CN108299450A - 一种检测生物硫醇的荧光探针 - Google Patents

一种检测生物硫醇的荧光探针 Download PDF

Info

Publication number
CN108299450A
CN108299450A CN201810070839.0A CN201810070839A CN108299450A CN 108299450 A CN108299450 A CN 108299450A CN 201810070839 A CN201810070839 A CN 201810070839A CN 108299450 A CN108299450 A CN 108299450A
Authority
CN
China
Prior art keywords
fluorescence
probe
biological thiol
present
fluorescence probe
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
CN201810070839.0A
Other languages
English (en)
Inventor
宋相志
苏远安
杨雷
熊海青
张帆
韩金量
张赟
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Central South University
Original Assignee
Central South University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Central South University filed Critical Central South University
Priority to CN201810070839.0A priority Critical patent/CN108299450A/zh
Publication of CN108299450A publication Critical patent/CN108299450A/zh
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/12—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains three hetero rings
    • C07D491/16—Peri-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
    • C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
    • C09K2211/10—Non-macromolecular compounds
    • C09K2211/1018—Heterocyclic compounds
    • C09K2211/1025—Heterocyclic compounds characterised by ligands
    • C09K2211/1029—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
    • C09K2211/10—Non-macromolecular compounds
    • C09K2211/1018—Heterocyclic compounds
    • C09K2211/1025—Heterocyclic compounds characterised by ligands
    • C09K2211/1088—Heterocyclic compounds characterised by ligands containing oxygen as the only heteroatom

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Immunology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Engineering & Computer Science (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Optics & Photonics (AREA)
  • Materials Engineering (AREA)
  • Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)

Abstract

本发明涉及一种检测生物硫醇的荧光探针，属于荧光探针领域。其分子结构如下：该探针分子没有荧光，与生物硫醇作用后发出蓝色荧光。本发明所述的探针分子不仅可以定性分析生物硫醇，而且可以实现快速、定量检测生物硫醇，在生物化学等领域具有重要的应用价值。

