凋亡显像剂~(99)mTc(CO)_3-Annexin V的制备及其在小鼠体内的生物学分布
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摘要
目的:使用放射性核素~(99m)Tc标记特异性靶向凋亡蛋白Annexin V,并研究标记化合物在小鼠体内的药代动力学特点。方法:采用Tricarbonyl kit制备标记前体三羰基锝化合物盐溶液以标记Annexin V;测定标记化合物的标记率、放化纯及稳定性。通过生物分布实验研究~(99m)Tc(CO)_3-Annexin V在健康昆明小鼠体内的药代动力学特点。结果:成功制备了~(99m)Tc(CO)_3-Annexin V,标记率为(57.21±2.83)%,比活度为(5.65±0.43)mCi/100μg,放化纯>90%,其在体外稳定性良好,至标记后6h放化纯仍大于>90%。~(99m)Tc(CO)_3-Annexin V在小鼠体内循环半衰期较短,注入体内2h后即可排出约40%,4h后排出约60%。肾脏和肝脏是~(99m)Tc(CO)_3-Annexin V的主要排泄器官。结论:本研究表明利用Tricarbonyl kit可以成功将放射性核素~(99m)Tc标记Annexin V,标记率高且方法简便可行,是一种具有广泛应用前景的凋亡显像剂。
Objective:To label the protein Annexin V for targeting apoptosis with radionuclide ~(99m)Tc,and to study pharmacokinetics of ~(99m)Tc(CO)_3-Annexin V in mice.Methods:The Tricarbonyl kit was used to prepare labelled precursor,and the Annexin V was labeled with carbonyl technetium.Then,the labeling yield,radiochemical purity and stability of ~(99m)Tc(CO)_3-Annexin V were determined.The metabolic characteristics of ~(99m)Tc(CO)_3-Annexin V were assessed in healthy Kunming mice by the biodistribution studies.Results:The ~(99m)Tc(CO)_3-Annexin V was prepared successfully,with labeling yield of(57.21±2.83)%,specific activity of(5.65±0.43)mCi/100μg,and radiochemical purity more than 90%.~(99m)Tc(CO)_3-Annexin V had excellent stability and the radiochemical purity was also more than 90% at 6 hin vitro.Biodistribution studies showed that the half-life of ~(99m)Tc(CO)_3-Annexin V was short in mice,manifested by about 40 and 60 percent of ~(99m)Tc(CO)_3-Annexin V that could be removed at 2 hand 4 hafter injection,respectively.In addition,the kidney and liver were the main excretory organs of ~(99m)Tc(CO)_3-Annexin Vin vivo.Conclusion:~(99m)Tc(CO)_3-Annexin V can be prepared successfully using Tricarbonyl kit with high labeling yield and simple method.It is a kind of apoptosis imaging agent with wide application prospect.
Objective:To label the protein Annexin V for targeting apoptosis with radionuclide ~(99m)Tc,and to study pharmacokinetics of ~(99m)Tc(CO)_3-Annexin V in mice.Methods:The Tricarbonyl kit was used to prepare labelled precursor,and the Annexin V was labeled with carbonyl technetium.Then,the labeling yield,radiochemical purity and stability of ~(99m)Tc(CO)_3-Annexin V were determined.The metabolic characteristics of ~(99m)Tc(CO)_3-Annexin V were assessed in healthy Kunming mice by the biodistribution studies.Results:The ~(99m)Tc(CO)_3-Annexin V was prepared successfully,with labeling yield of(57.21±2.83)%,specific activity of(5.65±0.43)mCi/100μg,and radiochemical purity more than 90%.~(99m)Tc(CO)_3-Annexin V had excellent stability and the radiochemical purity was also more than 90% at 6 hin vitro.Biodistribution studies showed that the half-life of ~(99m)Tc(CO)_3-Annexin V was short in mice,manifested by about 40 and 60 percent of ~(99m)Tc(CO)_3-Annexin V that could be removed at 2 hand 4 hafter injection,respectively.In addition,the kidney and liver were the main excretory organs of ~(99m)Tc(CO)_3-Annexin Vin vivo.Conclusion:~(99m)Tc(CO)_3-Annexin V can be prepared successfully using Tricarbonyl kit with high labeling yield and simple method.It is a kind of apoptosis imaging agent with wide application prospect.
