^(68)Ga标记靶向Nectin-4双环肽的制备及乳腺癌显像研究  

Synthesis of a ^(68)Ga-labeled bicyclic peptide targeting Nectin-4 and its application research in breast cancer imaging

作　　者：李励琦[1] 徐悦 潘栋辉[2] 严骏杰 王辛宇 陈重阳 王立振[2] 杨敏[2] 徐宇平[2] Li Liqi;Xu Yue;Pan Donghui;Yan Junjie;Wang Xinyu;Chen Chongyang;Wang Lizhen;Yang Min;Xu Yuping(Department of Thyroid and Breast Surgery,Affiliated Hospital of Jiangnan University,Wuxi 214122,China;NHC Key Laboratory of Nuclear Medicine,Jiangsu Key Laboratory of Molecular Nuclear Medicine,Jiangsu Institute of Nuclear Medicine,Wuxi 214063,China)

机构地区：[1]江南大学附属医院甲状腺乳腺外科,无锡214122 [2]国家卫生健康委员会核医学重点实验室、江苏省分子核医学重点实验室、江苏省原子医学研究所,无锡214063

出　　处：《中华核医学与分子影像杂志》2024年第12期741-747,共7页Chinese Journal of Nuclear Medicine and Molecular Imaging

基　　金：江苏省卫生健康委重点项目(ZD2021005);江苏省自然基金(BK20231141);无锡市"太湖之光"科技攻关(医疗卫生技术攻关)项目(Y20212050)。

摘　　要：目的制备一种新型^(68)Ga标记靶向脊髓灰质炎病毒受体相关蛋白4(PVRL4,又称Nectin-4)双环肽,并对其用于乳腺癌显像的可行性进行评价。方法合成生物素(Biotin)修饰的靶向Nectin-4双环肽(简写为BMIC)Biotin-BMIC,通过体外细胞染色实验对其靶向性进行初步评价。双环肽BMIC经1,4,7-三氮杂环壬烷-1,4-二乙酸(NODA)修饰制得标记前体NODA-BMIC,经一步法标记制备靶向Nectin-4探针^(68)Ga-NODA-BMIC。应用荷乳腺癌小鼠活体microPET显像结合体外实验对该探针显像性能进行考察。采用两独立样本t检验、重复测量方差分析处理数据。结果细胞荧光染色初步表明,荧光标记的双环肽Biotin-BMIC在Nectin-4阳性BT474乳腺癌细胞上较Nectin-4阴性MDA-MB-231乳腺癌细胞高度聚集。^(68)Ga-NODA-BMIC未校正产率可达(71.5±2.2)%,放化纯>95%。比活度>3 GBq/μmol。温育10、30、60和120 min后,BT474乳腺癌细胞较MDA-MB-231乳腺癌细胞均有高放射性摄取(F=1302.00,P<0.001)。荷瘤鼠^(68)Ga-NODA-BMIC microPET显像表明,BT474移植瘤较MDA-MB-231移植瘤显影清晰,且对比度良好。注射探针后10、30、60和120 min,BT474与MDA-MB-231移植瘤摄取差异有统计学意义(F=1826.00,P<0.001),其中注射后60 min BT474移植瘤的摄取值为(5.03±0.14)每克组织百分注射剂量率(%ID/g),显著高于MDA-MB-231移植瘤对应值[(0.19±0.04)%ID/g;t=79.40,P<0.001]。BT474肿瘤/肌肉(T/M)比值高于MDA-MB-231(F=222.00,P<0.001),其中注射后60 min前者T/M比值为24.75±3.10,显著优于后者对应值(1.30±0.15;t=14.31,P=0.002)。体内显像结果与离体免疫组织化学分析一致。结论新型靶向Nectin-4双环肽PET探针^(68)Ga-NODA-BMIC,合成简便,标记产率和放化纯满意。探针体内显像性能佳,靶组织显影清晰,可能在乳腺癌诊疗中发挥独特作用。ObjectiveTo prepare a novel ^(68)Ga-labeled bicyclic peptide targeting poliovirus receptor related protein 4(PVRL4,Nectin-4),and evaluate its feasibility for breast cancer imaging via in vitro and in vivo experiments.MethodsA Biotin-modified bicyclic peptide targeting Nectin-4,Biotin-BMIC,was synthesized,and its targeting properties were preliminarily evaluated by in vitro cell staining experiments.BMIC was modified by 1,4,7-triazonane-1,4-diacetic acid(NODA)and the labeling precursor NODA-BMIC was prepared.A potential PET probe targeting Nectin-4,^(68)Ga-NODA-BMIC was prepared by one-step labeling strategy.The imaging properties of the probe were investigated by in vivo microPET imaging and in vitro experiments in mice bearing breast tumors.Data were analyzed by independent-sample t test and repeated measures analysis of variance.ResultsFluorescence staining of the cells showed that the fluorescently labeled bicyclic peptide,Biotin-BMIC,was highly aggregated in Nectin-4 positive BT474 breast cancer cells compared to those in Nectin-4 negative MDA-MB-231 cells.The uncorrected yield of ^(68)Ga-NODA-BMIC was(71.5±2.2)%and the radiochemical purity was greater than 95%.The specific activity was greater than 3 GBq/μmol.After incubation 10,30,60 and 120 min,higher radioactivity uptakes were found in BT474 breast cancer cells compared to those in MDA-MB-231 breast cancer cells respectively(F=1302.00,P<0.001).MicroPET imaging showed that the BT474 xenograft tumors were clearly visible with favorable contrast.A significant statistical difference in uptakes between BT474 and MDA-MB-231 xenograft tumor uptake at 10,30,60,and 120 min after probe injection respectively was existed(F=1826.00,P<0.001).At 60 min postinjection,the uptake value of BT474 tumors was(5.03±0.14)percentage activity of injection dose per gram of tissue(%ID/g),which was significantly higher than that of MDA-MB-231 tumors((0.19±0.04)%ID/g;t=79.40,P<0.001).Meanwhile,the tumor-to-muscle ratios in the former were also greater than those in the latter(F=2

关 键 词：乳腺肿瘤 肽类 环 同位素标记 镓放射性同位素 肿瘤细胞 培养的 小鼠 

分 类 号：R737.9[医药卫生—肿瘤] R730.44[医药卫生—临床医学]