臺灣博碩士論文加值系統

近年來相關缺氧細胞造影劑之研究，18FMisonidazole(18FMISO)一直佔有相當重要之地位。此藥物對缺氧細胞有特異性積聚；故為一甚佳之缺氧造影劑，同時為一輻射增敏劑，亦可作為癌症治療藥物。18F為正子放射性同位素，必須利用迴旋加速器設施生產。且須使用價格昂貴的PET造影設備，使得18FMISO無法廣泛應用於臨床上。
[99mTc(CO)3(H2O)3]+，為近年來新興之鎝99m標誌前趨物，其高標誌效率與高穩定性，及可與雙牙基、三牙基之配位子相結合之特性頗受核醫界重視。於本研究中，試圖合成Misonidazole（MISO）配位子與[99mTc(CO)3(H2O)3]+標誌前趨物，並嘗試將 [99mTc(CO)3(H2O)3]+與MISO結合。藉由高效能液相層析儀（HPLC）、薄層層析法（TLC）、電泳（Electrophoresis）等分析方法以了解放射化學特性，再以缺氧細胞吸收試驗評估未來此藥物應用於SPECT核醫缺氧造影藥物之可能。
結果顯示，[99mTc(CO)3(H2O)3]+ 在HPLC滯留時間( Retention time)為4 分鐘，薄層層析法（TLC，MeOH/HCl (95/5 v/v)）Rf值為0.35，電泳顯示為帶正電，在細胞缺氧實驗中，富氧吸收比例較高；[99mTc(CO)3(H2O)3]+與MISO結合所形成之錯合物在HPLC滯留時間為10分鐘，薄層層析法（TLC，MeOH/HCl (95/5 v/v)）Rf值為0.7，電泳為電中性，在細胞缺氧實驗中，缺氧吸收比例較高。由以上結果証實fac-99mTc(CO)3已成功地標誌上MISO，並在缺氧細胞中有特異性吸收，對未來應用於SPECT 缺氧造影藥物之潛力，值得更進一步探討。

18FMISO has played an important role as a hypoxic tracer, presenting with a high specific accumulation in hypoxic cells. 18F is a position emitting nuclide and generally produced from a cyclotron. 18F labeled radiopharmaceuticals associated with expensive PET scan, would be restricted in clinical use for patients.
[99mTc(CO)3(H2O)3]+ was recently developed as an effective 99mTc labeling precursor, featured with high stability, highly labeling efficiency, and readily coordinating with bidentate or tridentate ligands with N, O and S donors. In this study, it has been attempted to label 99mTc in misonidazole (MISO), a hypoxic agent, using [99mTc(CO)3(H2O)3]+ as the precursor. Radiochemical characterization for the novel 99mTc labeled MISO complex was conducted via high performance liquid chromatography (HPLC)、 thin layer chromatography (TLC) ,and electrophoresis. Cell uptake test has been performed with tumor cells under aerobic and hypoxic conditions in order to assess the efficacy of 99mTcCO3  MISO as a hypoxic agent.
In HPLC , [99mTc(CO)3(H2O)3]+ is located at retention time = 4 min. In TLC, the 99mTc species is located at Rf = 0.3. In electrophoresis, it is prone to be positive. In cell uptake test, [99mTc(CO)3(H2O)3]+ presents higher uptake in tumor cells under aerobic condition than under hypoxic condition. For 99mTc(CO)3MISO species by labeling [99mTc(CO)3(H2O)3]+ in MISO, the results of radiochemical characterization are as follows. In HPLC, the 99mTc labeled MISO is located at retention time = 10 min. In TLC the 99mTc-complex species is located at Rf = 0.7. In electrophoresis, it presents to be neutral. In cell uptake test, 99mTc(CO)3MISO displays higher uptake under hypoxic condition than under aerobic condition.
In conclusion, this work has demonstrated the labeling of MISO with fac-99mTc(CO)3 for the first time. The novel 99mTc(CO)3MISO species exhibits the potential as a hypoxia tracer. It is worthy to evaluate furthermore the 99mTc labeled MISO complex as a hypoxia imaging agent for clinical use.

