【作者】 陈静；

【导师】 胡永洲；

【作者基本信息】 浙江大学 ， 药物化学， 2009， 博士

【摘要】 恶性肿瘤是严重危害人类健康的一大类疾病,已逐渐取代心脑血管疾病成为全球头号杀手。目前临床上常用的抗肿瘤药物普遍存在对实体瘤疗效较差、毒副作用较大、容易产生多药耐药等缺点。因此,研究开发更为高效低毒的新型抗肿瘤药物仍是新药研究的热点之一。抗有丝分裂剂是一类有效的抗肿瘤药物,而微管蛋白是抗有丝分裂药物的一个重要靶点。近年来,微管蛋白的秋水仙碱位点抑制剂（CSIs）由于结构简单而备受关注。本文根据文献报道的CSIs的共同结构特征,利用理性药物设计原理,结合该位点抑制剂中常出现的吲哚结构单元,引入具有抗肿瘤活性的平面结构咔啉和取代芳环分别作为两个疏水平面;以微管蛋白抑制剂中常出现的磺酰胺、酰胺、甲酰基等作为连接桥链,构建了该位点抑制剂的基本骨架。根据这一设计思想,共合成了三类112个全新的咔啉类衍生物,并对其进行了体外抗肿瘤活性筛选。结果表明,三种母核对活性的影响为:γ-咔啉＞β-咔啉＞α-咔啉;连接桥链一般以磺酰胺和甲酰基更有利于抗肿瘤活性的增强。进而选择其中活性较好的代表性化合物进行了初步的抗肿瘤作用机制研究:微管蛋白聚合抑制试验表明,14个受试化合物均能抑制微管蛋白的聚合;DNA插入实验表明,大部分γ-咔啉类化合物（1-34、1-37、1-74、1-87、1-109）对DNA均无明显作用,而在芳磺酰胺γ-咔啉类化合物1-34连接链的氮原子上引入乙基的化合物1-35,以及具有大平面结构的苯并[1,2-i]γ-咔啉类衍生物（1-115）和3位有碱性基团取代的β-咔啉类衍生物（1-161）均能通过插入DNA双螺旋碱基对置换出溴化乙锭（EB）,使EB-DNA结合物的荧光明显减弱;细胞周期影响实验表明,γ-咔啉类化合物1-34、1-74、1-87、1-109对人非小细胞肺癌细胞（A549）的G₂/M期有明显的阻滞作用。同时,基于1,2,3,4-四氢咔唑-1-酮类化合物具广泛的生物活性,尤其在抗肿瘤活性方面的作用,我们利用分子杂合原理,对其进行结构改造。通过Mannich反应在母核的2位引入取代甲氨基,合成了17个四氢咔唑-1-酮Mannich碱类化合物,以探讨2位Mannich碱的种类以及N⁹位不同取代基对活性的影响。初步体外抗肿瘤活性测试实验结果表明,在其2位引入二乙氨基甲基或4-甲基哌啶甲基,以及在N⁹位引入异戊烯基有利于活性的提高。代表化合物2-3的体外微管聚合抑制试验表明,该化合物也能抑制微管蛋白的聚合。本文还利用近年来发展较快的微波合成技术对Bischler吲哚合成法和Fischer吲哚合成法进行了改进,首次报道了微波促进的2-取代吲哚、1,2,3,4-四氢咔唑以及γ-咔啉等重要医药中间体的合成。为这三类化合物的合成提供了一种快速、方便、有效的方法,扩大了微波技术在杂环化合物合成中的应用。更多还原

【Abstract】 Cancer remains a significant health concem,and has gradually been the first killer in the world instead of cardiovascular diseases.The most clinically effective anticancer agents are associated with problems of low efficacy against solid tumors,high toxicity, and development of multidrug resistance.Therefore,it remains a hotspot for medicinal chemist to develop novel antieancer agents with high efficacy and low toxicity.Antimitotic agent is one of the most successful classes of anticancer drugs,and microtubule is recognized as an important target for the development of potential antimitotic agents.Recent years,the colchicine binding site inhibitors（CSIs）have attracted a great deal of attention because of their simple structures and potent activities. Based on the common structural features of CSIs reported in the literature,and the finding that indole moiety presented in many CSIs,we utilized rational drug design principle,introduced carboline,an planar antitumor moiey,and an aromatic ring as two hydrophobic planes.Sulfonamide,carbonyl,or amide group which presented in many potent tubulin polymerization inhibitors were used as the linker.Thus,we constructed the basic structure of CSIs,and synthesized three series of novel carboline derivatives. The total 112 compounds were tested for their cytotoxic activities in vitro against several human tumor cell lines.Primary SAR study indicated that the efficacy of three carboline moieties on antitumor activity was:γ-carboline＞β-carboline＞α-carboline. Utilizing of sulfonamide or carbonyl group as the linker was more beneficial for the activities.In order to gain insight on the mechanisms of action of these compounds,we chose the potent cytotoxic and representative compounds for further assay.Tubulin polymerization inhibition assay showed that all the 14 tested compounds exhibited potential inhibitory effect against microtubule.DNA intercalation assay indicated that most of theγ-carboline derivatives（1-34、1-37、1-74、1-87、1-109）could not interact with DNA except for compounds 1-35.Besides,benzo[1,2-i]γ-carboline derivatives （1-115）and 3-basic group substituted-β-carboline derivatives（1-161）could also intercalate into DNA and reduce the fluorescence of EB-DNA complex.We also tested the effects ofγ-carbolines 1-34、1-74、1-87、1-109 on cell cycle of A549 cell line.It clearly demonstrated that all these compounds arrested the cell cycle on G₂/M phase. Based on the extensive biological activities of 1,2,3,4-tetrahydrocarbazole-1-ones, especially their antitumor activity,we utilized the molecular heterozygosity principle and synthesized a series of 2-substituted aminomethyl-9-alkyl-1,2,3,4-tetrahydrocarbazole -1-ones through mannich reaction.The total 17 1,2,3,4-tetrahydrocarbazole -1-one mannich bases were test for their cytotoxic activities in vitro against several human tumor cell lines.The results implied diethylaminomethyl or 4-methylpiperidin methyl moiety was the most suitable mannich base in this series of compounds,and the presence of isoprenyl group at N⁹ was beneficial for the activities. Primary mechanism research of compound 2-3 indicated it exhibited a potential inhibitory effect against microtubule.In the last part of my dissertation,we described the application of microwave irradiation in organic synthesis.We found a practical and simple microwave enhanced Bischler reaction for the synthesis of 2-phenyl-indoles or 1,2,3,4-tetrahydrocarbazoles, and a microwave enhanced Fischer reaction for the synthesis ofγ-carbolines.These new approaches provided simple,fast,and efficacy methods for the construction of these important pharmaceutical active structures,and extended the microwave irradiation utility in heterocyclic chemistry.更多还原