An In Vivo Study Of PEG_2K-?Fmoc?lys-NLG₉₁₉ Loaded Doxorubicin Micelle Target Delivery For B Cell Lymphoma

Objective: To explore the target delivery of PEG2K-(Fmoc)lys-NLG919 loaded doxorubicin micelle for treatment of B cell lymphoma in vivo.Methods:1.10 4-weeks-old female balb/c mice were randomly divided into 2 group. Group A(n=5) were treated by 5mg/ml free doxorubicin i.v. Injection .Group B (n=5) were treated by PEG2K-(Fmoc)lys-NLG919 loaded doxorubicin micelle in same dox dose of Group A i.v injection.Evaluate the dox concentration of mice serum about these 2 group at the point of 3 min,5 min, 10 min,30 min,1 hour,2 hour,6 hour,8 hour after injection.2.25 4-weeks-old female balb/c mice ,A20 cell line were subcutaneously injected at right flank,when tumor size got to 150-300mm3,randomly divided into 5 group :control group(n=5),free dox group(n=5),doxil group(n=5),dox loaded micelle group(n=5),carrier alone group(n=5),Mice in five groups are treated by 3 times injection of Saline,free doxorubicin,doxorubicin lipsome,doxorubicin loaded micelle and free carrier every 3 days,and measure the tumor size and body weight every 3 days.Respectively,at the 21st day all mice were all sacrificed,the tumor weight was measured. The heart were examined by H&E staining to observe the changing of mycardial cells by microscope.3.15 4-weeks-old female balb/c mice,A20 cell line were subcutaneously injected at right flank,when tumor size got to about 500mm3,randomly divided into 5 group :control group(n=3),free dox group(n=3),doxil group(n=3),dox loaded micelle group(n=3), carrier alone group(n=3),Mice in five groups are treated by 3 times injection of Saline,free doxorubicin,doxorubicin lipsome,doxorubicin loaded micelle and free carrier every 3 days,after last injection, all mice were sacrificed and CD4+ T cell,CD8+ T cell, Treg cell proportion were examined by flow cytometric analysis.Results:1 .The every point serum dox concentration of PEG2k-(Fmoc)lys-NLG919 loaded doxorubicin micelle are higher than free dox group.(P <0.05)2.The relative tumor size and tumor weight of doxorubicin loaded micelle group were significantly smaller than other four group.(P <0.05).The pathological changes of HE staining showed the DOX-induced myocardial damage serious, but no obvious myocardial damage was observed in the dox loaded micelle group and doxil group.And carrier alone group shows no myocardial damage.3.Flow cytometric analysis shows that PEG2K-(Fmoc)lys-NLG919loaded doxorubicin micelle upgrade the proportion of CD4+ T cell and CD8+ T cell and obviously reduce the proportion of Treg cell inside the tumor.(P <0.05)Conclusion:1.PEG2K-(Fmoc)lys-NLG919 can keep doxorubicin for higher concentration and longer time in serum.2.PEG2K-(Fmoc)lys-NLG919 loaded doxorubicin can largely restrain the growth of lymphoma.And PEG2k-(Fmoc)lys-NLG919 micelle can obviously reduce the cardiactoxicity from doxorubicin.3.PEG2K-(Fmoc)lys-NLG919 loaded doxorubicin micelle can upgrade the positive T cell and reduce the Treg cell, make the immune response more sensitively to tumor cell.