CN108949161B - 一种用于检测次溴酸的比率型荧光探针化合物及其用途 - Google Patents

一种用于检测次溴酸的比率型荧光探针化合物及其用途 Download PDF

Info

Publication number
CN108949161B
CN108949161B CN201811142097.4A CN201811142097A CN108949161B CN 108949161 B CN108949161 B CN 108949161B CN 201811142097 A CN201811142097 A CN 201811142097A CN 108949161 B CN108949161 B CN 108949161B
Authority
CN
China
Prior art keywords
hypobromous acid
compound
npsn
lyso
detecting
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.)
Active
Application number
CN201811142097.4A
Other languages
English (en)
Other versions
CN108949161A (zh
Inventor
厉凯彬
韩得满
屈王波
陈彩霞
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.)
Taizhou University
Original Assignee
Taizhou 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 Taizhou University filed Critical Taizhou University
Priority to CN201811142097.4A priority Critical patent/CN108949161B/zh
Publication of CN108949161A publication Critical patent/CN108949161A/zh
Application granted granted Critical
Publication of CN108949161B publication Critical patent/CN108949161B/zh
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D221/00—Heterocyclic compounds containing six-membered rings having one nitrogen atom as the only ring hetero atom, not provided for by groups C07D211/00 - C07D219/00
    • C07D221/02—Heterocyclic compounds containing six-membered rings having one nitrogen atom as the only ring hetero atom, not provided for by groups C07D211/00 - C07D219/00 condensed with carbocyclic rings or ring systems
    • C07D221/04—Ortho- or peri-condensed ring systems
    • C07D221/06—Ring systems of three rings
    • C07D221/14—Aza-phenalenes, e.g. 1,8-naphthalimide
    • 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"
    • G01N21/643—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" non-biological material
    • 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
    • C09K2211/1003—Carbocyclic compounds
    • C09K2211/1007—Non-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
    • 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
    • C09K2211/1029—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
    • C09K2211/1033—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom with oxygen

Landscapes

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

Abstract

本发明属于分析化学的荧光探针领域，涉及一种新型用于检测次溴酸的比率型荧光探针化合物及其用途。所述合成方法通过将3‑氨基‑4‑溴‑1,8‑萘二甲酸酐与4‑(2‑氨基乙基)吗啉反应生成中间产物II，化合物II与2‑甲硫基苯硼酸发生偶联反应得到所述的探针分子。该探针稳定性好，制备方法简单，且对次溴酸的响应具有比率型、选择性高、快速灵敏的优点，其它活性小分子、离子和氨基酸对次溴酸的检测不产生干扰。该探针提供了一种新型的检测次溴酸的方法，具有很好的应用前景。

