甘露糖修饰提高瑞喹莫德脂质体对肿瘤的靶向和免疫治疗作用

路岳, 袁风娇, 郑书慧, 宋佳亮, 王菲菲, 孙若涵, 李军, 贾殿隆, 柳仁民

中国药学杂志 ›› 2022, Vol. 57 ›› Issue (22) : 1917-1925.

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中国药学杂志 ›› 2022, Vol. 57 ›› Issue (22) : 1917-1925. DOI: 10.11669/cpj.2022.22.008
论著

甘露糖修饰提高瑞喹莫德脂质体对肿瘤的靶向和免疫治疗作用

  • 路岳1, 袁风娇2, 郑书慧1, 宋佳亮1, 王菲菲1, 孙若涵1, 李军1, 贾殿隆1*, 柳仁民1*
作者信息 +

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*
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文章历史 +

摘要

目的 利用甘露糖(Mannase)修饰瑞喹莫德(R848)脂质体,赋予其对肿瘤相关巨噬细胞(tumor-associated macrophages,TAMs)的主动靶向能力,以提高其对小鼠移植瘤的免疫治疗作用。方法 用二硬脂酰磷脂酰乙醇胺-聚乙二醇-甘露糖(Mannase-PEG2000-DSPE)脂材掺入脂质体,并采用pH梯度主动载药法包封R848,制备获得甘露糖修饰的R848脂质体(M-LS/R848)。利用粒度仪、透射电镜等表征其理化性质,透析法分析其释药稳定性。体外考察其对小鼠巨噬细胞RAW264.7的靶向性,体内评估其对小鼠结肠癌细胞MC38移植瘤的靶向性和免疫治疗效果,并利用免疫荧光染色探讨其免疫治疗机制。结果 本研究制备的M-LS/R848大小均一,平均粒径为(123.43±2.43)nm,Zeta电位为(-0.33±0.24)mV,为球形或类球形粒子,R848包封率达(85.59±0.37)%,48 h R848释放率为(46.73±0.54)%,稳定性较高。体外靶向实验表明RAW264.7细胞对M-LS/R848的摄取率为(5.29±0.16)%,显著高于LS/R848的摄取率。体内靶向实验表明甘露糖修饰脂质体在MC38肿瘤中的累积显著高于非修饰脂质体。体内抑瘤实验表明M-LS/R848治疗组肿瘤平均体积和重量显著小于LS/R848组,甚至有两只小鼠肿瘤治愈并产生对MC38肿瘤的免疫记忆效应。免疫荧光分析显示M-LS/R848给药组肿瘤组织中M1型巨噬细胞和CD8阳性细胞较LS/R848组明显增多。结论 利用甘露糖修饰R848脂质体可有效提高其对肿瘤的靶向性并增强免疫治疗效果。

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.

关键词

肿瘤免疫治疗 / 甘露糖修饰 / 瑞喹莫德脂质体 / 肿瘤相关巨噬细胞

Key words

tumor immunotherapy / mannose modification / R848 liposome / tumor associated macrophage

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导出引用
路岳, 袁风娇, 郑书慧, 宋佳亮, 王菲菲, 孙若涵, 李军, 贾殿隆, 柳仁民. 甘露糖修饰提高瑞喹莫德脂质体对肿瘤的靶向和免疫治疗作用[J]. 中国药学杂志, 2022, 57(22): 1917-1925 https://doi.org/10.11669/cpj.2022.22.008
LU Yue, YUAN Feng-jiao, ZHENG Shu-hui, SONG Jia-liang, WANG Fei-fei, Sun Ruo-han, LI Jun, JIA Dian-long, LIU Ren-min. Mannose Modification Enhances Tumor Targeting and Immunotherapeutic Effects of Resiquimod Liposomes[J]. Chinese Pharmaceutical Journal, 2022, 57(22): 1917-1925 https://doi.org/10.11669/cpj.2022.22.008
中图分类号: R944   

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基金

国家自然科学基金项目资助(82000066);山东省自然科学基金项目资助(ZR2019PH006);山东省自然科学基金重点项目资助(ZR2020KH019);山东省自然科学基金创新发展联合基金项目资助(ZR2021LSW001)
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