CN110407821A - 一种基于fret机制的次氯酸荧光探针 - Google Patents
一种基于fret机制的次氯酸荧光探针 Download PDFInfo
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Abstract
本发明公开了一种基于FRET机制的次氯酸的荧光探针,该探针以香豆素荧光团为能量供体，硫甲基苯为能量受体，通过FRET机制可实现比率型检测。该探针能选择性与HClO作用，随着HClO浓度的增加，其荧光强度在480 nm处逐渐增强，在550 nm处逐渐减弱。该探针能够快速与HClO响应,在生物化学领域具有重要的应用价值。
Description
技术领域
本发明涉及分析化学领域，涉及一种基于FRET机制的次氯酸的荧光探针的制备。
背景技术
活性氧种类（ROS），如次氯酸（HClO）、过氧化氢（H2O2）、羟基（•OH）、超氧阴离子（O2•-）是在氧气代谢过程中形成为一种单电子还原产物，在各种生理和病理过程中发挥关键作用。其中HClO在氧化应激和信号转导中起着重要的作用，控制着广泛的生理功能。更重要的是，因其强大的氧化作用，HClO还是一种有效的抗菌药物，是天然免疫系统中的一种防御工具，可以应对细菌入侵并杀死各种病原体。在免疫细胞中，内源性次氯酸盐主要由氯离子(Cl-)和过氧化氢(H2O2)在髓过氧化物酶(MPO)催化下产生。此外，由于HClO具有高氧化性、非特异性和扩散性，过量产生HClO会导致氧化应激, 从而导致不可逆转的组织损伤和疾病，包括神经元退化、心血管疾病、骨关节炎、哮喘、囊性纤维化和动脉粥样硬化。此外，次氯酸盐在日常生活中得到了广泛的应用，如纸张和纺织品的漂白，饮用水的消毒除臭。在水中残留的次氯酸会引起水体污染，对人体健康存在潜在危害。因此，高效、灵敏地检测实际水体和生物体系中次氯酸的浓度已成为一个重要的研究课题。
荧光探针具有技术简单、灵敏度高、选择性好、时空分辨率高、易于可视化、生物相容性好等优点，引起广泛关注。而基于FRET机制比率型荧光探针因其能够有效消除环境、探针浓度及激发光强度的干扰更是备受青睐。因此，开发新的基于FRET机制的次氯酸比率荧光探针意义重大。
发明内容
本发明目的之一是提供一种基于FRET机制的次氯酸荧光探针的合成方法；目的之二是提供一种基于FRET机制的次氯酸的比率型荧光探针。
为实现上述目的，本发明采用如下技术方案。
一种基于FRET机制的次氯酸荧光探针，其分子结构如下：。具体合成路线如下：。具体合成方法如下：（1）将4-(二乙胺)水杨醛和丙二酸二乙酯溶于甲苯乙腈(1：2)混合溶剂中，加入少量哌嗪催化，将反应溶液加热回流10 h。减压除去溶剂，经柱层析分离(石油醚/乙酸乙酯，4：1) 得到黄色油状物化合物1。（2）将化合物1和3-氨基-1-丙醇溶于无水乙醇中，加热回流36 h。减压除去大部分溶剂，至即将析出固体，缓慢滴加水，室温搅拌10 min后，过滤干燥，经柱层析分离（石油醚：乙酸乙酯 = 1 ：1）得黄色固体化合物2。（3）将三苯基膦和咪唑溶于CH2Cl2中，缓慢加入碘单质，室温搅拌10 min后，缓慢加入化合物2，室温搅拌24 h。减压除去溶剂，加入少量乙酸乙酯溶解，缓慢滴加大量石油醚析出固体，搅拌30 min，过滤干燥，经柱层析分离（二氯甲烷），得黄色固体化合物3。（4）将化合物3和4-甲基吡啶溶于乙腈中，加热回流过夜。减压除去溶剂，经柱层析分离（二氯甲烷：甲醇= 20 ：1）得黄色固体化合物4。（5）将化合物4和4-（甲基硫基）苯甲醛溶于无水乙醇中，加入少量哌嗪催化，加热回流24 h。减压除去溶剂，经柱层析分离（二氯甲烷：甲醇= 20 ：1）得探针CMS。
本发明荧光探针在检测次氯酸的应用。
所述荧光探针在检测次氯酸的应用中，激发波长为400 nm，检测波段为450-750nm。
本发明具有以下优点：
