臺灣博碩士論文加值系統

本論文主要的研究是合成短鍊型硫代氨基甲酸類化合物做為乙醯膽鹼酯酵素 (AChE)、丁醯膽鹼酯酵素 (BChE)、膽固醇酯酵素 (CEase)、青黴素酶、脂肪酵素和凝血酶六種絲氨酸水解酵素，利用酵素催化三元組中絲氨酸所做的親核性攻擊反應機制，探討硫代氨基甲酸類化合物對於不同酵素抑制效果的研究，藉由停時驗析得知此類化合物皆屬類受質不可逆抑制劑。透過動力學實驗分析反應速率常數 (kc)、解離常數 (Ki)及總抑制常數(ki)，可以得到化合物對於絲氨酸水解酵素的抑制結果。實驗中，由氨基甲酸類化合物、硫代氨基甲酸類化合物和市售藥物對乙醯膽鹼酯酵素等六種不同酵素之抑制常數，進一步可以比較抑制效果的差異。
實驗中，就本文所合成出的O,O''-propane-1,3-diyl bis(butylcarbamothioate) (化合物８)而言，對酵素的抑制效果由好到壞依序為，脂肪酵素、丁醯膽鹼酯酵素、膽固醇酯酵素、乙醯膽鹼酯酵素、青酶素酶和凝血酶；就化合物８和氨基甲酸類化合物比較，對乙醯膽鹼酯酵素、丁醯膽鹼酯酵素、膽固醇酯酵素和脂肪酵素而言，抑制效果則是化合物８較好；對青黴素酶和凝血酶而言，氨基甲酸類化合物則有較好的抑制效果。

1-1　酵素特性 1
1-2　酵素介紹 3
1-2-1　乙醯膽鹼酯酵素 3
1-2-2　丁醯膽鹼酯酵素 6
1-2-3　膽固醇酯酵素 8
1-2-4　脂肪酵素 10
1-2-5　青黴素酶 12
1-2-6　凝血酶 14
第二章　酵素動力學原理 16
2-1　原理 16
2-2　不可逆抑制劑 18
第三章　材料與方法 30
3-1　儀器設備 30
3-2　試藥溶劑 31
3-3　抑制劑的合成與光譜數據 33
3-3-1　化合物的合成 33
3-3-2　化合物光譜數據 34
3-3-2-1　3-hydroxypropyl ethylcarbamothioate 34
3-3-2-2　3-hydroxypropyl propylcarbamothioate 35
3-3-2-3　3-hydroxypropyl butylcarbamothioate 36
3-3-2-4　3-hydroxypropyl tert-butylcarbamothioate 37
3-3-2-5　3-hydroxypropyl benzylcarbamothioate 38
3-3-2-6　O,O''-propane-1,3-diyl bis(ethylcarbamothioate) 39
3-3-2-7　O,O''-propane-1,3-diyl bis(propylcarbamothioate) 40
3-3-2-8　O,O''-propane-1,3-diyl bis(butylcarbamothioate) 41
3-3-2-9　O,O''-propane-1,3-diyl bis(tert-butylcarbamothioate) 42
3-3-2-10　O,O''-propane-1,3-diyl bis(benzylcarbamothioate) 43
3-4　酵素動力學的材料與方法 44
3-4-1　乙醯膽鹼酯酵素 44
3-4-2　丁醯膽鹼酯酵素 45
3-4-3　膽固醇酯酵素 46
3-4-4　脂肪酵素 47
3-4-5　青黴素酶 48
3-4-6　凝血酶 49
3-5　動力學實驗 50
3-5-1　停時驗析法 50
3-5-2　不可逆型抑制劑動力學反應 51
第四章 結果與討論 54
4-1 停時驗析法 54
4-2　動力學實驗結果 58
4-2-1　類受質抑制劑動力學中吸收度對時間作圖 58
4-2-1-1　乙醯膽鹼酯酵素 58
4-2-1-2 丁醯膽鹼酯酵素 63
4-2-1-3　膽固醇酯酵素 68
4-2-1-4 脂肪酵素 73
4-2-1-5　青黴素酶 78
4-2-1-6 凝血酶 83
4-2-2 類受質抑制劑動力學中Kapp對時間作圖 88
4-2-2-1 乙醯膽鹼酯酵素 88
4-2-2-2 丁醯膽鹼酯酵素 94
4-2-2-3 膽固醇酯酵素 99
4-2-2-4 脂肪酵素 104
4-2-2-5 青黴素酶 109
4-2-2-6 凝血酶 114
4-3　酵素動力學反應相關數據 119
4-3-1　乙醯膽鹼酯酵素 119
4-3-2　丁醯膽鹼酯酵素 121
4-3-3　膽固醇酯酵素 123
4-3-4　脂肪酵素 125
4-3-5　青黴素酶 127
4-3-6　凝血酶 129
第五章 結論 131
附錄　抑制劑光譜圖 136
參考文獻 156

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