【作者】 李伯玉；

【导师】 李志良； 李志斌；

【作者基本信息】 重庆大学 ， 分析化学， 2003， 硕士

【摘要】 组蛋白通常以复合物形式参与遗传信息转录激活和转录抑制，是重要癌变靶标，通常以乙酰化和去乙酰化两形式存在。组蛋白乙酰化由组蛋白乙酰化酶(HAT)引发并与转录激活有关；相反组蛋白去乙酰化由组蛋白去乙酰酶(HDAC)催化并与转录抑制有关。抑制组蛋白去乙酰酶便成为一种新颖干扰细胞周期控制方法，故组蛋白去乙酰酶抑制剂对治疗细胞增生疾病如癌症很有潜力。迄今已报道许多类型HDAC抑制剂如丁酸钠，Trichostain A和Trapoxin等，具诱发癌细胞系分化作用进而抑制癌细胞增生。本论文以开发新颖的组蛋白去乙酰酶抑制剂类抗癌药物为主线，依据先导物MS-275设计了四个系列的苯甲酰胺衍生物，设计了酶结合区内不同芳基取代、芳环上不同电性取代，酶抑制活性区内不同活性官能团取代等。经结构简化、筛选设计并合成分析了四系列苯甲酰胺类衍生物，深入讨论分子结构与HDAC抑制性和诱变细胞功能活性之间的关系。主要内容及结论说明如下：第一部分为苯甲酰胺衍生物合成与分析。前三系列以芳甲醇为起始原料，与1,1’-碳酰二咪唑在0~10℃温度下搅拌反应得到酰基咪唑胺活性中间体，再与胺交换制得4-[N-(取代芳基－甲氧基碳酰基)-氨甲基]苯甲酸中间体，收率60～84.1%。继与氯甲酸已酯及三乙胺在0~5℃温度下搅拌反应得活性酸酐中间体或与1,1’-碳酰二咪唑在0~10℃下搅拌反应制活性酰基咪唑胺中间体；两种活性中间体都可不经分离直接分别与邻苯二胺或邻氟苯胺或1,2-二氨基环己烷在室温条件下反应约10h，经硅胶色谱柱分离制得目标产物，收率在18-59％。通过质谱、紫外光谱、红外光谱及核磁共振氢谱对新合成化合物结构进行分析鉴定。第四系列则经三步反应得到：首先采用克努文反应生成丙稀酸衍生物，后两步类同前三系列合成。第二部分为分子结构与生物活性的构效关系分析。结构与活性关系研究发现在酶抑制区内活性官能团2’-氨基苯基被2’-氟苯基或2’-氨基环己烷基取代后导致抑制活性降低和丧失，表明2’-氨基苯基在酶抑制区的活性点部位起氢键或其它电性作用，该基团是产生抑制活性的必需条件。在疏水区芳基上取代基的电子效应对HDAC抑制活性影响甚小；大多数化合物在细胞中诱发组蛋白乙酰化能力与之对酶抑制活性能力密切相关。在此类苯甲酰胺类衍生物中，系列四中N-(2-氨基苯基)-4-[N-(吡啶-3-丙烯酰基)氨甲基]苯甲酰胺表现出很好的离体HDAC抑制活性和多种癌细胞杀伤活性，选作深入开发。更多还原

【Abstract】 Histone possesses two functions of both acetylation and deacetylation: the former is caused by histone acetylase (HAT) often associated with activated transcription, whereas the latter is catalyzed by histone deacetylase (HDAC) related to transcriptional silencing. Inhibition of HDAC activity represents a novel approach for intervening in cell cycle regulation; thus HDAC inhibitors possess great therapeutic potential in treatment of cell proliferative diseases. Several classes of HDAC inhibitors such as sodium butyrate, trichostain and trapoxin were reported to induce differentiation of several cancer cell lines and suppress cell proliferation. In this M.Sc. thesis taking innovation of innovate novel HDAC inhibitors as the mainline, four series of structurally modified benzamide derivatives with a postulated general structure were designed according to the lead compound MS-275. Four new series of benzamides were prepared through variation of substituted aryls, aryl substitutions, chain connections and functional groups. Both synthesis and analysis of these target compounds were done and the structure-activity relationships (SAR) were stated for HDAC inhibition and alterations in cellular function. The main contents and some results are illustrated as following:Four series of benzamide derivatives were synthesized and analyzed. The intermediate 4-[N-(substituted aryl-methoxycarbonyl)aminomethyl]-benzoic acids were prepared from substituted aryl methanols under the temperature of 0~10℃ with 60-84.1% yields, then converted to the active anhydrides with ethyl chloroformate and triethylamine under 0~5℃, or to carboxyl imidiazole with 1,1’-carbonylimidiazlole under 0~10℃, and then reacted with o-phenlenediamine or 2-fluorophenylamine, under room temperature for about 10h without further separation, or cyclohexyl-1,2-diamine separated by silica gel column chromatography to give target compounds in yield of 18-59%. Structural analysis was done by mass spectrometry, nuclear magnetic resonance, ultraviolet and infrared spectroscopy. The forth series of benzamides were synthesized by a green chemical strategy. Structure-activity relationship on the functional group moiety in HDAC inhibitors showed that substitution of 2’-amino(fluoro)phenyl or 2’-aminocyclohexyl resulted in decreasing or losing inhibitory activity. Results indicate that the 2’-aminophenyl group<WP=6>in enzyme inhibiting regions which might act as a hydrogen-bonding or electrostatic interaction, was indispensable for inhibitory activity. The electronic influence of the aryl substituent in the hydrophobic moiety seemed to play a small role for HDAC inhibitory activity. In addition, the ability of most compounds to induce histone acetylation in cells correlated largely with their enzyme inhibitory activities in vitro against HDAC1. N-(2-aminophenyl)-4-[N-(pyridin-3-acryl aminomethyl)- benzamide in series 4 showed good HDAC inhibitory activity and pretty potent antiproliferative activities in vitro against various human cancer cell for further development.更多还原