以EGFR为靶点的抗肿瘤化合物的设计、合成及生物活性评价

发布时间：2018-02-24 01:10

本文关键词： EGFR抑制剂抗癌分子模拟细胞迁移构效分析吡唑环噻唑环萘酚环 1 4二氧八环咖啡酸酰胺　出处：《南京大学》2014年博士论文　论文类型：学位论文

【摘要】：癌症是世界上首位导致死亡的因素,在美国每一年因为癌症死亡的人数大约占总死亡人数的25%,不仅如此每年新增癌症患者的人数也不断增加。随分子生物学的研究发展,对于癌症的发病机理和其在产生和发展的过程中起到重要作用的特异性的酶有了更加深入的了解,使得这些特异酶作为研制抗癌药物的分子靶标成为了可能。表皮生长因子受体EGFR (epidermal growth factor receptor)在肿瘤细胞的增殖、血管生成、肿瘤侵袭、转移及细胞凋亡的抑制等方面起到重要的作用,大量的研究表明EGFR可作为治疗癌症的分子靶标,像EGFR抑制剂埃罗替尼和吉非替尼就是非常成功的抗癌药物。本篇论文我们设计了两个系列共45个全新的化合物,并对它们进行了抗增殖、抑制EGFR、毒性、凋亡和抑制细胞迁移等生物活性的检测,研究表明这些化合物可能是通过抑制抑制EGFR,起到抗肿瘤效果的。 (1)由于吡唑、噻唑和萘酚环具有低毒和显著地生物活性,我们在此基础上合成出同时含有吡啶、噻唑和萘酚环的化合物(la-7a,1b-7b,1c-7c,1d-7d)通过元素分析、H-NMR、ESI-MS分析确定了这些新化合物的结构,通过单晶X-射线结构分析确定了1d和5d化合物的三维空间结构。抗增殖和抑制EGFR的活性结果显示这些化合物较好的抗癌活性,其中化合物7d抗Hela癌细胞和抑制EGFR活性最好,对应的IC50值为0.86μM和0.12μM,同时构效分析表明苯环上的取代基种类对化合物的生物活性影响较大。实验结果显示EGFR抑制活性和抗增殖活性具有一定的相关性,表明这个系列化合物的抗增殖活性可能是通过抑制EGFR的活性产生的。另外细胞毒性和抑制Hela细胞迁移测试表明7d没有毒性并且能够较好地抑制Hela细胞迁移。分子模拟显示7d中的萘环与EGFR中的氨基酸LYS721形成了两个p-π键,提高了抗癌活性。 (2)合成了含有苯并1,4二氧八环的咖啡酸酰胺,并对化合物进行结构鉴定,包括1H NMR、ESI-MS、元素分析以及晶体结构测定,合成出的17个化合物均为首次报道。对合成出的化合物的抗增殖和抑制EGFR活性检测显示大部分的化合物具有很强的抗癌活性,尤其是D9(IC50=0.79μM对HepG2和IC50=0.36μM对EGFR)。构效分析表明苯并1,4二氧八环结构和苯胺上苯环的对位的取代基种类对于化合物的生物活性影响很大,同时在细胞凋亡、细胞毒性和抑制HepG2细胞的测试中化合物均显示出很好的活性。分子模拟显示D9与EGFR中的氨基酸MET793和LEU1001形成了两个氢键,增加了粘合力提高了抗癌活性。这两个系列化合物的研究表明7d和D9有望成为以EGFR为靶点的新型抗癌药物先导化合物。
[Abstract]:Cancer is the first cause of death in the world. Cancer accounts for about 25 of the total deaths in the United States every year, and the number of new cancer patients increases every year. We have a deeper understanding of the pathogenesis of cancer and the specific enzymes that play an important role in the development and development of cancer. This makes it possible for these specific enzymes to be used as molecular targets for the development of anticancer drugs. Epidermal growth factor receptor, EGFR epidermal growth factor receptor, proliferates, angiogenesis, and invades tumor cells. Metastasis and inhibition of apoptosis play an important role. A large number of studies have shown that EGFR can be used as a molecular target for the treatment of cancer. Erotini and gefitinib, EGFR inhibitors, are very successful anticancer drugs. In this paper, we designed two series of 45 new compounds, and carried out anti-proliferation, inhibition of EGFR, toxicity. The detection of biological activities such as apoptosis and inhibition of cell migration suggests that these compounds may have antitumor effects by inhibiting EGFR. (1) due to the low toxicity and remarkable biological activity of pyrazole, thiazole and naphthol rings, we have synthesized compounds containing pyridine, thiazole and naphthol ring, which are composed of pyridine, thiazole and naphthol ring. The structures of these compounds have been determined by elemental analysis with H-NMRESI-MS. The three-dimensional spatial structures of 1-d and 5-d compounds were determined by single crystal X-ray structure analysis. The results of anti-proliferation and inhibition of EGFR activity showed that these compounds had better anticancer activity, of which 7d had the best antitumor activity against Hela cancer cells and the best inhibition of EGFR activity. The corresponding IC50 values were 0.86 渭 M and 0.12 渭 M. meanwhile, the structural activity analysis showed that the substituents on the benzene ring had a great influence on the bioactivity of the compounds. The experimental results showed that the inhibitory activity of EGFR and the anti-proliferation activity of the compounds were related to each other. The results indicated that the antiproliferative activity of this series of compounds may be produced by inhibiting the activity of EGFR. In addition, the cytotoxicity and inhibition of Hela cell migration test showed that the antiproliferative activity of this series of compounds was not toxic at 7 days and could inhibit the migration of Hela cells well. The simulation shows that the naphthalene ring in 7 d and the amino acid LYS721 in EGFR form two p- 蟺 bonds. The anti-cancer activity was improved. The caffeic acid amide containing benzo 1o 4 dioxo octamer was synthesized and its structure was identified, including 1H N MRN ESI-MS, elemental analysis and crystal structure determination. For the first time, 17 compounds were synthesized. The detection of the antiproliferation and inhibition of EGFR activity of the synthesized compounds showed that most of the compounds had strong anticancer activity. In particular, the structure-activity analysis of HepG2 and IC50=0.36 渭 M in D9 / IC50 / 0.79 渭 M and IC50=0.36 渭 M showed that the structure of benzo 1 / 4 dioxo 8 ring and the type of p-substituents of benzene ring on aniline had a great influence on the bioactivity of the compounds, and apoptosis was also observed in the cells. Molecular simulation showed that D9 formed two hydrogen bonds with amino acid MET793 and LEU1001 in EGFR and increased adhesion to enhance anticancer activity. The studies of these two series of compounds indicate that 7d and D9 are expected to be the leading compounds of new anticancer drugs targeting EGFR.
【学位授予单位】：南京大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R914

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