臺灣博碩士論文加值系統

具有八個胺基酸的環型胜肽octreotide，是一種體抑素類似物，對體抑素受體具有高度親和力。本研究旨在發展99mTc(I)-HYNIC-octreotide為一種有效的體抑素受體造影劑。本研究參考核能研究所的方法，以固相胜肽合成法製備HYNIC-octreotide。並參考文獻方法合成HYNIC及[99mTc(CO)3(OH2)3]+。以[99mTc(CO)3(OH2)3]+為前驅物，於75℃及中性條件下進行標誌HYNIC-octreotide得99mTc(I)-HYNIC-octreotide。根據實驗結果得知[99mTc(CO)3(OH2)3]+標誌於HYNIC-octreotide標誌產率可達90%以上。HYNIC-octreotide在體抑素受體競爭實驗中，有效地抑制125I-Tyr11-Somatostatin的結合，證實HYNIC-octreotide具有對體抑素受體的結合能力。經由活體動物實驗，注入99mTc(I)-HYNIC-octreotide於種有AR42J腫瘤之SCID小鼠，其腫瘤位置顯現清楚並且持續累積的影像，證實99mTc(I)-HYNIC-octreotide對具體抑素受體之腫瘤有造影之潛力。

Octreotide, a cyclopeptide with eight amino acids, is a somatostatin analog with the affinity to somatostatin receptors. The aim of the study is to develop 99mTc(I)-HYNIC-octreotide as radiopharmaceutical for somatostatin receptor scintigraphy. HYNIC-octreotide is synthesizd by the method of solid-phase peptide synthesis with disulfide bonding developed by Institute of Nuclear Energy Research. HYNIC and [99mTc(CO)3(OH2)3]+ were prepared respectively according to the previous reports. Octreotide was labeled with [99mTc(CO)3(OH2)3]+ via HYNIC as a bifunctional chelator. 99mTc(I)-HYNIC-octreotide was prepared by mixing the [99mTc(CO)3(OH2)3]+ solution with HYNIC-octreotide in saline at 75℃. According to the results, the radiochemical purity for 99mTc(I)-HYNIC-octreotide was greater than 90%. The labeling technique for 99mTc(I)-HYNIC-octreotide has been successfully established in this study. The HYNIC-octreotide efficiently inhibited the binding ability of 125I-Tyr11-somatostatin with somatostatin receptor regarding to the retention of its receptor-binding affinity. In the animal model, 99mTc(I)-HYNIC-octreotide was uptaken by the pancreas tumor cells (AR42J) implanted into the SCID mice, and the result showed by showing a clear image for the tumor. These results demonstrated that 99mTc(I)-HYNIC-octreotide is potential for imaging pancreas tumors via somatostatin receptors .

謝誌 I
中文摘要 II
英文摘要 III
目錄 IV
附表目錄 VIII
附圖目錄 IX
第一章 緒論 1
1-1 核醫藥物簡介 1
1-2 標誌方法介紹 3
1-3 體抑素及其衍生物 4
1-4 固相胜肽合成法介紹 5
1-5 研究方向及目的 6
第二章 有機合成實驗 11
2-1 試藥 11
2-2 儀器 11
2-3 配位子合成 12
2-3.1 Hydrazine nicotinic acid (HYNIC)之合成 12
2-3.2 6-Boc-hydrazinopyridine-3-carboxylic acid (HYNIC-Boc) 之合成 13
2-4 結果與討論 14
第三章 生物分子耦合實驗 23
3-1 試藥 23
3-2儀器 24
3-3 HYNIC-octreotide合成 24
3-3.1 胜肽合成 25
3-3.2 裂解反應 25
3-3.3 環化反應 25
3-4 高效能液相層析儀分析條件 26
3-5 結果與討論 26
第四章 [99mTc(CO)3(OH2)3]+標誌HYNIC-octreotide 32
4-1 試藥 32
4-2 儀器與材料 32
4-3 [99mTc(CO)3(OH2)3]+標誌HYNIC-octreotide 33
4-3.1 緩衝液配製 33
4-3.2 [99mTc(CO)3(OH2)3]+ 之合成 33
4-4 標誌效率分析 34
4-4.1 薄層層析法 34
4-4.2 高效能液相層析法 34
4-4.2.1 高效能液相層析儀分析條件 35
4-5 穩定度試驗 35
4-5.1 以Sep-Pak C18 cartridge分離純化99mTc(I)-HYNIC-octreotide 35
4-5.2 結構穩定度試驗 35
4-5.3 生物穩定度試驗 36
4.6 結果與討論 36
第五章 AR42J細胞受體結合實驗 43
5-1 細胞培養基及相關試劑 43
5-2 器材 43
5-3 儀器與設備 44
5-4 實驗溶液製備 45
5-5 細胞培養 46
5-6 受體結合實驗 46
5-6.1 細胞膜蛋白之製備 46
5-6.2 蛋白質濃度之測定 47
5-6.3 受體結合實驗 47
5-7結果與討論 49
第六章 生物分佈與腫瘤造影 52
6-1 試藥 52
6-2 儀器與材料 52
6-3 實驗動物 52
6-3.1 動物模式建立 52
6-3.2 生物分佈 53
6-3.3 動物造影 53
6-4 結果與討論 53
第七章 結論 60
第八章 參考文獻 61

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