Description

一种用于检测次溴酸的比率型荧光探针化合物及其用途
技术领域
本发明涉及一种新型的用于检测次溴酸的比率型荧光探针化合物及其用途。
背景技术
次溴酸是生物体中含有的一种活性氧化物，其具有强氧化能力。在生物体中，次溴酸是在血红素过氧化物酶的催化下由溴离子与过氧化氢经过过氧化反应产生的，血红素过氧化物酶包括髓过氧化物酶(MPO)或嗜酸性粒细胞过氧化物酶(EPO)。次溴酸可以与许多生物分子发生反应，例如氨基酸、蛋白质，抗氧化剂(包括硫醇)，碳水化合物，脂质和DNA等，所以其对于免疫系统具有重要的作用。然而，过量的次溴酸会造成人体的组织损伤，这与人类的很多炎性疾病具有紧密的关系，包括心血管疾病，肾脏疾病，肠道炎症，神经退行性疾病，类风湿性关节炎，炎症组织损伤和癌症。因此，对次溴酸浓度快速、灵敏检测非常重要。
然而，目前检测次溴酸的方法还十分缺少，因此开发一种新型用于检测次溴酸的方法具有重要意义。
发明内容
本发明涉及一种新型的用于检测次溴酸的比率型荧光探针化合物及其用途。所述合成方法包括：将3-氨基-4-溴-1,8-萘二甲酸酐与4-(2-氨基乙基)吗啉在乙醇中回流发生取代反应生成中间产物II，然后在氮气中，化合物II与2-甲硫基苯硼酸在Pd(dppf)2Cl2·CH2Cl2的催化下发生偶联反应生成目标化合物Lyso-NpSN。该探针稳定性好，制备方法简单，且对次溴酸的响应具有比率型、选择性高、快速灵敏的优点。其表现为与次溴酸响应后，紫外光谱和荧光发射都有比率型变化，分别伴有40nm和55nm红移。此外，动力学实验结果表明，该化合物与次溴酸反应时间少于1s，具有快速检测的能力。在选择性测试中，其它活性小分子、离子和氨基酸对次溴酸的检测不产生干扰。因此，本发明所述的化合物可作为检测次溴酸的荧光探针应用。
本发明的一个目的在于提供一种萘二甲酰亚胺衍生化合物，其为式Lyso-NpSN所述化合物(简记为化合物Lyso-NpSN，下同)：
Figure BDA0001816028820000021
化合物Lyso-NpSN的合成路线如下：
Figure BDA0001816028820000031
其中，化合物I的合成步骤参见文献Journal ofPhysical Chemistry B,2008,112(49),15636-15645。
本发明的另一个目的在于揭示上述萘二甲酰亚胺衍生化合物(式Lyso-NpSN所述化合物)的一种用途，即式Lyso-NpSN所述化合物作为检测次溴酸的荧光探针的应用。
附图说明
图1化合物Lyso-NpSN的紫外吸收光谱随次溴酸浓度变化曲线；
图2在475nm激发波长下，化合物Lyso-NpSN的荧光发射光谱随次溴酸浓度变化曲线；
图3在475nm激发波长下，化合物Lyso-NpSN在610nm处与555nm处的荧光强度比值与次溴酸浓度(12-22μM)的关系图；
图4化合物Lyso-NpSN对活性小分子干扰物的选择性图；
图5化合物Lyso-NpSN对常见离子和氨基酸干扰物的选择性图；
图6化合物Lyso-NpSN与次溴酸反应的动力学曲线；
具体实施方式
以下将结合实施例对本发明做进一步说明，本发明的实施例仅用于说明本发明的技术方案，并非限定本发明。
实施例1
准确称取3-氨基-4-溴-1,8-萘二甲酸酐化合物I(120mg,0.41mmol)，4-(2-氨基乙基)吗啉(106mg,0.82mmol)，溶解在10mL乙醇中，80℃回流搅拌反应6小时。TLC检测反应完全，停止反应，冷却至室温，抽滤，并用适量乙醇进行冲洗，干燥后得到淡黄色固体(化合物II)133mg，产率80.2％。1H NMR(400MHz,d6-DMSO):δ8.20(d,J＝8.4Hz,1H),8.15(d,J＝7.2Hz,1H),8.11(s,1H),7.76(t,J＝7.8Hz,1H),6.32(s,2H),4.14(t,J＝7.0Hz,2H),3.53(t,J＝4.4Hz,4H),2.55(t,J＝6.8Hz,2H),2.46(s,4H)；13C NMR(400MHz,d6-DMSO):δ163.7,163.5,145.8,132.0,128.9,126.3,122.7,122.4,122.3,121.7,107.1,66.7,56.0,53.9,37.3；HR-ESI-MS m/z:[M+H]+calcd.for 404.0610 found 404.0609。
实施例2
