Mannose Modification Enhances Tumor Targeting and Immunotherapeutic Effects of Resiquimod Liposomes
LU Yue1, YUAN Feng-jiao2, ZHENG Shu-hui1, SONG Jia-liang1, WANG Fei-fei1, Sun Ruo-han1, LI Jun1, JIA Dian-long1*, LIU Ren-min1*
1. School of Pharmaceutical Sciences, Liaocheng University, Liaocheng 252059, China; 2. Joint Laboratory for Translational Medicine Research, Liaocheng People′s Hospital, Liaocheng 252000, China
Abstract：OBJECTIVE To modify the resiquimod(R848) liposomes with mannose to improve its immunotherapeutic effect on mouse tumors through endowing it with active targeting ability to tumor-associated macrophages(TAMs). METHODS The mannose-modified R848 liposomes(M-LS/R848)were prepared by mixing Mannase-PEG2000-DSPE into lipid and encapsulating R848 with a pH gradient driving drug loading method. The physicochemical properties of M-LS/R848 were characterized by laser particle sizer and transmission electron microscope, and the R848 release was analyzed by a dialysis method. In vitro, the targeting of M-LS/R848 to mouse macrophage RAW 264.7 was analysed. In vivo, the tumor-homing and immunotherapy effect of M-LS/R848 were evaluated on MC38 tumor models, and its therapeutic mechanism was investigated by immunofluorescence staining. RESULTS The M-LS/R848 liposomes prepared in this study were uniform spherical particles, with an average diameter of (123.43±2.43) nm and Zeta potential of(-0.33±0.24) mV. The R848 encapsulation rate of M-LS/R848 is (85.59±0.37)% with a release rate of 46.8% in 48 h. In vitro targeting experiments showed that the uptake rate of M-LS/R848 by RAW264.7 cells was (5.29±0.16)%, which significantly higher than that of LS/R848 . In vivo tumor targeting analysis revealed that the accumulation of mannose-modified liposomes in MC38 tumors was significantly higher than that of non-modified liposomes. The antitumor experiments showed that the average tumor volume and weights of the M-LS/R848-treated group were significantly smaller than that of the LS/R848-treated group, and even two mice were cured and immune to the reinoculation of MC38 tumors. Immunofluorescence analysis revealed that M1-type macrophages and CD8 positive cells in tumor tissues of M-LS/R848-treated group were significantly increased compared with the LS/R848 group. CONCLUSION Modifying R848 liposome with mannose could effectively improve its tumor targeting ability and enhance its immunotherapy effect.
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2022, Vol. 57 
