Study On The Hyaluronic Acid Coated Chinese Medicine Multicomponents Loaded Nanostructured Lipid Carriers

In today's medical technology is highly developed cancer remains a serious threat to human health and life.As drug carrier for transportation of drug therapy is the most promising treatments.This topic to build more Chinese traditional medicine ingredients hyaluronic acid modified load of nanostructured lipid carrier system,first of all,nanostructured lipid carriers used to wrap mount three difficult soluble active ingredients in ginseng oleanolic acid and ursolic acid,ginseng saponin Rg3,increase the solubility of drugs;Hyaluronic acid and then used as the target factor,the charge adsorption to targeted modification of nanostructured lipid carriers,achieve the goal of active targeting tumors.Explore the hyaluronic acid modified load as the carrier of traditional Chinese medicine composition more nanometer system coating and drug release rules,through the study in vivo and in vitro in order to achieve within the tumor cells to medicine.1.OUR-NLC craft and prescription optimizationBy nanoparticles size,coating rate,drug-polymer interactions,such as for evaluation index,using the single factor investigation method with star design-response surface method to optimize craft and prescription.Optimization results show that the NLC as carrier material and CTAB as the emulsifier,the solvent by ultrasonic dispersing OUR-NLC appearance of light blue opalescence,total envelopment rate and total drug loadings,respectively(45.668 + 1.14)%and(8.76 + 0.95)%.2.The preparation of HA-OUR-NLC and process optimizationCharge adsorption of hyaluronic acid prepared by the modified take Chinese traditional medicine composition more nanostructured lipid carriers(HA-OUR-NLC).Hyaluronic acid containing carboxyl burden of electricity,OUR-NLC lotus positively charged,charge adsorption method is used to make the HA adsorption in OUR-surface of NLC,gain the initiative targeted nanostructured lipid carriers.Prescription and process factors.To determine the optimal prescription for:HA concentration of 0.5 mg/ml,HA and the quality of the NLC ratio of 3:1,dropping speed of 1 ml/min,stirring speed of 30 r/min.Freeze-drying technology was studied by comparative experiment method,the results showed that the concentration of 5%mannitol as HA-OUR freeze-NLC loose appearance of producing is white powder,soluble good.Three batch of product display,good reproducibility,the technology is feasible.3.HA-OUR-NLC nanoparticle characteristics researchWith transmission electron microscope HA-OUR-the morphology of NLC,Zetasizer Nano-ZS90 laser particle size analyzer to determine particle size of the nanoparticles,PDI and Zeta potential.Results show that the HA-OUR-NLC roundness,uniform distribution,particle size and PDI value were 165.15 +/-3.84 nm and 0.227 +/-0.01,Zeta potential of 22.87 +/-0.97 mV.Dynamic dialysis method was used to pH7.4 slightly the PBS for drug release medium,37 ? temperature away from light under constant speed mixing HA-OUR-NLC drug release in vitro experiment.,according to the results of the HA-OUR-NLC conforms to Higuchi equation,in vitro drug release curve fitting coefficient R2 were 0.976,0.985 and 0.988,proved that HA-OUR-NLC has certain sustained-release effect.,through the study of drug release kinetics HA-OUR differences-the release of the three components in NLC,small molecular weight,small volume of more than the molecular weight,larger volume of saponins released quickly,at the same point in time,the accumulation of OA and UA to release a quantity greater than Rg3.4.HA-OUR-NLC in rats in vivo pharmacokinetic studyTo OUR normal saline solution as the control group,the HA-OUR-the pharmacokinetic parameters of NLC,in the design of the timing of the rat eyes take blood,HPLC method for determination of OA and UA and Rg3 blood drug concentration,with DAS2.0 software for data processing,fitting of pharmacokinetic parameters.Results show that the control group with medication in process of nanoparticles group were in line with the second chamber model of dynamic characteristics.Fruit showed that HA-OUR-NLC changed its pharmacokinetic parameters,three components after nanoparticlization to significantly prolong the half-life of drugs in mice and residence time in the body,significantly lower clearance,the AUC value increase,extended the function of the liver,improve the bioavailability of the drugs,has played an effective role.5.HA-OUR-NLC