【作者】 叶青；

【导师】 胡永洲；

【作者基本信息】 浙江大学 ， 药物化学， 2009， 博士

【摘要】 恶性肿瘤严重威胁人类健康,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、毒副作用大、容易产生多药耐药性等缺点。因此,研究疗效好,副作用小的抗肿瘤药物仍然是新药研究的热点之一。抗肿瘤药物的作用靶点有许多,如拓扑异构酶I,蛋白激酶C和细胞周期蛋白依赖性激酶等。蛋白激酶C(PKC)是一大类磷脂依赖的丝/苏氨酸激酶,它在细胞的增殖调控、恶性转化以及癌变过程中起重要作用,是抗肿瘤药物设计的一个重要靶点。目前已有多个PKC抑制剂如UCN-01,CGP 41251和Enzastaurin作为抗肿瘤药物已进入临床试验。据文献报道,双吲哚马来酰亚胺类化合物具有显著的PKC抑制活性。为了获得具有更强的PKC抑制活性和抗肿瘤活性的新化合物,本文以双吲哚马来酰亚胺类化合物为先导化合物,对其进行结构改造,共合成了四类86个具有全新结构的二氮杂芳基马来酰亚胺类化合物,其中包括51个7-氮杂吲唑-吲哚马来酰亚胺类化合物,17个4-氮杂吲哚-吲哚-马来酰亚胺类化合物,8个3-(吡咯-3-基)-4-芳基马来酰亚胺类化合物和10个3-氯-4-吲哚马来酰亚胺类化合物,并对它们进行了体外抗肿瘤活性筛选。结果表明,7-氮杂吲唑-吲哚-马来酰亚胺类化合物和3-氯-4-吲哚马来酰亚胺类化合物与4-氮杂吲哚-吲哚-马来酰亚胺类化合物和3-(吡咯3-基)-4-芳基马来酰亚胺类化合物相比显示了较强的抗肿瘤活性。进而我们选择了部分代表性的化合物进行了初步的抗肿瘤机制的研究,结果表明:受试的大部分化合物显示了中等强度的PKC抑制活性,但活性顺序与它们的抗肿瘤活性并不一致,说明化合物除了PKC抑制作用,可能存在着其它作用机制。对代表性化合物a-14的进一步研究表明,促进肿瘤细胞的凋亡是其具有抗肿瘤活性的另一作用机制。糖原合成酶-3p(GSK-3p)是双吲哚马来酰亚胺类化合物的另一个重要的作用靶点。GSK-3B在非胰岛素依赖型糖尿病(NIDDM)中,其活性会异常升高,从而抑制细胞糖原合成酶活性,降低细胞糖原合成水平。GSK-3β在阿尔茨海默尔氏病中,会导致Tau蛋白过度磷酸化,引起神经元纤维缠结。GSK-3β抑制剂对糖尿病和阿尔茨海默尔氏病(AD)的治疗都有重要的价值。本文对前期合成的部分二氮杂芳基马来酰亚胺类化合物进行了GSK-3β抑制活性的测试,并根据初步的构效关系进一步设计合成了16个结构全新的二氮杂芳基马来酰亚胺类化合物。结果表明,7-氮杂吲唑-吲哚-马来酰亚胺类化合物,4-氮杂吲哚-吲哚-马来酰亚胺类化合物和3-(吡咯-3-基)-4-芳基马来酰亚胺类化合物中的大部分化合物均显示了很强的GSK-3β抑制活性。其中部分化合物a-36,a-38～a-40,a-49,a-50,b-18,b-23,c-2和c-3能够显著的减少A-beta淀粉样蛋白诱发神经元tau蛋白的过度磷酸化,在细胞水平显示了GSK-3B的抑制活性。更多还原

【Abstract】 As a significant health concern to human beings, cancer may soon displaced cardiovascular diseases as the leading killer. Although great efforts have been devoted to the discovery of anticancer agent, even the most widely used anticancer agents bear drawbacks, such as low efficacy against solid tumors, high toxicity, and development of multidrug resistance. Thus, research on anti-tumor drugs with higher efficacy, lower toxicity and minimum side effects has been a hotspot in the field of drug discovery.Various targets including topoismerase I, protein kinase C and Cyclin-dependent Kinase have been validated for cancer chemotherapy. Among them, protein kinase C (PKC), a family of serine/threonine kinases, is regarded as a suitable target in anticancer drug design for its close connection with neoplasias. PKC inhibitors such as UCN-01, CGP 41251 and Enzastaurin are in clinical trials for their potential role in cancer therapy. Bisindolylmaleimides were reported to exhibit remarkable PKC inhibition. In this paper, we designed and synthesized 86 novel bisazaarylmaleimides, which could be group into four series: 51 7-azaindazole-indole-maleimides, 17 4-azaindole-indole-maleimides, 8 3-(pyrolle-3-yl)-4-arymaleimides and 10 3-chloro-4-indolemaleimides, in an attempt to discover novel PKC inhibitors with improved activity. All 86 synthesized compounds were tested for their in vitro antiproliferative activities against several human tumor cell lines, the result of which showed that 7-azaindazole-indole-maleimides and 3-chloro-4-indolemaleimides exhibited more potent antiproliferative activity than 4-azaindole-indole-maleimides and 3-(pyrolle-3-yl )-4-arymaleimides. Other biological evaluation studies involving some representive compounds were conducted to probe the mechanisms of these compounds. The fact that moderate PKC inhibitory activity for most tested compounds were observed was not consistent with the result of antiproliferative activity against human cancer cell lines, indicating that the tested compounds might have other mechanism against human cancer cell lines besides PKC inhibition. Further mechanism studies revealed that the antiproliferative activity of compound a-14 in tumor cells might be explained by its connection with the mitochondria-mediated apoptosis pathway.GSK-3βis another target for bisindolylmaleimides. The high level of GSK-3βin NIDDM inhibits the activity of glycogen synthase kinase. GSK-3βleads to hyperphosphorylation of Tau protein, inducing neurofibrillary tanglesn. Thus, GSK-3βinhibitors are valuable agents for the treatment of diabetes and Alzerheim’s Disease. Guided by the GSK-3B inhibitorary activity tests on some bisazaarylmaleimides synthesized in the preliminary phase, 16 novel inhibitors were designed based on preliminary QSAR information. The results showed that most 7-azaindazole-indole-maleimides, 4-azaindole-indole-maleimides and 3-(pyrolle-3-y 1 )-4-arymaleimides exhibited potent inhibitory activity against GSK-3β, among which compounds a-36, a-38～a-40, a-49, a-50, b-18, b-23, c-2 and c-3 significantly reduced Aβ-induced Tau hyperphosphorylation, demonstrating GSK-3βinhibitory activity at cell level.更多还原