CN110305111A - 一种新的亚铜离子近红外探针及其制备方法和应用 - Google Patents
一种新的亚铜离子近红外探针及其制备方法和应用 Download PDFInfo
- Publication number
- CN110305111A CN110305111A CN201910727683.3A CN201910727683A CN110305111A CN 110305111 A CN110305111 A CN 110305111A CN 201910727683 A CN201910727683 A CN 201910727683A CN 110305111 A CN110305111 A CN 110305111A
- Authority
- CN
- China
- Prior art keywords
- cuprous ion
- probe
- preparation
- formula
- organic solvent
- 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.)
- Granted
Links
- VMQMZMRVKUZKQL-UHFFFAOYSA-N Cu+ Chemical compound [Cu+] VMQMZMRVKUZKQL-UHFFFAOYSA-N 0.000 title claims abstract description 67
- 239000000523 sample Substances 0.000 title claims abstract description 55
- 238000002360 preparation method Methods 0.000 title claims abstract description 19
- 239000007850 fluorescent dye Substances 0.000 claims abstract description 14
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 6
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 16
- 238000001514 detection method Methods 0.000 claims description 16
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 15
- 239000003960 organic solvent Substances 0.000 claims description 13
- 238000006243 chemical reaction Methods 0.000 claims description 11
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 8
- 235000019441 ethanol Nutrition 0.000 claims description 6
- 229910052736 halogen Inorganic materials 0.000 claims description 6
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 5
- 125000004123 n-propyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])* 0.000 claims description 5
- 239000001632 sodium acetate Substances 0.000 claims description 5
- 235000017281 sodium acetate Nutrition 0.000 claims description 5
- 150000001298 alcohols Chemical group 0.000 claims description 4
- 150000002367 halogens Chemical class 0.000 claims description 4
- 229910052739 hydrogen Inorganic materials 0.000 claims description 4
- 239000001257 hydrogen Substances 0.000 claims description 4
- 230000035484 reaction time Effects 0.000 claims description 4
- 239000000126 substance Substances 0.000 claims description 4
- 150000001875 compounds Chemical class 0.000 claims description 3
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims description 2
- 150000008378 aryl ethers Chemical class 0.000 claims description 2
- 239000003153 chemical reaction reagent Substances 0.000 claims description 2
- 125000005843 halogen group Chemical group 0.000 claims description 2
- 125000004435 hydrogen atom Chemical group [H]* 0.000 claims description 2
- 239000000243 solution Substances 0.000 abstract description 10
- 230000004044 response Effects 0.000 abstract description 9
- 150000002500 ions Chemical class 0.000 abstract description 8
- 239000012043 crude product Substances 0.000 abstract description 5
- 230000002452 interceptive effect Effects 0.000 abstract description 4
- 230000035945 sensitivity Effects 0.000 abstract description 4
- 208000037273 Pathologic Processes Diseases 0.000 abstract description 2
- 230000009054 pathological process Effects 0.000 abstract description 2
