CN108129500A - 一种识别过氧化氢的比值型荧光探针的合成与应用 - Google Patents
一种识别过氧化氢的比值型荧光探针的合成与应用 Download PDFInfo
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Abstract
本发明涉及一种用于检测过氧化氢(H2O2)的荧光探针的制备方法和应用，属于化学分析检测技术领域。其分子结构如下：该探针分子最大发射波长在666nm，探针分子与过氧化氢(H2O2)作用后，荧光光谱蓝移至594nm处，实现了比值红光检测过氧化氢(H2O2)，提高检测的灵敏度；发射波长在红光区能够减少探针检测过程中的背景荧光和活细胞的光损伤，增强生物对组织的穿透能力。本发明所述的探针分子在一定时间与浓度范围具有良好线性，对过氧化氢(H2O2)识别能力强，选择性好、抗干扰能力强，能够准确定位线粒体，实现线粒体内检测过氧化氢，该类探针在生物化学等领域具有重要的应用价值。
Description
本发明属于化学分析检测技术领域，具体涉及一种线粒体内检测过氧化氢的新型比值红光型荧光探针的制备方法以及其在体外和活细胞内部检测过氧化氢(H2O2)方面的应用。
背景技术
活性氧(ROS)类在生物***的生理和病理过程中扮演着重要的角色。作为重要的ROS之一，过氧化氢在几乎所有的氧化过程中产生，并参与了对生物活性的调控。内源性过氧化氢(H2O2)作为一种“信号分子”，可以激活信号传导，刺激细胞的增殖、分化等生理活动。过量的内源性过氧化氢(H2O2)通过细胞内酶的催化刺激产生，从而产生氧化压、破坏蛋白质、核酸、损害生物分子等脂质分子，被认为是许多疾病的触发器，如糖尿病、癌症和老年痴呆症。此外，大量的实验结果表明，线粒体呼吸过程中产生的过量的内源性过氧化氢(H2O2)可能会引起强烈的氧损害，进一步引发线粒体肿胀和细胞凋亡。因此，有必要设计一种有效的方法，实现亚细胞器中过氧化氢局部可视化的检测，特别是在线粒体内。
发明内容
本发明目的之一在于提供一种简便高效的荧光探针合成方法；本发明之另一目的是提供一种选择性好，抗干扰能力强，具有比值红光发射波长，能够实现对体外或者活细胞内部特别是线粒体内过氧化氢(H2O2)检测的荧光探针。
本发明解决问题采取的技术方案为，一种比值型识别过氧化氢新型荧光探针，其分子结构式如下：
合成路线如下：
具体合成方法如下：(a)将化合物1(200.0g,1.4mmol)和4-溴甲基苯硼酸频哪醇酯(620.0g,2.1mmol)溶于2.0mL无水甲苯于密闭反应器中110℃搅拌6h。反应过程中析出大量白色固体，停止反应冷却至室温抽滤，滤饼用甲苯洗涤，得到纯的产物2(180.0mg，29.3％)。(b)在室温下将化合物2(160.0g,0.46mmol)和化合物3(100.0g,0.4mmol)溶于2.0mL乙醇中的溶液中，加入哌啶(5.0μL，0.05mmol)。在室温下搅拌5分钟后，将所得溶液回流6小时。然后将反应溶液减压浓缩，柱层析分离得橙红色固体的化合物4(60.0mg，22.6％产率)。
本发明的荧光探针的作用机理如下，探针分子通过正电荷的引入强化了分子内ICT效应而发射近红外光(666nm)。过氧化氢能够氧化探针分子中溴代硼酸酯部分，水解释放出染料6，减弱了分子内ICT过程而导致发射波长蓝移而发射红光(594nm)，从而实现特异性检测过氧化氢(H2O2)的目的。探针分子的响应过程如下：
本发明的荧光探针具有比值红光发射，其与过氧化氢(H2O2)作用前发射近红外荧光在666nm处，作用后荧光发射峰在594nm处。
本发明的荧光探针具有线粒体靶向位点，表明该探针能够实现线粒体内过氧化氢(H2O2)的检测。
本发明的荧光探针选择性好。探针分子的测试体系为pH为7.0的10mM的PBS缓冲溶液包含30％乙腈，室温下测量。探针分子本身发射近红外荧光，在加入40倍当量过氧化氢(H2O2)之后，在最大发射波长666nm处荧光强度发生了蓝移至594nm处。而在加入其他活性氧物质(ROO.,.OH,NO.,TBHP,NaClO,.O2,KO2,.OtBu,ONOO-)后，荧光发射峰没有变化。
本发明的荧光探针抗干扰能力强，其他活性氧物质(ROO.,.OH,NO.,TBHP,NaClO,.O2,KO2,.OtBu,ONOO-)的存在不影响探针分子与过氧化氢(H2O2)的作用。
本发明的荧光探针在加入40倍当量的过氧化氢(H2O2)作用后，荧光发射峰蓝移至594nm，在2h时594nm处荧光达到最大值，666nm处的发射峰消失。
本发明的荧光探针表现出良好的动力学现象，探针分子与过氧化氢(H2O2)选择性识别后在，荧光强度比值与时间表现出良好的线性关系。
