- 學位論文
設計與合成喹唑啉酮類化合物做為組蛋白去乙醯酶抑制劑
Design and Synthesis of Quinazolinone Derivatives as Potential Histone Deacetylase (HDAC) Inhibitors
摘要
本論文主旨為設計與合成喹唑啉酮衍生物作為組蛋白去乙醯酶抑制劑進而作為具有潛能的抗癌試劑，甚至神經退化性疾病。 第一部分以喹唑啉-2,4-酮作為核心結構，並在其上加入該酵素表面作用官能基及鋅離子螫合基團。首先將鄰氨基苯甲酸或靛红酸酐經羰基化及環化反應後得到帶有酵素表面作用官能基的喹唑啉-2,4-酮主結構，之後將該主結構依序透過親核取代反應以及海克(Heck)偶合反應合成4-甲基肉桂酸的衍生物，最後含羧酸類官能基的中間體再被轉換成帶有酰基羥胺的目標化合物7a-e, 14a-g和18a-c；同時也利用了微波反應使得實驗進行得更有效率且方便。 根據酵素抑制實驗結果顯示，喹唑啉-2,4-酮衍生化合物7a-e, 14a-g和18a-c皆對第一、八、十一亞型的組蛋白去乙醯酶達到μM的抑制活性，且對第六亞型的組蛋白去乙醯酶有稍微的選擇性在sub-μM的效果，其相對於第一型組蛋白去乙醯酶多出8至107倍的抑制活性。接著透過細胞毒性的測試發現，喹唑啉-2,4-酮衍生化合物7a-e, 14a-g和18a-c對不同種類的癌細胞(胸、肺、胰臟、白血球)可達到μM的毒殺效果，化合物14c和14f更是可以達到sub-μM的毒殺效果。另外透過細胞轉移實驗，發現喹唑啉-2,4-酮衍生化合物可降低癌細胞的轉移能力且不影響正常細胞傷口癒合的情性。接著透過半胱氨酸天冬氨酸蛋白酶-3實驗發現喹唑啉-2,4-酮衍生化合物可以增加半胱氨酸天冬氨酸蛋白酶-3的活性。 第二部分根據Tubastatin A(高選擇性第六亞型之組蛋白去乙醯酶抑制劑)的結構，我們以咪唑喹唑啉-2-酮這種三環結構作為主架構。之後將該主結構透過親核取代反應與帶有酯類官能基的側鏈接上，再將酯類官能基轉換成帶有酰基羥胺的目標化合物26。 根據酵素抑制與細胞毒性實驗，化合物26表現出顯著的第六亞型組蛋白去乙醯酶抑制活性 (HDAC6 IC50 = 3.5 nM, HDAC1 IC50 = 1540 nM) 及相較於第一亞型組蛋白去乙醯酶多出446倍的選擇性且化合物26的IC50甚至低於Tubastatin A，另外透過細胞毒性測試化合物26對許多不同種癌細胞具低毒殺效果，此外化合物26在PC12細胞上也具有出促進神經軸突生長能力的效果。
並列摘要
Base on general structure of HDAC inhibitors, the aim of this thesis is the design and synthesis of quinazolinone derivatives as HDAC inhibitors for the treatment of variety of cancers such as breast, lung, pancreas and leukemia, and even neurodegenerative disorder. In the first part, quinazolin-2, 4-dione core structure with different substitutions and length were connected with para-methyl styrene as the linker to attach hydroxamic acid group, finally afforded the target compounds 7a-e, 14a-g and 18a-c. Besides, microwave irradiation was used to make experiments more convenient and more efficient. From the enzyme inhibitory data, compounds 7a-e, 14a-g and 18a-c showed micromolar (μM) range inhibition against HDAC1, 8, 11. However, all the compounds revealed the selectivity to HDAC6 at sub-μM range and the ratio against HDAC6 over HDAC1 is about 8 to 107-fold. In addition, compounds 7a-e, 14a-g and 18a-c were also tested on cytotoxicity assay. Through the MTT assay with different cancer cell lines, compounds 7a-e, 14a-g and 18a-c showed μM or sub-μM range activity against variety of cancer cells. Furthermore, compounds 7a, 14a, 14e-g and 18a-c were tested on the anti-migration analysis and Caspase-3 study, the data showed that our compounds can inhibit the migration on A549 cancer cells without affecting the wound healing on normal cell and increase the activation of Caspase-3, respectively. In the second part, based on the studies of Tubastatin A, a highly selective HDAC6 inhibitor, It was designed and synthesized this kind of tricyclic structure with hydroxamic acid moiety as a novel potent selective HDAC 6 inhibitor. In this study, starting from imidazo [1,2-c]quinazolinone core structure which was previously defined in our lab. Further it was connected with benzyl group as the linker and attached with hydroxamate moiety to obtain our target compound 26. From enzyme inhibitory data, compound 26 showed a high selectivity to HDAC6 at nanomolar (nM) range and the ratio for HDAC6 over HDAC1 is up to 446-fold, and the IC50 for HDAC6 is even lower than Tubastatin A. Further, the cytotoxicity data showed that compound 26 revealed low cytotoxic effect on a variety of cancers. In addition, compound 26 can significantly stimulated the neurite outgrowth in PC12 cell.
參考文獻
延伸閱讀
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