- 239000007864 aqueous solution Substances 0.000 abstract 1
- 238000004451 qualitative analysis Methods 0.000 abstract 1
- 125000006853 reporter group Chemical group 0.000 abstract 1
- 239000002904 solvent Substances 0.000 abstract 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract 1
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 20
- 239000010949 copper Substances 0.000 description 10
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 8
- 229910052802 copper Inorganic materials 0.000 description 8
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 5
- 238000005160 1H NMR spectroscopy Methods 0.000 description 5
- 238000005481 NMR spectroscopy Methods 0.000 description 5
- 230000005284 excitation Effects 0.000 description 5
- JPVYNHNXODAKFH-UHFFFAOYSA-N Cu2+ Chemical compound [Cu+2] JPVYNHNXODAKFH-UHFFFAOYSA-N 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 4
- 238000000862 absorption spectrum Methods 0.000 description 4
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 4
- 229910052751 metal Inorganic materials 0.000 description 4
- 239000002184 metal Substances 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 239000007853 buffer solution Substances 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 230000007613 environmental effect Effects 0.000 description 3
- 238000003384 imaging method Methods 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- HBENZIXOGRCSQN-VQWWACLZSA-N (1S,2S,6R,14R,15R,16R)-5-(cyclopropylmethyl)-16-[(2S)-2-hydroxy-3,3-dimethylpentan-2-yl]-15-methoxy-13-oxa-5-azahexacyclo[13.2.2.12,8.01,6.02,14.012,20]icosa-8(20),9,11-trien-11-ol Chemical compound N1([C@@H]2CC=3C4=C(C(=CC=3)O)O[C@H]3[C@@]5(OC)CC[C@@]2([C@@]43CC1)C[C@@H]5[C@](C)(O)C(C)(C)CC)CC1CC1 HBENZIXOGRCSQN-VQWWACLZSA-N 0.000 description 2
- HZAXFHJVJLSVMW-UHFFFAOYSA-N 2-Aminoethan-1-ol Chemical compound NCCO HZAXFHJVJLSVMW-UHFFFAOYSA-N 0.000 description 2
- 208000024827 Alzheimer disease Diseases 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 206010002026 amyotrophic lateral sclerosis Diseases 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 150000001768 cations Chemical class 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 229910001431 copper ion Inorganic materials 0.000 description 2
- 231100000135 cytotoxicity Toxicity 0.000 description 2
- 230000003013 cytotoxicity Effects 0.000 description 2
- 229960003180 glutathione Drugs 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 description 2
- 238000010898 silica gel chromatography Methods 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- FYSNRJHAOHDILO-UHFFFAOYSA-N thionyl chloride Chemical compound ClS(Cl)=O FYSNRJHAOHDILO-UHFFFAOYSA-N 0.000 description 2
- 239000012224 working solution Substances 0.000 description 2
- IVSZLXZYQVIEFR-UHFFFAOYSA-N 1,3-Dimethylbenzene Natural products CC1=CC=CC(C)=C1 IVSZLXZYQVIEFR-UHFFFAOYSA-N 0.000 description 1
- CSDSSGBPEUDDEE-UHFFFAOYSA-N 2-formylpyridine Chemical compound O=CC1=CC=CC=N1 CSDSSGBPEUDDEE-UHFFFAOYSA-N 0.000 description 1
