含硫脲吡唑类衍生物的合成及生物活性评价

发布时间：2018-01-28 07:27

本文关键词： CDKs 分子模拟细胞周期吡唑硫脲抗肿瘤　出处：《南京大学》2014年硕士论文　论文类型：学位论文

【摘要】：细胞周期是细胞生命活动的重要环节,细胞周期调控的失调与细胞癌变密切相关。研究发现多种CDK(周期素依赖性蛋白激酶)在肿瘤组织中都处于失调状态；究其原因,可能是由于CDK和Cyclin基因处于高表达状态或者CDK抑制因子失活造成的。因此,选择性的抑制肿瘤细胞中的CDK活性可能是控制肿瘤增殖的一条有效途径。现在农药及医药界对越来越多的硫脲结构的化合物也给予了极大的关注；此外,吡唑类衍生物具有良好的抗癌活性已被报道许久,根据活性叠加的原理,我们将吡唑和硫脲结构进行有机结合并设计了一系列含硫脲结构的吡唑类衍生物,并推测此类化合物具有良好的抗癌活性。为了进一步验证上述推测,我们借助于计算机辅助药物设计技术,运用Discovery studio分子对接软件对我们设计的批量的吡唑类衍生物进行筛选。本实验根据分子对接软件的筛选结果,设计合成了20个新型的含硫脲的吡唑类衍生物(6a-10d),采用元素分析、1H-NMR、13C-NMR、MS分析等手段对所有化合物进行表征并确定其结构；其中,通过单晶X-射线结构分析确定了化合物7a的三维空间结构。因为这20个化合物均没有活性方面的报道,我们用MTT法测试了所有化合物对3株肿瘤细胞A549,H460和MCF-7抗增殖活性,结果显示多数化合物表现出良好的抗肿瘤活性：其中化合物10b活性最好,对A549的半数抑制率(IC50值)仅为0.75μM；远优于阳性化合物Olomoucine的130.00μM。根据上述的抗增值活性数据我们初步总结了化合物的构效关系,即发现与硫脲骨架相连的供电子取代基表现出强的抗增值活性。为进一步验证此构效关系,采用Elisa法测试了部分化合物对周期素依赖性蛋白激酶的抑制活性,酶抑制活性结果显示具有较高抗肿瘤增值活性的化合物l0b同样具有最好的CDKs抑制活性(ICso值为25 nM),故而推测目标化合物10b的抗增值活性可能是通过抑制CDKs的活性表现出来的。与此同时,流式实验结果也显示出化合物10b在体外可强烈诱导A549阻滞于细胞周期G1期,进一步证实了目标化合物10b具有良好的生物活性,其可能是一类具有较好研究前景的先导化合物。
[Abstract]:Cell cycle is an important part of cell life. The imbalance of cell cycle regulation is closely related to cell carcinogenesis. Many kinds of CDK (cyclin dependent protein kinase) have been found to be disordered in tumor tissues. The reason may be due to the high expression of CDK and Cyclin genes or the inactivation of CDK suppressor. Selective inhibition of CDK activity in tumor cells may be an effective way to control tumor proliferation. In addition, pyrazole derivatives with good anticancer activity have been reported for a long time, according to the principle of activity superposition. We combined pyrazole with thiourea structure and designed a series of pyrazole derivatives containing thiourea structure and speculated that these compounds had good anticancer activity. We use computer-aided drug design techniques. Discovery studio molecular docking software was used to screen the batch of pyrazole derivatives designed by us. The results of this experiment were based on the results of molecular docking software. Twenty new pyrazole derivatives containing thiourea were designed and synthesized. The 1H-NMR-13C-NMR spectra were analyzed by elemental analysis. All the compounds were characterized and their structures were determined by MS analysis. Among them, the three-dimensional spatial structure of compound 7a was determined by single crystal X-ray structure analysis, because none of the 20 compounds were active. The antiproliferative activities of all the compounds against three tumor cell lines A549 H460 and MCF-7 were tested by MTT assay. The results showed that most of the compounds showed good antitumor activity: the activity of compound 10b was the best, and the IC50 of A549 was only 0.75 渭 m; Far better than the positive compound Olomoucine 130.00 渭 M. according to the above anti-value-added activity data, we preliminarily summarized the structure-activity relationship of the compound. It was found that the electron donor group attached to the thiourea skeleton showed strong anti-increment activity. The inhibitory activity of some compounds on cyclin dependent protein kinase was measured by Elisa method. The result of enzyme inhibitory activity showed that the compound 10b with high anti-tumor increment activity also had the best CDKs inhibitory activity (ICso = 25 nm). Therefore, we speculate that the anti-increment activity of target compound 10b may be expressed by inhibiting the activity of CDKs. The results of flow cytometry also showed that compound 10b could strongly induce A549 to block in G1 phase of cell cycle in vitro, which further confirmed that the target compound 10b had good biological activity. It may be a kind of leading compounds with good research prospect.
【学位授予单位】：南京大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R914.5

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