我們已知α-亞甲基-γ-五環內醯胺化合物(α-methylene-γ-
lactam)證實對癌症細胞有毒殺效果，在生物活性上也擁有廣泛的應用，像是消炎、抗菌、抗黴菌…等。為了瞭解此化合物能否作為腎癌細胞的毒殺藥物，我們必須進一步探討其對腎臟細胞之毒性高低。我們利用實驗室合成出來的八種α-亞甲基-γ-五環內醯胺化合物作為試驗藥品，選用腎臟可發綠螢光之基因轉殖斑馬魚Tg(wt1b:EGFP)為生物模型，藉此探討其對腎臟造成的影響，以用來評估化合物之安全性。我們將24 hpf的斑馬魚胚胎浸泡於六組不同的條件中，分別是三種濃度(1 nM、10 nM、100 nM)與兩個時間（24-36 hpf、24-72 hpf）之組合，發現腎臟的畸形率會與濃度成正比(1nM，10-100%；10nM，16.7-100%；100nM，16.7-100%)，發現在長時間浸泡下的斑馬魚胚胎，其嚴重畸形與輕微畸形的比例下降，推測可能是隨著胚胎的成熟，腎臟自我修復的能力增強所導致。存活率分析的部分，除了編號5的α-亞甲基-γ-五環內醯胺化合物在長時間的浸泡下存活率會低於80%，其餘實驗結果則皆達100%存活率，因此，α-亞甲基-γ-五環內醯胺化合物具有潛力作為腎癌細胞毒殺藥物的可行性。

The α-methylene-γ-lactams have known as antitumor agents and own lower cytotoxicity which compared to α-methylene-γ-lactaones, and present many biological activities, such as anti-inflammatory, antibacterial, and antifungal…etc. In order to understand these compounds may serve as antitumor drugs for kidney cancer cells, This experiment synthesized eight kindsα-methylene-γ-lactam compound by our laboratory, and used transgenic zebrafish Tg(wt1b:EGFP) as the toxicity investigation model to investigated their toxicities for normal cells. The stage of 24 hpf zebrafishes were treated six different protocols, which comprised combinations of three concentrations(1, 10, 100 nM) and two exposure methods [methods I, 24-36 hpf(hours post-
fertilization); II, 24-72 hpf]. The results showed that the malformation rate of kidney is proportion to the concentration of introducing γ-lactam(1nM, 10-100%；10nM,16.7-100%；100nM，16.7-100%), the longer time soaking of embryo with α-methylene-γ-lactam decrease the severe and mild ratio of embryo’s malformation rate, this observation may cause by kidney self-repair mechanism which increased with embryo mature; All testing γ-lactams exhibit normal cell survival rate reach 100%, except the No.5 compound. Therefore, the α-methylene-γ-lactam as renal cancer medication candidate may be achieved after more studies.

目錄
中文摘要 ……………………………………………………………… I
英文摘要 ………………………………………………………………II
目錄 ………………………………………………………………… IV
第一章	緒論…………………………………………………………… 1
1-1	α-亞甲基-γ-五環內醯胺衍生物的生物活性………………1
1-2	α-亞甲基-γ-五環內醯胺的合成方式………………………7
1-3	癌症起源與治療對策…………………………………………11
1-4	生物活性檢測-斑馬魚……………………………………… 26
1-4-1 斑馬魚的優勢…………………………………………26
1-4-2 斑馬魚的腎臟發育……………………………………27
1-4-3基因轉殖魚的介紹…………………………………… 31
第二章 材料與方法……………………………………………………33
2-1 合成α-亞甲基-γ-五環內醯胺化合物	……………………33
2-1-1 亞胺化合物之合成……………………………………33
2-1-2 α-亞甲基-γ-五環內醯胺之合成………………… 34
2-2 斑馬魚的飼養與胚胎收集………………………………… 36
2-2-1 實驗用斑馬魚之飼養…………………………………36
2-2-2 斑馬魚胚胎收集 …………………………………… 37
2-3 浸泡條件…………………………………………………… 39
2-3-1 製備α-亞甲基-γ-五環內醯胺溶液 ………………39
2-3-2 觀察斑馬魚腎臟發育與畸形率的統計………………41
2-4 儀器及測試方式 …………………………………………… 43
2-5 溶劑乾燥 …………………………………………………… 44
2-6 螢光顯微鏡、照相機以及其他儀器…………………………46
第三章 結果與討論……………………………………………………47
3-1 化合物濃度對存活率之影響……………………………… 47
3-2 浸泡濃度對腎臟的影響…………………………………… 49
3-3 浸泡時間對腎臟的影響…………………………………… 57
3-4 結論………………………………………………………… 59
第四章 光譜……………………………………………………………62
第五章 參考資料………………………………………………………68

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