- 206010067484 Adverse reaction Diseases 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 241000675108 Citrus tangerina Species 0.000 description 1
- 108010024636 Glutathione Proteins 0.000 description 1
- 101710094503 Metallothionein-1 Proteins 0.000 description 1
- 208000018737 Parkinson disease Diseases 0.000 description 1
- 102000029797 Prion Human genes 0.000 description 1
- 108091000054 Prion Proteins 0.000 description 1
- 229910006124 SOCl2 Inorganic materials 0.000 description 1
- 102000008221 Superoxide Dismutase-1 Human genes 0.000 description 1
- 108010021188 Superoxide Dismutase-1 Proteins 0.000 description 1
- WWFMINHWJYHXHF-UHFFFAOYSA-N [6-(hydroxymethyl)pyridin-2-yl]methanol Chemical compound OCC1=CC=CC(CO)=N1 WWFMINHWJYHXHF-UHFFFAOYSA-N 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 238000001460 carbon-13 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 239000012930 cell culture fluid Substances 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 231100000263 cytotoxicity test Toxicity 0.000 description 1
- 229960000935 dehydrated alcohol Drugs 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- QGBSISYHAICWAH-UHFFFAOYSA-N dicyandiamide Chemical compound NC(N)=NC#N QGBSISYHAICWAH-UHFFFAOYSA-N 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 238000000295 emission spectrum Methods 0.000 description 1
- 239000005447 environmental material Substances 0.000 description 1
- 229960004756 ethanol Drugs 0.000 description 1
- 229940031098 ethanolamine Drugs 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 231100000636 lethal dose Toxicity 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 150000002739 metals Chemical class 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 230000004770 neurodegeneration Effects 0.000 description 1
- 208000015122 neurodegenerative disease Diseases 0.000 description 1
- 230000002018 overexpression Effects 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 229910000027 potassium carbonate Inorganic materials 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000011833 salt mixture Substances 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 235000017557 sodium bicarbonate Nutrition 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000004577 thatch Substances 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
-
- 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
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/33—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using ultraviolet light
-
- 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
- G01N21/6428—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
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Optics & Photonics (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Materials Engineering (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
本发明设计并合成了一种在水溶液体系里对亚铜离子特异性响应,具有全新受体结构(N3O)的近红外荧光探针,主要解决了亚铜离子近红外荧光探针选择性较差、灵敏度低和的问题。其制备方法包括:中间体B与中间体5在乙醇溶剂中反应,粗品经柱层析纯化得到该具有全新识别基团的亚铜离子近红外荧光探针。本发明以N3O结构作为新型亚铜离子识别基团,qCy7为报告基团合成了该具有N3O结构的全新亚铜离子近红外探针,其对亚铜离子表现出了高灵敏度、高选择性,对其他干扰离子几乎没有响应。该具有N3O结构的全新亚铜离子近红外探针可作为水溶液中定量和活细胞中定性分析亚铜离子的工具,具有应用于亚铜离子及其相关生理、病理过程的前景。
