Studies On The Tumor-targeted Hyaluronic Acid-mPEG Modified Cantharidin Loaded Nanostructuted Lipid Carriers Nanoparticles

[Objective]Cancer is a major disease that endangers human health.In particular,the mortality rate of liver cancer in China is extremely high,and serious threats to people’s health.Cantharidin is the main component of Mylabris anticancer.It has good inhibitory effects on many cancers such as liver cancer,colon cancer and breast cancer.Because of its toxicity,oral administration can cause various toxic and side effects,leading to its limited by clinical application.Therefore,the development of a novel nano-drug delivery system targeting cantharidin should be a research hotspot.This article selected cantharidin as a model drug,and prepared a biodegradable nano-lipid carrier harboring cantharidin,and modified with hyaluronic acid(HA)to enhance its tumor targeting.[Methods and Results](1)Synthesis of HA-mPEG by amidation,and fourier-infrared and nuclear magnetic profiling were performed and the results showed that the target has been successfully synthesized.The HA-mPEG-CTD-NLC was prepared by emulsion-ultrasonic dispersion method.Based on the single factor test,the optimal prescription was determined by orthogonal test(Central Composite Design-Response Surface Methodology).The HA-mPEG-CTD-NLC prepared by the optimized method has a light blue opalescence,a particle size was 119.3 nm,potential was 37.3 mV,PDI was 0.157,entrapment efficiency was 56.27%,and drug loading was 1.58%.The lyophilization agent of nanoparticles was examined by comparative test method.The surface of the lyophilized powder was flat,did not collapse,and the entire block could be detached without breaking up as the indicator.5%mannitol was selected as a lyoprotectant for nanoparticles,and the reconstitution of the sample after lyophilization was good,the particle size is 139.3nm,PDI and potential values were almost constant before.Dynamic dialysis technique was used to investigate the in vitro release of HA-mPEG-CTD-NLC.Mathematical parameters were fitted to the results.The results showed that the drug release was in accordance with the Weibull equation and had a good sustained release effect.(2)The CTD solution was used as the control group to investigate the pharmacokinetic parameters of CTD-NLC and HA-mPEG-CTD-NLC in rats.At a specific time point,the rats were subjected to eyelid sampling.The content of CTD was measured by HPLC and DAS 2.0 was used to software processing data.The results showed that compared with the CTD solution group,HA-mPEG-CTD-NLC and CTD-NLC changed their pharmacokinetic parameters,and all met the two-compartment model.The drug half-life of HA-mPEG-CTD-NLC was 12.56 and 2.20 times that of the CTD solution group,the AUC value was 2.28 times,and the plasma clearance rate was only 0.41 of the CTD solution,which significantly prolonged the residence time of the drug in vivo.(3)Using nude mice as the research object,Optical In Vivo Imaging technology was used to evaluate the tissue distribution and targeting of CTD-NLC and HA-mPEG-CTD-NLC in vivo,and its pharmacodynamics was verified by the size of tumors.The results showed that the fluorescence of tumor cells in HA-mPEG-CTD-NLC group was significantly stronger than that of CTD-NLC group.Through the determination of the drug content in the tissue,we can see that the modified nanoparticles can significantly increase the RUE value of the drug in the tumor,reaching 4.75,also reduced the RUE value in the heart and spleen,the value can be as low as 0.93 and 1.07,both proved that have a good HA-mPEG-CTD-NLC targeting.(4)The tumor-bearing mice were administered for a prolonged period of time drug,and the inhibitory effects of the normal saline group,the CTD solution group,the CTD-NLC group,the HA-mPEG-CTD-NLC group,and the positive control group(5FU)on the tumor cells were compared.According to the size of the tumor,HA-mPEG-CTD-NLC significantly inhibited the growth of tumor-bearing mice.The tumor inhibition rate was 65.96%.In vitro tumor inhibition experiments were performed by MTT cytotoxicity assays.The results showed that CTD-NLC and HA-mPEG-CTD-NLC inhibited the proliferation of SMMC-7721 cells,and the inhibitory rate increased with the increase of drug concentration.The IC50 value of HA-mPEG-CTD-NLC for drugs was lower than that of CTD-NLC group,indicating that HA-mPEG-CTD-NLC had a certain sustained release effect.[Conclusion]This article successfully prepared HA-mPEG-CTD-NLC,a nano-administration system that actively targets tumors.It not only has the effect of sustained release,but also overcomes the disadvantages of poor carrier targeting,that reduces the toxicity of drugs,improves the efficacy and anticancer index of cantharidin drugs,and improves their bioavailability.It provides scientific data for the study of cantharidin active targeting nano-systems.