Rhodol类生物活性小分子荧光探针的构建以及应用研究

发布时间：2018-02-11 06:28

本文关键词： 荧光探针 Rhodol荧光团次氯酸硫化氢生物成像　出处：《西北大学》2017年硕士论文　论文类型：学位论文

【摘要】：近年来荧光成像技术因其时间和空间分辨率好,操作安全、简便?经发展成为监测各种生理过程中生物分子变化最重要的手段之一,而选择光谱性能优良的荧光分子对成像分析至关重要。本文从荧光分子结构改进入手,首次设计并合成了一种新型的Rhodol类荧光染料ROA,并以此为荧光团设计合成了三种不同类型的荧光探针。研究内容如下:(1)以ROA为荧光骨架,设计合成了一种高选择性次氯酸根荧光探针RO610。该探针分子由于碳氮双键的异构化作用荧光被猝灭,而与次氯酸反应之后,碳氮双键被氧化、水解为甲酰基,异构化作用消失,分子荧光显著增强。利用这种荧光信号的变化,我们已将RO610成功应用于活细胞及小鼠体内外源性次氯酸根的检测。(2)首次用肼基修饰ROA上的甲酰基,合成了一种同时具有C=N双键和联氨基的氧杂蒽类次氯酸根荧光探针R0585。此探针实现了缓冲溶液测试体系中次氯酸的实时检测,且不受其它活性氧类物质的干扰。对其检测机理进行研究发现次氯酸首先作用于此分子的联氨基团,将其氧化为重氮基后经过分子内质子转移形成一种稳定的结构RON,并已经通过核磁滴定与质谱分析验证了此机理。这一设计策略也为次氯酸荧光探针的设计提供了一种新思路。(3)以化合物ROA为母体,设计、合成了一种H2S荧光探针R0587。我们将Cu2+与RO587络合得到RO587-Cu2+,因Cu2+的顺磁性效应,R0587的荧光信号被猝灭。因为Cu2+与S2-之间具有更强的亲和作用,所以我们利用S2-将配合物的Cu2+夺出,使得RO587荧光得到恢复,从而实现对S2-的检测。此方法具有反应迅速,灵敏度高等优点并能选择性地检测硫化氢而不受其他阴离子及含巯基的生物小分子的影响,为专一性检测硫化氢提供了一种新的思路和方法。
[Abstract]:In recent years, fluorescence imaging technology because of its time and space resolution, safe operation, convenient? Become changes in various physiological processes in molecular biological monitoring is one of the most important means and the choice of fluorescent molecular spectra with excellent performance is of vital importance to image analysis. This paper starts from the fluorescence molecular structure is improved, and a new design for the first time in Rhodol type of fluorescent dye ROA was synthesized, and then three different types of fluorescent probes were synthesized fluorophore design. The contents are as follows: (1) using the ROA fluorescent skeleton, a highly selective fluorescent probe for hypochlorite RO610. the probe molecule due to isomerization of carbon and nitrogen double bonds were designed and synthesized. The fluorescence quenching, and with hypochlorous acid after the reaction, the carbon nitrogen double oxidation, hydrolysis of formyl, isomerization disappeared, molecular fluorescence was significantly enhanced. The change of the fluorescence signal, we have RO6 Detection of 10 successful application in living cells and in mice with exogenous hypochlorite. (2) for the first time with hydrazide modified ROA formyl, and has a C=N double bond and amino xanthene fluorescent hypochlorite probe R0585. the probe was synthesized to achieve the real-time detection of buffer solution test system in hypochlorous acid, and is not interfered by other reactive oxygen species. The study found that hypochlorite in the role of the first combination of this molecular amino group on the detection mechanism of the oxidation of diazo after intramolecular proton transfer to form a stable structure of RON, and has been through the analysis of NMR titration and mass spectrometry to verify the mechanism. This design strategy also provides a new idea for the design of hypochlorous acid fluorescent probe. (3) design to compound ROA as the matrix, a H2S fluorescent probe R0587. was synthesized by complexation with RO587 Cu2+ we will get RO587-Cu2+ for Paramagnetic effects of Cu2+, R0587 fluorescence was quenched. Because the affinity has a stronger effect between Cu2+ and S2-, so we will use S2- complexes of Cu2+ took out, makes the RO587 fluorescence recovery, so as to realize the detection of S2-. This method has the advantages of rapid response, high sensitivity and selective detection of hydrogen sulfide not affected by other anions and sulfhydryl molecules, provides a new idea and method for the specific detection of hydrogen sulfide.

【学位授予单位】：西北大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O657.3

【相似文献】