Description
技术领域
本发明涉及一种具有全新识别基团的亚铜离子近红外荧光探针及其制备方法和应用。
背景技术
铜是所有金属中使用最广泛的金属之一,并且是许多行业和技术的支柱之一,此外它还是国内管道中使用最广泛的材料之一。同时,铜元素是体内必不可少的微量元素之一,其氧化态(Cu2+,铜离子)和还原态(Cu+,亚铜离子)在生物体内都具有非常重要的意义。当其发生失衡,含量过高或者过低都会扰乱生命系统的正常功能,引起系列不良反应。在突变型超氧化物歧化酶-1所致肌萎缩性侧索硬化小鼠模型中,细胞核与细胞内铜转运有关,Cu+储存蛋白金属硫蛋白-1的过表达减弱了铜失调的停滞并延长了寿命。铜稳态的改变还与神经退行性疾病(阿尔茨海默氏症、朊病毒、帕金森病和肌萎缩侧索硬化)有关。在AD(阿尔茨海默氏症)中,铜分布不当,并在细胞外积累,这可能解释了AD影响的中枢神经系统组织铜缺乏的观察。因此对环境和生物体内的铜元素的检测显得尤为重要。Cu的特异性探针逐渐备受关注。
目前检测Cu+的探针相对较少;且通常在水溶液环境下选择性差、灵敏度低、易受其它离子的干扰。Cu+检测受到诸多限制。如果需要对活的组织细胞内进行Cu+检测,还要求探针具有较好的生物相容性,但现有的很多探针由于具有毒性,不能达到该要求。现有的Cu+检测探针还具有合成复杂、难以纯化的缺点。
发明内容
为了解决上述问题,本发明提供了一种新型亚铜离子检测探针及其制备方法和应用。
本发明的技术方案包括:
一种具有高选择性的亚铜离子近红外荧光探针,其特征在于,它具有N3O结构,该结构可以识别亚铜离子;
所述N3O结构为:
式中Y表示氢或者卤素。
如前述的探针,其分子结构如式1所示:
其中R1、R2选自C1-3烷基;X为卤素。
优选地,式1中,R1为正丙基;和/或,R2为正丙基;和/或,X为Br。
前述探针的制备方法,它包括以下步骤:
(1)中间体A与中间体N3O在有机溶剂中反应成芳香醚,得中间体5;
中间体A的结构式为:
中间体N3O的结构式为:
式中Y为氢或卤素;
中间体5为4-羟基取代的间苯二甲醛,其结构式为:
(2)中间体B与中间体5在有机溶剂中反应,得到式1所示化合物;
中间体B为1-取代的2,3,3-三甲基吲哚盐,其结构式为:
式中R表示C1-3的烷基;
步骤(1)所述有机溶剂为醇类、DMF、THF或甲苯;
步骤(2)所述有机溶剂为醇类、DMF、THF或甲苯。
如前述的制备方法,步骤(1)中中间体N3O与中间体A的摩尔比是1∶(2~4);
和/或,反应温度是50~100℃。
如前述的制备方法,步骤(1)中的有机溶剂为N,N-二甲基甲酰胺;和/或,步骤(2)中的有机溶剂为乙醇。
如前述的制备方法,步骤(2)中中间体5与中间体B的摩尔比是1∶(2~4);
和/或,反应温度是50~100℃;
和/或,反应时间是2~6小时。
如前述的制备方法,步骤(2)的反应还需加入醋酸钠,醋酸钠的物质的量为中间体5的1~3倍。
前述探针在亚铜离子检测中的用途。
前述探针在制备亚铜离子检测试剂中的用途。
本发明具有如下有益效果:
1)本发明的探针能够在水溶液环境中实现有效检测。
2)本发明的探针特异性高,其对亚铜离子响应的荧光强度是其他干扰离子响应强度的7-35倍。
3)本发明的探针灵敏度高,能有效检测浓度低至1μM的亚铜离子。
4)本发明的探针适用于活体检测,其对细胞的半致死浓度高达200μM,且实验证明能够有效结合细胞内的亚铜离子,产生明显的阳性检测信号。
5)本发明的探针制备简单,纯化方便,得率较高。
本发明在生物化学、环境化学和生物医学等多个领域都有广泛的应用前景。
附图说明
图1为本发明探针的核磁共振氢谱;
图2为本发明探针的核磁共振碳谱;
图3为本发明探针的高分辨质谱;
图4为本发明探针检测亚铜离子的紫外吸收光谱曲线变化;
图5为本发明探针检测亚铜离子的3D荧光图谱变化(图A为本发明的50μM探针本身荧光性质的二维和三维图谱,x轴显示其最大激发波长为~420nm,y轴显示其最大发射波长为~560nm,z轴显示对应的荧光强度;图B为本发明的50μM探针与500μM亚铜离子进行孵育60min后荧光性质的二维和三维图谱,x轴显示其最大激发波长为~560nm,y轴显示其最大发射波长为~710nm,z轴显示对应的荧光强度);
图6为本发明探针检测亚铜离子荧光强度随反应时间的变化曲线;
图7为本发明探针检测亚铜离子荧光强度随Cu+浓度的变化曲线;
图8为本发明探针检测不同金属阳离子的荧光强度对比图谱;
图9为本发明探针的细胞成像激光共聚焦图。
具体实施方式
为更好地理解本发明,下面结合实施例进一步阐述本发明的内容,但本发明并不仅仅局限于下面的实施例。
本发明实施例中所用的原料均为已知化合物,可由市场购得,或按照现有的方法合成得到。
实施例1制备中间体N3O
a.将SOCl2(15ml)缓慢滴加至2,6-吡啶二甲醇中搅拌1~3h后升温回3~8h,加入甲苯稀释反应液,调节pH=7析出固体并洗涤干燥得中间体1a,产率87.3%。中间体1a的核磁共振数据为:1H NMR(400MHz,DMSO-d6)δ:7.89(t,J=8.0Hz,1H),7.51(d,J=8.0Hz,2H),4.77(s,4H).13C NMR(100MHz,DMSO-d6)δ156.63,139.02,123.23,46.94.
b.冰盐浴下将乙醇胺与2-吡啶甲醛(1∶1~1.5,n∶n)均匀混合,随后室温反应2~6小时,分批加入NaBH4,粗品用DCM:MeOH(5~12%)体系纯化得化合物1b,产率98.70%。化合物1b的核磁共振数据为:1H NMR(400MHz,DMSO-d6)δ:8.49(ddd,J=5.2,2.0,1.2Hz,1H),7.74(td,J=8.0,2.0Hz,1H),7.41(dt,J=8.0,1.2Hz,1H),7.23(ddd,J=8.0,5.2,1.2Hz,1H),3.80(s,2H),3.48(t,J=5.6Hz,2H),2.60(t,J=5.6Hz,2H).13C NMR(100MHz,DMSO-d6)δ160.66,149.19,136.89,122.31,122.25,60.81,54.92,51.69.
c.上述化合物1b与碳酸氢钠(1∶0.4~1,n∶n)均匀分散在乙腈中预先于50~60℃下搅拌30~90分钟,再加入中间体1c至反应结束,粗产品用MeOH/EA从2-10%进行纯化得橘色中间体,产率77.8%。中间体的核磁共振数据为:1H NMR(400MHz,DMSO-d6)δ:8.47(d,J=5.2Hz,1H),7.81(t,J=8.0Hz,1H),7.75(t,J=8.0Hz,1H),7.55(dd,J=8.0,5.2Hz,2H),7.40(d,J=8.0Hz,1H),7.24(dd,J=8.0,5.2Hz,1H),4.74(s,2H),4.53(s,1H),3.80(s,4H),3.52(t,J=6.4Hz,2H),2.60(t,J=6.4Hz,2H).13C NMR(100MHz,DMSO-d6)δ160.12,159.83,155.79,149.05,138.04,136.83,123.01,122.42,122.33,121.76,60.52,60.45,59.46,56.57,47.25.
