Preparation And Anti-hepatocellular Carcinoma Activity Of Double-loaded Liver Targeting Liposomes Comodified With SP94 And TAT

Hepatoma,as one of the common malignant tumors,is preferably treated with chemotherapy.In the chemotherapy of liver cancer,there are problems such as large toxic and side effects caused by non-targeted drugs,and easy drug resistance due to a single drug,which seriously affects the therapeutic efficiency of chemotherapy.Objective:Liposomes are ideal drug carriers,whose hydrophilic and lipophilic properties can be leveraged to co-encapsulate doxorubicin(DOX)and docetaxel(DTX)with favourable synergistic ability in liposomes to prepare DTX/DOX-LP,so as to realize simultaneous drug administration and play the synergistic effect of DOX and DTX.Then,SP94 and TAT are modified together on DTX/DOX-LP by utilizing the characteristics of SP94 peptide that can specifically bind to liver cancer cells and the good ability of TAT peptide in mediating carrier transmembrane transport to prepare SP94/TAT-DTX/DOX-LP.In this way,the ability of the drug to target liver cancer cells and penetrate liver cancer cells can be achieved.Finally,the optimum preparation path of SP94/TTA-DTX/DOX-LP is screened by process optimization,and the physical and chemical properties and anti-liver cancer activity of various liposomes in vitro are investigated.Methods:(1)DTX in vitro assay was established by high performance liquid chromatography(HPLC),and DOX in vitro assay was established by fluorescence spectrophotometry.(2)The encapsulation rates of DTX and DOX were determined by dialysis method,and DTX and DOX were encapsulated in liposomes by film dispersion method and ammonium sulfate gradient method,respectively.TAT and SP94 were modified on liposomes by organic reaction method and insertion method,respectively,and the optimal dosage and preparation route of SP94/TTA-DTX/DOX-LP were screened out by single factor investigation.(3)Liposomes prepared according to the optimal route and prescription dosage were characterized,including morphology,particle size,PDI,Zeta potential,and the serum stability,in vitro drug release ability and storage stability of different liposomes at different temperatures were investigated.(4)MTT assay was employed to investigate the cytotoxicity of different drug-loaded liposomes on Hep G2 cells.Results:(1)Satisfactory DTX and DOX in vitro assays were established.The R~2 of the standard curve was greater than 0.9996,and the intra-day precision,inter-day precision and recovery rate all met the analytical requirements.(2)The formulation dosage and preparation route of SP94/TAT-DTX/DOX-LP were determined.The DTX encapsulation rate of the liposomes prepared by the optimal process was 80.33±0.96%,the DOX encapsulation rate was 92.97±1.73%.(3)The prepared SP94/TAT-DTX/DOX-LP was spherical and spheroid in structure,with a particle size of about 120 nm,a PDI of about 0.15,slightly negative,and a Zeta potential of about-10 m V.Various liposomes showed good stability at 4℃,but poor stability at 37℃.The serum stability test showed that good serum stability was maintained within 24 h.The in vitro release experiments showed that all liposomes have obvious sustained release ability.(4)The cytotoxicity of SP94/TAT-DTX/DOX-LP on Hep G2 cells were significantly stronger than that of DTX/DOX-LP,SP94-DTX/DOX-LP,TAT-DTX/DOX-LP,and other liposomes,and the single-ligand modification was stronger than that of unmodified liposomes.The dual-drug strategy is superior to the single-drug strategy.Conclusions:In this paper,a drug delivery system for liver cancer is constructed.SP94 and TAT have good synergy,The prepared SP94/TAT-DTX/DOX-LP boasts satisfactory physical and chemical properties as well as excellent anti-liver cancer activity in vitro.