Caspase-3抑制剂的设计、合成及活性评价

发布时间：2018-01-24 21:31

本文关键词： 1 2-苯并异噻唑-3-酮 CaspaSe-3 抑制剂细胞凋亡　出处：《天津科技大学》2014年硕士论文　论文类型：学位论文

【摘要】：细胞凋亡,又称细胞程序性死亡,是组织发育和体内平衡重要的生理过程。细胞异常凋亡会引起很多疾病,例如老年痴呆、帕金森、亨廷顿舞蹈症,细胞凋亡被认为是针对此类疾病的重要靶点。Caspase家族在细胞凋亡的启动和执行过程中起到重要作用,尤其是作为效应因子的Caspase-3,在细胞凋亡模型中的不同信号通路都起到至关重要的作用。因此,Caspase-3被认为是治疗细胞凋亡异常类疾病的潜在治疗靶点。本实验室在前期研究中通过高通量筛选发现1,2-苯并异噻唑-3-酮对Caspase-3有很好的抑制作用,同时报道了一系列具有高度亲和性的Caspase-3抑制剂,其半数抑制率达到31 nmol/L。通过分子对接研究发现了这一类抑制剂只能填充入S1,S2,S3口袋,在S4口袋并没有氢键形成。而通过对比已报道的抑制剂的共晶结构发现,S4口袋是比较重要的。因此,进一步的工作希望通过延长小分子化合物的长度使其与蛋白分子更好的结合。本论文中共合成30个1,2-苯并异噻唑-3-酮类衍生物,其中28个为未见文献报道的新化合物。对这些化合物进行体外酶水平活性评估,包括对Caspase-3和Caspase-7的抑制作用。大部分化合物对Caspase-3的抑制作用达到纳摩级别,其中化合物5j对Caspase-3的抑制活性达到1.15 nmol/L。在此基础上,对化合物进行细胞水平上的生物评价的结果表明化合物5t对喜树碱诱导的Jurkat T细胞的凋亡表现很好的保护作用。为了进一步了解化合物的抑制作用,对化合物5j和13a进行了分子对接研究。通过对比5j和13a分子对接模型,可以看出通过引入苯环-杂环的双环结构来延长分子长度,提高了小分子化合物与蛋白分子的亲和力,能够改善抑制活性。这一点正好与之前的预想相吻合。此外,由于之前合成的一系列抑制剂水溶性不好,本课题的另一个目的是提高这一类化合物的水溶性,其中化合物5a-5f、5t、9、13水溶性有很大改善,为进一步的细胞活性和动物模型方面研究提供可能。
[Abstract]:Apoptosis, also known as programmed cell death, is an important physiological process in tissue development and balance in vivo. Abnormal apoptosis can cause many diseases, such as Alzheimer's disease, Parkinson's disease and Huntington's disease. Apoptosis is considered to be an important target for this disease. Caspase family plays an important role in the initiation and implementation of apoptosis, especially as an effector of Caspase-3. Different signaling pathways play a crucial role in the apoptosis model. Caspase-3 is considered to be a potential therapeutic target for the treatment of apoptosis-like diseases. 2-benzoisothiazole-3-one has a good inhibitory effect on Caspase-3, and a series of Caspase-3 inhibitors with high affinity have been reported. Its half inhibition rate was 31 nmol / L. by molecular docking study, it was found that this kind of inhibitor could only be filled into the S1 / S2 / S3 pocket. There is no hydrogen bond formation in the S4 pocket, and it is important to compare the eutectic structure of the inhibitors reported. Further work was carried out to improve the binding of small molecular compounds to protein molecules by lengthening their length. In this paper, 30 1h2benzoisothiazole-3-one derivatives were synthesized. 28 of them were new compounds which had not been reported in the literature. The enzyme activity of these compounds was evaluated in vitro. The inhibition of most compounds on Caspase-3 reached the Nama level, including the inhibition of Caspase-3 and Caspase-7. The inhibitory activity of compound 5j on Caspase-3 reached 1.15 nmol / L on this basis. The results of biological evaluation of the compounds at the cell level showed that the compound 5t could induce camptothecin induced Jurkat. The apoptosis of T cells showed a good protective effect. In order to further understand the inhibitory effect of compounds. The molecular docking of compounds 5j and 13a was studied. By comparing the molecular docking models of 5j and 13a, we can see that the molecular length can be prolonged by introducing the double ring structure of benzene-heterocyclic. It improves the affinity of small molecular compounds to protein molecules and improves the inhibitory activity. This is exactly as expected. In addition, a series of previously synthesized inhibitors are not water-soluble. Another purpose of this paper is to improve the water solubility of this kind of compounds, in which the water solubility of compound 5a-5f5 TX 913 has been greatly improved. It provides the possibility for further study on cell activity and animal model.
【学位授予单位】：天津科技大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R914;O626

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