实施例2制备一种具有全新亚铜离子识别基团的近红外荧光探针(式1)
a.将实施例1中的中间体与对羟基间二甲苯(1∶1~3n∶n)和无水碳酸钾混合分散在无水DMF(N,N-二甲基甲酰胺)中,50~100℃下反应至结束,冷却至室温后用有机溶剂萃取,经洗涤,干燥,过滤,浓缩的粗品经硅胶柱层析纯化得中间体5,产率72.5%。中间体5的核磁共振数据为:1H NMR(400MHz,DMSO-d6)δ10.47(s,1H),9.95(s,1H),8.46(d,J=4.8Hz,1H),8.26(d,J=2.4Hz,1H),8.14(dd,J=8.0,2.4Hz,1H),7.83(t,J=8.0Hz,1H),7.73(m,1H),7.53(m,4H),7.23(t,J=8.0Hz,1H),5.46(s,2H),4.53(s,1H),3.80(s,4H),3.50(t,J=6.4Hz,2H),2.59(t,J=6.4Hz,2H).13C NMR(100MHz,DMSO-d6)δ191.64,189.12,164.65,162.66,159.88,159.84,154.95,149.04,137.94,136.80,130.70,129.87,124.95,122.95,122.39,122.33,120.23,115.14,71.66,60.53,60.41,59.49,56.55.
b.将上述中间体5和中间体B(1∶2~4,n∶n)溶于无水乙醇中,再加入醋酸钠(1~3当量),50~100℃下回流反应2~6小时,粗品经硅胶柱层析纯化得该全新亚铜离子识别基团的近红外荧光探针,产率61.9%。
所得近红外荧光探针的化学式如式I所示:
式中,R1、R2均为正丙基,X为Br。
产物的核磁共振氢谱和碳谱数据为:
1H NMR(400MHz,DMSO-d6)δ:8.62(dd,J=16.0,8.0Hz,2H),8.59-8.54(m,1H),8.45(d,J=5.2Hz,1H),8.15(d,J=16.0Hz,1H),8.03-7.96(m,2H),7.95-7.85(m,4H),7.78-7.72(m,1H),7.66(dd,J=12.0,5.6Hz,5H),7.61(d,J=12.0Hz,1H),7.55(d,J=8.0Hz,4H),7.25-7.20(m,1H),5.55(s,2H),4.83(t,J=7.4Hz,2H),4.75(t,J=7.4Hz,2H),4.58(s,1H),3.83(s,4H),3.53(t,J=6.4Hz,2H),2.61(t,J=6.4Hz,2H),1.96-1.82(m,11H),1.75(s,5H),1.05(t,J=7.2Hz,3H),0.96(t,J=7.2Hz,3H).13C NMR(100MHz,DMSO-d6)δ:182.51,182.36,162.29,160.47,159.96,154.79,153.02,149.17,147.45,144.40,144.22,141.29,141.27,138.12,137.84,136.99,134.02,130.24,129.93,129.75,129.62,128.51,124.09,123.61,123.58,123.06,122.68,122.58,121.18,116.17,115.87,115.08,114.99,112.56,72.39,63.25,60.64,59.59,56.62,52.77,52.75,48.80,48.34,26.56,26.37,22.41,22.36,11.26,11.23.m/z[M+H]+:386.6984,calcd,386.7334.
