CN107840829A - 一种特异性识别硫化氢的荧光探针 - Google Patents

一种特异性识别硫化氢的荧光探针 Download PDF

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CN107840829A
CN107840829A CN201711062732.3A CN201711062732A CN107840829A CN 107840829 A CN107840829 A CN 107840829A CN 201711062732 A CN201711062732 A CN 201711062732A CN 107840829 A CN107840829 A CN 107840829A
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hydrogen sulfide
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宋相志
苏远安
杨雷
齐风佩
廖立德
谭倩
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Central South University
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Abstract

本发明公开了一种特异性识别硫化氢的荧光探针，其分子结构式如下：

Description

一种特异性识别硫化氢的荧光探针
技术领域
本发明涉及的是化学分析检测技术领域，具体涉及一种可以特异性识别硫化氢的荧光探针，并研究了它的制备方法以及该荧光探针在体外和活细胞内检测硫化氢方面的应用。
背景技术
硫化氢(H2S)被认为是除一氧化氮(NO)、一氧化碳(CO)之外的第三种气体信号分子。在人体内，胱硫醚β-合酶、胱硫醚γ-裂解酶以及3-巯基丙酮酸硫基转移酶参与的酶促反应产生硫化氢。硫化氢在舒张血管、神经调节、细胞凋亡、消炎、抗氧化、抑制胰岛素信号等生理方面有重要作用。研究表明，体内硫化氢含量异常与唐氏综合征、糖尿病、高血压、阿尔茨海默氏症相关。因此，它们在生物体内含量变化可以作为这些疾病诊断的依据。目前为止应用于测定硫化氢的方法有多种，包括电化学方法，比色法，气相色谱法，这些方法只能够在在体外检测，不能够进行实时生物体内分析。荧光探针由于其灵敏度高、检测限低、选择性好，抗干扰性强，能够在活细胞中可视化检测分析物，所以研究者们正致力于将荧光探针应用于体外或细胞内硫化氢的监测或细胞成像。目前报道的反应型硫化氢荧光探针机理有：还原叠氮基(-N3)成氨基反应、亲核加成反应、迈克尔加成反应。生物体内的硫化氢检测会受到生物硫醇的干扰，硫化氢作为一种强还原剂可以特异还原叠氮基减少生物硫醇的干扰。近红外荧光染料由于能量小、对组织伤害小、组织穿透能力强、背景干扰小在细胞成像方面有很大的应用前景，研发近红外荧光探针检测硫化氢有重要意义。
发明内容
本发明目的之一是提供一种合成路线简单、反应条件温和、成本较低、操作简单的荧光探针合成方法；目的之二是提供一种灵敏度高、选择性好、抗干扰能力强、斯托克斯位移大、发射波长在近红外、能够在生物体外及生物体内检测硫化氢的近红外荧光探针。
本发明解决问题采取的技术方案为，一种可以特异性识别硫化氢的新型荧光探针，其分子结构式如下：具体合如下：
具体成方法如下：
(1)将1.0g对甲苯胺加入到4.6mL的盐酸溶液(V盐酸：V水＝1:1)，0℃搅拌5分钟。再将0.63g亚硝酸钠溶于2.6mL冷水中，逐滴加入到反应液中，最后将0.61g叠氮化钠溶于7.8mL水中，逐滴加入反应液中，继续保持冰水浴12小时。向反应液中加入50mL二氯甲烷摇匀分液，有机相用30mL×2水洗涤，旋干溶剂得到1.0g棕黄色油状物为化合物3，产率81％。(2)将1.0g化合物3溶于30mLCCl4中，加入1.48g NBS、210mg BPO，氩气保护，回流反应12小时。旋干溶剂，柱层析得到1.2g黄色油状物为化合物4，产率70.2％。(3)将234mg化合物5，424mg化合物4，溶于20mL丙酮，加入560mg碳酸钾，氩气保护，回流反应8小时。抽滤，旋干溶剂，柱层析，得到330mg红色固体为化合物6，产率90.4％。(4)将120mg化合物6，30mg丙二腈溶于10mL无水乙醇，加入20μL哌啶，氩气保护，室温反应5小时。旋干溶剂，柱层析得到110mg红色粉末为探针1，产率78.0％。
本发明的荧光探针测试方法如下，将探针分子溶解在pH为7.4的HEPES(20.0mMHEPES，1.0mM CTAB)缓冲溶液中，室温下进行测试。可以对硫化氢进行定量检测，具体实施方法在实施实例中详细介绍。
本发明的荧光探针的响应机理如下：硫化氢与探针分子响应后，叠氮基还原成氨基，脱去一个3-亚氨基-6-亚甲基-1，4-环己二烯分子，产生一个氧负离子中间体，经历一次分子内合环，形成一个亚胺香豆素染料。溶液从无荧光变成发红光，从而实现了检测硫化氢过程。探针分子的响应过程如下：
本发明的荧光探针与硫化氢响应后的荧光发射峰在610nm处，荧光增强了9.3倍。
本发明所述的探针分子合成路线简单，操作简单，成本较低，对硫化氢的选择性好、抗干扰能力强，斯托克斯位移大，发射波长在近红外，该荧光探针在生物诊断、分析化学、环境科学等领域具有实际的应用价值。
附图说明
图1为本发明荧光探针的选择性，荧光探针(1.0×10-5mol/L)在pH为7.4的HEPES(20.0mM HEPES，1.0mM CTAB)缓冲溶液中，与不同物质响应后的荧光光谱，横坐标为波长，纵坐标为荧光强度。