本发明涉及的一种基于FRET机制的次氯酸荧光探针，灵敏度高，选择性好，响应快，在生物化学领域具有重要的应用价值。
附图说明
图1为探针分子结构图。
图2为探针的1H NMR谱。
图3为探针的13C NMR谱。
图4为探针对不同浓度次氯酸响应的荧光光谱。
图5为探针对次氯酸响应时间图。
图6为探针抗干扰性图。
具体实施方式
实施例1 探针的合成。
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（1）将4-(二乙胺)水杨醛(5.0 g，25.9mmol)和丙二酸二乙酯(4.97g，31.0 mmol)溶于甲苯乙腈(1：2，100 ml)混合溶剂中，加入少量哌嗪催化，将反应溶液加热回流10 h。减压除去溶剂，经柱层析分离(石油醚/乙酸乙酯，4：1) 得到黄色油状物化合物1(7.18 g，96%)。（2）将化合物1（2.15 g，7.43 mmol）和3-氨基-1-丙醇（0.84 g，11.18 mmol）溶于20mL无水乙醇中，加热回流36 h。减压除去大部分溶剂，至即将析出固体，缓慢滴加20 mL水，室温搅拌10 min后，过滤干燥，经柱层析分离（石油醚：乙酸乙酯 = 1 ：1）得黄色固体化合物2（2.30 g，7.22 mmol，97.2%）。（3）将三苯基膦（2.46 g，9.38 mmol）和咪唑（0.64 g，9.40mmol）溶于30 mL CH2Cl2中，缓慢加入碘单质（2.38 g，9.38 mmol），室温搅拌10 min后，缓慢加入化合物2（2.30 g，7.22 mmol），室温搅拌24 h。减压除去溶剂，加入少量乙酸乙酯溶解，缓慢滴加大量石油醚析出固体，搅拌30 min，过滤干燥，经柱层析分离（二氯甲烷），得黄色固体化合物3（1.68 g，3.92 mmol，54.3%）。（4）将化合物3（1.68 g，3.92 mmol）和4-甲基吡啶（0.55 g，5.91 mmol）溶于乙腈中，加热回流过夜。减压除去溶剂，经柱层析分离（二氯甲烷：甲醇= 20 ：1）得黄色固体化合物4（1.02 g，1.96 mmol，50%）。（5）将化合物4（0.5 g，0.96 mmol）和4-（甲基硫基）苯甲醛（0.15 g，0.99 mmol）溶于10 mL无水乙醇中，加入少量哌嗪催化，加热回流24 h。减压除去溶剂，经柱层析分离（二氯甲烷：甲醇= 20 ：1）得探针CMS（0.56 g，0.83 mmol，86.5%）。探针1H NMR谱和13C NMR谱如图2和3：1H NMR (600 MHz,CDCl3) δ 9.15 (d, J = 6.7 Hz, 2H), 8.89 (t, J = 6.1 Hz, 1H), 8.60 (s, 1H),8.05 (d, J = 6.7 Hz, 2H), 7.65 (d, J = 16.2 Hz, 1H), 7.53 (d, J = 8.4 Hz,2H), 7.43 (d, J = 9.0 Hz, 1H), 7.19 (d, J = 8.4 Hz, 2H), 7.12 (d, J = 16.2Hz, 1H), 6.60 (dd, J = 9.0, 2.3 Hz, 1H), 6.39 (d, J = 2.2 Hz, 1H), 4.81 (t, J= 6.4 Hz, 2H), 3.53 (dd, J = 12.1, 6.1 Hz, 2H), 3.40 (q, J = 7.1 Hz, 4H),2.49 (s, 3H), 2.42 – 2.32 (m, 2H), 1.20 (t, J = 7.1 Hz, 6H). 13C NMR (151 MHz,CDCl3) δ 125.77, 124.00, 96.47, 77.27, 77.06, 76.85, 58.48, 45.12, 35.82,31.84, 14.98, 12.44.