实施例3一种具有全新亚铜离子识别基团的近红外荧光探针的体外分析检测
缓冲溶液的配制:用购买的PBS混合盐配置成浓度为25mM的溶剂体系(pH 7.2),含2mM的GSH(谷胱甘肽)用于防止亚铜离子在空气中被氧化。
浓储的配制:本发明中所用的探针均用实施例2所制备的一种具有全新亚铜离子识别基团的近红外荧光探针(下文称“本发明的探针”)均以DMSO(二甲基亚砜)配置成10mM浓储,各种金属阳离子均用缓冲溶液配置成10mM浓储,本实施例3所用的浓度均为用缓冲液由浓储稀释而来。
1.紫外吸收光谱检测
将终浓度为50μM的本发明的探针溶液与终浓度500μM的亚铜离子溶液混合,检测探针与亚铜离子混合前后的紫外吸收光谱。
结果表明:本发明的探针自身在缓冲体系中的紫外最大吸收波长为415nm,而与底物亚铜离子反应后的体系的最大吸收波长达到560nm,发生近150nm的红移(图4)。
2.荧光图谱
将终浓度为50μM的本发明的探针溶液与终浓度500μM的亚铜离子溶液混合,检测探针与亚铜离子混合前后的吸收和发射光谱的变化。
结果表明:本发明的探针自身的最大激发和发射波长分别为~420nm和~565nm,与亚铜离子反应后的体系的最大激发和发射波长均发生~150nm的红移,分别达到了~560nm和~710nm。
3.荧光强度与亚铜离子反应时间的关系
将终浓度为50μM的本发明的探针溶液与终浓度500μM的亚铜离子溶液混合,孵育不同时间,检测荧光信号。
结果表明:其荧光强度在反应15min内迅速攀升,接下来荧光强度继续增加,,在反应90min后荧光强度趋于稳定,达到对应探针浓度下最大响应信号。总的来说,探针结合亚铜离子后的0~240min内的荧光强度都大大高于未结合亚铜离子的情形。
4.荧光强度与亚铜离子浓度的关系
将终浓度50μM的本发明的探针与终浓度0、2.5、5、10、20、30、40、50、100、200、400、500、800、1000μM的亚铜离子共孵育60min,使用560nm波长的激发光,于710nm波长处检测的荧光强度。
结果表明:荧光强度随着亚铜离子浓度升高而递增,当该亚铜离子近红外探针浓度为≥1μM时其荧光强度为空白组荧光强度的2~40倍,对亚铜离子表现出较优的灵敏度(图7)。
5.本发明的探针对不同金属离子的选择性
使用终浓度50μM的本发明的探针分别与终浓度为500μM的不同的离子溶液混合,离子包括:Li+、Na+、K+、Ca2+、Mg2+、Cd2+、Cr2+、Fe2+、Fe3+、Hg2+、Ni2+、Zn2+、Cu2+、Co2+、Mn2+、Ba2+、Gd2+和Pb2+,以超纯水为对照(Control)。
结果显示:该亚铜离子近红外探针对其他干扰离子在发射波长为710nm处的响应值近乎可以忽略,但对亚铜离子响应的荧光强度是其他干扰离子响应强度的7-35倍(图8)。
实施例4一种具有全新亚铜离子识别基团的近红外荧光探针的细胞毒性及细胞成像
根据实施例3所述的一种具有全新亚铜离子识别基团的近红外荧光探针能对水溶液环境中的亚铜离子高灵敏度、高选择性地响应,为了拓展其应用,发明人将本发明的亚铜离子探针进一步应用于分析检测细胞层面亚铜离子的水平方面的能力进行了探究。
a.细胞毒性:用MTT法进行细胞毒性测试,本实例中所用的探针均用细胞培养液将浓储梯度稀释成不同浓度工作液,且保证工作液里DMSO含量不会对细胞的存活率有影响。根据本发明的探针当浓度高达200μM时,细胞存活率为≥50%,对细胞存活率影响极低,可用于进一步应用于细胞水平亚铜离子的探测。
b.细胞成像:根据实施例2所描述的最适激发和发射波长下,亚铜离子共孵育的细胞组细胞成像发出鲜艳的红色荧光,而没有亚铜离子参与的空白对照组则是仅有微弱的光(图9)。
本实施例表明,本发明的探针可以适用于活细胞等活体的亚铜离子检测。
综上,本发明的一种具有全新亚铜离子识别基团的近红外荧光探针能对亚铜离子进行高选择性、高灵敏度的响应,并且可以低毒性应用于细胞水平亚铜离子的检测和分析,为生物化学、环境化学和生物医学等各领域提供有力的分析工具,在学习和探究亚铜离子含量及其生理、病理过程方面具有重大应用前景。
以上所述仅为本发明的优选实施方式,对于本领域的普通技术人员来说,在不脱离本发明创造构思的前提下,还可以做出若干改进和变换,均属于本发明的保护范畴。
Claims (11)
1.一种具有高选择性的亚铜离子近红外荧光探针,其特征在于,它具有N3O结构;
所述N3O结构为:
式中Y表示氢或卤素。
2.如权利要求1所述的探针,其特征在于,其分子结构如式1所示:
其中R1、R2选自C1-3烷基;X为卤素。
3.如权利要求2所述的探针,其特征在于,式1中,R1为正丙基;和/或,R2为正丙基;和/或,X为Br。
4.权利要求2所述探针的制备方法,其特征在于,它包括以下步骤:
(1)中间体A与中间体N3O在有机溶剂中反应成芳香醚,得中间体5;
中间体A的结构式为:
中间体N3O的结构式为:
式中Y为氢或卤素;
中间体5为4-羟基取代的间苯二甲醛,其结构式为:
(2)中间体B与中间体5在有机溶剂中反应,得到式1所示化合物;
中间体B为1-取代的2,3,3-三甲基吲哚盐,其结构式为:
式中R表示C1-3的烷基;
步骤(1)所述有机溶剂为醇类、DMF、THF或甲苯;
步骤(2)所述有机溶剂为醇类、DMF、THF或甲苯。