图2为本发明荧光探针的抗干扰能力，硫化氢与其它干扰物共存时，与荧光探针(1.0×10-5mol/L)在pH为7.4的HEPES(20.0mM HEPES，1.0mM CTAB)缓冲溶液中与硫化氢响应后的荧光强度比值(I/I0)柱状图。
图3为本发明的荧光探针(1.0×10-5mol/L)在pH为7.4的HEPES(20.0mM HEPES，1.0mM CTAB)缓冲溶液中，与不同浓度硫化氢响应后的荧光光谱变化，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(1.0×10-5mol/L)pH为7.4的HEPES(20.0mM HEPES，1.0mMCTAB)缓冲溶液中，与硫化氢浓度的线性关系，横坐标为波长，纵坐标为荧光强度。
图5为本发明的荧光探针(1.0×10-5mol/L)在pH为7.4的HEPES(20.0mM HEPES，1.0mM CTAB)缓冲溶液中，与硫化氢响应过程中荧光强度随时间的变化，横坐标为时间，纵坐标为荧光强度。
图6为本发明的荧光探针(1.0×10-5mol/L)在不同pH值缓冲溶液中，与硫化氢响应前后的荧光强度，横坐标为pH，纵坐标为荧光强度。
图7为本发明的荧光探针(1.0×10-5mol/L)在不同条件下的细胞(Hela细胞)成像图(a)探针和硫化氢溶液处理的细胞中明场成像(b)探针和硫化氢溶液处理的细胞中复合场成像(c)探针和硫化氢溶液处理的细胞中荧光成像中荧光成像(d)探针在细胞中明场成像(e)探针在细胞中复合场成像(f)探针在细胞中荧光成像。
具体实施实例
实施例1：中间产物3的合成
将1.0g对甲苯胺加入到4.6mL的盐酸溶液(V盐酸：V水＝1:1)，0℃搅拌5分钟。再将0.63g亚硝酸钠溶于2.6mL冷水中，逐滴加入到反应液中，最后将0.61g叠氮化钠溶于7.8mL水中，逐滴加入反应液中，继续保持冰水浴12小时。向反应液中加入50mL二氯甲烷摇匀分液，有机相用30mL×2水洗涤，旋干溶剂得到1.0g棕黄色油状物为化合物3，产率81％。
实施例2：中间产物4的合成
将1.0g化合物3溶于30mLCCl4中，加入1.48g NBS、210mg BPO，氩气保护，回流反应12小时。旋干溶剂，柱层析得到1.2g黄色油状物为化合物4，产率70.2％。
实施例3：中间产物6的合成
将234mg化合物5，424mg化合物4，溶于20mL丙酮，加入560mg碳酸钾，氩气保护，回流反应8小时。抽滤，旋干溶剂，柱层析，得到330mg红色固体为化合物6，产率90.4％。HRMS(EI)m/z:理论值C20H23N5O2[M+Na]+,388.1749；实验值，388.1451。1H NMR(500MHz,CDCl3)δ10.21(s,1H),7.42(d,J＝8.4Hz,2H),7.04(d,J＝8.4Hz,2H),7.00(s,1H),6.04(s,1H),5.09(s,2H),3.52–3.47(m,2H),3.33(dq,J＝21.3,7.1Hz,4H),3.18–3.14(m,2H),1.16(q,J＝7.0Hz,6H)。13C NMR(101MHz,CDCl3)δ186.51,157.54,143.40,139.61,133.80,129.14,128.82,119.15,114.25,108.32,94.41,70.58,47.81,45.72,45.20,44.69,10.67,9.96。
实施例4：探针1的合成
将120mg化合物6，30mg丙二腈溶于10mL无水乙醇，加入20μL哌啶，氩气保护，室温反应5小时。旋干溶剂，柱层析得到110mg红色粉末为探针1，产率78.0％。HRMS(EI)m/z:理论值C23H23N7O[M]+,413.1964；实验值,413.1568。1H NMR(500MHz,CDCl3)δ8.01(s,1H),7.55(s,1H),7.39(d,J＝8.4Hz,2H),7.09(d,J＝8.4Hz,2H),6.00(s,1H),5.07(s,2H),3.59(t,J＝4.8Hz,2H),3.37(dq,J＝28.5,7.0Hz,4H),3.25–3.17(m,2H),1.20(dt,J＝27.8,7.1Hz,6H)。13C NMR(101MHz,CDCl3)δ149.68,129.06,119.45,93.38,70.59,48.23,46.08,45.52,44.51,10.92,9.76,8.70。
实施例5：本发明:荧光探针的应用
将探针溶于7.4的HEPES(20.0mM HEPES,1.0mM CTAB)缓冲溶液中配制成1.0×10- 5mol/L的溶液，向溶液中加其它选择项物质(Na+、K+、Ca2+、Mg2+、PO4 3-、S2O3 2-、SCN-、SO4 2-、Cl-、CO3 2-、HCO3 -、NO2 -、Br-、SO3 2-、HSO3 -、NO3 -、AcO-、F-、N3 -、I-、H2O2、ClO-、Cys、Hcy、GSH)，加人硫化氢导致溶液荧光强度变化，该荧光探针对硫化氢表现出高灵敏性、高选择性的识别。当硫化氢与干扰物质(Na+、K+、Ca2+、Mg2+、PO4 3-、S2O3 2-、SCN-、SO4 2-、Cl-、CO3 2-、HCO3 -、NO2 -、Br-、SO3 2-、HSO3 -、NO3 -、AcO-、F-、N3 -、I-、H2O2、ClO-、Cys、Hcy、GSH)共存时，探针对硫化氢响应表现出来很强的抗干扰能力。该探针分子与硫化氢响应速度快，半个小时内可以观察到荧光的变化。探针分子在pH为7至10的范围内都可以对硫化氢选择性识别，可以在细胞内检测硫化氢。