引文
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[11]王荣福,刘萌,张春丽,等.荷瘤小鼠单次化疗后活体探测肿瘤细胞凋亡的实验研究[J].中华医学杂志,2005,85(44):3 155-3 157.Wang RF,Liu M,Zhang CL,et al.Experimental study on the dection of tumor cell apoptosis in tumor bearing mice received single chemotherapy[J].Natl Med J China,2005,85(44):3 155-3 157.
[12]吕中伟,朱承谟,李彪,等.99mTc-Annexin V血栓显像及其体内分布的初步实验研究[J].上海医学影像杂志,2000,9(3):137-139.Lv ZW,Zhu CM,Li B,et al.Thrombosis imaging with 99mTc labeled Annexin V and its distribution in vivo[J].Shanghai Med Imaging,2000,9(3):137-139.
[2]Hardy JW,Levashova Z,Schmidt TL,et al.99mTcAnnexin V-128SPECT monitoring of splenic and disseminated listeriosis in mice:a model of imaging sepsis[J].Mol Imaging Biol,2015,17(3):345-354.
[3]Lu C,Jiang Q,Hu M,et al.Preliminary biological evaluation of18 F-FBEM-Cys-Annexin V a novel apoptosis imaging agent[J].Molecules,2015,20(3):4 902-4 914.
[4]Lahorte CM,van de Wiele C,Bacher K,et al.Biodistribution and dosimetry study of 123I-rh-annexin V in mice and humans[J].Nucl Med Commun,2003,24(8):871-880.
[5]贾支俊,申景涛,郭万华,等.99Tcm标重组H-Annexin V在动物血栓显像中的研究[J].中华核医学杂志,2007,27(5):291-294.Jia ZJ,Shen JT,Guo WH,et al.An experimental study on thrombosis imaging with 99 Tcm-H-annexin V[J].Chin J Nucl Med,2007,27(5):291-294.
[6]van de Wiele C,Lahorte C,Vermeersch H,et al.Quantitative tumor apoptosis imaging using technetium-99m-HYNIC Annexin V single photon emission computed tomography[J].J Clin Oncol,2003,21(18):3 483-3 487.
[7]Vanderheyden JL,Liu G,He J,et al.Evaluation of99m Tc-MAG3-annexin V:influence of the chelate on in vitro and in vivo properties in mice[J].Nucl Med Biol,2006,33(1):135-144.
[8]Waibel R,Alberto R,Willuda J,et al.Stable one-step technetium-99m labeling of His-tagged recombinant proteins with a novel Tc(I)-carbonyl complex[J].Nat Biotechnol,1999,17(9):897-901.
[9]Li C,Wen B,Wang L,et al.99mTc-labeled single-domain antibody EG2in targeting epidermal growth factor receptor:an in-vitro and mouse model in-vivo study[J].Nucl Med Commun,2015,36(5):452-460.
[10]He J,Wang Y,Feng J,et al.Targeting prostate cancer cells in vivo using a rapidly internalizing novel human single-chain antibody fragment[J].J Nucl Med,2010,51(3):427-432.
[11]王荣福,刘萌,张春丽,等.荷瘤小鼠单次化疗后活体探测肿瘤细胞凋亡的实验研究[J].中华医学杂志,2005,85(44):3 155-3 157.Wang RF,Liu M,Zhang CL,et al.Experimental study on the dection of tumor cell apoptosis in tumor bearing mice received single chemotherapy[J].Natl Med J China,2005,85(44):3 155-3 157.
[12]吕中伟,朱承谟,李彪,等.99mTc-Annexin V血栓显像及其体内分布的初步实验研究[J].上海医学影像杂志,2000,9(3):137-139.Lv ZW,Zhu CM,Li B,et al.Thrombosis imaging with 99mTc labeled Annexin V and its distribution in vivo[J].Shanghai Med Imaging,2000,9(3):137-139.