将化合物II(80mg,0.198mmol)，2-甲硫基苯硼酸(50mg,0.297mmol)，Pd(dppf)2Cl2·CH2Cl2(9mg,0.01mmol)，K2CO3(4M,2mL)，EtOH(0.6mL)，甲苯(5mL)加入到50mL圆底烧瓶中，在氮气保护下，80℃搅拌反应24小时。TLC检测反应完全，停止反应，减压蒸馏除去溶剂，使用柱层析分离提纯得到灰黄色固体产物(化合物Lyso-NpSN)75.4mg，产率85.1％。1HNMR(400MHz,CDCl3):δ8.25(d,J＝7.2Hz,1H),8.07(s,1H),7.47-7.32(m,4H),7.27-7.11(m,2H),4.28(t,J＝7.0Hz,2H),3.92(s,2H),3.64(t,J＝4.2Hz,4H),2.65(t,J＝7.0Hz,2H),2.56(s,4H),2.29(s,3H)；13C NMR(400MHz,CDCl3):δ164.6,164.2,142.8,139.4,132.6,132.2,130.8,130.3,129.6,127.3,127.2,125.6,125.0,123.6,123.2,122.7,122.5,122.2,67.0,56.2,53.8,37.1,15.0；HR-ESI-MS m/z:[M+H]+calcd.for 448.1695found 448.1697。
将实施例中制备的化合物Lyso-NpSN应用于次溴酸的检测，其具体操作方法及结果如下应用实例：
应用实例1
分别配制2mM化合物Lyso-NpSN的DMF溶液，1mM次溴酸溶液备用。向2mL的10mMPBS-CH3CN(v/v,3:2)的缓冲体系中加入10μL配制好的化合物Lyso-NpSN溶液，得到浓度10μM的化合物Lyso-NpSN溶液。分别加入不同量的次溴酸溶液，并测定了加入0-28μM次溴酸的紫外吸收光谱变化曲线(图1)。从图1可以看出，随着体系中次溴酸浓度的增加，425nm处的紫外吸收峰逐渐减弱，同时伴随着465nm处紫外吸收峰的增强，在440nm处出现等吸收点，说明次溴酸能够使探针的紫外光谱发生红移，并伴随比率型变化。
应用实例2
分别配制2mM化合物Lyso-NpSN的DMF溶液，1mM次溴酸溶液备用。向2mL的10mMPBS-CH3CN(v/v,3:2)的缓冲体系中加入10μL配制好的化合物Lyso-NpSN溶液，得到浓度10μM的化合物Lyso-NpSN溶液。分别加入不同量的次溴酸溶液，并测定了加入0-28μM次溴酸的荧光发射光谱变化曲线(图2)。从图2可以看出，随着体系中次溴酸浓度的增加，555nm处的荧光发射峰逐渐减弱，同时在610nm处出现新的荧光峰并逐渐增强，说明次溴酸能使探针的荧光发射光谱发生红移，并伴随比率型变化。从图3可以看出，在12-22μM次溴酸浓度范围，610nm处与555nm处荧光强度比值与次溴酸具有较好的线性依赖关系，线性方程式：y＝0.05859x+0.1459,R2＝0.9993，说明探针Lyso-NpSN可用于次溴酸的定量检测。
应用实例3
分别配制2mM化合物Lyso-NpSN的DMF溶液，1mM次溴酸溶液备用。向2mL的10mMPBS-CH3CN(v/v,3:2)的缓冲体系中加入10μL配制好的化合物Lyso-NpSN溶液，得到浓度10μM的化合物Lyso-NpSN溶液。再分别加入60μM的HOBr，3mM的GSH，500μM的Cys、Hcy、Vc，75μM的HOCl，300μM的HS-，1O2，NO2 -，NO3-，·OH，H2O2，并测定荧光发射光谱曲线(图4)。图4为常见活性小分子的选择性图，探针Lyso-NpSN只与次溴酸响应，并产生光谱红移现象，而其他活性小分子并不产生干扰，说明探针Lyso-NpSN对次溴酸具有很高的选择性。
应用实例4
分别配制2mM化合物Lyso-NpSN的DMF溶液，1mM次溴酸溶液备用。向2mL的10mMPBS-CH3CN(v/v,3:2)的缓冲体系中加入10μL配制好的化合物Lyso-NpSN溶液，得到浓度10μM的化合物Lyso-NpSN溶液。再分别加入60μM的HOBr，300μM的K+、Na+、Cu2+、Zn2+、Fe2+、Fe3+、CO3 2-、Ac-、SO4 2-、S2O3 2-、Br-、met、pro、his、ala、thr、leu，并测定荧光发射光谱曲线(图5)。图5为常见离子和氨基酸的选择性图，探针Lyso-NpSN只与次溴酸响应，并产生光谱红移现象，而常见的离子和氨基酸并不产生干扰，说明探针Lyso-NpSN对次溴酸具有很高的选择性。
应用实例5
分别配制2mM化合物Lyso-NpSN的DMF溶液，1mM次溴酸溶液备用。向2mL的10mMPBS-CH3CN(v/v,3:2)的缓冲体系中加入10μL配制好的化合物Lyso-NpSN溶液，得到浓度10μM的化合物Lyso-NpSN溶液，每隔0.2min，测定一次610nm和555nm处的荧光强度，在3.2min加入60μM次溴酸，继续每隔0.2min测定一次610nm和555nm处的荧光强度。以610nm处与555nm处的荧光强度比值为纵坐标，时间(min)为横坐标，做点线图6。由图6可见，化合物Lyso-NpSN在激发光很稳定，当加入次溴酸后，I610/I555在1秒内发生突变，之后保持稳。说明化合物Lyso-NpSN对次溴酸能快速、灵敏地响应。

