本文選題：熒光探針 + 喹啉　； 參考：《中國科學(xué)技術(shù)大學(xué)》2017年博士論文

【摘要】：生物體內(nèi)活性硫和活性氮物種是兩類重要的生物活性分子。三種生物硫醇和硫化氫屬于活性硫物種;一氧化氮是極具代表性的活性氮物種。大多數(shù)細(xì)胞中含有生物硫醇,濃度約在毫摩爾/升。硒代半胱氨酸(Sec)作為半胱氨酸(Cys)類似物,是構(gòu)成硒蛋白(SePs)的基本成分。硫化氫(H_2S)和一氧化氮(NO)是細(xì)胞內(nèi)重要的信號分子,因此,這些生物活性分子在細(xì)胞中的濃度的在線檢測對生理和病理研究的意義重大。本論文以這幾種生物活性分子為檢測對象,開展了熒光探針的設(shè)計(jì)合成,傳感性質(zhì)研究以及生物體系中熒光成像等研究工作。具體為:.1.設(shè)計(jì)合成了兩類取代的喹啉-2-甲撐丙二酸(QMAs)及其乙酯型(QMEs)的硫醇熒光探針。QMAs和QMEs與硫醇的邁克爾加成反應(yīng)有著截然不同的pH效應(yīng):QMAs不同于QMEs以及大多數(shù)微堿性條件下的邁克爾型探針,能夠在酸性環(huán)境(pHpKa3)中活化,與硫醇發(fā)生反應(yīng)。通過對不同取代基的QMAs篩選,兩種QMAs(取代基:NEt2,OH)能夠定位于溶酶體內(nèi),與硫醇反應(yīng)得到溶酶體的熒光成像,是首例反應(yīng)型的溶酶體探針。2.以疊氮基修飾QME,設(shè)計(jì)合成了硫醇和硫化氫雙輸入型探針QME-N_3。該探針有較大的生物親和性,能夠分別響應(yīng)硫醇和硫化氫這兩種化學(xué)性質(zhì)相近的活性硫物種,且識(shí)別產(chǎn)物是具有雙光子性質(zhì)的熒光產(chǎn)物、可進(jìn)行雙光子成像。與單一檢測物的識(shí)別相比,雙輸入型熒光探針能夠在更復(fù)雜的生物體系中參與生理和病理研究,是更具價(jià)值的生物傳感器。3.設(shè)計(jì)合成了一種新的紅色熒光發(fā)色團(tuán)(B-BODIPY),以此構(gòu)建了高靈敏、高選擇性的硒醇探針Sel-p1和Sel-p2。實(shí)驗(yàn)表明,探針Sel-p2能夠檢測細(xì)胞中外源性Sec和亞硒酸鹽誘導(dǎo)的內(nèi)源性Sec,以及進(jìn)行小鼠體內(nèi)Sec的熒光成像,有望應(yīng)用于探測體內(nèi)硒化合物代謝,為研究硒化合物在癌癥治療中的作用提供了一條途徑。4.設(shè)計(jì)合成了兩種NO熒光探針NO-a5和NO-a6,都具有響應(yīng)快、選擇性高的特點(diǎn)。NO-a5與NO發(fā)生識(shí)別后呈現(xiàn)熒光打開的響應(yīng)模式、具有雙光子吸收性質(zhì);NO-a6與檢測物NO反應(yīng)前后,熒光呈比率變化,這一性質(zhì)在這類熒光探針中屬首次發(fā)現(xiàn)。探針NO-a5和NO-a6都能實(shí)現(xiàn)在活細(xì)胞中外源和內(nèi)源性NO的檢測。
[Abstract]:Active sulfur and nitrogen species are two important bioactive molecules. Three biological mercaptan and hydrogen sulfide belong to active sulfur species and nitric oxide is a representative active nitrogen species. Most cells contain biological mercaptan at concentrations of about ml / l. Selenocysteine (Sec), as a cysteine (Cys) analogue, is the basic component of selenium protein (SePs). Hydrogen sulfide (H _ 2S) and nitric oxide (no) are important signaling molecules in cells. Therefore, on-line detection of the concentrations of these bioactive molecules in cells is of great significance to physiological and pathological studies. In this paper, the design and synthesis of fluorescent probe, the study of sensing properties and the study of fluorescence imaging in biological system were carried out. Specifically:. 1. Two kinds of substituted quinoline-2-methylene malonic acid (QMAs) and their ethyl ester type (QMEs) mercaptan fluorescence probe. QMAs and QMEs have been designed and synthesized. The Michael addition reaction between QMEs and mercaptan has a different pH effect: QMAs is different from QMEs and most microbasic strips. The Michael probe under the device, It can be activated in acidic environment (pH pKa3) and react with mercaptan. Through the screening of QMAs with different substituents, two kinds of QMAs can be located in lysosomes and react with mercaptan to obtain the fluorescence imaging of lysosomes. These two QMAs are the first reaction type lysosomal probes. The double input probe QME-N _ 3 was designed and synthesized with azide modified QME-N _ 3. The probe has great biological affinity and can respond to two kinds of active sulfur species with similar chemical properties, mercaptan and hydrogen sulfide, respectively, and the recognition product is a fluorescence product with two-photon properties, which can be used for two-photon imaging. Compared with single detection, dual-input fluorescent probes can participate in physiological and pathological studies in more complex biological systems and are more valuable biosensors. A new red fluorescent chromophore (B-BODIPY) was designed and synthesized to construct selenol probes Sel-p1 and Sel-p2 with high sensitivity and selectivity. The results showed that the probe Sel-p2 could detect the endogenous Secs induced by exogenous Sec and selenite, as well as the fluorescence imaging of SECs in mice, which could be used to detect the metabolism of selenium compounds in vivo. It provides a way to study the role of selenium compounds in cancer treatment. 4. 4. Two kinds of no fluorescence probes, NO-a5 and NO-a6, were designed and synthesized. Both have the characteristics of fast response and high selectivity. The fluorescence of NO-a6 and NO-a6 showed a ratio change before and after the reaction between NO-a6 and no. This property is first found in this kind of fluorescent probe. The probes NO-a5 and NO-a6 can detect both exogenous and endogenous no in living cells.
【學(xué)位授予單位】：中國科學(xué)技術(shù)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號】：O657.3

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本文編號：2092894