CN104119263B - A kind of organic compound based on cyanine and application thereof - Google Patents
A kind of organic compound based on cyanine and application thereof Download PDFInfo
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- CN104119263B CN104119263B CN201410294697.8A CN201410294697A CN104119263B CN 104119263 B CN104119263 B CN 104119263B CN 201410294697 A CN201410294697 A CN 201410294697A CN 104119263 B CN104119263 B CN 104119263B
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- halfcystine
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- cyanine
- organic compound
- fluorescent probe
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- QGKMIGUHVLGJBR-UHFFFAOYSA-M (4z)-1-(3-methylbutyl)-4-[[1-(3-methylbutyl)quinolin-1-ium-4-yl]methylidene]quinoline;iodide Chemical compound [I-].C12=CC=CC=C2N(CCC(C)C)C=CC1=CC1=CC=[N+](CCC(C)C)C2=CC=CC=C12 QGKMIGUHVLGJBR-UHFFFAOYSA-M 0.000 title claims abstract description 19
- 150000002894 organic compounds Chemical class 0.000 title claims abstract description 19
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 claims abstract description 62
- 238000001514 detection method Methods 0.000 claims abstract description 28
- 239000007850 fluorescent dye Substances 0.000 claims abstract description 27
- 239000000523 sample Substances 0.000 claims description 27
- 201000010099 disease Diseases 0.000 claims description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 3
- 150000001875 compounds Chemical class 0.000 abstract description 29
- 230000008859 change Effects 0.000 abstract description 18
- 238000000034 method Methods 0.000 abstract description 5
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 abstract description 4
- 235000018417 cysteine Nutrition 0.000 abstract description 4
- 238000009825 accumulation Methods 0.000 abstract description 3
- 230000009471 action Effects 0.000 abstract description 3
- 238000011835 investigation Methods 0.000 abstract description 3
- 230000007246 mechanism Effects 0.000 abstract description 3
- 230000008569 process Effects 0.000 abstract description 3
- 210000004027 cell Anatomy 0.000 description 12
- 230000000694 effects Effects 0.000 description 9
- 239000000243 solution Substances 0.000 description 9
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 8
- 239000007787 solid Substances 0.000 description 8
- 238000006243 chemical reaction Methods 0.000 description 7
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 6
- 239000012530 fluid Substances 0.000 description 6
- 238000002360 preparation method Methods 0.000 description 6
- 238000000921 elemental analysis Methods 0.000 description 5
- 238000002189 fluorescence spectrum Methods 0.000 description 5
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 5
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 4
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 4
- FFFHZYDWPBMWHY-VKHMYHEASA-N L-homocysteine Chemical compound OC(=O)[C@@H](N)CCS FFFHZYDWPBMWHY-VKHMYHEASA-N 0.000 description 4
- 210000001175 cerebrospinal fluid Anatomy 0.000 description 4
- 230000003834 intracellular effect Effects 0.000 description 4
- 230000004044 response Effects 0.000 description 4
- 238000004088 simulation Methods 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 description 3
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 3
- 239000004475 Arginine Substances 0.000 description 3
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- 239000004471 Glycine Substances 0.000 description 3
- 239000007995 HEPES buffer Substances 0.000 description 3
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 3
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 3
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 3
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 description 3
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 3
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 3
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 3
- 239000004473 Threonine Substances 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 3
- 239000006035 Tryptophane Substances 0.000 description 3
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 3
- 239000003480 eluent Substances 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N ethyl acetate Substances CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- 229930182817 methionine Natural products 0.000 description 3
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 3
- 238000001556 precipitation Methods 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 238000010992 reflux Methods 0.000 description 3
- 229960004799 tryptophan Drugs 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 235000009328 Amaranthus caudatus Nutrition 0.000 description 2
- 240000001592 Amaranthus caudatus Species 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- RWSOTUBLDIXVET-UHFFFAOYSA-N Dihydrogen sulfide Chemical compound S RWSOTUBLDIXVET-UHFFFAOYSA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 235000012735 amaranth Nutrition 0.000 description 2
- 239000004178 amaranth Substances 0.000 description 2
- 235000001014 amino acid Nutrition 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 229910021529 ammonia Inorganic materials 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 229960004132 diethyl ether Drugs 0.000 description 2
- 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 2
- 229910000037 hydrogen sulfide Inorganic materials 0.000 description 2
- 239000005457 ice water Substances 0.000 description 2
- XHXFXVLFKHQFAL-UHFFFAOYSA-N phosphoryl trichloride Chemical compound ClP(Cl)(Cl)=O XHXFXVLFKHQFAL-UHFFFAOYSA-N 0.000 description 2
- 239000000741 silica gel Substances 0.000 description 2
- 229910002027 silica gel Inorganic materials 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- SKDHHIUENRGTHK-UHFFFAOYSA-N 4-nitrobenzoyl chloride Chemical compound [O-][N+](=O)C1=CC=C(C(Cl)=O)C=C1 SKDHHIUENRGTHK-UHFFFAOYSA-N 0.000 description 1
- LWLKGUVXXNOKHI-MPZVPXKJSA-N CCC(Cc1c(C(C)(C)C(/C=C/C(CCC/C2=C\C=C(/C3(C)C)\N(CC)c4c3cccc4)=C2OC(c(cc2)ccc2[N+]([O-])=O)=O)=C)cccc1)=[IH] Chemical compound CCC(Cc1c(C(C)(C)C(/C=C/C(CCC/C2=C\C=C(/C3(C)C)\N(CC)c4c3cccc4)=C2OC(c(cc2)ccc2[N+]([O-])=O)=O)=C)cccc1)=[IH] LWLKGUVXXNOKHI-MPZVPXKJSA-N 0.000 description 1