Claims (2)

1.一种双通道检测次溴酸的比率型荧光探针化合物，其为式Lyso-NpSN所述化合物：
Figure DEST_PATH_IMAGE002
。
2.一种如权利要求1所述的化合物在制备检测次溴酸的比率型荧光探针中的应用。
CN201811142097.4A 2018-09-28 2018-09-28 一种用于检测次溴酸的比率型荧光探针化合物及其用途 Active CN108949161B (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201811142097.4A CN108949161B (zh) 2018-09-28 2018-09-28 一种用于检测次溴酸的比率型荧光探针化合物及其用途

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201811142097.4A CN108949161B (zh) 2018-09-28 2018-09-28 一种用于检测次溴酸的比率型荧光探针化合物及其用途

Publications (2)

Publication Number Publication Date
CN108949161A CN108949161A (zh) 2018-12-07
CN108949161B true CN108949161B (zh) 2021-03-16

Family

ID=64472283

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201811142097.4A Active CN108949161B (zh) 2018-09-28 2018-09-28 一种用于检测次溴酸的比率型荧光探针化合物及其用途

Country Status (1)

Country Link
CN (1) CN108949161B (zh)

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111333643B (zh) * 2018-12-18 2021-11-09 中国科学院大连化学物理研究所 一类高亮度、高光稳定性、环境不敏感的细胞核荧光探针
CN109705055B (zh) * 2019-01-12 2022-05-13 湖南城市学院 苯并噻唑类化合物及其制备方法及作为次溴酸探针的应用
CN110878085B (zh) * 2019-12-13 2020-11-13 山东省科学院生物研究所 一种快速高选择性次溴酸荧光探针、制备方法与应用

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108148573A (zh) * 2018-02-01 2018-06-12 济南大学 一种检测溶酶体pH的荧光探针及其合成方法和应用

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108148573A (zh) * 2018-02-01 2018-06-12 济南大学 一种检测溶酶体pH的荧光探针及其合成方法和应用

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
A New Lysosome Targetable Turn-On Fluorogenic Probe for Carbon Monoxide Imaging in Living Cells;Koushik Dhara et al.，;《Anal. Chem.》;20180122;第90卷(第4期);第1-21页 *
High-Quantum-Yield Mitochondria-Targeting Near-Infrared Fluorescent Probe for Imaging Native Hypobromous Acid in Living Cells and in Vivo;Xiaojun Liu et al.，;《Anal. Chem.》;20170105;第89卷(第3期);第1787-1792页 *

Also Published As

Publication number Publication date
CN108949161A (zh) 2018-12-07

Similar Documents

Publication Publication Date Title
Fang et al. BODIPY-based naked-eye fluorescent on-off probe with high selectivity for H2S based on thiolysis of dinitrophenyl ether
CN108949161B (zh) 一种用于检测次溴酸的比率型荧光探针化合物及其用途
Yang et al. A turn-on near-infrared fluorescence probe with aggregation-induced emission based on dibenzo [a, c] phenazine for detection of superoxide anions and its application in cell imaging
CN111423423B (zh) 一种比率型荧光探针在检测过氧亚硝基阴离子中的应用
Yadav et al. A novel colorimetric and fluorogenic chemosensor for selective detection of Cu 2+ ions in mixed aqueous media
Wang et al. A β-diketonate–europium (III) complex-based fluorescent probe for highly sensitive time-gated luminescence detection of copper and sulfide ions in living cells
Li et al. A novel optical probe for Hg2+ in aqueous media based on mono-thiosemicarbazone Schiff base
CN109336815B (zh) 一种检测细胞内质网内次氯酸的双光子荧光探针
Xie et al. A simple BODIPY–imidazole-based probe for the colorimetric and fluorescent sensing of Cu (II) and Hg (II)
CN105349135B (zh) 一种检测铅离子的荧光探针及制备方法
Yang et al. Colorimetric and Highly Selective Fluorescence" Turn‐on" Detection of Cr3+ by Using a Simple Schiff Base Sensor
Wu et al. A visible-light-excited Eu 3+ complex-based luminescent probe for highly sensitive time-gated luminescence imaging detection of intracellular peroxynitrite
CN113264954A (zh) 一种用于检测过氧化氢的荧光探针分子及其制备方法
CN109206351B (zh) 一种基于花菁结构测钯离子的近红外荧光探针、其制备方法及应用
JP6958781B2 (ja) 発蛍光性化合物又はその塩、イオン性化合物の検出剤及びイオン性化合物の検出方法
Dalkilic et al. An AIE active fluorescence sensor for measuring Fe 3+ in aqueous media and an iron deficiency anemia drug
CN111423439B (zh) 一种检测Fe(Ⅲ)的香豆素类共轭杂环荧光探针
KR20130017879A (ko) 시아나이드 음이온의 선택적 검출이 가능한 형광센서, 그 제조방법 및 이를 이용한 시아나이드 음이온의 선택적 검출방법
Zhao et al. Construction of a novel ESIPT and AIE-based fluorescent sensor for sequentially detecting Cu2+ and H2S in both living cells and zebrafish
Mei et al. A novel fluorescence probe for the selective detection of cysteine in aqueous solutions and imaging in living cells and mice
Xu et al. Development of a two-photon ratiometric fluorescent probe for Glutathione and its applications in living cells
CN114539181A (zh) 一种用于铝离子和锌离子快速检测的荧光探针及其制备方法
CN110078772B (zh) 基于铱配合物的荧光探针及其制备方法与在次氯酸检测中的应用
CN109705055B (zh) 苯并噻唑类化合物及其制备方法及作为次溴酸探针的应用
CN109020988B (zh) 一种三氮唑类化合物的罗丹明荧光探针在Hela细胞成像中的应用

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
GR01 Patent grant
GR01 Patent grant