可逆荧光探针的设计、合成及其在药物研究中的应用

发布时间：2018-04-21 09:16

本文选题：荧光探针 + 谷胱甘肽　；参考：《延边大学》2017年博士论文

【摘要】：细胞内的谷胱甘肽(GSH)是含量最多的非蛋白生物硫醇,浓度在1-10mM之间,通过还原型GSH与氧化型GSSG维持细胞内的氧化还原平衡。因此,实时检测GSH的浓度,有助于了解细胞内的氧化还原态的变化。目前开发的荧光探针只能对GSH定性,不能进行定量或者浓度的动态检测。基于此,本研究设计合成了一类基于α,β-不饱和酮可逆加成机理的新型比率荧光探针,可视化研究了体外和细胞内GSH含量的变化,实现了在分子和细胞层次上对GSH原位、实时、动态荧光成像分析。通过对结构的优化,调控了探针与GSH的反应活性。其中,氰基丙烯酰胺类化合物4d与GSH发生可逆加成反应,解离常数为2.59mM,利用该探针定量了 Hela细胞内GSH(5.40±0.87mM)。氰基丙烯酸酯类化合物4c可以特异性的识别亚硫酸氢根,是一种利用率极高的可再生性荧光探针,利用该探针对亚硫酸盐的荧光检测,可以全程跟踪亚硫酸盐引发的疾病状态以及治疗过程,对了解发病的途径和治疗的效果带来巨大的帮助。将齐墩果酸与可逆性GSH荧光探针相偶联,荧光成像研究揭示了齐墩果酸作用过程中GSH的实时变化,有助于研究GSH与药效、细胞凋亡之间的联系。将齐墩果酸与指示型荧光团相连,利用荧光的性能观察了药物在细胞内的特异性定位和药物刺激后细胞内的应激变化。本文主要探索了可逆荧光探针定量以及实时检测GSH的问题,同时将其与药物偶联,利用药物的抗癌活性和探针的定位、识别能力,探索了药物的作用机理。
[Abstract]:Intracellular glutathione glutathione (GSH) is the most abundant non-protein biological mercaptan. The concentration of glutathione is between 1-10mM and redox equilibrium is maintained by reductive GSH and oxidized GSSG. Therefore, real-time detection of GSH concentration is helpful to understand the changes of redox state in cells. The fluorescence probe developed at present can only determine GSH qualitatively, but not quantitatively or dynamically. Based on this, a novel fluorescent probe based on the reversible addition mechanism of 伪, 尾 -unsaturated ketones was designed and synthesized. The changes of GSH content in vitro and in cells were visualized, and the GSH in situ and in real time were realized at molecular and cellular levels. Dynamic fluorescence imaging analysis. The reactivity of the probe and GSH was regulated by optimizing the structure of the probe. The cyanoacrylamide compounds reacted with GSH for 4 d and the dissociation constant was 2.59mM.The probe was used to quantify GSH(5.40 卤0.87mM-1 in Hela cells. Cyanoacrylates (4c) can specifically recognize hydrogen sulfite, which is a highly efficient renewable fluorescent probe for the fluorescence detection of sulfites. It can be used to track the state of the sulfite-induced disease and the course of treatment all the time. It is helpful to understand the way of the disease and the effect of treatment. By coupling oleanolic acid with reversible GSH fluorescence probe, fluorescence imaging revealed the real time change of GSH during the process of oleanolic acid action, which was helpful to study the relationship between GSH and pharmacodynamics and apoptosis. Oleanolic acid was linked to the indicative fluorescence group. The specific localization of the drug in the cells and the changes of the intracellular stress after stimulation were observed by using the fluorescence properties. In this paper, the quantitative and real-time detection of GSH with reversible fluorescence probe was studied. At the same time, the mechanism of drug action was explored by using the anti-cancer activity of the drug and the location of the probe, as well as the recognition ability of the probe.
【学位授予单位】：延边大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R91;O657.3

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