含嘧啶类骨架蛋白抑制剂的设计、合成及生物活性评价
发布时间：2018-08-26 15:01
【摘要】：嘧啶是重要的六元氮杂环化合物,具有极强的生物学意义。在形成遗传物质DNA和RNA的5种碱基中,有3种是嘧啶类衍生物：胞嘧啶,胸腺嘧啶,尿嘧啶。嘧啶类化合物具有抗代谢、抗肿瘤、抗病毒等多种生理活性,是目前药物研究的热点。随着药物化学的发展和技术手段的日新月异,越来越多的靶向蛋白逐渐被人们了解认识。针对靶向蛋白在信号通路的角色,设计合成具有特异性选择靶向蛋白的抑制剂,以达到增强选择性,减少对正常组织和细胞的损伤的目的。结合计算机药物辅助设计技术,引入嘧啶骨架,本论文设计合成了1个系列β-酮脂酰-酰基载体蛋白合成酶Ⅲ(E. coli FabH)蛋白抑制剂和1个系列组蛋白去乙酰化酶(HDAC)蛋白抑制剂。通过元素分析、1H-NMR、13C-NMR、MS等手段确定了这些新化合物的结构。对所有的化合物进行了系统的活性筛选和构效关系研究,其中多数化合物具有良好的抗菌、抗癌活性。简述如下：以甲硝唑希夫碱类衍生物作为基本构架,引入嘧啶骨架合成系列化合物。采用1H-NMR、ESI-MS和元素分析对所有化合物进行表征鉴定。使用MTT法研究了所有化合物对4株细菌(E. coli ATCC 35218、 P. aeruginosa ATCC 13525、B. subtilis ATCC 6633、S. aureus ATCC 6538)的抑制活性。抗菌活性筛选结果表明,化合物10q表现优于阳性对照药物青霉素,对E. coli ATCC 35218、P. aeruginosaATCC 13525、B. subtilis ATCC 6633、S. aureus ATCC 6538等4株菌的最小抑制浓度MIC分别为1.56μg/ml,3.13μg/ml,1.56μg/ml,3.13μg/ml。通过激酶抑制试验,测试了全部化合物对大肠杆菌p-酮脂酰-酰基载体蛋白合成酶Ⅲ的抑制作用,化合物10q优于阳性对照药物,半数抑制浓度IC50=2.6883μM。构效关系表明,在甲硝唑希夫碱类衍生物引入嘧啶骨架有利于抗菌活性和抗靶向蛋白活性的提高。组蛋白去乙酰化酶是针对染色质组蛋白乙酰化修饰的重要靶向蛋白,在表观遗传分子基础的研究中占有重要地位。目前处于临床阶段的组蛋白去乙酰化酶抑制剂已有数十种,从中选取17种药物,然后进行碎片切割,重新整合建立小分子数据库。通过已有组蛋白去乙酰化酶的蛋白结构,运用分子对接技术和同源性建模技术,筛选出表现突出的化合物进行合成。在已有药物结构的基础上,获得药效更优,半衰期更长的潜在先导化合物。采用1H-NMR, ESI-MS和元素分析对所有化合物进行表征鉴定。使用MTT法研究化合物在浓度梯度和时间梯度对5种细胞(Hela, A549, HCT116, HepG2, MCF-7)的增值抑制活性。抗肿瘤实验表明,新型化合物在抑制活性和作用时间上优于阳性模板药物。进一步通过细胞周期和凋亡实验探究了药物作用触发的细胞凋亡机制,新型化合物表现优于阳性模板药物。体外稳定性实验证实新型药物的稳定性优于模板阳性药物SAHA,能够在体内作用更长时间。激酶抑制实验和免疫荧光实验结果表明在靶向酶抑制方面新型化合物优于阳性模板药物。根据所有的生物活性评价,本论文中所陈述化合物是在一代模板药物的基础上进一步优化获得的药效和作用时间更长的极具潜力的组蛋白去乙酰化酶抑制剂类先导药物。
[Abstract]:Pyrimidine is an important six-membered heterocyclic compound with strong biological significance. Among the five bases of DNA and RNA, three of them are pyrimidine derivatives: cytosine, thymine, uracil. Pyrimidine compounds have many physiological activities, such as anti-metabolism, anti-tumor, anti-virus and so on, which are the focus of drug research. With the development of pharmacochemistry and the development of technology, more and more targeted proteins have been recognized by people. In view of the role of targeted proteins in signaling pathways, inhibitors with specific selective targeting proteins are designed and synthesized to enhance selectivity and reduce damage to normal tissues and cells. A series of protein inhibitors of beta-ketolipid-acyl carrier protein synthase III (E.coli FabH) and a series of protein inhibitors of histone deacetylase (HDAC) were designed and synthesized by introducing pyrimidine framework. The structures of these new compounds were determined by elemental analysis, 1H-NMR, 13C-NMR, MS and so on. Some compounds have been systematically screened and their structure-activity relationships have been studied. Most of them have good antimicrobial and anticancer activities. The following are briefly described: a series of compounds have been synthesized from pyrimidine skeleton using metronidazole Schiff base derivatives as basic framework. All compounds have been characterized by 1H-NMR, ESI-MS and elemental analysis. Inhibitory activity of all compounds against four strains of bacteria (E. coli ATCC 35218, P. aeruginosa ATCC 13525, B. subtilis ATCC 6633, S. aureus ATCC 6538) was studied by MTT assay. Antimicrobial activity screening showed that compound 10q was superior to penicillin, and it was superior to E. coli ATCC 35218, P. aeruginosa ATCC 13525, B. subtilis A. The minimal inhibitory concentration MICs of TCC 6633, S. aureus ATCC 6538 were 1.56 ug/ml, 3.13 ug/ml, 1.56 ug/ml and 3.13 ug/ml, respectively. The inhibitory effect of all compounds on Escherichia coli p-keto-lipoyl-acyl carrier protein synthase III was tested by kinase inhibition test. Compound 10q was superior to the positive control drug, with half inhibitory concentration IC50=2. The structure-activity relationship of 6883 mu M. showed that the introduction of pyrimidine skeleton into metronidazole Schiff base derivatives was conducive to the enhancement of antibacterial activity and anti-targeting protein activity. Histone deacetylase is an important targeting protein for chromatin histone acetylation modification and plays an important role in the study of epigenetic molecular basis. There are dozens of histone deacetylase inhibitors in the stage, 17 of which are selected, then fragmented and re-integrated to establish a small molecular database. The compounds were characterized by 1H-NMR, ESI-MS and elemental analysis. The inhibitory activities of the compounds on 5 cell lines (Hela, A549, HCT116, HepG2, MCF-7) at concentration gradient and time gradient were studied by MTT method. The results showed that the new compounds were superior to the positive template drugs in inhibition activity and action time. The mechanism of cell apoptosis triggered by the drug action was further explored by cell cycle and apoptosis experiments. The new compounds were superior to the positive template drugs. The stability of the new drugs was better than that of the template positive drug SAHA in vitro. Kinase inhibition test and immunofluorescence assay showed that the new compounds were superior to the positive template drugs in targeting enzyme inhibition. According to all bioactivity evaluations, the compounds described in this paper were further optimized on the basis of the first generation template drugs to achieve more potency and duration of action. Long potential drugs for histone deacetylase inhibitors.