targeted researchIn female BALB/C-nu pure nude mice as experimental animals for targeted research.By caudal vein injection dosage,high performance liquid chromatography(HPLC)method is used to determine the content of drugs in mice,to OUR normal saline solution for reference,study the HA-OUR-NLC in the distribution of a tumor-burdened mice,relative to drugs targeting rate(RTE)and the relative evaluation of drug-loading nanoparticles uptake rate(RUE)distribution in mice after the treatment.Experiments show that OUR-NLC and HA-OUR-NLC group comparison shows that the nanoparticles and target the tumor targeting more obvious after head,and liver and kidney containing the dosage is slightly lower,in other words at the same time reduce the liver and kidney toxicity.Give the HA-after OUR-NLC,NLC systemic rapid enrichment in tumor and liver,and in the blood and other tissues distribution is less,the HA-OUR-NLC group in cancer drug accumulation increased,concluded that HA may combine with the tumor cell surface receptor CD44 specificity,mediated drug transport into the cell,thus increasing the accumulation of the drug in swollen nodule.Mice tail intravenous FITC labeled HA-OUR-NLC solution and FITC solution,using fluorescence endoscopic laser confocal imaging system,in mice in vivo real-time level observation,cell size whether nanoparticles into the tumor cells.The results showed that the group without fluorescence,FITC solution exclude the interference of FITC.HA-FITC signs on OUR-NLC group,with the extension of time,the nanoparticles by cells gradually into the cell,outside the quantity also gradually to increase,has a positive correlation.Therefore,HA--OUR NLC can be targeted liver cells.Mice tail intravenous DiR tag HA-OUR-NLC solution and DiR solution,the small animal in vivo imaging system,on the distribution of the target organs of real-time location tracking drugs and imaging.The results showed that the physiological saline group without fluorescence,DiR-HA-OUR-NLC group fluorescence intensity in six time point of the tumor site were significantly stronger than the DiR-OUR-NLC group.All the DiR packages in the two groups in 3 h in tumor volume,the largest of the DiR-HA-OUR-NLC group from the trend of the volume will decrease after 3 h,12 h after the organization of the fluorescence intensity of the weak.6.HA-OUR-NLC in vivo pharmacodynamics researchTumor-burdened SMMC-7721 mice as model,evaluated the different preparations after delivery,according to the results of the inhibitory rate of tumor growth rapidly in addition to the blank control group,the rest of the group all has inhibitory effect on tumor,inhibiting effect from weak to strong,respectively for OUR solution,OUR-NLC group,positive control group,the HA-OUR-NLC group.Further,using the method of HE staining,the results show that the groups are different degrees of cell shrinkage,which HA-OUR-NLC group are smaller,the nuclei are hyperchromatic,chromatin condensed and presents the typical apoptotic morphological changes.7.HA-OUR-NLC cell toxicity studiesDetermined by MTT method is used to investigate the HA-OUR-NLC on SMMC-7721 cell toxicity.Results show that the free drug group with nanoparticles(OUR-NLC,HA-OUR-NLC)has inhibitory effect on SMMC-7721 cell proliferation,and increases with the increase of drug concentration inhibition rate;The same concentration,the groups of drug effect on SMMC-7721 cell inhibition rate with the extension of time;Free drugs with nanoparticles group compared with inhibition rate fell slightly,but the cells are still killing;Nanoparticles in OUR-NLC group compared with the HA-OUR-NLC group inhibition rate fell slightly,showed that after modified hyaluronic acid nanoparticles to cells inhibition effect is better;24 h inhibition rate nanoparticles group were lower,after 48 h inhibition rate increased to achieve the inhibition of free drug levels,show that nanoparticles group has obviously sustained-release effect.In conclusion,this topic is the preparation of HA-OUR-NLC biodegradable,stability is good.Have active tumor targeting effect,pharmacodynamics in vitro and in vivo pharmacodynamics results show that the HA-OUR-NLC has obvious tumor suppression effect,therefore,this topic of the preparation of HA-NLC has the advantages of high efficiency,low toxicity and tumor active targeting,paper difficult soluble more ingredients of traditional Chinese medicines give medicine research provides methods and techniques of the system.