臺灣博碩士論文加值系統

本研究旨在探討鎵-67及鎵-68標誌含胺基酸大環化合物─ 1,4,7,10-tetraazacyclododecane N,N’,N”,N’”-tetraacetic acid-glutamine（DOTA-Gln）之放射化學性質及其腫瘤細胞吸收與生物分佈。首先以液相及固相合成方法合成DOTA-Gln，比較兩種方法之產率，進而與鎵-67及鎵-68進行錯合反應。經薄層層析法及電泳動分析法予以分析探討其標誌效率、親脂性及穩定度。接著再進一步做此藥物對纖維瘤細胞的細胞存活試驗、毒性測試、吸收試驗及動物器官組織分佈實驗，以評估其作為腫瘤PET造影之潛力。
在標誌方面，DOTA-Gln與68Ga (acac)3或67Ga-citrate於室溫下超音波振盪20分鐘經置換反應即可完成標誌，標誌效率高達90％以上，經由放射化學分析結果證實68Ga-DOTA-Gln為一帶負電且親水性高之化合物。以67Ga-DOTA-Gln測試其穩定度發現經72小時後穩定度仍然很高。在細胞試驗結果中發現67Ga-DOTA-Gln對纖維瘤細胞無化學及放射毒性，且其吸收量隨著時間及細胞數增加，顯示此藥物具有腫瘤造影診斷之潛力。
接著以67Ga-DOTA-Gln進一步做動物器官組織分佈實驗及SPECT造影。由生物分佈結果顯示，藥物主要以腎臟及泌尿道為其代謝途徑，腫瘤吸收量隨時間增加，於1小時達到最大值8.54％（％ ID/g organ），約為第一時間點的6倍且為肌肉的1.3倍。而其在脾臟的滯留現象，亦有做更進一步探討的價值。

In this study, synthesis, radiochemical characteristic, tumor cell uptake, and biodistribution of 67Ga and 68Ga labeled 1,4,7,10-tetra- azacyclododecane N,N’,N”,N’”-tetraacetic acid-Glutamine（DOTA-Gln）were investigated. First, the DOTA-Gln couple was prepared by two different synthetic methods, in liquid phase (traditional) and in solid phase. Subsequently, both 68Ga and 67Ga were labeled to this couple to get the radiolabeled conjugate. Labeling efficiency, lipophilicity, and stability of 68Ga-DOTA-Gln and 67Ga-DOTA-Gln were surveyed by the radioanalytical methods including thin layer chromatography and electrophoresis. Finally, animal biodistribution study, cell survival test, toxicity test, and cell uptake trials were undertaken to assess the potential of the 68Ga-DOTA-Gln as a PET imaging agent.
For labeling, 68Ga-DOTA-Gln and 67Ga-DOTA-Gln were acquired by ligand-ligand exchange of DOTA-Gln with 68Ga (acac)3 or 67Ga-citrate; the reaction was facilitated via supersonication at room temperature for 20 min. Labeling efficiency of either 68Ga or 67Ga labeled DOTA-Gln could reach ca. 90%. From radiochemical analysis including TLC and electrophoresis, the radiolabeled conjugate was evidenced to be anionic and hydrophilic. The stability of 67Ga-DOTA-Gln was found to be high even storage for 72h. In cell uptake experiment, 67Ga-DOTA-Gln showed negligible chemical toxicity and radiation damage for spontaneous and non-immunogenic fibrosarcoma cells (Nfsa). The cell uptake increased with increasing cell numbers and elongating the elapsed time. From the results, it is indicated that 68Ga-DOTA-Gln has the potential as a PET imaging agent for diagnosing tumors.
In the biodistribution study, 67Ga-DOTA-Gln was found to excrete mainly via kidney and urinary tract. Tumor uptake increased with the elapsed time, reaching a maximum at 8.54% (% ID/g organ) at 1h postinjection. The uptake ratio of tumor to muscle was ca. 1.3, suggesting that 67Ga-DOTA-Gln possesses the potential as a diagnostic agent for tumors. In addition, the high uptake and retention of 67Ga-DOTA-Gln in the spleen may reveal another purpose pertaining to nuclear medicine.

中文摘要-------------------------------------------------------I
英文摘要-----------------------------------------------------III
謝誌-----------------------------------------------------------V
目錄---------------------------------------------------------VII
表目錄---------------------------------------------------------X
圖目錄--------------------------------------------------------XI
第一章 緒論-------------------------------------------------1
1-1 正子造影-------------------------------------------------1
1-2 鎵之化學特性及應用---------------------------------------5
1-3 放射性金屬錯合物之應用-----------------------------------6
1-4 研究方向及目的-------------------------------------------9
第二章 有機合成--------------------------------------------13
2-1 試藥--------------------------------------------13
2-2 儀器與材料--------------------------------------14
2-3 實驗步驟----------------------------------------15
2-3-1 DOTA 之合成--------------------------------15
2-3-2 DOTA-Gln之液相合成-------------------------16
2-3-3 DOTA-Gln之固相合成-------------------------16
2-4 結果與討論--------------------------------------19
第三章 鎵-68與鎵-67螯合物之標誌及特性分析-------------------22
3-1 試藥--------------------------------------------22
3-2 儀器--------------------------------------------22
3-3 鎵-67與鎵-68錯合物之標誌------------------------23
3-3-1 原理---------------------------------------23
3-3-2 實驗步驟-----------------------------------24
3-4 化學特性分析------------------------------------24
3-4-1 電泳動分析---------------------------------24
3-4-2 薄層層析法---------------------------------26
3-5 67Ga-DOTA-Gln之穩定度試驗-----------------------27
3-6 結果與討論--------------------------------------27
第四章 腫瘤細胞與小鼠對67Ga-DOTA-Gln之吸收與生物分佈試驗----29
4-1 藥品--------------------------------------------29
4-2 儀器與材料--------------------------------------29
4-3 細胞培養----------------------------------------30
4-3-1 細胞及其培養條件---------------------------30
4-3-2 纖維瘤細胞之生長曲線測定-------------------30
4-3-3 細胞對DOTA-Gln藥物之存活試驗 (MTT assay)
--------------------------------------------31
4-3-4 67Ga-DOTA-Gln對細胞之毒性測試--------------32
4-3-5 細胞對67Ga-DOTA-Gln之吸收試驗--------------32
4-4 動物器官組織分佈實驗----------------------------33
4-4-1 實驗動物-----------------------------------33
4-4-2 皮下注射-----------------------------------33
4-4-3 生物分佈試驗-------------------------------34
4-5 SPECT造影---------------------------------------34
4-6 結果與討論--------------------------------------34
第五章 結論-------------------------------------------------38
參考文獻-----------------------------------------------------40
附錄---------------------------------------------------------44

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