本发明的荧光探针能够实现细胞内外检测过氧化氢(H2O2)，通过共定位试剂罗丹明123证实探针具有好的线粒体靶向性能，重叠系数达到了94％，表明该探针能实现线粒体内检测过氧化氢(H2O2)。
本发明所述的探针分子对过氧化氢(H2O2)响应前的发射波长在近红外，响应后发射蓝移而呈现红色荧光并且对活性氧类具有良好的选择性和抗干扰能力，并且具有好的灵敏度，具有较宽的应用范围。长的比值发射波长有强的组织穿透能力，该荧光探针在生物与化学等领域具有实际的应用价值。
附图说明
图1为本发明的荧光探针(10.0×10-6mol/L)在PBS(10mM，pH＝7.0)/CH3CN30％溶液中，与过氧化氢(H2O2)作用前后的光谱变化图，横坐标为波长，纵坐标分别为吸收/荧光强度。
图2为本发明的荧光探针(10.0×10-6mol/L)在PBS(10mM，pH＝7.0)/CH3CN30％中的荧光发射谱图随加入过氧化氢(H2O2)量的变化。
图3为本发明的荧光探针(10.0×10-6mol/L)在PBS(10mM，pH＝7.0)/CH3CN30％中选择性测试的荧光谱图变化，横坐标为波长，纵坐标为荧光强度。
图4为本发明的荧光探针(10.0×10-6mol/L)在PBS(10mM，pH＝7.0)/CH3CN30％中选择性的条形图，纵坐标为I594/I666强度的变化。
图5为本发明的荧光探针(10.0×10-6mol/L)在PBS(10mM，pH＝7.0)/CH3CN30％中检测过氧化氢(H2O2)存在(ROO.,.OH,NO.,TBHP,NaClO,.O2,KO2,.OtBu,ONOO-)等活性氧物质时抗干扰性的条形图，纵坐标为I594/I666强度的变化。
图6为本发明的荧光探针(10.0×10-6mol/L)在PBS(10mM，pH＝7.0)/CH3CN30％中，与过氧化氢(H2O2)作用过程中比值荧光强度与时间的线性关系研究。
图7为本发明的荧光探针(10.0×10-6mol/L)在PBS(10mM，pH＝7.0)/CH3CN30％中，与过氧化氢(H2O2)作用前后，I594/I666强度随时间变化的曲线。
图8为本发明的荧光探针细胞适用性的毒性研究实验，横坐标为探针浓度，纵坐标为细胞存活率。
图9为本发明的荧光探针在细胞内检测内源性、外源性过氧化氢(H2O2)的成像实验，第一横排为探针本身(10μM)在细胞内的成像情况；第二横排为加入探针(10μM)和外源性过氧化氢(H2O2)(0.5mM)后的成像情况；第三横排为加入内源性刺激药物PMA(1μg/mL)和探针(10μM)后检测过氧化氢(H2O2)的成像情况，使用488nm作为激发波长得到的实验结果。
图10为本发明的荧光探针的共定位成像实验，b为共定位试剂罗丹明123(2.0μM)的成像图，c为探针(10μM)的成像实验图，a为b和c的叠加效果图，d为明场效果图，使用488nm作为激发波长得到的实验结果。
图11为本发明的荧光探针共定位实验得到的重叠系数，说明探针和共定位试剂能很好重叠，从而验证探针能准确定位线粒体。
具体实施实例
实施例1：探针分子的合成
在室温下将化合物2(160.0g,0.46mmol)和化合物3(100.0g,0.4mmol)溶于2.0mL乙醇中的溶液中，加入哌啶(5.0μL，0.05mmol)。在室温下搅拌5分钟后，将所得溶液回流6小时。然后将反应溶液减压浓缩，以二氯甲烷/甲醇(v/v＝15/1)为洗脱剂进行柱层析分离得橙红色固体的化合物4(60.0mg，22.6％产率)。HRMS(EI)m/z:calcd for C37H34BN2O3S+(M+H)+597.23777,found597.23822.1H NMR(400MHz,DMSO)δH 12.35(s,1H),9.61(s,1H),9.12(s,1H),8.77(s,1H),8.68(s,1H),8.46(d,J＝15.1Hz,2H),8.33(s,3H),8.19(s,2H),8.12(s,2H),8.00(s,1H),7.79(d,J＝6.2Hz,1H),7.59(s,1H),7.49(s,1H),7.33(s,2H),6.28(s,2H),1.23(s,13H).13C NMR(101MHz,DMSO)δc 163.98,160.12,159.12,154.31,151.85,149.78,148.53,144.18,138.51,137.27,136.62,135.61,135.14,132.04,131.72,130.05,129.72,127.97,127.58,127.25,127.01,126.36,125.68,123.47,122.60,120.00,118.45,116.54,60.00,59.43,31.59,30.26,29.36,20.37.