頁次
謝誌 i
中文摘要 ii
英文摘要 iv
目錄 v
圖表目錄 vii
第一章 緒論 1
1-1. 核醫藥物簡介 1
1-2. 鎝之基礎特性 2
1-3. 鎝三羰基錯合物 4
1-4. 細胞缺氧造影藥物 6
1-5. 本研究的方向及目的 10
第二章 實驗部分 12
2-1 材料與儀器 12
2-2 實驗步驟 15
2-2-1 Tricarbonyl technetium(I) complex 15
2-2-1-1 Tricarbonyl technetium(I) complex製備 15
2-2-1-2 瓶產生效率測試 15
2-2-2 MISO(misodazole)化合物之製備 16
2-2-3 [99mTc(CO)3(H2O)3]+標誌MISO 17
2-2-3-1 標準步驟 17
2-2-3-2 溫度對標誌之影響 17
2-2-3-3 不同pH值對標誌之影響 17
2-2-3-4 穩定度測試 18
2-2-4 腫瘤細胞吸收測試 18
2-2-4-1 [99mTc(CO)3(H2O)3]+缺氧吸收測試 18
2-2-4-2 fac-99mTc(CO)3MISO缺氧吸收測試 19
2-3 分析方法 20
2-3-1 高效能液相層析儀 20
2-3-2 薄層層析法 20
2-3-3 電泳 21
2-3-4 萃取效率 21
2-3-5 溶點測定儀 21
2-3-6 核磁共振儀 21
第三章 實驗結果 22
3-1 鎝三羰基錯合物 22
3-2 MISO化合物之製備 23
3-3 [99mTc(CO)3(H2O)3]+標誌MISO 23
3-4 腫瘤細胞吸收測試 24
第四章 討論 25
4-1 鎝三羰基錯合物 25
4-2 MISO化合物之製備 26
4-3 [99mTc(CO)3(H2O)3]+標誌MISO 26
4-4 腫瘤細胞吸收測試 28
第五章 結論 30
第六章 參考文獻 31
圖表 37
圖（一）[99mTcO4] 之典型HPLC圖 37
圖（二）[99mTc(CO)3(H2O)3]+ 之典型HPLC圖 37
圖（三）[99mTcO4] 以MeOH/HCl展開之TLC圖 38
圖（四）[99mTc(CO)3(H2O)3]+ MeOH/HCl展開之TLC圖
38
圖（五）[99mTcO4]以MEK展開之TLC圖 39
圖（六）[99mTc(CO)3(H2O)3]+以MEK展開之TLC圖39
圖（七）[99mTcO4] 之電泳圖 40
圖（八）[99mTc(CO)3(H2O)3]+之電泳圖 40
圖（九）第一週配製[99mTc(CO)3(H2O)3]+之HPLC圖
41
圖（十）第二週配製[99mTc(CO)3(H2O)3]+之HPLC圖
41
圖（十一）第三週配製[99mTc(CO)3(H2O)3]+之HPLC圖
42
圖（十二）第四週配製[99mTc(CO)3(H2O)3]+之HPLC圖
42
圖（十三）用pH=11buffer流洗[99mTcO4]配製[99mTc(CO)3(H2O)3]+之HPLC圖 43
圖(十四) [99mTc(CO)3(H2O)3]+與MISO振盪後之HPLC圖 44
圖(十五) [99mTc(CO)3(H2O)3]+與MISO加熱後之HPLC圖 44
圖(十六) 99mTc(CO)3MISO之TLC圖 45
圖(十七) 99mTc(CO)3MISO之電泳圖 46
圖(十八) 不同pH值對99mTc(CO)3標誌MISO之效率影響圖 47
圖(十九) 不同pH值對99mTc(CO)3標誌MISO之效率影響圖 48
圖(二十) 99mTc(CO)3標誌MISO穩定度圖 49
圖(二十一) [99mTc(CO)3(H2O)3]+之缺氧細胞吸收圖 50
圖(二十二) 99mTc(CO)3MISO之缺氧細胞吸收圖 50
圖(二十三) MISO之1H-NMR圖 51
附圖(一) MISODAZOLE與FLUOROMISONIDAZOLE結構圖 52
附圖(二) A)圖是nitromidazole的生物反應式；B)圖是FMISO在細胞內的生物反應式。 53
表1 不同氧化態之鎝錯合物及其幾何形狀、配位數與磁矩 54

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