【学位授予单位】：南京大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R914
本文编号：2205234
[Abstract]:Pyrimidine is an important six-membered heterocyclic compound with strong biological significance. Among the five bases of DNA and RNA, three of them are pyrimidine derivatives: cytosine, thymine, uracil. Pyrimidine compounds have many physiological activities, such as anti-metabolism, anti-tumor, anti-virus and so on, which are the focus of drug research. With the development of pharmacochemistry and the development of technology, more and more targeted proteins have been recognized by people. In view of the role of targeted proteins in signaling pathways, inhibitors with specific selective targeting proteins are designed and synthesized to enhance selectivity and reduce damage to normal tissues and cells. A series of protein inhibitors of beta-ketolipid-acyl carrier protein synthase III (E.coli FabH) and a series of protein inhibitors of histone deacetylase (HDAC) were designed and synthesized by introducing pyrimidine framework. The structures of these new compounds were determined by elemental analysis, 1H-NMR, 13C-NMR, MS and so on. Some compounds have been systematically screened and their structure-activity relationships have been studied. Most of them have good antimicrobial and anticancer activities. The following are briefly described: a series of compounds have been synthesized from pyrimidine skeleton using metronidazole Schiff base derivatives as basic framework. All compounds have been characterized by 1H-NMR, ESI-MS and elemental analysis. Inhibitory activity of all compounds against four strains of bacteria (E. coli ATCC 35218, P. aeruginosa ATCC 13525, B. subtilis ATCC 6633, S. aureus ATCC 6538) was studied by MTT assay. Antimicrobial activity screening showed that compound 10q was superior to penicillin, and it was superior to E. coli ATCC 35218, P. aeruginosa ATCC 13525, B. subtilis A. The minimal inhibitory concentration MICs of TCC 6633, S. aureus ATCC 6538 were 1.56 ug/ml, 3.13 ug/ml, 1.56 ug/ml and 3.13 ug/ml, respectively. The inhibitory effect of all compounds on Escherichia coli p-keto-lipoyl-acyl carrier protein synthase III was tested by kinase inhibition test. Compound 10q was superior to the positive control drug, with half inhibitory concentration IC50=2. The structure-activity relationship of 6883 mu M. showed that the introduction of pyrimidine skeleton into metronidazole Schiff base derivatives was conducive to the enhancement of antibacterial activity and anti-targeting protein activity. Histone deacetylase is an important targeting protein for chromatin histone acetylation modification and plays an important role in the study of epigenetic molecular basis. There are dozens of histone deacetylase inhibitors in the stage, 17 of which are selected, then fragmented and re-integrated to establish a small molecular database. The compounds were characterized by 1H-NMR, ESI-MS and elemental analysis. The inhibitory activities of the compounds on 5 cell lines (Hela, A549, HCT116, HepG2, MCF-7) at concentration gradient and time gradient were studied by MTT method. The results showed that the new compounds were superior to the positive template drugs in inhibition activity and action time. The mechanism of cell apoptosis triggered by the drug action was further explored by cell cycle and apoptosis experiments. The new compounds were superior to the positive template drugs. The stability of the new drugs was better than that of the template positive drug SAHA in vitro. Kinase inhibition test and immunofluorescence assay showed that the new compounds were superior to the positive template drugs in targeting enzyme inhibition. According to all bioactivity evaluations, the compounds described in this paper were further optimized on the basis of the first generation template drugs to achieve more potency and duration of action. Long potential drugs for histone deacetylase inhibitors.
【学位授予单位】：南京大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R914
【参考文献】
相关期刊论文 前1条
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