Study On Anti-tumor Activity Of Quercetin Nano-suspension With Different Particle Size

Objective:At present,malignant tumor is still a serious threat to human health,there are a variety of clinical diagnosis and treatment for tumor treatment,in which chemotherapy has gradually become one of the main treatment methods to inhibit tumor.In recent years,with the continuous development of pharmaceutical research,great progress has been made in drug research,and the therapeutic effect of tumor has also been significantly improved.However,the side effects of chemotherapy come one after another.the toxic and side effects of immunosuppression caused by chemotherapeutic drugs are difficult to overcome,which has caused great pain to cancer patients,so this problem needs to be solved urgently.Quercetin is a natural drug with a wide range of pharmacological effects..However,its solubility in water is very small,which leads to its bioavailability in vivo is also greatly reduced,so insoluble in water greatly hinders its clinical development and utilization.With the continuous innovation and development of pharmaceutical preparations,nanosuspensions have attracted more and more attention because of their ability to greatly improve the solubility of insoluble drugs.It is a submicron colloidal dispersion system prepared with polymer or surfactant as stabilizer.It has the characteristics of improving the solubility of insoluble drugs,increasing bioavailability in vivo,good biocompatibility and so on.Therefore,the preparation of drugs into nano-suspension has obvious advantages.Methods:In this paper,using amphiphilic polyethylene glycol monomethyl ether-deoxycholic acid?mPEG-DCA?as stabilizer,three kinds of quercetin nanosuspensions with different particle sizes were prepared by micro precipitation-high pressure homogenization method with different homogenization pressure and homogenization times.Firstly,its Size,Zeta potential and PDI were studied,and its storage stability and plasma stability were investigated.In addition,the crystal form of nanosuspension was characterized by DSC,DTG and XRD.Then the content of quercetin in vitro was determined by UV spectrophotometer,the content and dissolution of quercetin in vitro were investigated.The toxic effect of quercetin injection blank solvent on 7702 human normal hepatocytes was detected by MTT assay,and the toxic effects of injection,injection blank solvent and three kinds of nanosuspensions on MCF-7 cells were studied.To establish the method of plasma sample treatment and the determination of plasma sample content in vivo,to study its pharmacokinetics in vivo,to establish the model of H₂₂2 tumor-bearing mice and the model of immunosuppression,To investigate its inhibitory effect on tumor and immunomodulatory function in tumor-bearing mice.Results:The particle sizes of the three quercetin nanosuspensions were 30 nm,160 nm and330 nm,respectively,and the particle size distribution was narrow and the stability was good.The results of DSC,DTG and XRD showed that the particle size of quercetin nanosuspension with different particle size changed only,and the crystal form of quercetin nanosuspension did not change significantly.The wavelength scanning was carried out by ultraviolet spectrophotometer.the optimum wavelength was 373 nm.Under this wavelength,the standard curve was established and the content was determined.The dissolution in vitro was carried out by intelligent dissolution instrument.The results showed that the dissolution performance of nanosuspension was significantly improved compared with that of API,and the dissolution rate was accelerated with the decrease of particle size.The effective boundary surface characteristic analyzer was used to verify the rationality of the dissolution phenomenon.the particle size decreased and the surface area increased,so the dissolution was accelerated.MTT assay was used to investigate the cytotoxicity of quercetin injection.The results showed that the blank solvent of quercetin injection had been killing effect on human normal hepatocytes,so it had certain toxicity,and the quercetin nanosuspension with different particle size was compared with the injection.The results showed that 330nm quercetin nanosuspension had better curative effect on tumor cells.To study the pharmacokinetics of quercetin nano-suspension in rats,to investigate the pharmacokinetics of quercetin nanosuspension in rats,and to establish a method for the determination of HPLC in plasma samples.the results showed that the preparation of nanosuspension had greater bioavailability.With the increase of particle size,the elimination rate of the drug in rats decreased,which indicated that the nanosuspension group had a longer blood retention time in vivo.The T_1/2,AUC and MRT of 330nm quercetin nanosuspension were significantly better than those of the other two preparations.The antitumor activity and immunomodulatory effect of different particle size preparations and injections in vivo were studied.There was no significant difference in antitumor effect between three kinds of quercetin nanosuspension with different particle size and injection.However,the results of immune index showed that the immune index of 30,160 and 330nm quercetin nanosuspensions group was higher than that of cyclophosphamide group,and better than that of quercetin injection group,indicating that quercetin nano-suspension could enhance the immunity of mice to a certain extent..The body weight of tumor-bearing mice and the H&E staining physiological sections of some tissues showed that some tissue damage occurred in the injection and cyclophosphamide groups,but no obvious toxicity was observed in the three different particle size groups.