[1]刘鹏,董文涛,苏新辉,等.靶向TSPO显像剂99Tcm-DTPA-CB86的制备及其对关节炎症的SPECT/CT显像研究[J].国际放射医学核医学杂志,2018,(1):47-52.[doi:10.3760/cma.j.issn.1673-4114.2018.01.009]
　Liu Peng,Dong Wentao,Su Xinhui,et al.Preparation and imaging of arthritis of 99Tcm-DTPA-CB86 for TSPO targeted imaging[J].International Journal of Radiation Medicine and Nuclear Medicine,2018,(1):47-52.[doi:10.3760/cma.j.issn.1673-4114.2018.01.009]
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靶向TSPO显像剂99Tcm-DTPA-CB86的制备及其对关节炎症的SPECT/CT显像研究(/HTML)
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《国际放射医学核医学杂志》[ISSN:1673-4114/CN:12-1381/R]

卷:
期数:
2018年第1期
页码:
47-52
栏目:
基础研究
出版日期:
2018-03-20

文章信息/Info

Title:
Preparation and imaging of arthritis of 99Tcm-DTPA-CB86 for TSPO targeted imaging
作者:
刘鹏1;  董文涛1;  苏新辉1; 4;  黄剑全2;  王亮亮1;  蒋怡臻1;  郭志德3;  马超1;  苏福1
1. 361004, 厦门大学附属中山医院核医学科;
2. 366000, 福建省三明市第二医院核医学科;
3. 361102, 厦门大学分子影像暨转化医学研究中心;
4. 350001 福州, 福建医科大学附属协和临床医学院
Author(s):
Liu Peng1;  Dong Wentao1;  Su Xinhui1; 4;  Huang Jianquan2;  Wang Liangliang1;  Jiang Yizhen1;  Guo Zhide3;  Ma Chao1;  Su Fu1
1. Department of Nuclear Medicine, Zhongshan Hospital Xiamen University, Xiamen 361004, China;
2. Department of Nuclear Medicine, the Second Hospital of Sanming in Fujian Province, Sanming 366000, China;
3. Center for Molecular Imaging and Translational Medicine, Xiamen University, Xiamen 361102, China;
4. Union Clinical Medical College of Fujian Medical University, Fuzhou 350001, China
关键词:
转运蛋白; 关节炎; 同位素标记; 锝放射性同位素; 体层摄影术; 发射型计算机; 单光子; 体层摄影术; X线计算机
Keywords:
Translocator protein; Arthritis; Isotope labeling; Technetium radioisotope; Tomography; emission-computed; single-photon; Tomography; X-ray computed
DOI:
10.3760/cma.j.issn.1673-4114.2018.01.009
摘要:
目的 制备99Tcm标记的转运蛋白（TSPO）配体CB86[99Tcm-DTPA（二亚乙基三胺五乙酸）-CB86]，探讨其作为TSPO靶向关节炎症显像新型分子探针的可行性。方法 通过偶联双功能螯合剂制备DTPA-CB86，进行99Tcm标记，经高效液相色谱纯化，测定其放化纯度和体外稳定性。选用巨噬细胞RAW264.7进行体外细胞结合实验，测定99Tcm-DTPA-CB86的结合率和外排率。采用弗氏佐剂建立左踝关节炎症小鼠模型，对其行99Tcm-DTPA-CB86 Micro SPECT/CT显像，并观察探针的体内分布情况。采用SPSS 18.0统计软件对符合正态分布及方差齐性的数据进行t检验。结果 99Tcm-DTPA-CB86的标记率为（95.86 ±2.45）%，放化纯度为（97.45 ±0.69）%，其在室温下的磷酸盐缓冲液中的稳定性良好，放置4 h后，其标记率仍>90%。99Tcm-DTPA-CB86能与巨噬细胞RAW264.7特异性结合，3 h的摄取率达到最高峰[（36.45 ±2.18）%]，在加入过量未标记的DTPA-CB86后，RAW264.7细胞对99Tcm-DTPA-CB86的摄取明显受到抑制[（10.43 ±2.01）%]，与未阻断时相比差异具有统计学意义（t=6.217，P < 0.05）；RAW264.7细胞对99Tcm-DTPA-CB86外排较少，摄取率从4.5 h时的（33.31 ±2.34）%到8 h时的（19.32 ±2.01）%，减少了13.99%。Micro SPECT/CT显像结果显示，99Tcm-DTPA-CB86在小鼠左踝关节炎症部位清晰可见，且能被过量的DTPA-CB86明显抑制；体内生物学分布结果表明，其具有较好的炎症靶向性；注射后3 h踝关节炎症部位的摄取仍可达（2.35 ±0.10）% ID/g。结论 99Tcm标记CB86易于制备，具有较好的理化性质及体内代谢学性质，同时具有较好的关节炎症摄取，有望发展成新的TSPO靶向SPECT关节炎症显像分子探针。
Abstract:
Objective To develope a novel radiolabeled translocator protein(TSPO) ligand CB86 targeting agent 99Tcm-diethylene-triaminepentaacetic acid (DTPA)-CB86 and evaluate its biological properties. Methods DPTA-CB86 was prepared by coupling with a bifunctional chelating agent, and then labeled with 99Tcm to obtain 99Tcm-DTPA-CB86. The labeling efficiency, radiochemical purity, and stability were determined in vitro. In vitro cellular uptake and efflux were performed using RAW264.7 macrophage cells. Biodistribution and micro-SPECT/CT images were investigated on Freund’s adjuvant-induced left arthritis in mice. SPSS 18.0 analysis software(t-test) was used to fit the normal distribution and homogeneity of variance. Results The labeling yields and radiochemical purity of 99Tcm-DTPA-CB86 were (95.86±2.45)% and (97.45±0.69)%, respectively. 99Tcm-DTPA-CB86 displayed good stability, with a radiochemical purity of more than 90%, in phosphate-buffered solution(PBS) at 4 h. It also exhibited high specific TSPO binging in RAW264.7 macrophage cells in vitro. The highest uptake ratio was (36.45±2.18)% at 3 h after incubation, which then decreased significantly[(10.43 ±2.01)%; t=6.217, P < 0.05)] after adding excessive unlabeled DTPA-CB86. The difference was significant. Cell efflux analysis showed that 99Tcm-DTPA-CB86 had good cell retention by RAW264.7 cells, with only about 13.99%[decreased from (33.31±2.34)% to (19.32±2.01)% of total input radioactivity] of 99Tcm-DTPA-CB86 efflux observed during 4.5 h to 8 h of incubation. Biodistribution and SPECT/CT imaging demonstrated that the uptake of 99Tcm-DTPA-CB86 in the left arthritic ankles was significantly increased compared with that in contralateral normal ankles. Uptake in the arthritic ankles could be largely blocked by an excess of DTPA-CB86. Conclusion 99Tcm-DTPA-CB86 can be readily synthesized and clearly visualize arthritis with low background, thus demonstrating its potential as a promising molecular probe targeting TSPO for arthritic SPECT imaging.

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备注/Memo

备注/Memo:
收稿日期:2017-11-19。
基金项目:国家自然科学基金面上项目（81571707）；福建省自然科学基金（2015J01519）；福建省卫生系统中青年骨干人才培养项目（2014-ZQN-ZD-35）；人社部留学人员科技活动项目择优资助（2014-240）；教育部留学回国人员科研启动基金（2014-1685）；福建省卫计委青年科研课题（2015-2-46）；厦门市科技项目（2015S0288）
通讯作者:苏新辉,Email:suxinhuii@163.com
更新日期/Last Update: 2018-03-20