实施例2 荧光探针对次氯酸的响应
在DMSO中配制浓度为1 mM探针储备液。配制不同浓度的次氯酸缓冲溶液DMSO/PBS（1/1，v/v，PBS 10 mM，pH=7.4）（浓度梯度为5 eq的0-60 eq），其中探针含量为10 µM，进行荧光检测（λex=400 nm），记录480 nm与550 nm处的荧光强度，如图4所示。分析I480nm/I550nm处的荧光强度与次氯酸浓度的线性关系，如图所示：随着次氯酸浓度的增加，480 nm处的吸收峰逐渐增加550 nm处的吸收峰逐渐降低，当次氯酸浓度达到60 eq时，反应体系吸收峰值基本不再发生变化。
实施例3 荧光探针的响应时间
取出30 µL探针母液于3 mL测试体系DMSO/PBS（1/1，v/v，PBS 10 mM，pH=7.4）中，然后加入40 eq次氯酸，摇晃均匀后立即用荧光光谱仪测试其荧光强度变化。由图5可知，20 s内荧光强度比值达到最大值，探针与次氯酸迅速响应，可用于实时分析。
实施例4 荧光探针的选择性
配制浓度为 20 mM的ROS，阴离子和硫醇（NaClO, TBHP, H2O2, ONOO-, .OH, F-, Cl-,Br-, I-, NO3 -, NO2 -, AcO-, PO4 3-, SO4 2-, HSO3 -, HCO3 -, CO3 2-, Na2S, Na2S2O3, Cys,Hcy, GSH ）溶液。
将NaClO和不同干扰物质的溶液与探针配制成缓冲溶液DMSO/PBS（1/1，v/v，PBS10 mM，pH=7.4），干扰物质终浓度为400 μM，探针终浓度为10 μM，摇匀后进行荧光检测（λex=400 nm），建立荧光强度比I480/I550与各干扰物质的柱状图，如图5所示。由图可知，荧光探针对其他干扰物质响应值低，对NaClO的I480/I550比值高，特异性好、抗干扰性能好。其他干扰对探针的荧光比值几乎没有影响，能够选择性的识别次氯酸。
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1.一种检测次氯酸的荧光探针，其结构为：。
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| CN106518860A (zh) * | 2016-11-07 | 2017-03-22 | 山东大学 | 一种基于荧光共振能量转移机理的靶向线粒体的次氯酸比例荧光探针及其应用 |
| CN109053700A (zh) * | 2018-07-16 | 2018-12-21 | 山东大学 | 一种次氯酸比率荧光探针及其应用 |
| CN109796966A (zh) * | 2019-02-25 | 2019-05-24 | 山东大学 | 一种次氯酸比率荧光探针及其应用 |
| CN109912581A (zh) * | 2019-03-28 | 2019-06-21 | 山东大学 | 基于香豆素与苯乙烯吡啶鎓的次氯酸荧光探针及其应用 |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106518860A (zh) * | 2016-11-07 | 2017-03-22 | 山东大学 | 一种基于荧光共振能量转移机理的靶向线粒体的次氯酸比例荧光探针及其应用 |
| CN109053700A (zh) * | 2018-07-16 | 2018-12-21 | 山东大学 | 一种次氯酸比率荧光探针及其应用 |
| CN109796966A (zh) * | 2019-02-25 | 2019-05-24 | 山东大学 | 一种次氯酸比率荧光探针及其应用 |
| CN109912581A (zh) * | 2019-03-28 | 2019-06-21 | 山东大学 | 基于香豆素与苯乙烯吡啶鎓的次氯酸荧光探针及其应用 |
Non-Patent Citations (3)
| Title |
|---|
| LI-JIE ZHANG，等: "A new water-soluble and mitochondria-targeted fluorescence probe for ratiometric detection of hypochlorous acid in living cells", 《SENSORS & ACTUATORS: B. CHEMICAL》 * |
| YE-HAO YAN，等: "A new FRET-based ratiometric fluorescence probe for hypochlorous acid and its imaging in living cells", 《TALANTA》 * |
| YE-HAO YAN，等: "A ratiometric fluorescence probe based on a novel recognition mechanism for monitoring endogenous hypochlorite in living cells", 《ANALYTICA CHIMICA ACTA》 * |
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