臺灣博碩士論文加值系統

本論文主要以抑制劑對蛇毒、蜂毒及豬胰臟磷脂&;#37238;酵素( PLA2 )反應中各步驟的速率常數進行定量結構活性關係(QSAR)分析，利用酵素-受質複合體結合常數( KS )、酵素-抑制劑複合體抑制常數( KI )、酵素-抑制劑中間體正反應常數( k2 )與酵素-抑制劑中間體逆反應常數( k-2 )，四種不同的動力學常數對不同取代基之電子效應常數( σ* )、立體效應常數( ES )以及疏水性常數( π )作線性迴歸分析，並探討不同取代基的抑制劑對三種磷脂&;#37238;酵素的影響因素及抑制效果，了解芳香族氨基甲酸類化合物對三種不同磷脂&;#37238;酵素的抑制機理，讓我們對此類抑制劑的抑制機理有更深一層的研究，並依此探討三-醯氧-一-氮-正辛烷氨基甲酸苯類化合物在三種不同磷脂&;#37238;酵素中的差異。
利用理論計算軟體(Gaussian 03)計算三-醯氧-一-氮-正辛烷氨基甲酸苯類化合物的能量大小、電荷數據與氨基甲酸上的化學位移( δ )，將其進行定量結構活性關係分析；最後，藉由電荷、化學位移以及能量與動力學數據作線性迴歸分析，可以推測出三種不同磷脂&;#37238;酵素對此類抑制劑的反應快慢與安定性。
最後，我們根據分析結果得知，抑制劑抑制效果的好壞，還是要從由定量活性結構關係與動力學數據的結果來做判斷，當取代基的推電子能力越大時，此抑制劑對三種磷脂&;#37238;酵素(PLA2)的抑制效果越好，抑制性也越佳。

目錄

摘要-------------------------------------------------------i
抑制劑的結構與名稱----------------------------------------ii
目錄-----------------------------------------------------iii
表目錄----------------------------------------------------vi
圖目錄---------------------------------------------------vii

第一章 緒論------------------------------------------------1
1-1 研究動機與目的-----------------------------------------1
1-1.1 磷脂酶部分-------------------------------------------2
1-1.2 抑制劑部分-------------------------------------------4
1-2 酵素介紹-----------------------------------------------6
1-2.1磷脂酶酵素(Phospholipase A2, PLA2)--------------------6

第二章 實驗原理-------------------------------------------16
2-1 酵素催化水解受質的抑制反應----------------------------16
2-2 可逆抑制劑--------------------------------------------19
2-2.1 可逆競爭型抑制劑------------------------------------20
2-2.2 KI值的判斷------------------------------------------25
2-2.3 可逆非競爭性抑制劑----------------------------------31
2-3 定量結構活性關係--------------------------------------36
2-3.1 原理------------------------------------------------36
2-3.2 疏水性 ( Hydrophobicity )---------------------------38
2-3.3 電子效應 ( Electronic effects )---------------------40
2-3.4 立體性質 ( Steric factors)--------------------------42
2-3.5 Hammett Equation------------------------------------44
2-3.6 Taft equation---------------------------------------49

第三章 實驗材料與方法步驟---------------------------------52
3-1 試藥及溶劑--------------------------------------------52
3-2 儀器設備----------------------------------------------54
3-3 樣品的配置--------------------------------------------55
3-4 抑制劑的合成步驟--------------------------------------56
3-4.1 芳香族氨基甲酸類抑制劑------------------------------58
3-4.2 芳香族氨基甲酸類化合物合成範例----------------------60
3-5 動力學實驗步驟----------------------------------------62
3-5.1 磷脂酶停時驗析法(Stop Time Assay)之操作步驟---------63

