本文選題：表皮生長(zhǎng)因子受體 + 抑制劑��； 參考：《蘇州大學(xué)》2014年碩士論文

【摘要】：本論文內(nèi)容主要分為兩個(gè)部分。 第一部分為新型EGFR抑制劑的合成及其抗癌活性研究。 表皮生長(zhǎng)因子受體酪氨酸激酶（Epidermal growth factor receptor tyraminekinase, EGFR-TK）是細(xì)胞激酶家族的一員，通常在癌變細(xì)胞中有異常表達(dá)，是腫瘤治療的重要靶點(diǎn)。 論文中合成了丙烯酸酰胺類、二胺類、羅丹寧衍生物類三個(gè)系列共34個(gè)在6位胺基上取代的4-苯胺基喹唑啉衍生物作為新型的表皮生長(zhǎng)因子受體(Epidermalgrowth factor receptor, EGFR)抑制劑。并進(jìn)行了MTT細(xì)胞水平活性，酶抑制分子水平活性，分子對(duì)接模擬實(shí)驗(yàn)分析，細(xì)胞凋亡實(shí)驗(yàn)的研究。部分化合物顯示出良好的抗細(xì)胞增殖活性以及對(duì)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，表現(xiàn)最為突出。其對(duì)HepG2,A549的IC50分別為3.11μM及0.82μM。分子對(duì)接模擬分析顯示這些化合物能夠很好的與EGFR的細(xì)胞內(nèi)ATP結(jié)合區(qū)域競(jìng)爭(zhēng)性結(jié)合。赫斯特細(xì)胞形態(tài)染色實(shí)驗(yàn)表明這些化合物能夠有效的引起A549細(xì)胞的凋亡。研究為獲得更有效的EGFR小分子抑制劑提供了實(shí)驗(yàn)依據(jù)。 第二部分為萊克多巴胺單克隆抗體的制備。 萊克多巴胺(Ractopamine, RAC)是一種含有雙苯烷胺類結(jié)構(gòu)的β-腎上腺素受體激動(dòng)劑，與瘦肉精屬同類化合物。萊克多巴胺被非法用于動(dòng)物飼養(yǎng)，目的是提高瘦肉率，大量食用含有萊克多巴胺殘留的肉類或內(nèi)臟后，可能引發(fā)中毒癥狀，，嚴(yán)重威脅到消費(fèi)者的生命健康。目前萊克多巴胺的檢測(cè)方法主要依靠色譜分析方法，色譜分析方法雖然比較靈敏，準(zhǔn)確度高，但需要大型儀器支持，需要繁瑣的樣品前處理以及專業(yè)的人員操作，導(dǎo)致不能適應(yīng)市場(chǎng)對(duì)分析方法快速、簡(jiǎn)便的要求。因此，建立一種能夠快速、簡(jiǎn)便、準(zhǔn)確檢測(cè)萊克多巴胺在肉類食品中殘留的分析方法顯得至關(guān)重要。 本論文合成了萊克多巴胺的類似物作為半抗原，并使其與BSA偶聯(lián)成完全抗原后用于Balb/c小鼠的免疫。經(jīng)過(guò)一定的免疫程序后通過(guò)間接ELISA實(shí)驗(yàn)對(duì)受免小鼠血清進(jìn)行分析，篩選適宜用于細(xì)胞融合的小鼠進(jìn)行細(xì)胞融合，制備抗萊克多巴胺單克隆抗體，從而建立用于檢測(cè)食品中萊克多巴胺藥物殘留的免疫分析方法。細(xì)胞融合及后續(xù)實(shí)驗(yàn)正在進(jìn)行中。 所有目標(biāo)化合物及部分中間體經(jīng)1H NMR、13C NMR、HR-MS分析確證。
[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.

【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R914.3;R965

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