The Preparation And Vitro Activity Evaluation Of Liver-targeted Oxaliplatin And Sorafenib Co-loaded Liposomes

Sorafenib(SOR),a multi-kinase inhibitor,is the first oral multikinase inhibitor approved by FDA for advanced HCC therapy.In clinics,the combination use of sorafenib with other cytotoxic drugs is a promising strategy to enhance the anticancer effect.However,these two drugs with completely different pharmacokinetic profiles and tissue distribution have limited therapeutic success.In this study,oxaliplatin(OXA)and sorafenib were co-loaded into the glycyrrhetinic acid(GA)mediated liver targeted liposomes to improve the therapy effects of HCC.The new liver targeted molecule GA-DSPE was designed and synthesised.The synergistic effect of OXA and SOR was verified by CI index methods.The GA-OS-Lip was prepared by the thin film hydration method.And methods orthogonal experiment was carried out to optimize the formulation.Laser diffraction-based particle size analyzer was used to determine the mean diameter,polydispersity index and Zeta potential of the liposomes.The morphology of the liposomes was characterized by transmission electronic microscopy.The encapsulation efficiency of OXA and SOR were established by HPLC and UV-Vis,respectively.The differential scanning calorimeter(DSC)was employed to study the thermodynamic characteristics of the liposomes.The in vitro release properties were accessed using dialysis bag with diffusion method.The stability of GA-OS-Lip in different media was evaluated.In vitro cytotoxicity study and apoptosis study were tested in HepG2 cells using CCK-8 assay and Annexin V/PI apoptosis assay,respectively.The cellular internalization of the liposomes in HepG2 was monitored using CLSM and quantified directly by flow cytomete.Real-time near infrared fluorophore imaging was applied to monitor the tissue distribution of the liposomes in vivo.The maximum synergistic effect of SOR/SOR was obtained at 1/1 molar ratio.The morphology of GA-OS-Lip was spherical and the mean particle size,PDI and Zeta potential were(119.0±2.15)nm,(0.196±0.021)and(-34.82±2.84)mV,respectively.The encapsulation efficiency and drug loading of OXA and SOR were(33.8±3.72)% and(70.80±6.80)%,(1.45±0.14)% and(1.61±0.22)%,respectively.Good stability of GA-OS-Lip was displayed in the tested media including pH 7.4 PBS,cell culture media and 10% plasma at 37?.The liposomes showed controlled-release of both drugs.90% of the OXA and 70% of the SOR were released in 48 h.In vitro cytotoxicity study and apoptosis study,GA-OS-Lip(IC50=3.242/3.242)showed the highest cell cytotoxicity in comparison with the free drugs(IC50=5.057/5.057)and OS-Lip(IC50=5.079/5.079).The surface modification of GA on liposomes effectively altered the uptake of HepG2 in vitro and the body distribution in vivo.In conclusion,GA-OS-Lip could achieve better synergistic effect by co-delivery of two drugs.And GA modified liposomes showed more uptaking by cells and more accumulation in the liver.The targeted co-loaded liposomes of OXA and SOR showed great potential for HCC treatment.