新型EGFR抑制剂喹唑啉衍生物的合成及抗癌活性研究

发布时间：2018-04-30 13:07

本文选题：表皮生长因子受体 + 抑制剂　；参考：《苏州大学》2014年硕士论文

【摘要】：本论文内容主要分为两个部分。第一部分为新型EGFR抑制剂的合成及其抗癌活性研究。表皮生长因子受体酪氨酸激酶（Epidermal growth factor receptor tyraminekinase, EGFR-TK）是细胞激酶家族的一员，通常在癌变细胞中有异常表达，是肿瘤治疗的重要靶点。论文中合成了丙烯酸酰胺类、二胺类、罗丹宁衍生物类三个系列共34个在6位胺基上取代的4-苯胺基喹唑啉衍生物作为新型的表皮生长因子受体(Epidermalgrowth factor receptor, EGFR)抑制剂。并进行了MTT细胞水平活性，酶抑制分子水平活性，分子对接模拟实验分析，细胞凋亡实验的研究。部分化合物显示出良好的抗细胞增殖活性以及对EGFR的有效抑制作用。其中，N6-((5-溴噻吩-2)甲基)-N4-(3-氯苯基)喹唑啉-4,6-二胺，即化合物11e，表现最为突出。其对HepG2,A549的IC50分别为3.11μM及0.82μM。分子对接模拟分析显示这些化合物能够很好的与EGFR的细胞内ATP结合区域竞争性结合。赫斯特细胞形态染色实验表明这些化合物能够有效的引起A549细胞的凋亡。研究为获得更有效的EGFR小分子抑制剂提供了实验依据。第二部分为莱克多巴胺单克隆抗体的制备。莱克多巴胺(Ractopamine, RAC)是一种含有双苯烷胺类结构的β-肾上腺素受体激动剂，与瘦肉精属同类化合物。莱克多巴胺被非法用于动物饲养，目的是提高瘦肉率，大量食用含有莱克多巴胺残留的肉类或内脏后，可能引发中毒症状，，严重威胁到消费者的生命健康。目前莱克多巴胺的检测方法主要依靠色谱分析方法，色谱分析方法虽然比较灵敏，准确度高，但需要大型仪器支持，需要繁琐的样品前处理以及专业的人员操作，导致不能适应市场对分析方法快速、简便的要求。因此，建立一种能够快速、简便、准确检测莱克多巴胺在肉类食品中残留的分析方法显得至关重要。本论文合成了莱克多巴胺的类似物作为半抗原，并使其与BSA偶联成完全抗原后用于Balb/c小鼠的免疫。经过一定的免疫程序后通过间接ELISA实验对受免小鼠血清进行分析，筛选适宜用于细胞融合的小鼠进行细胞融合，制备抗莱克多巴胺单克隆抗体，从而建立用于检测食品中莱克多巴胺药物残留的免疫分析方法。细胞融合及后续实验正在进行中。所有目标化合物及部分中间体经1H NMR、13C NMR、HR-MS分析确证。
[Abstract]:The content of this paper is divided into two parts.
The first part is the synthesis and anticancer activity of new EGFR inhibitors.
Epidermal growth factor receptor tyrosine kinase (Epidermal growth factor receptor tyraminekinase, EGFR-TK) is a member of the cell kinase family. It is usually abnormal in cancerous cells and is an important target for cancer treatment.
In this paper, acrylic amide, two amines, Luo Danning derivatives, three series of Luo Danning derivatives and 34 substituted aniline quinazoline derivatives on 6 amino groups were synthesized as a new type of Epidermalgrowth factor receptor (EGFR) inhibitor. The level activity of MTT cells, the enzyme inhibition molecular level activity, and the molecular level activity were carried out. Some compounds showed good anti cell proliferation activity and effective inhibition to EGFR. Among them, N6- ((5- bromo thiophene -2) methyl) -N4- (3- chlorophenyl) quinazoline -4,6- two amine, that is, compound 11e, was the most prominent. The IC50 of HepG2, A549 was 3.11 mu M and 0.82 micron respectively. Molecular docking simulation analysis showed that these compounds were able to compete well with the intracellular ATP binding region of EGFR. Hearst cell morphology staining experiments showed that these compounds could effectively induce apoptosis of A549 cells. The study provided an experimental basis for a more effective EGFR small molecule inhibitor.
The second part is the preparation of monoclonal antibody against ractopamine.
Ractopamine (RAC) is a beta adrenoceptor agonist containing a diphenyl amine structure, a similar compound with lean meat. Lek dopamine is illegally used in animal breeding to improve lean meat rates. A large number of meat or viscera containing leecdopamine residues may cause toxic symptoms, Yan Zhongwei. At present, the methods of detecting the life and health of the consumers are mainly depended on the method of chromatographic analysis. Although the chromatographic analysis method is more sensitive and accurate, it needs large instrument support. It needs tedious sample pretreatment and professional personnel operation, which can not adapt to the rapid and simple requirement of the analysis method. Therefore, it is very important to establish a rapid, simple and accurate method for the determination of ractopamine residues in meat products.
In this paper, the analogue of ractopamine was synthesized as a semi antigen, and it was used as a complete antigen with BSA to be used to immunization of Balb/c mice. After a certain immunization program, the serum of the mice was analyzed by indirect ELISA test, and the cells suitable for cell fusion were screened for cell fusion and the preparation of RC dopamine was prepared. Monoclonal antibodies have been established to establish an immunoassay for the detection of ractopamine residues in foods. Cell fusion and subsequent experiments are in progress.
All target compounds and some intermediates were confirmed by 1H NMR, 13C NMR and HR-MS analysis.

【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R914.3;R965

【参考文献】