基于氟硼吡咯衍生物的荧光分子探针的合成及其性能研究

发布时间：2019-05-15 14:03

【摘要】：近年来,将荧光探针与荧光成像技术进行结合并运用到对一些特定物质的检测已成为极其重要的科学命题,同时因其操作方便快捷,灵敏度和选择性都较高,也被普遍应用到食品、环境等各个领域。氟硼吡咯(BODIPY)类荧光染料因具有高的摩尔消光系数、良好的光稳定性、高的荧光量子产率等优点可作为荧光分子探针的理想染料,因此,本论文以BODIPY为荧光母体,设计合成了BODIPY类衍生物B1和B2,检测试验结果显示该探针不仅能特异性识别半胱氨酸(Cys)、同型半胱氨酸(Hcy),而且能进一步通过反应动力学上的差异,又可以将Cys与Hcy区别开来。具体工作内容如下:第一章:介绍了本论文的研究背景。介绍了荧光探针的组成及其识别机理,根据探针与生物硫醇的反应机理进行简单归纳分类,为本论文中生物硫醇小分子探针的设计、合成与应用奠定文献基础。第二章:设计并合成了两个BODIPY类衍生物B1和B2,并且分别用飞行时间质谱(TOP-MS)、氢谱(1H-NMR)和碳谱(13C-NMR)和元素分析方法,对其结构进行了表征。第三章:重点介绍了用于检测实际样品中Cys的“关-开型”BODIPY类荧光分子探针B1。此探针表现出对Cys高度选择性,并不受Hcy和谷胱甘肽(GSH)的干扰。此分子探针与Cys在0.2-30μM保持良好的线性关系,检测限为0.05μM。在pH=6.0-8.0的中性条件下,B1对Cys的响应不受pH的影响,并且B1对Cys响应迅速,响应时间在5 min之内。再者,分子探针B1对Cys选择性响应机理(PET机理)通过质谱,核磁和高斯密度泛函理论(DFT)计算进行了充分的验证。最后,将此分子探针应用到活细胞成像试验中,并得到了满意的结果。第四章:重点介绍了用于检测实际样品中Cys的“关-开型”近红外BODIPY类荧光分子探针B2。此探针的发射在708 nm处,应用在近红外区域对Cys有高度的选择性。此分子探针与Cys在0.2-30μM保持良好的线性关系,检测限为0.05μM。实验结果显示,探针B2对Cys的检测在中性条件(pH=6.0-9.0)下基本不受pH的干扰且响应迅速(少于5 min)。再者,分子探针B2对Cys选择性响应机理(PET机理)通过质谱,核磁和高斯密度泛函理论(DFT)计算进行了充分的验证。最后,将此分子探针应用到活细胞成像和组织试验中,得到了令人满意的结果。
[Abstract]:In recent years, it has become an extremely important scientific proposition to combine fluorescence probe with fluorescence imaging technology and apply it to the detection of some specific substances. At the same time, because of its convenient and rapid operation, high sensitivity and selectivity. It is also widely used in food, environment and other fields. Fluoroboron polypyrrole (BODIPY) fluorescent dyes can be used as ideal dyes for fluorescent molecular probes because of their high molar extinction coefficient, good photostability and high fluorescence quantum yield. Therefore, BODIPY is used as fluorescent parent in this paper. BODIPY derivatives B1 and B2 were designed and synthesized. the results showed that the probe could not only specifically recognize cysteine (Cys), homocysteine (Hcy), but also further pass the difference in reaction kinetics. You can also distinguish Cys from Hcy. The specific work is as follows: the first chapter: introduces the research background of this paper. The composition and recognition mechanism of fluorescent probe are introduced. According to the reaction mechanism between probe and biomercaptan, the composition and recognition mechanism of fluorescent probe are briefly summarized and classified, which lays a literature foundation for the design, synthesis and application of biomercaptan small molecular probe in this paper. In chapter 2, two BODIPY derivatives B1 and B2 were designed and synthesized, and their structures were characterized by time-of-flight mass spectrometry (TOP-MS), hydrogen spectroscopy (1H-NMR), carbon spectrum (13C-NMR) and elemental analysis, respectively. In chapter 3, the "off-open" BODIPY fluorescent molecular probe B1, which is used to detect Cys in practical samples, is introduced in detail. The probe showed high selectivity to Cys and was not interfered by Hcy and Hcy. The molecular probe has a good linear relationship with Cys at 0.2-30 渭 M, and the detection limit is 0.05 渭 M. Under the neutral condition of pH=6.0-8.0, the response of B1 to Cys is not affected by pH, and B1 responds quickly to Cys, and the response time is within 5 min. Furthermore, the selective response mechanism of molecular probe B1 to Cys (PET mechanism) was fully verified by mass spectrometry, nuclear magnetic field and Gao Si density functional theory (DFT) calculation. Finally, the molecular probe was applied to living cell imaging test, and satisfactory results were obtained. Chapter 4: the "off-open" near-infrared BODIPY fluorescent molecular probe B2, which is used to detect Cys in practical samples, is introduced in detail. The probe emits at 708 nm and is highly selective to Cys in the near infrared region. The molecular probe has a good linear relationship with Cys at 0.2-30 渭 M, and the detection limit is 0.05 渭 M. The experimental results show that the detection of Cys by probe B2 is basically undisturbed by pH under neutral condition (pH=6.0-9.0) and has a rapid response (less than 5 min). Furthermore, the selective response mechanism of molecular probe B2 to Cys (PET mechanism) was fully verified by mass spectrometry, nuclear magnetic field and Gao Si density functional theory (DFT) calculation. Finally, the molecular probe was applied to living cell imaging and tissue test, and satisfactory results were obtained.
【学位授予单位】：湘潭大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O657.3

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