喹唑啉酮類化合物作為組蛋白去乙醯脢抑制劑之設計 合成及其生物活性研究

Design, Synthesis, and Biological Evaluation of Quinazolinone Derivatives as Histone Deacetylase (HDAC) Inhibitors

余兆武(Chao-Wu Yu)

指導教授 ： 陳基旺

國立臺灣大學/醫學院/藥學研究所/博士(2012年)

https://doi.org/10.6342/NTU.2012.10898

摘要

本論文主旨為設計與合成喹唑啉酮衍生物作為選擇性第六型組蛋白去乙醯酶抑制劑及其治療退化性神經疾病如阿茲海默症、巴金森氏症及杭丁頓舞蹈症等之可行性之研究。
以喹唑啉-2-酮及喹唑啉-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。首先將鹵化的苯胺或鄰氨基苯甲酸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-酮及喹唑啉-2,4-酮主結構。之後將該主結構透過海克(Heck)及根岸(Negishi) 等偶合反應與帶有酯類官能基的側鏈接上。最後含有酯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。
根據酵素抑制實驗，化合物3f (HDAC6 IC50 = 29 nM, HDAC1 IC50 = 1880 nM), 3i, (HDAC6 IC50 = 11 nM, HDAC1 IC50 = 1190 nM), 3j (HDAC6 IC50 = 33 nM, HDAC1 IC50 = 3200 nM), and 7d (HDAC6 IC50 = 7 nM, HDAC1 IC50 = 6970 nM) 等顯現出一位至二位數nM的第六型組蛋白去乙醯酶抑制活性及60至1000倍相對於第一型組蛋白去乙醯酶的選擇性。經過構效關係研究後發現，帶有鋅離子螫合基的側鏈在喹唑啉-2-酮的第六及第七位置或在喹唑啉-2,4-酮的第一位置時表現出較好的第六型組蛋白去乙醯酶抑制活性及相對於其他亞型去醯酶的選擇性。當乙基及苯乙基分別在喹唑啉-2-酮的第二及第三位置時顯現出相對於其他官能基較好的抑制活性。另一方面，化合物系列3也被發現具有抑制Aβ聚合的能力。
最後，目標化合物也經過癌細胞及神經細胞的篩選，發現目標化合物並沒有顯著的細胞毒性或神經毒性。而且，化合物系列3也帶有兩位數或一位數μM以下的誘導神經細胞軸突生長的能力。其中化合物3c及3f被進一步地在Aβ病變的小鼠動物模式上試驗，並顯著地改善其在滾筒式跑步機 (rotar rod)上的表現。
另外，在合成目標化合物時，也發現並建立了針對氯苯衍生物以微波加速的新的鈀金屬催化的羥基化反應。

關鍵字

組蛋白去乙醯脢 ； 抑制劑

並列摘要

The aim of this thesis is design and synthesis of quinazolinone derivatives as selective HDAC6 inhibitors for the potential treatment of neurodegenerative diseases such as Alzheimer, Parkinson, and Huntington diseases.
The quinazolin-4-one and quinazolin-2, 4-dione were employed as core structure tethered with surface contacting cap group and zinc binding group (ZBG). Halogenated aniline or anthranilic acid derivatives were carbonylated and cyclized to give quinazolin-4-one and quinazolin-2, 4-dione core structures with surface contacting cap group. Then these core structures were coupled with various side chain containing ester group through Heck, Negishi, or substitution reactions. Finally, the ester groups at the end of side chain were transformed to hydroxamic acid as ZBG to give target compounds.
From the results of enzyme based assay, compound 3f (HDAC6 IC50 = 29 nM, HDAC1 IC50 = 1880 nM), 3i, (HDAC6 IC50 = 11 nM, HDAC1 IC50 = 1190 nM), 3j (HDAC6 IC50 = 33 nM, HDAC1 IC50 = 3200 nM), and 7d (HDAC6 IC50 = 7 nM, HDAC1 IC50 = 6970 nM) showed one to two digit nM activity against HDAC6 and 60 to 1000 times selectivity over HDAC1. From SAR studies, the zinc binding side chain at C-6 and C-7 position of quinazolin-4-one or at N-1 position of quinazolin-2, 4-dione provided better activity against HDAC6 and selectivity over other HDACs. Ethyl group at C-2 position and phenethylene group at N-3 position gave optimal activity against HDAC6. On the other hand, compounds of series 3 also showed anti-amyloid aggregation activity in Zn dependent and Zn independent conditions.
Furthermore, compounds of series 3 showed two digit to sub-micromolar EC50 in neurite outgrowth assay of PC12 and SH-SY5Y cells, and most of them showed no significant cytotoxicity or neurotoxicity. Compound 3c and 3f were tested with Aβ-lesioned mice on rotar-rod performance model and displayed significant improvement on rotar-rod performance after treatment of two weeks.
In addition, during the synthetic study, a new microwave accelerated palladium-catalyzed hydroxylation of aryl chlorides was also developed.

並列關鍵字

Histone deacetylase ； Inhibitor

參考文獻

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