^(64)Cu标记单链DNA及其体内分布的初步研究  

Preliminary study on ^(64)Cu labeled single-stranded DNA and its distribution in vivo

作　　者：段晓燕[1] 都义日[1] 翁兆平 王相成[1] 王春梅[1] 王涛[1] 李剑波[1] DUAN Xiaoyan;DU Yiri W;ENG Zhaoping;WANG Xiangcheng;WANG Chunmei;WANG Tao;LI Jianbo(Inner Mongolia Medical University Affiliated Hospital,Hohhot 010050,China)

机构地区：[1]内蒙古医科大学附属医院,呼和浩特010050

出　　处：《核技术》2021年第2期30-34,共5页Nuclear Techniques

基　　金：国家自然科学基金(No.81360227);内蒙古自然科学基金(No.2018LH08044);内蒙古自治区科技计划项目(No.2019GG106)资助。

摘　　要：本文探索^(64)Cu标记单链脱氧核糖核酸(Deoxyribo Nucleic Acid,DNA)方法及小鼠体内分布情况。将单链DNA(ssDNA)与螯合剂p-SCN-Bn-NOTA偶联,得到NOTA-ssDNA,加入64Cu孵育一定时间,利用薄层色谱监测反应,NAP-5脱盐柱纯化,得到64Cu-ssDNA。静脉注射进入昆明鼠体内后,研究不同时间点感兴趣器官或脏器的放射性摄取量。NOTA-ssDNA与64Cu反应1 h后,放化标记率为(66.4±2.85)%;纯化后,放化纯度大于95%。体外稳定实验显示,^(64)Cu-ssDNA在PBS(Phosphate Buffer Saline)或小鼠血清中至少能稳定存在6 h。^(64)Cu-ssDNA通过静脉注射到小鼠体内后,在胃肠道有一定的摄取,而肾脏和膀胱出现高放射性摄取量。注射180 min后,膀胱中的放射性摄取量达到了(63.84±5.86)%ID∙g^(−1)。结果表明:64Cu-ssDNA主要通过泌尿系统代谢排出体外。^(64)Cu-ssDNA的成功制备,为基于DNA多面体的PET分子探针的制备解决了关键一步;其体内分布实验为研究^(64)Cu标记DNA多面体的体内生物分布提供了实验对照信息。[Background]DNA(DeoxyriboNucleic Acid)polyhedral structure has great potential in molecular imaging probe with good properties in model animals such as low toxicity,low immunogenicity and excellent in vivo stability.[Purpose]This article aims to use positron emission computed tomography(PET)radionuclide ^(64)Cu to label DNA single strands instead of SPECT radionuclide ^(99)mTc,and to study the distribution and metabolism of ^(64)Cu labeled DNA single strands in Kunming mice in vivo.[Methods]Single strand DNA(ssDNA)was coupled with chelating agent p-SCN-Bn-NOTA and incubated with ^(64)Cu for a certain time to obtain NOTA-ssDNA.The labeling reaction was monitored by thin layer chromatography,and purified by NAP-5 desalting column to obtain ^(64)Cu-ssDNA.After injection into Kunming mice,the radioactive uptakes of organs of interest at different time were studied.[Results]After NOTA-ssDNA reacted with ^(64)Cu for 1 h,the radiochemical labeling rate is(66.4±2.85)%.After purification,the radiochemical purity is>95%.The in vitro stability experiments show that ^(64)Cu-ssDNA can stably exist in PBS and mouse serum for at least 6 h.There was a certain uptake in the gastrointestinal tract and high radioactive uptake in both the kidney and bladder after ^(64)Cu-ssDNA is injected into the mice,indicating that ^(64)Cu-ssDNA is mainly excreted through the urinary system.[Conclusions]Successful preparation of ^(64)Cu-ssDNA is a critical step for the preparation of PET molecular probes based on DNA polyhedral.In vivo distribution experiment provides experimental control data for studying the in vivo biological distribution of ^(64)Cu labeled DNA polyhedra.

关 键 词：单链DNA ^(64)Cu标记 体内分布 

分 类 号：TL99[核科学技术—核技术及应用]