Description

一种检测生物硫醇的荧光探针
本发明涉及一种基于香豆素型生物硫醇荧光探针的制备及其在检测生物硫醇方面的研究，属于荧光探针领域。
背景技术
生物硫醇是生物体内含巯基化合物的总称，广泛地存在于生物体内，最具代表性的生物硫醇为三种巯基化合物：半胱氨酸(Cys)、同型半胱氨酸(Hcy)和谷胱甘肽(GSH)。巯基化合物是生物体酶发挥作用后和生理活动中的重要信号分子，同时也可以调节细胞的正常氧化还原状态，在生物体的生理活动中具有重要的功能。生物体内巯基化合物的浓度异常与许多种疾病有关，Cys的缺乏与生长缓慢、肝损伤、毛发褪色、皮肤病、昏睡以及水肿等疾病有关；血浆中Hcy浓度过高与心血管疾病、阿兹海默症和骨质疏松症有关；GSH能够保护机体免受氧化及自由基介导的氧应激损伤，其浓度与白血病、癌症及艾滋病等有关。因此，检测生物体内巯基化合物浓度对研究细胞功能和进行疾病早期诊断具有重要的意义。相对于其它传统的检测技术，荧光分析法具有操作简单、选择性好、灵敏度高等优点，尤其可以用于细胞及生物活体成像，因此近年来利用荧光探针来检测离子及生物分子的研究受到了广泛的关注。
发明内容
本发明目的在于提供一种检测生物硫醇的荧光探针，其结构式如下：合成路线如下：
具体合成方法如下：(a)在25mL的圆底烧瓶中加入400mg化合物1、295mg化合物2，再加入10mL甲苯，氩气保护回流反应24小时，抽滤，用10mL石油醚洗涤滤饼，烘干得到210mg灰白色粉末为化合物3，产率：54.5％。(b)在25mL圆底烧瓶中加入50mg化合物3、三乙胺35μL冰水浴30分钟，再将63.8mg 2,4-二硝基苯磺酰氯加入到烧瓶中，继续保持冰水浴，2小时后旋干溶剂进行柱层析，得到34.2mg黑色粉末为探针，产率35.1％。
本发明的荧光探针测试方法如下，将探针分子溶解在pH＝7.4PBS(10.0mM)与乙腈混合溶液(V:V＝7：3)中，室温下进行测试。可以对生物硫醇进行定性、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：生物硫醇与探针分子作用后，生物硫醇的巯基进攻磺酰酯基，探针分子脱去一个二氧化硫分子、2,4-二硝基苯硫醇分子，释放出染料分子。溶液从黄色变成无色，从没有荧光变成有荧光，实现检测生物硫醇。探针分子的响应过程如下：
本发明的荧光探针没有荧光，与生物硫醇响应后的荧光发射峰在409nm处。探针与生物硫醇完全响应后409nm处荧光强度显著增强。
本发明所述的探针分子合成路线简单，操作简单，成本较低，可以实现快速检测生物硫醇。
附图说明
图1为本发明的荧光探针在氘代DMSO中核磁共振氢谱，横坐标化学位移，纵坐标为强度。
图2为本发明的荧光探针在氘代DMSO中核磁共振碳谱，横坐标化学位移，纵坐标为强度。
图3为本发明的荧光探针(1.0×10-5mol/L)在pH＝7.4PBS(10.0mM)与乙腈混合溶液(V:V＝7：3)中，与浓度均为50μM三种不同生物硫醇作用后荧光强度与时间关系，横坐标为时间，纵坐标为荧光强度。
图4为本发明的荧光探针(1.0×10-5mol/L)在pH＝7.4PBS(10.0mM)与乙腈混合溶液(V:V＝7：3)中，与不同浓度半胱氨酸作用后荧光光谱图，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＝7.4PBS(10.0mM)与乙腈混合溶液(V:V＝7：3)中，与不同浓度半胱氨酸作用后在409纳米处荧光强度与半胱氨酸浓度关系，横坐标为半胱氨酸浓度，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pH＝7.4PBS(10.0mM)与乙腈混合溶液(V:V＝7：3)中，与不同浓度半胱氨酸作用后在409纳米处荧光强度与半胱氨酸浓度线性关系，横坐标为半胱氨酸浓度，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pH＝7.4PBS(10.0mM)与乙腈混合溶液(V:V＝7：3)中，与不同浓度同型半胱氨酸作用后荧光光谱图，横坐标为波长，纵坐标为荧光强度。
图8为本发明的荧光探针(1.0×10-5mol/L)在pH＝7.4PBS(10.0mM)与乙腈混合溶液(V:V＝7：3)中，与不同浓度同型半胱氨酸作用后在409纳米处荧光强度与同型半胱氨酸浓度关系，横坐标为同型半胱氨酸浓度，纵坐标为荧光强度。
图9为本发明的荧光探针(1.0×10-5mol/L)在pH＝7.4PBS(10.0mM)与乙腈混合溶液(V:V＝7：3)中，与不同浓度同型半胱氨酸作用后在409纳米处荧光强度与同型半胱氨酸浓度线性关系，横坐标为同型半胱氨酸浓度，纵坐标为荧光强度。
图10为本发明的荧光探针(1.0×10-5mol/L)在pH＝7.4PBS(10.0mM)与乙腈混合溶液(V:V＝7：3)中，与不同浓度谷胱甘肽作用后荧光光谱图，横坐标为波长，纵坐标为荧光强度。
图11为本发明的荧光探针(1.0×10-5mol/L)在pH＝7.4PBS(10.0mM)与乙腈混合溶液(V:V＝7：3)中，与不同浓度谷胱甘肽作用后在409纳米处荧光强度与谷胱甘肽浓度关系，横坐标为谷胱甘肽浓度，纵坐标为荧光强度。
图12为本发明的荧光探针(1.0×10-5mol/L)在pH＝7.4PBS(10.0mM)与乙腈混合溶液(V:V＝7：3)中，与不同浓度谷胱甘肽作用后在409纳米处荧光强度与谷胱甘肽浓度线性，横坐标为谷胱甘肽浓度，纵坐标为荧光强度。
具体实施实例
实施例1：化合物3的合成
在25mL的圆底烧瓶中加入400mg化合物1、295mg化合物2，再加入10mL甲苯，氩气保护回流反应24小时，抽滤，用10mL石油醚洗涤滤饼，烘干得到210mg灰白色粉末为化合物3，产率，54.5％。
实施例2：探针的合成
在25mL圆底烧瓶中加入50mg化合物3、三乙胺35μL冰水浴30分钟，再将63.8mg 2,4-二硝基苯磺酰氯加入到烧瓶中，继续保持冰水浴，2小时后旋干溶剂进行柱层析，得到34.2mg黑色粉末为探针，产率35.1％。1H NMR(400MHz,DMSO)δ9.13(d,J＝1.8Hz,1H),8.66(dd,J＝8.7,1.9Hz,1H),8.55(d,J＝8.7Hz,1H),6.98(s,1H),5.85(s,1H),3.29(s,4H),2.71(t,J＝6.2Hz,2H),2.65(t,J＝5.9Hz,2H),1.92–1.79(m,4H).13C NMR(101MHz,DMSO)δ161.21(s),158.19(s),152.24(s),151.18(s),148.46(s),147.55(s),134.31(s),130.99(s),128.30(s),121.76(s),119.85(s),119.22(s),106.06(s),101.65(s),97.28(s),49.75(s),49.20(s),27.35(s),20.97(s),20.31(s),20.04(s).
实施案例3：本发明荧光探针检测生物硫醇研究
将探针溶pH＝7.4PBS(10.0mM)与乙腈混合溶液(V:V＝7：3)中,配制成1.0×10-5mol/L的溶液，加入三种生物硫醇可以迅速观测到409纳米处的荧光变化，检测时间短，可以实现定性、定量检测生物硫醇。

Claims (1)

1.一种检测生物硫醇的荧光探针，其结构为：
CN201810070839.0A 2018-01-25 2018-01-25 一种检测生物硫醇的荧光探针 Withdrawn CN108299450A (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201810070839.0A CN108299450A (zh) 2018-01-25 2018-01-25 一种检测生物硫醇的荧光探针

