一类含磺胺哌嗪骨架的吲哚类衍生物作为微管蛋白聚合抑制剂的设计、合成及生物活性评价

发布时间：2018-06-24 20:37

本文选题：吲哚 + 微管蛋白聚合抑制剂　；参考：《南京大学》2016年硕士论文

【摘要】：微管作为纺锤体的重要组成部分,在细胞有丝分裂中扮演着重要的角色。在肿瘤细胞中某些与微管相关的蛋白表达异常,使其有丝分裂过程频繁且细胞周期变短,细胞增殖迅速。由此可见,微管蛋白在肿瘤细胞中起到了很大的作用,若破坏微管的动态循环,使得纺锤体形成受阻,肿瘤细胞有丝分裂过程中断且细胞周期受到阻滞,从而可以诱导肿瘤细胞的凋亡,达到抗肿瘤的目的。因此微管蛋白作为一个重要的靶点,越来越受到药学家们的关注,而且微管蛋白抑制剂已经成为临床治疗有效的抗肿瘤药物。吲哚类似物是一类含有苯并五元杂环结构的化合物,在对其进行大量的药物研发和生物活性研究中已证实具有抗肿瘤、抗菌抗病毒、止吐、降血压、抗炎抗抑郁等多种生物活性,因而吲哚类化合物在药物研究中受到广泛关注。在抗肿瘤作用机制研究中发现,吲哚类衍生物通过抑制或促进微管蛋白的聚合,干扰细胞的有丝分裂过程,从而达到抗肿瘤目的,并且某些化合物可通过抑制肿瘤调控激酶的活性而起到抗肿瘤效果。据研究磺胺类药物除具有广谱的抗菌活性,在抗肿瘤方面表现较好的生物活性。目前,部分磺胺类化合物作为治疗直肠癌、乳腺癌等的抗癌药物已经进入临床肿瘤治疗的研究中。哌嗪是分子结构中含有两个氮原子的六元杂环,为很多药物的重要组成部分。研究表明,作为增效基团,哌嗪骨架的引入可以使药物的药代动力学性质得到改善,并且能提高药物的生物活性及其水溶性。大部分的哌嗪类药物具有起效快、毒性低、副反应小等特点,可用于抗菌、抗氧化、抑制心血管、抗肿瘤等疾病。同时,哌嗪类化合物以其突出的生物活性而在作为抗肿瘤药物的研究中越来越受关注。本论文基于吲哚类、磺胺类及哌嗪类化合物的活性优势,设计、合成了一系列含磺胺哌嗪骨架的吲哚类衍生物作为抗肿瘤药物。我们运用Discovery Studio分子模拟对接软件对批量的含磺胺哌嗪骨架的吲哚衍生物进行筛选。从中我们筛选并且成功合成了24个目标化合物(7-30),所有的化合物结构都通过1H-NMR、13C-NMR、MS、单晶衍射等现代分析手段进行表征。我们对其生物活性进行了评价,通过细胞增殖抑制实验进行初筛,实验表明大部分的化合物具有良好的抑制活性,其中化合物18对HepG2、Hela、A549和MCF-7的增殖抑制活性最强,IC50值分别为0.49、0.24、0.35和0.59μM,优于对照药物秋水仙碱,显示了较好的抗肿瘤活性。并且对正常细胞肾上皮细胞293T进行了增殖抑制测试,得到CC50值。结果显示化合物的细胞毒性普遍较小,低于阳性对照药—秋水仙碱的CC50值。同时,体外微管蛋白聚合抑制实验的结果显示化合物18对微管蛋白聚合抑制效果最好,其IC50为1.82μM,优于对照药—秋水仙碱的抑制率。进一步细胞凋亡与周期实验显示化合物18能有效的诱导Hela细胞凋亡诱导并能够将细胞周期阻滞在G2/M期并且呈现剂量与时间的依赖性。免疫荧光实验证实化合物18可作用于微管蛋白并抑制其聚合。此外,用DS软件进行分子对接模拟的结果显示,化合物18和微管蛋白通过1个氢键和众多的分子间作用力结合,表现出最好的亲和性,这也在分子结构机制上为生物活性结果提供了一定的支持。
[Abstract]:Microtubule, as an important component of the spindle, plays an important role in cell mitosis. In the tumor cells, some proteins associated with microtubules are abnormal, which make the mitosis process frequent and the cell cycle shorten and the cell proliferation is rapid. Thus, microtubule egg white plays a great role in the tumor cells. The dynamic cycle of bad microtubules causes the formation of the spindle to be blocked, the cell mitosis is interrupted and the cell cycle is blocked, which can induce the apoptosis of the tumor cells and achieve the purpose of anti-tumor. Therefore, microtubulin, as an important target, has attracted more and more attention from pharmacists and microtubulin inhibitors already. It is an effective antitumor drug for clinical treatment. Indole analogues are a class of compounds containing five membered heterocyclic heterocyclic structures, which have been proved to have many biological activities, such as antitumor, antiviral, antiemetic, antiemetic, anti inflammatory and antidepressant, and indole compounds in drugs. In the study of the anti-tumor mechanism, it is found that indole derivatives interfere with the mitosis process of cells by inhibiting or promoting the polymerization of microtubule proteins, thus achieving anti-tumor purposes, and some compounds can inhibit tumor effects by inhibiting the activity of tumor regulated kinases. At present, some of the sulfonamides have entered the research of clinical cancer treatment as the treatment of rectal cancer and breast cancer. Piperazine is a six membered heterocyclic ring containing two nitrogen sources in the molecular structure, which is an important component of many drugs. Part. As a synergist, the introduction of piperazine framework can improve the pharmacokinetic properties of the drug and improve the bioactivity and water solubility of the drug. Most piperazine drugs have the characteristics of rapid onset, low toxicity and minor side effects, which can be used for antiseptic, antioxidation, cardiovascular, anti-tumor and other diseases. At the same time, piperazine compounds are becoming more and more concerned in the study of antitumor drugs with their prominent biological activities. Based on the active advantages of indoles, sulfonamides and piperazine compounds, a series of indole derivatives containing sulfamethazine skeleton are designed and synthesized as antitumor drugs. We use Discovery Studio We screened and screened 24 target compounds (7-30), and all the compounds were characterized by modern analytical methods such as 1H-NMR, 13C-NMR, MS, and single crystal diffraction. Preliminary screening of cell proliferation inhibition experiments showed that most of the compounds had good inhibitory activity, of which compound 18 had the strongest inhibitory activity on HepG2, Hela, A549 and MCF-7, IC50 was 0.49,0.24,0.35 and 0.59 M respectively, superior to the control drug colchicine, and showed good antitumor activity. The proliferation inhibition test and CC50 value were obtained in the epithelial cell 293T. The results showed that the cytotoxicity of the compound was generally small and lower than the CC50 value of the positive control drug colchicine. At the same time, the results of microtubule polymerization inhibition experiment in vitro showed that compound 18 had the best inhibition effect on microtubulin polymerization, and its IC50 was 1.82 mu M, superior to the control drug. The inhibition rate of colchicine. Further cell apoptosis and cycle experiments showed that compound 18 could induce apoptosis induction of Hela cells effectively and could block cell cycle in G2/M phase and show dependence on dose and time. The results of the row molecular docking simulation show that the combination of compound 18 and microtubulin through the combination of 1 hydrogen bonds and numerous intermolecular forces shows the best affinity, which provides a certain support for the biological activity results on the molecular structure.
【学位授予单位】：南京大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R91

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