含嘧啶類骨架蛋白抑制劑的設計、合成及生物活性評價
發(fā)布時間：2018-08-26 15:01
【摘要】：嘧啶是重要的六元氮雜環(huán)化合物,具有極強的生物學意義。在形成遺傳物質DNA和RNA的5種堿基中,有3種是嘧啶類衍生物：胞嘧啶,胸腺嘧啶,尿嘧啶。嘧啶類化合物具有抗代謝、抗腫瘤、抗病毒等多種生理活性,是目前藥物研究的熱點。隨著藥物化學的發(fā)展和技術手段的日新月異,越來越多的靶向蛋白逐漸被人們了解認識。針對靶向蛋白在信號通路的角色,設計合成具有特異性選擇靶向蛋白的抑制劑,以達到增強選擇性,減少對正常組織和細胞的損傷的目的。結合計算機藥物輔助設計技術,引入嘧啶骨架,本論文設計合成了1個系列β-酮脂酰-�；d體蛋白合成酶Ⅲ(E. coli FabH)蛋白抑制劑和1個系列組蛋白去乙�；�(HDAC)蛋白抑制劑。通過元素分析、1H-NMR、13C-NMR、MS等手段確定了這些新化合物的結構。對所有的化合物進行了系統(tǒng)的活性篩選和構效關系研究,其中多數化合物具有良好的抗菌、抗癌活性。簡述如下：以甲硝唑希夫堿類衍生物作為基本構架,引入嘧啶骨架合成系列化合物。采用1H-NMR、ESI-MS和元素分析對所有化合物進行表征鑒定。使用MTT法研究了所有化合物對4株細菌(E. coli ATCC 35218、 P. aeruginosa ATCC 13525、B. subtilis ATCC 6633、S. aureus ATCC 6538)的抑制活性�？咕钚院Y選結果表明,化合物10q表現優(yōu)于陽性對照藥物青霉素,對E. coli ATCC 35218、P. aeruginosaATCC 13525、B. subtilis ATCC 6633、S. aureus ATCC 6538等4株菌的最小抑制濃度MIC分別為1.56μg/ml,3.13μg/ml,1.56μg/ml,3.13μg/ml。通過激酶抑制試驗,測試了全部化合物對大腸桿菌p-酮脂酰-�；d體蛋白合成酶Ⅲ的抑制作用,化合物10q優(yōu)于陽性對照藥物,半數抑制濃度IC50=2.6883μM。構效關系表明,在甲硝唑希夫堿類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。組蛋白去乙酰化酶是針對染色質組蛋白乙�；揎椀闹匾邢虻鞍�,在表觀遺傳分子基礎的研究中占有重要地位。目前處于臨床階段的組蛋白去乙酰化酶抑制劑已有數十種,從中選取17種藥物,然后進行碎片切割,重新整合建立小分子數據庫。通過已有組蛋白去乙酰化酶的蛋白結構,運用分子對接技術和同源性建模技術,篩選出表現突出的化合物進行合成。在已有藥物結構的基礎上,獲得藥效更優(yōu),半衰期更長的潛在先導化合物。采用1H-NMR, ESI-MS和元素分析對所有化合物進行表征鑒定。使用MTT法研究化合物在濃度梯度和時間梯度對5種細胞(Hela, A549, HCT116, HepG2, MCF-7)的增值抑制活性�？鼓[瘤實驗表明,新型化合物在抑制活性和作用時間上優(yōu)于陽性模板藥物。進一步通過細胞周期和凋亡實驗探究了藥物作用觸發(fā)的細胞凋亡機制,新型化合物表現優(yōu)于陽性模板藥物。體外穩(wěn)定性實驗證實新型藥物的穩(wěn)定性優(yōu)于模板陽性藥物SAHA,能夠在體內作用更長時間。激酶抑制實驗和免疫熒光實驗結果表明在靶向酶抑制方面新型化合物優(yōu)于陽性模板藥物。根據所有的生物活性評價,本論文中所陳述化合物是在一代模板藥物的基礎上進一步優(yōu)化獲得的藥效和作用時間更長的極具潛力的組蛋白去乙�；敢种苿╊愊葘幬�。
[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.
【學位授予單位】：南京大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R914
本文編號：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.
【學位授予單位】：南京大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R914
【參考文獻】
相關期刊論文 前1條
1 楊彩玲;韓新光;于春亞;;鹽酸�？颂婺釋θ松圜[狀細胞癌Tca8113細胞周期及Cy-clinD1、P27表達的影響[J];鄭州大學學報(醫(yī)學版);2011年01期
，本文編號：2205234
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