Preparation Of Tamoxifen-tranilast Compound Liposomes And Anti-breast Cancer Effects

Objective:Tamoxifen（Tam）is a common drug for breast cancer,but long-term use can develop drug resistance and reduce the effectiveness of treatment.Tranilast（Tra）is an analogue of tryptophan metabolites with a variety of antitumor activity,which can produce synergistic effects in combination with classical anticancer drugs.In this study,we intend to construct Tam-Tra co-supported liposomes,and study their preparation process and physicochemical properties.Its anti-tumor effect was evaluated by its proliferation inhibition effect on 4T1,MDA-MB-231 and MCF-7 tumor cells,as well as the tumor inhibition rate of BALB/c tumor-bearing mice charged with 4T1 tumor cells.Methods:（1）Cell proliferation inhibition experiments were used to determine the synergistic effect between Tam and Tra.（2）Establish high performance liquid chromatography to determine the content of Tam and Tra in compound liposomes,and calculate the encapsulation rate of the two drugs.（3）The prescription and process of compound liposomes were optimized by single-factor experiment and response surface effect method,and the surface modification conditions of liposomes were optimized,and freeze-drying conditions were determined.（4）The properties of compound liposomes,such as particle size,zeta potential and stability,were investigated,and their in vitro release was investigated by dynamic membrane dialysis.（5）The in vitro antitumor activity of compound liposomes was evaluated by CCK-8 method,and the antitumor activity of compound liposomes in vivo was evaluated by tumor suppression rate by establishing a tumor-bearing mouse model.Results:（1）The Jin’s formula verified that Tam and Tra had a synergistic effect on the inhibition of breast cancer cells,and when 2μM Tam was combined with 10μM Tra,the synergistic effect was the strongest,and the q values for 48 h of 4T1,MDA-MB-231 and MCF-7 breast cancer cell lines were 1.79,1.26 and 1.37,respectively.（2）The method for determining the content and encapsulation rate of compound liposomes was successfully established,and the linear relationship between the two drugs within 0.2-25.6μg/mL remained good,and the RSD value of the sampling transmittance of the encapsulation rate was less than 3%by membrane method.（3）The particle size of compound liposomes prepared by thin film dispersion method was 200.94 nm±16.03nm,and the encapsulation rate of both drugs was above 90%;The optimal prescription lipid ratio was 1:25,the phospholipid-cholesterol ratio was 9:1,and the phospholipid concentration was 20 mg/mL;And successfully prepared polyethylene glycol modified liposomes（PEG-Lips）and hyaluronic acid modified liposomes（HA-Lips）and hyaluronic acid modified liposomes（HA-Lip）and hyaluronic acid and polyethylene glycol modified liposomes（HA-PEG-Lip）;The liposomal lyophilized powder had good stability,and the encapsulation rate of both drugs remained above 85%within one month.（4）The release of drugs in liposomes conformed to the first-order kinetic process,Tra was released completely within 24 h,and Tam was completely released within 72 h,which had obvious sustained release effect.（5）The IC₅₀ values of liposomes on 4T1,MDA-MB-231 and MCF-7 breast cancer cells for 48 h were 10.75,15.48 and 5.61μg/mL,respectively,which were better than those in the Free Drug（FD）group;The tumor suppression rate of mouse breast cancer model was 37.74%,which was higher than that of the FD group,and the tumor suppression rate was further improved after the surface modification of liposomes.Among them,the tumor suppression rates of PEG-Lip group,HA-Lip group and HA-PEG-Lip group on mouse breast cancer models were 42.38%,49.01%and 59.60%,respectively.Conclusion:In this study,Tam-Tra co-loaded liposomes were successfully prepared,which had good physicochemical properties.It has a significant tumor inhibitory effect on three tumor cell lines of breast cancer 4T1,MDA-MB-231 and MCF-7 and BALB/c tumor-bearing mice with 4T1 breast cancer cells.