5.如权利要求4所述的制备方法,其特征在于:步骤(1)中中间体N3O与中间体A的摩尔比是1∶(2~4);
和/或,反应温度是50~100℃。
6.如权利要求4所述的制备方法,其特征在于:步骤(1)中的有机溶剂为N,N-二甲基甲酰胺;
和/或,步骤(2)中的有机溶剂为乙醇。
7.如权利要求4所述的制备方法,其特征在于:步骤(2)中中间体5与中间体B的摩尔比是1∶(2~4);
和/或,反应温度是50~100℃。
8.如权利要求4所述的制备方法,其特征在于:步骤(2)的反应时间是2~6小时。
9.如权利要求4所述的制备方法,其特征在于:步骤(2)的反应还需加入醋酸钠,醋酸钠的物质的量为中间体5的1~3倍。
10.权利要求1-3任一所述探针在亚铜离子检测中的用途。
11.权利要求1-3任一所述探针在制备亚铜离子检测试剂中的用途。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201910727683.3A CN110305111B (zh) | 2019-08-07 | 2019-08-07 | 一种亚铜离子近红外探针及其制备方法和应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201910727683.3A CN110305111B (zh) | 2019-08-07 | 2019-08-07 | 一种亚铜离子近红外探针及其制备方法和应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN110305111A true CN110305111A (zh) | 2019-10-08 |
CN110305111B CN110305111B (zh) | 2021-06-15 |
Family
ID=68082096
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201910727683.3A Active CN110305111B (zh) | 2019-08-07 | 2019-08-07 | 一种亚铜离子近红外探针及其制备方法和应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN110305111B (zh) |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1844119A (zh) * | 2006-04-26 | 2006-10-11 | 山东师范大学 | 检测细胞内锌离子的近红外荧光探针及合成方法和用途 |
CN107266929A (zh) * | 2017-06-21 | 2017-10-20 | 四川大学 | 一类以菁染料荧光基团为母体骨架结构的近红外荧光染料及其制备方法与应用 |
CN109020955A (zh) * | 2018-08-02 | 2018-12-18 | 深圳大学 | 一种分子探针、制备方法及其应用 |
-
2019
- 2019-08-07 CN CN201910727683.3A patent/CN110305111B/zh active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1844119A (zh) * | 2006-04-26 | 2006-10-11 | 山东师范大学 | 检测细胞内锌离子的近红外荧光探针及合成方法和用途 |
CN107266929A (zh) * | 2017-06-21 | 2017-10-20 | 四川大学 | 一类以菁染料荧光基团为母体骨架结构的近红外荧光染料及其制备方法与应用 |
CN109020955A (zh) * | 2018-08-02 | 2018-12-18 | 深圳大学 | 一种分子探针、制备方法及其应用 |
Non-Patent Citations (3)
Title |
---|
DEBABRATA ET AL.: ""Reaction-based probes for Co(II) and Cu(I) with dual output modes: fluorescence live cell imaging"", 《RSC ADV.》 * |
DEBABRATA MAITY ET AL.: ""Reactive Probes for Ratiometric Detection of Co2+ and Cu+ Based on Excited-State Intramolecular Proton Transfer Mechanism"", 《ORGANIC LETTERS》 * |
陈华: ""新型吲哚三碳菁衍生物的设计合成及其光谱性能研究"", 《中国优秀硕士学位论文全文数据库 工程科技I辑》 * |
Also Published As
Publication number | Publication date |
---|---|
