination of docetaxel nanoparticles by high performance liquid chromatography was established.The mobile phase was acetonitrile:water=55:45,the flow rate was 1.0 m L/min,and the detection wavelength was 230 nm.Methodological validation showed that the peak area of docetaxel had a good linear relationship with the concentration in the range of 0.2000-3.2000 mg/m L,R2=0.9998.The specificity,intra-day difference and inter-day difference as well as the recovery rate were all meet the requirements with RSD<5 %.The drug was dissolved in organic solvents as O phase,2% Tween 20 as W phase,and O/W emulsion was evaporated to remove organic solvents.Nanoparticles were obtained.The entrapment efficiency,drug loading,particle size and Zeta potential were used as indicators to screen the prescription through single factor investigation.Ganoderma lucidum polysaccharides were extracted by water extraction and alcohol precipitation,and Ganoderma lucidum polysaccharide GLP-1 was separated by Sephadex Sephadex G-75 column chromatography.GLP-1 was charged negatively by Zeta potential,and octadecylamine was selected to prepare positive charge drug-loaded nanoparticles.The polysaccharide molecule was adsorbed to the surface of the nanoparticles due to the positive and negative charge attraction,and the drug-loaded nanoparticles were formed by the adsorption of Ganoderma lucidum polysaccharide.Results: The stability of nanoparticles under high temperature and light was studied with docetaxel injection as the control.The results showed that the degradation rate of nanoparticles was lower than that of commercial preparations,and it needed to be preserved in low temperature and avoiding light.PBS solution with 0.5 % Tween-80 p H of 7.2 was used as release medium in vitro.The cumulative drug release percentage was determined.The results showed that nanoparticles had no burst release and sustained release and the release rate was about 70%.Docetaxel nanoparticle solution was used as injection to evaluate the safety of human umbilical vein endothelial cell Huvec.MTT cytotoxicity results showed that GLP-PEG-Dox Nps nanoparticle solution was safe for human umbilical vein endothelial cell Huvec,and PEG-Dox Nps nanoparticle solution was safe for human umbilical vein endothelial cell Huvec.Docetaxel injection is less safe for human umbilical vein endothelial cell Huvec.The nanoparticles was validated the anti-tumor effect on human breast cancer cell MCF-7 in vitro,GLP-PEG-Dox NPs nanoparticles solution was the best,followed by PEG-Dox NPs nanoparticles solution,and finally docetaxel injection.A mice breast cancer model was established to evaluate the anti-tumor effect in vivo.Docetaxel injection inhibited the growth of tumors were 64.53 %,PEG-Dox Nps were 71.45 % and GLP-PEG-Dox Nps were 78.21 %.The inhibition rates of tumor growth were more than 40 %,compared with the docetaxel injection P < 0.05,with statistical difference.Conclusion: The prescription composition of Ganoderma lucidum polysaccharide adsorbed drug-loaded nanoparticles was docetaxel 6.0 mg,lecithin 80.0 mg,octadecylamine 6.0 mg,DSPE-PEG2000 15.0 mg,GLP-1 15.0 mg.The nanoparticles were clear and transparent emulsion with an average particle size of 126.4 nm,a Zeta potential of-6.25 m V,a encapsulation efficiency of 91.6 %,and a drug loading of 5.14 %.The Ganoderma lucidum polysaccharide adsorbed docetaxel nanoparticles has the advantages of enhancing the efficacy and prolonging the drug time compared with the commercial preparation,which makes the foundation for the far research of targeted nanoparticles.