^(64)Cu-NOTA-Herceptin的设计、活性测定及肿瘤靶向分子显像研究  

Design and Bio-evaluation of ^(64)Cu-NOTA-Herceptin for Tumor Targeted Micro-PET Imaging

作　　者：朱华[1] 李一林[2] 赵传科[3] 解清华 刘菲[1] 韩雪迪 高静[1] 夏传琴[4] 沈琳[2] 杨志[1] 

机构地区：[1]北京大学肿瘤医院暨北京市肿瘤防治研究所核医学科,北京100142 [2]北京大学肿瘤医院暨北京市肿瘤防治研究所消化肿瘤内科,北京100142 [3]北京大学肿瘤医院暨北京市肿瘤防治研究所生化与分子生物学研究室恶性肿瘤发病机制及转化研究教育部重点实验室,北京100142 [4]四川大学化学学院,成都610041

出　　处：《高等学校化学学报》2016年第12期2132-2137,共6页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金(批准号:81371592;81401467;81501519;81301323;81571705);北京市自然科学基金(批准号:7154188;7162041)资助~~

摘　　要：利用双功能螯合剂2-[(4-异硫氰基苯基)甲基]-1,4,7-三氮杂环九烷-1,4,7-三乙酸(NCS-Bz-NOTA)对Herceptin单抗表面的氨基进行修饰获得了NOTA-Herceptin,通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF)对该偶联物进行了表征.利用酶联免疫吸附测定了偶联前后Herceptin抗体效价的改变.利用新型正电子核素^(64)Cu标记,获得可用于肿瘤放射靶向精准诊疗的^(64)Cu-NOTA-Herceptin探针,其标记率为90%,放化纯度>98%,比活度185 MBq/nmol.分别进行了该探针在HER2过表达胃癌细胞NCI-N87及HER2低表达胃癌细胞BGC823等肿瘤细胞中的摄取实验,测定了该探针的肿瘤特异性.建立了荷人胃癌BGC823裸鼠模型,通过微型正电子断层显像(Micro-PET)设备观察了探针在模型动物体内的代谢情况:在静脉注射7.4 MBq^(64)Cu-NOTA-Herceptin探针后,分别于4和60 h进行正电子断层显像(PET)的显像,观察到其在肿瘤部位的摄取有所富集,且随着代谢时间的延长,肝脏部位摄取得到明显降低.研究结果表明,^(64)Cu-NOTAHerceptin探针有望应用于肿瘤放射性靶向诊疗.The precursor compound 2-（p-thiocyanatobenzyl）-1,4,7-triazacyclononane-1,4,7-triacetic acidHerceptin（NOTA-Herceptin） was synthesized by the nucleophilic addition reaction of Herceptin and bi-functional chelator NCS-Bz-NOTA. The original Herceptin and NOTA- Herceptin conjugate were investigated by UV-Vis and MALDI-TOF mass spectra. The average number of chelators per Herceptin for the conjugated used in this study was 5. 4. The enzyme-linked immunosorbent assay（ELISA） was conducted to compare the biological activity of original Herceptin and NOTA-Herceptin. NOTA- Herceptin kept high immunoreactivity towards HER2 antigen. Then,the PET radio-nuclide64Cu（T1/2= 12. 7 h） was labeled to got the novel tumor Immuno-Theranostics probe64Cu-NOTA-Herceptin. The labeling efficiency of64Cu-NOTA-Herceptin was tested by Radio-TLC/HPLC. The radiolabeling yield was over 90%,the radio chemical purity was over 98% after PD-10 column purification and the specific activity was 185 MBq/nmol. The immune reactivity and specific activity of radiolabeled Herceptin with HER2 antigen were performed by HER2 positive NCI-N87 cell line and HER2 negative BGC823 cell line. Micro- PET imaging of BGC823 tumor-bearing nude mice revealed that tumor uptake of64Cu-NOTA-Herceptin got a gradual accumulation from 4 h to 60 h after intravenous injection of 7. 4 MBq radiolabeled Herceptin. Uptake in the tumor was clearly visualized by emission computed tomography.64Cu-NOTA-Herceptin owns a great potential for molecular imaging of PET for diagnosis and follow-up of HER2 expression.

关 键 词：曲妥珠单抗 64Cu 免疫活性 分子显像 肿瘤靶向诊疗 

分 类 号：O614[理学—无机化学] O628[理学—化学]