三苯胺基衍生物荧光探针的合成、性质及生物学应用研究

发布时间：2018-03-18 13:08

本文选题：三苯胺衍生物　切入点：有机小分子荧光探针　出处：《安徽大学》2017年硕士论文　论文类型：学位论文

【摘要】：有机小分子荧光探针因其分子量小、合成简单、易获取等优点被广泛应用于检测水体、土壤、空气、生物体中的金属离子含量,其中应用于活体细胞显影的有机小分子荧光探针,为科研工作者在生物医疗和药物研发等问题上提供了一个不可或缺的工具。本论文以桨状结构的三苯胺为功能团,对其进行结构修饰和优化,利用Vilsmeier-Haack、Witting等反应合成了一系列三苯胺基衍生物。基于化合物的结构特点,我们系统地研究其识别性能,结合体外实验结果,进一步探索了它们在生物学方面的应用。主要研究内容如下:1、以三苯胺基为给体中心,C=N及苯乙烯基为π桥,引入羟基基团,合成了三苯胺基希夫碱衍生物SASB。其中多羟基的引入不仅增加有机小分子SASB的水溶性,而且还与-C=N-构筑半空腔的类冠醚型结构,实现对主族元素的响应。实验结果表明,在乙醇中,SASB对主族元素镁离子有特异性识别,且不受同主族钙离子的干扰。SASB在生物细胞内毒性低、光稳定性高、细胞膜穿透性好,不仅对宫颈癌细胞的亚细胞器--溶酶体上过量镁离子有显色响应;而且可以穿透拟南芥幼苗的细胞壁,通过脉管传送到植物叶片的其他部位并伴有镁离子的浓度依赖性荧光增强响应。2、在上一章基础上,改变末端官能团,制备了能靶向定位细胞内亚细胞器线粒体的席夫碱PBSB。研究结果表明,PBSB在乙腈溶液中对重金属铜离子有比率型荧光增强的选择性识别,利用荧光滴定、核磁滴定实验对其识别机理进行了探究。体内实验结果表明,探针能靶向性定位细胞内亚细胞器线粒体部位,且能够通过细胞比率显影来反应细胞内铜离子的分布情况。3、考虑到席夫碱易于水解,限制了其应用,而C-N相较于C=N易于旋转,利于结合金属离子,进而我们对分子结构进行微调,合成了一种柔性的有机小分子仲胺类化合物PBSA。实验结果表明,PBSA具有以下优异特性:1)在乙腈溶液中能特异性识别重金属镍离子;2)不受其它金属离子的干扰;3)检测限度低;4)时效性强;5)可循环性检测。利用荧光滴定、Job's曲线及核磁滴定实验确定了 PBSA与金属镍离子的作用模式。生物学研究表明,探针PBSA特异性定位HepG2细胞的线粒体部位,且能在HepG2细胞内循环使用,有利于检测线粒体相关的疾病。
[Abstract]:Because of its advantages of low molecular weight, simple synthesis and easy to obtain, organic small molecule fluorescent probes are widely used to detect metal ions in water, soil, air and organisms. The fluorescence probe of organic small molecules used in the development of living cells provides an indispensable tool for researchers in biomedicine and drug development. In this paper, the propeller structure of trianiline was used as functional group. A series of triphenylamino derivatives were synthesized by Vilsmeier-Haack-Witting reaction. Based on the structural characteristics of the compounds, the recognition properties of the compounds were systematically studied and combined with the results of in vitro experiments. Their biological applications were further explored. The main research contents were as follows: 1. The hydroxyl group was introduced with triphenylamino as donor center Con N and styrene as 蟺 bridge. The triphenylamino Schiff base derivative SASBs was synthesized. The introduction of polyhydroxyl groups not only increased the water solubility of small organic molecules SASB, but also constructed crown ether-like structures of semicircular cavities with Cn- to achieve the response to the main group elements. In ethanol, SASB has specific recognition for magnesium ions, and it is not interfered by calcium ions of the same group. SASB has low cytotoxicity, high photostability and good cell membrane penetration. It can not only react to the excessive magnesium ion on the lysosome of cervical cancer cells, but also penetrate the cell wall of Arabidopsis thaliana seedlings. The vasculature is transported to other parts of the plant's leaves with a concentration-dependent fluorescence enhancement response of magnesium. 2. Based on the previous chapter, the terminal functional groups are changed. Schiff base PBSBs capable of targeting intracellular suborganelle mitochondria were prepared. The results showed that PBSB had selective recognition of heavy metal copper ions in acetonitrile solution and was titrated by fluorescence. The results of in vivo experiments showed that the probe could target the mitochondria of suborganelles in cells. Moreover, the distribution of copper ions in cells can be reflected by cell ratio development. Considering that Schiff base is easy to hydrolyze, which limits its application, C-N is easier to rotate than Con N, which is favorable for the binding of metal ions. And then we fine-tune the molecular structure, A flexible small organic secondary amine compound PBSA3 was synthesized. The experimental results show that PBSA has the following excellent properties: 1) in acetonitrile solution, PBSA can specifically recognize heavy metal nickel ion 2) and is not interfered by other metal ions. The interaction mode between PBSA and nickel ion was determined by fluorescence titration curve and nuclear magnetic titration experiment. The probe PBSA can specifically locate the mitochondrial site of HepG2 cells and can be circularly used in HepG2 cells, which is helpful for the detection of mitochondrial diseases.
【学位授予单位】：安徽大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O657.3

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