基于BODIPY的汞离子及生物硫醇分子荧光探针的设计、合成与应用

发布时间：2018-05-18 06:36

本文选题：荧光探针 + BODIPY　；参考：《河南大学》2016年硕士论文

【摘要】：荧光探针作为一种新型的生物检测技术,深受市场和研究者的关注,相较于传统的检测手段,有以下优点:操作简单、选择性好、灵敏性高、花费低,并且容易应用在环境和生物系统中。1968年,Treibs小组发现了氟硼二吡咯甲川(BODIPY)类染料,由于其具有众多优点,如高摩尔消光系数、高荧光量子产率、良好的光化学稳定性、分子结构易于修饰、荧光光谱半峰宽窄、对溶剂极性和p H的耐受性好等被广泛应用在标记试剂、荧光开关、化学传感器及激光染料等领域。BODIPY的8位作为敏感的位置,可以通过改变其连接取代基的电子特性,对探针的最大发射波长及荧光量子产率产生很大的影响。基于课题组以往的研究和文献调研,我们将利用O-,N-,S-等基团在不对称和对称BODIPY的8位修饰,以期望改变BODIPY的光物理特性,并研究其在探针方面的应用。在第二章中,对Cl-BODIPY进行修饰,合成了3个meso-Cl的探针,并着重研究了取代基对探针溶解性、反应活性和稳定性的影响。从紫外吸收和荧光发射图谱可知,当BDP2与半胱氨酸(Cys)相遇时,首先Cys的巯基进攻正电性较高的BODIPY 8位碳原子,同时离去一分子氯化氢,生成动力学控制的巯基取代的产物,然后,Cys的氨基环化连接到BODIPY 8位碳原子上,生成氨基取代的热力学控制的产物,释放蓝色荧光。谷胱甘肽(GSH)由于其分子骨架较大,氨基离巯基较远,其与BDP2反应的产物停留在巯基取代阶段。同时,探针的比例式荧光行为大大地增加了荧光测试的动态范围,能够尽可能地减少实验中的误差。探针在血浆中对氨基酸的检测,更展示了其在生物方面的潜在应用。其次,以BODIPY为母体,对硝基偶氮基团为识别团,设计合成了一个BODIPY-偶氮共轭的荧光探针。探针利用双波段对半胱氨酸/同型半胱氨酸和谷胱甘肽显示出良好地选择性,并能区分检测出半胱氨酸/同型半胱氨酸和谷胱甘肽。最后,设计合成了以BODIPY为荧光团,苯硫脲基团为官能团的荧光增强型探针。首先,我们用乙二胺和苯异硫氰酸酯合成了N-氨基乙基-N'-苯硫脲,在8-Cl-BODIPY的作用下,合成探针。探针对汞离子显示了极好地选择性,据实验结果推断可能的检测机制是汞离子导致的环化和相继的水解作用。另外,探针在人体肿瘤细胞SMMC-7721内实现了对汞离子的检测。
[Abstract]:As a new biological detection technology, fluorescent probe has attracted much attention from market and researchers. Compared with traditional methods, fluorescent probe has the following advantages: simple operation, good selectivity, high sensitivity and low cost. In 1968, the Treibs group discovered fluoroboron dipyrrolidine-pyrrolidene BODIPY) dyes because of their many advantages, such as high molar extinction coefficient, high fluorescence quantum yield, good photochemical stability, and good photochemical stability, due to their many advantages, such as high molar extinction coefficient, high fluorescence quantum yield, and good photochemical stability. The molecular structure is easy to modify, the half peak of fluorescence spectrum is narrow, the solvent polarity and pH tolerance are good, and so on are widely used in labeling reagent, fluorescent switch, chemical sensor and laser dye. The maximum emission wavelength and fluorescence quantum yield of the probe can be greatly affected by changing the electronic properties of the connected substituents. Based on the previous research and literature investigation, we will use the 8-position modification of O-N- and S- in asymmetric and symmetric BODIPY in order to change the photophysical properties of BODIPY and study its application in probe. In chapter 2, Cl-BODIPY was modified and three meso-Cl probes were synthesized. The effects of substituents on the solubility, reaction activity and stability of the probes were studied. The UV absorption and fluorescence emission spectra show that when BDP2 meets cysteine, the sulfhydryl group of Cys attacks the positively charged 8 carbon atom of BODIPY and leaves a molecule of hydrogen chloride at the same time to form the product of mercapto substitution controlled by kinetics. Then the amino cyclization of Cys is connected to the carbon atom at the BODIPY 8 position to produce a thermodynamically controlled product of amino substitution, which releases blue fluorescence. Glutathione (GSH) has a large molecular skeleton and the amino group is far away from the sulfhydryl group. At the same time, the proportional fluorescence behavior of the probe greatly increases the dynamic range of fluorescence measurement, and can reduce the errors in the experiment as much as possible. The detection of amino acids in plasma by the probe shows its potential biological application. Secondly, using BODIPY as parent and p-nitroazo group as recognition group, a BODIPY- azo conjugated fluorescence probe was designed and synthesized. The probe showed good selectivity for cysteine / homocysteine and glutathione, and could distinguish cysteine / homocysteine and glutathione. Finally, a fluorescence enhanced probe with BODIPY as fluorescence group and benzene thiourea as functional group was designed and synthesized. First, we synthesized N-aminoethyl-N-thiourea with ethylenediamine and phenylisothiocyanate. The probe was synthesized under the action of 8-Cl-BODIPY. The probe showed excellent selectivity for mercury ions. According to the experimental results, the possible detection mechanism is the cyclization and successive hydrolysis induced by mercury ions. In addition, the probe can detect mercury ions in SMMC-7721 of human tumor cells.
【学位授予单位】：河南大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：O657.3

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