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201810070839.0A CN108299450A (zh) 2018-01-25 2018-01-25 一种检测生物硫醇的荧光探针

Publications (1)

Publication Number Publication Date
CN108299450A true CN108299450A (zh) 2018-07-20

Family

ID=62866506

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201810070839.0A Withdrawn CN108299450A (zh) 2018-01-25 2018-01-25 一种检测生物硫醇的荧光探针

Country Status (1)

Country Link
CN (1) CN108299450A (zh)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108912134A (zh) * 2018-07-17 2018-11-30 商丘师范学院 一种高灵敏检测半胱氨酸的荧光探针及制备方法、应用
CN109912612A (zh) * 2019-04-04 2019-06-21 郑州大学 一种基于分子逻辑门识别Cys、GSH和HOCl荧光探针的合成、制备及其应用
CN110615786A (zh) * 2019-07-17 2019-12-27 浙江工业大学 用于检测粘度的近红外荧光化合物及其制备与应用
CN112209923A (zh) * 2020-10-14 2021-01-12 中南大学 一种高选择性检测Cys/Hcy和GSH的近红外荧光探针的合成和应用

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
XIAO-BO WANG，等: "A novel reaction-based fluorescent turn-on probe for biothiols and itsapplication in cell imaging", 《SENSORS AND ACTUATORS B》 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108912134A (zh) * 2018-07-17 2018-11-30 商丘师范学院 一种高灵敏检测半胱氨酸的荧光探针及制备方法、应用
CN109912612A (zh) * 2019-04-04 2019-06-21 郑州大学 一种基于分子逻辑门识别Cys、GSH和HOCl荧光探针的合成、制备及其应用
CN109912612B (zh) * 2019-04-04 2021-05-11 郑州大学 一种基于分子逻辑门识别Cys、GSH和HOCl荧光探针的合成、制备及其应用
CN110615786A (zh) * 2019-07-17 2019-12-27 浙江工业大学 用于检测粘度的近红外荧光化合物及其制备与应用
CN112209923A (zh) * 2020-10-14 2021-01-12 中南大学 一种高选择性检测Cys/Hcy和GSH的近红外荧光探针的合成和应用

Similar Documents

Publication Publication Date Title
Chen et al. A novel imidazo [1, 5-α] pyridine-based fluorescent probe with a large Stokes shift for imaging hydrogen sulfide
CN108299450A (zh) 一种检测生物硫醇的荧光探针
Yin et al. Near-infrared fluorescent probe with rapid response and large Stokes shift for imaging peroxynitrite in living cells, zebrafish and mice
CN106220640B (zh) 一类汞离子荧光探针及其制备方法和应用
CN111807993B (zh) 一种用于特异性检测肼的近红外荧光化合物及制备方法
CN107602502B (zh) 一种用于生物硫醇检测的esipt型荧光探针及应用
Yang et al. Detection and differentiation of Cys, Hcy and GSH mixtures by 19F NMR probe
CN108997326A (zh) 一种硫醇荧光探针及其制备方法和应用
CN110563689A (zh) 一种特异性检测活细胞中的半胱氨酸的长波长发射荧光探针及其制备方法和应用
Yang et al. A red-emitting luminescent probe for sequentially detecting Cu2+ and cysteine/histidine in aqueous solution and its imaging application in living zebrafish
CN105712964A (zh) 一种基于香豆酰肼的硫醇荧光探针的制备方法及应用
CN111072648A (zh) 一种检测溶酶体内生物硫醇的荧光探针及其制备方法和应用
Ge et al. A novel NIR fluorescence probe with cysteine-activated structure for specific detection of cysteine and its application in vitro and in vivo
CN114773288A (zh) 一种区别检测半胱氨酸和同型半胱氨酸/谷胱甘肽的荧光探针及其制备方法和应用
CN108003173A (zh) 一种特异性识别水合肼的荧光探针
Cuiling et al. A dicyanoisophorone-based near-infrared fluorescent probe with fast detection for H2S in living cells and zebrafish
Yang et al. Real-time monitoring of endogenous cysteine in LPS-induced oxidative stress process with a novel lysosome-targeted fluorescent probe
Yan et al. Simultaneous discrimination of Cys/Hcy and GSH with simple fluorescent probe under a single-wavelength excitation and its application in living cells, tumor tissues, and zebrafish
CN115215833B (zh) 化学发光探针及其应用
Zhang et al. A novel quinoline-based fluorescent probe for real-time monitoring of Cys in glioma
CN112939963A (zh) 一种苯并吡喃酸酯衍生物及其合成方法和应用
CN108383774A (zh) 一种基于端基炔酮的半胱氨酸荧光探针及其制备和应用
CN114736199B (zh) 一种基于亚甲基蓝的近红外荧光探针及其合成方法和应用
CN114634473B (zh) 一种能够快速高效检测生物硫醇的荧光探针及其制备方法
Rong et al. Long-term thiol monitoring in living cells using bioorthogonal chemistry

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
WW01 Invention patent application withdrawn after publication
WW01 Invention patent application withdrawn after publication

Application publication date: 20180720