苯甲基氟化磷酸酯類化學探針之開發

Development of Activity Probes based on Benzyl Fluorophosphonate

廖敏翔(Min-Shiang Liao)

指導教授 ： 羅禮強

國立臺灣大學/理學院/化學研究所/碩士(2009年)

https://doi.org/10.6342/NTU.2009.10325

摘要

龐大且具多樣性的蛋白質族群在許多人體生理反應中扮演著舉足輕重的角色,蛋白質失調經常牽涉到許多疾病的發生,例如癌症或糖尿病等等，而化學探針的發展則為蛋白質體學研究提供了有效的工具。
氟化磷酸酯是目前已知能有效標示絲胺酸水解酶的化學探針,其屬於親電性試劑,利用辨識端來進入特定蛋白質族群的受質結合區,使之能與其附近蛋白質上的親核基作用而在蛋白質催化活性部位形成共價鍵結,達到標示的目的。在本論文中,我們以苯甲基氟化磷酸酯為基礎,於其上進行一系列烷基之修飾，為了探討烷基長度與空間立體性質對於探針活性及選擇性的影響,我們合成了包含乙烷基、丁烷基、辛烷基以及環己烷基修飾的苯甲基氟化磷酸酯辨識端,並藉由具較高親水性的聚乙二醇衍生連接橋結合三種不同的發報端,分別有疊氮基團、生物素及螢光基團來構成不同的化學探針組合,這一系列苯甲基氟化磷酸酯類化學探針將可被應用於不同的生物測試策略。在此同時,我們也合成了由相同聚乙二醇衍生連接橋鏈結生物素發報端的氟化磷酸酯,進而與苯甲基氟化磷酸酯類化學探針進行對照比較。

關鍵字

化學探針 ； 苯甲基氟化磷酸酯 ； 烷基 ； 連接橋 ； 疊氮基團 ； 生物素 ； 螢光團

並列摘要

Numerous and diverse protein families play a very important role in a lot of human physiological processes. Disorder of proteins always involves in many diseases, such as cancer and diabetes, etc.. Therefore, the development of chemical probes has offered efficient tools for studying protein functions and structures.
It is known that fluorophosphonate is an effective chemical probe for serine hydrolases, which belongs to an electrophilic reagent and enters substrate binding site of specific protein with its recognition head. When being attacked by nearby nucleophile on the protein, there would form the covalent binding at protein active site to achieve labeling. In this thesis, we did modifications of alkyl groups upon benzyl fluorophosphonate. In order to study the influence of alkyl length and steric effect on activity and selectivity of probes, we synthesized benzyl fluorophosphonate recognition heads with modifications including ethyl, butyl, octyl and cyclohexyl. By way of connecting different kinds of tag like azido group, biotin and rhodamine with recognition heads separately through a higher hydrophilic ethylene glycol derived linker, a series of benzyl fluorophosphonate probes were made up. And they will apply to various biological testing strategies. At the same time, we also synthesized the known fluorophosphonate with biotin tag through an identical ethylene glycol derived linker. And then carry on contrast to compare with the benzyl fluorophosphonate chemical probes.

並列關鍵字

chemical probes ； benzyl fluorophosphonate ； alkyl groups ； linker ； azido group ； biotin ； rhodamine

參考文獻

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