第四章 實驗結果與討論-------------------------------------64
4-1磷脂酶酵素的動力學數據探討-----------------------------64
4-2磷脂酶酵素(PLA2 from Naja mossambica mossambica)的QSAR數據探討與結果----------------------------------------------67
4-2.1 磷脂酶(Naja mossambica mossambica)的QSAR數據處理----67
4-2.2 抑制劑1-9 抑制磷脂酶動力學數據與抑制劑QSAR常數之單參數線性迴歸分析----------------------------------------------68
4-2.3 抑制劑1-9 抑制磷脂酶動力學數據與抑制劑QSAR常數之雙參數線性迴歸分析----------------------------------------------78
4-2.4 抑制劑1-9 抑制磷脂酶動力學數據與抑制劑QSAR常數之三參數線性迴歸分析----------------------------------------------93
4-3磷脂酶酵素(PLA2 from Bee Venom)的動力學數據探討--------96
4-3.1 磷脂酶(Bee Venom)的QSAR數據處理---------------------96
4-3.2 抑制劑1-9 抑制磷脂酶動力學數據與抑制劑QSAR常數之單參數線性迴歸分析----------------------------------------------97
4-3.3 抑制劑1-9 抑制磷脂酶動力學數據與抑制劑QSAR常數之雙參數線性迴歸分析---------------------------------------------107
4-3.4 抑制劑1-9 抑制磷脂酶動力學數據與抑制劑QSAR常數之三參數線性迴歸分析---------------------------------------------121
4-4磷脂酶酵素(PLA2 from Porcine Pancreas)的動力學數據探討124
4-4.1 磷脂酶(Porcine Pancreas)的QSAR數據處理-------------124
4-4.2 抑制劑1-9 抑制磷脂酶動力學數據與抑制劑QSAR常數之單參數線性迴歸分析---------------------------------------------125
4-4.3 抑制劑1-9 抑制磷脂酶動力學數據與抑制劑QSAR常數之雙參數線性迴歸分析---------------------------------------------135
4-4.4 抑制劑1-9 抑制磷脂酶動力學數據與抑制劑QSAR常數之三參數線性迴歸分析---------------------------------------------149
4-5 芳香族氨基甲酸類抑制劑1-9抑制磷脂酶之QSAR實驗數據製表152
4-5.1抑制劑1-9抑制磷脂酶(Naja)之QSAR實驗數據製表---------152
4-5.2抑制劑1-9抑制磷脂酶(Bee)之QSAR實驗數據製表----------154
4-5.3抑制劑1-9抑制磷脂酶(Porcine)之QSAR實驗數據製表------155
4-6 芳香族氨基甲酸類抑制劑抑制磷脂酶(PLA2)之機理探討-----157

第五章 利用理論計算方法應用在結構上的分析----------------159
5-1 理論計算的研究動機與目的-----------------------------159
5-2 理論化學原理介紹-------------------------------------160
5-2.1 量子化學的發展-------------------------------------160
5-2.2 計算化學的理論-------------------------------------163
5-2.2.1 第一原理方法-------------------------------------164
5-2.2.2 半經驗方法---------------------------------------168
5-2.2.3 分子力學方法-------------------------------------170
5-2.3 密度泛函理論---------------------------------------172
5-2.3.1 B3LYP理論----------------------------------------178
5-2.4 基底函數組-----------------------------------------179
5-2.5 Gaussian 程式發展史--------------------------------186
5-2.6 模組正確性-----------------------------------------189
5-2.7 Gaussian 03 模組中建立計算-------------------------190
5-2.7.1 單點能計算(Single Point Energy)------------------190
5-2.7.2 幾何優化(Geometry Optimization)------------------191
5-2.7.3 振動頻率分析(Frequency Analysis)-----------------191
5-3 抑制劑之理論化學計算結構分析-------------------------193
5-3.1 13C NMR的化學位移與能量的分析結果------------------195
5-3.1.1芳香族氨基甲酸類抑制劑理論計算數據與電子效應常數之關係圖-----------------------------------------------------195
5-3.1.2芳香族氨基甲酸類抑制劑最低能量與QSAR常數之關係圖--198
5-3.2.1芳香族氨基甲酸類抑制劑之動力學數據與理論計算數據之關係圖-----------------------------------------------------202
5-3.3 理論計算數據與動力學數據的結果討論-----------------209

第六章 結論與未來展望-----------------------------------210
6-1 結論-------------------------------------------------210
6-2 未來展望---------------------------------------------216

參考文獻-------------------------------------------------217

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