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1.一种特异性识别硫化氢的荧光探针，其结构为：
CN201711062732.3A 2017-11-02 2017-11-02 一种特异性识别硫化氢的荧光探针 Pending CN107840829A (zh)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108676012A (zh) * 2018-08-01 2018-10-19 华侨大学 一种亚胺香豆素类化合物的制备方法
CN109180716A (zh) * 2018-09-19 2019-01-11 中南大学 一种多信号比率型区分检测h2o2和h2s的荧光探针的设计、合成及应用
CN109593087A (zh) * 2018-09-19 2019-04-09 中南大学 一种多通道同时区分检测Cys/Hcy，GSH和H2S的荧光探针的设计、合成及应用
CN110724114A (zh) * 2018-07-16 2020-01-24 北京工商大学 一种检测亚硫酸氢根及硫化氢的荧光探针

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104804728A (zh) * 2015-04-27 2015-07-29 苏州罗兰生物科技有限公司 一种荧光增强型苯硫酚荧光探针的制备及应用
CN105884713A (zh) * 2016-04-26 2016-08-24 济南大学 一种荧光增强型硫化氢分子荧光探针及其制备方法和应用
CN106946801A (zh) * 2016-01-06 2017-07-14 中南大学 一种特异性识别半胱氨酸的新型荧光探针的制备与应用

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104804728A (zh) * 2015-04-27 2015-07-29 苏州罗兰生物科技有限公司 一种荧光增强型苯硫酚荧光探针的制备及应用
CN106946801A (zh) * 2016-01-06 2017-07-14 中南大学 一种特异性识别半胱氨酸的新型荧光探针的制备与应用
CN105884713A (zh) * 2016-04-26 2016-08-24 济南大学 一种荧光增强型硫化氢分子荧光探针及其制备方法和应用

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
K. J. JARAG，等: "Synthesis, Photophysical Properties, and Antimicrobial Activity of Novel Styryl Colorants Derived from 7-Methoxy-1,4-diphenethyl-1,2,3,4-tetrahydroquinoxaline-6-carbaldehyde", 《JOURNAL OF HETEROCYCLIC CHEMISTRY》 *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110724114A (zh) * 2018-07-16 2020-01-24 北京工商大学 一种检测亚硫酸氢根及硫化氢的荧光探针
CN110724114B (zh) * 2018-07-16 2022-10-21 北京工商大学 一种检测亚硫酸氢根及硫化氢的荧光探针
CN108676012A (zh) * 2018-08-01 2018-10-19 华侨大学 一种亚胺香豆素类化合物的制备方法
CN109180716A (zh) * 2018-09-19 2019-01-11 中南大学 一种多信号比率型区分检测h2o2和h2s的荧光探针的设计、合成及应用
CN109593087A (zh) * 2018-09-19 2019-04-09 中南大学 一种多通道同时区分检测Cys/Hcy，GSH和H2S的荧光探针的设计、合成及应用
CN109180716B (zh) * 2018-09-19 2020-11-13 中南大学 一种多信号比率型区分检测h2o2和h2s的荧光探针的设计、合成及应用

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