实施例2：本发明的荧光探针的应用
将探针分子溶于PBS(10mM，pH＝7.0)/CH3CN30％中配制成10.0×10-6mol/L的溶液，向溶液中加入各种活性氧物质(ROO.,.OH,NO.,TBHP,NaClO,.O2,KO2,.OtBu,ONOO-)后没有引起探针本身发射峰的明显变化，当过氧化氢(H2O2)与干扰物质(ROO.,.OH,NO.,TBHP,NaClO,.O2,KO2,.OtBu,ONOO-)共存时，能够发生响应而导致本身的发射消失而发生蓝移生成新的发射峰，探针不受干扰因素的影响，表现出来很强的抗干扰能力。该探针分子与过氧化氢(H2O2)响应在一定时间及浓度范围内都具良好的线性关系。通过细胞成像试验表明探针能用于内源性、外源性过氧化氢(H2O2)的检测，共定位实验结果证实探针能准确定位线粒体，表现出了良好的生物适应能力，从而实现线粒体内过氧化氢(H2O2)的检测。
Claims (2)
1.一种识别过氧化氢(H2O2)荧光探针的合成，其结构为：
2.一种在细胞内检测过氧化氢(H2O2)比值型红光发射荧光探针的应用。
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108727362A (zh) * | 2018-08-01 | 2018-11-02 | 中南大学 | 一类固体荧光小分子的合成与应用 |
| CN108752371A (zh) * | 2018-07-13 | 2018-11-06 | 济南大学 | 一种基于喹啉的双光子过氧化氢荧光探针 |
| CN110172070A (zh) * | 2019-06-05 | 2019-08-27 | 商丘师范学院 | 一种检测粘度与过氧化氢的荧光探针及其合成方法与应用 |
| CN110894201A (zh) * | 2019-12-13 | 2020-03-20 | 安徽大学 | 一种用于线粒体过氧化氢、蛋白和核酸同时超分辨成像的单分子荧光探针及其制备和应用 |
| CN112358508A (zh) * | 2020-12-01 | 2021-02-12 | 南京工业大学 | 一种通过光调控精确检测生物体内h2o2的荧光探针及其制备方法和应用 |
| CN113387973A (zh) * | 2021-05-24 | 2021-09-14 | 云南师范大学 | 一种双识别荧光探针分子及其制备方法与用途 |
| CN114736223A (zh) * | 2021-01-07 | 2022-07-12 | 湖南超亟检测技术有限责任公司 | 一种新型近红外荧光检测试剂的制备及其体外诊断应用 |
| CN115112656A (zh) * | 2022-07-08 | 2022-09-27 | 四川大学华西医院 | 一种结合免疫荧光染色的黏膜***整体成像的方法 |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105038766A (zh) * | 2015-06-25 | 2015-11-11 | 中国科学院合肥物质科学研究院 | 一种可视可逆的比率荧光探针及其制备方法与应用 |
| CN106749359A (zh) * | 2016-11-30 | 2017-05-31 | 中南大学 | 一种检测过氧化氢新型荧光探针的合成与应用 |
-
2018
- 2018-01-11 CN CN201810028472.6A patent/CN108129500A/zh active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105038766A (zh) * | 2015-06-25 | 2015-11-11 | 中国科学院合肥物质科学研究院 | 一种可视可逆的比率荧光探针及其制备方法与应用 |
| CN106749359A (zh) * | 2016-11-30 | 2017-05-31 | 中南大学 | 一种检测过氧化氢新型荧光探针的合成与应用 |
Non-Patent Citations (2)
| Title |
|---|
| JIAN XU 等: "Mitochondria-Targeted Fluorescent Probe for Imaging Hydrogen Peroxide in Living Cells", 《ANAL. CHEM.》 * |
| YAJIAO ZHANG等: "Reversible Fluorescent Probe for Selective Detection and Cell Imaging of Oxidative Stress Indicator Bisulfite", 《ANAL. CHEM.》 * |
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108752371A (zh) * | 2018-07-13 | 2018-11-06 | 济南大学 | 一种基于喹啉的双光子过氧化氢荧光探针 |
| CN108752371B (zh) * | 2018-07-13 | 2020-06-30 | 济南大学 | 一种基于喹啉的双光子过氧化氢荧光探针 |
| CN108727362A (zh) * | 2018-08-01 | 2018-11-02 | 中南大学 | 一类固体荧光小分子的合成与应用 |
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