CN110305111B (zh) | 2021-06-15 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Li et al. | A benzothiazole-based fluorescent probe for efficient detection and discrimination of Zn2+ and Cd2+, using cysteine as an auxiliary reagent | |
CN106220640B (zh) | 一类汞离子荧光探针及其制备方法和应用 | |
Huo et al. | The synthesis, characterization of three isomers of rhodamine derivative and their application in copper (II) ion recognition | |
Li et al. | A new fluorescent chemosensor for Zn2+ with facile synthesis:“Turn-on” response in water at neutral pH and its application for live cell imaging | |
CN106967053B (zh) | 二价铜离子荧光探针及其制备方法和用途 | |
CN107459483B (zh) | 一种细胞膜靶向h2s荧光探针及其制备方法和应用 | |
CN109608414A (zh) | 检测过氧亚硝酸根的荧光探针及其制备方法和应用 | |
CN102993207A (zh) | 一类罗丹明b-三嗪化合物及其制备方法和应用 | |
CN102344449B (zh) | 并杂环萘酰亚胺及其制备方法与应用 | |
Zeng et al. | A near-infrared fluorescent sensor with large Stokes shift for rapid and highly selective detection of thiophenols in water samples and living cells | |
Liu et al. | Rhodamine based turn-on fluorescent sensor for Hg2+ and its application of microfluidic system and bioimaging | |
CN110229165A (zh) | 上转换荧光探针罗丹明衍生物及其应用 | |
CN114591633A (zh) | 一类氧杂蒽-半花菁近红外荧光染料,其合成方法及应用 | |
Cui et al. | A rhodamine B-based turn on fluorescent probe for selective recognition of mercury (II) ions | |
CN105906619B (zh) | 一种双光子荧光探针及其制备方法和用途 | |
CN102093270A (zh) | 细胞内银离子检测用二苯代乙烯双光子荧光探针 | |
CN107382977B (zh) | 基于吡嗪联吡唑酰腙衍生物的荧光探针及其制备方法和应用 | |
CN106977585B (zh) | 一种线粒体定位用于光动力疗法的双光子荧光探针库及其应用 | |
CN106554770B (zh) | 一种三唑衍生物金属离子荧光探针及其制备方法和应用 | |
CN110305111A (zh) | 一种新的亚铜离子近红外探针及其制备方法和应用 | |
CN115261015B (zh) | 一种基于ICT原理检测N2H4和Cu2+的双通道荧光探针及其制备方法和应用 | |
CN105968098A (zh) | 一种含咔唑、苯并咪唑取代的喹啉衍生物及其制备方法和应用 | |
CN104946237A (zh) | 一种钯离子荧光探针化合物及其应用 | |
CN102633789B (zh) | 含有双罗丹明b的化合物及其制备方法与应用 | |
Wang et al. | Easily accessible and highly selective “Turn-on” fluorescent sensor for imaging cadmium in living cells |
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 | ||
CB03 | Change of inventor or designer information | ||
CB03 | Change of inventor or designer information |
Inventor after: Zhang Yuwei Inventor after: Ke Bowen Inventor after: Qi Qingrong Inventor after: Kang Ting Inventor before: Ke Bowen Inventor before: Zhang Yuwei Inventor before: Qi Qingrong Inventor before: Kang Ting |
|
GR01 | Patent grant | ||
GR01 | Patent grant |