- MIJQBFLKIMCJPS-UHFFFAOYSA-N CCN(/C(/C1(C)C)=C/C=C(\CCC2)/C(OC(c(cc3)ccc3[N+]([O-])=O)=O)=C2/C=C/C(C2(C)C)=[N+](CC)c3c2cccc3)c2c1cccc2 Chemical compound CCN(/C(/C1(C)C)=C/C=C(\CCC2)/C(OC(c(cc3)ccc3[N+]([O-])=O)=O)=C2/C=C/C(C2(C)C)=[N+](CC)c3c2cccc3)c2c1cccc2 MIJQBFLKIMCJPS-UHFFFAOYSA-N 0.000 description 1
- 108010024636 Glutathione Proteins 0.000 description 1
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 1
- 239000006004 Quartz sand Substances 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- NSOXQYCFHDMMGV-UHFFFAOYSA-N Tetrakis(2-hydroxypropyl)ethylenediamine Chemical compound CC(O)CN(CC(C)O)CCN(CC(C)O)CC(C)O NSOXQYCFHDMMGV-UHFFFAOYSA-N 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- SMWDFEZZVXVKRB-UHFFFAOYSA-N anhydrous quinoline Natural products N1=CC=CC2=CC=CC=C21 SMWDFEZZVXVKRB-UHFFFAOYSA-N 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- OSVXSBDYLRYLIG-UHFFFAOYSA-N chlorine dioxide Inorganic materials O=Cl=O OSVXSBDYLRYLIG-UHFFFAOYSA-N 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 239000012043 crude product Substances 0.000 description 1
- JHIVVAPYMSGYDF-UHFFFAOYSA-N cyclohexanone Chemical compound O=C1CCCCC1 JHIVVAPYMSGYDF-UHFFFAOYSA-N 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000000840 electrochemical analysis Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 244000144992 flock Species 0.000 description 1
- 229960003180 glutathione Drugs 0.000 description 1
- -1 gsh Chemical compound 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- HVTICUPFWKNHNG-UHFFFAOYSA-N iodoethane Chemical compound CCI HVTICUPFWKNHNG-UHFFFAOYSA-N 0.000 description 1
- 230000000302 ischemic effect Effects 0.000 description 1
- 238000012417 linear regression Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 210000005229 liver cell Anatomy 0.000 description 1
- 230000003908 liver function Effects 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000002438 mitochondrial effect Effects 0.000 description 1
- 239000012046 mixed solvent Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000012452 mother liquor Substances 0.000 description 1
- 239000010813 municipal solid waste Substances 0.000 description 1
- 230000002107 myocardial effect Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 230000036542 oxidative stress Effects 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 210000002381 plasma Anatomy 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 230000002787 reinforcement Effects 0.000 description 1
- 230000010410 reperfusion Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 238000011896 sensitive detection Methods 0.000 description 1
- 229960004249 sodium acetate Drugs 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- LMBFAGIMSUYTBN-MPZNNTNKSA-N teixobactin Chemical group C([C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(=O)N[C@H](CCC(N)=O)C(=O)N[C@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(=O)N[C@H]1C(N[C@@H](C)C(=O)N[C@@H](C[C@@H]2NC(=N)NC2)C(=O)N[C@H](C(=O)O[C@H]1C)[C@@H](C)CC)=O)NC)C1=CC=CC=C1 LMBFAGIMSUYTBN-MPZNNTNKSA-N 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
- 230000003313 weakening effect Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
- C07D209/04—Indoles; Hydrogenated indoles
- C07D209/10—Indoles; Hydrogenated indoles with substituted hydrocarbon radicals attached to carbon atoms of the hetero ring
- C07D209/12—Radicals substituted by oxygen atoms
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
- C09B23/00—Methine or polymethine dyes, e.g. cyanine dyes
- C09B23/10—The polymethine chain containing an even number of >CH- groups
- C09B23/107—The polymethine chain containing an even number of >CH- groups four >CH- groups
-
- 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"
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- 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
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- 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
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Optics & Photonics (AREA)
- Biochemistry (AREA)
- Materials Engineering (AREA)
- Engineering & Computer Science (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The present invention relates to fluorescent probe, specifically a kind of organic compound based on cyanine and application thereof. Based on the organic compound of cyanine, structural formula is such as formula shown in I, this compounds of halfcystine fluorescent probe of the present invention, fluorescence intensity generation considerable change corresponding under halfcystine exists, can be used for the detection of halfcystine, and can greatly reduce the interference of external detection condition, it is to increase accuracy of detection. This compounds of this invention halfcystine fluorescent probe, when halfcystine exists also there is considerable change in uv-absorbing, can take into account naked eyes with ultraviolet scene luminosity simultaneously detect. This compounds can be used for the detection of intraor extracellular cysteine levels as fluorescent probe, this, to the kinetics mechanism of the processes such as the further investigation generation of halfcystine in organism, conveying and accumulation, especially studies halfcystine and has important biomedical meaning in plastosome physiological action.
Description
Technical field
The present invention relates to fluorescent probe, specifically a kind of organic compound based on cyanine and application thereof.
Background technology
Halfcystine is a kind of amino acid containing sulfydryl, is one of important physiologically active substance. It can increase the growing amount of gsh as the substrate of reductive glutathione, thus the stability of Cell protection film, alleviate the damage of myocardial cell in ischemic reperfusion. Cys participates in the phospholipid metabolism process in the reduction of cell and liver, has protection liver cell, promotes the effect of liver function. In addition, Cys also has stimulates pre-T lymphocyte be divided into the effect of ripe lymphocyte and increase human body to the resistibility of some toxin. In organism, the content of halfcystine and Metabolic disorder all can cause the generation of some diseases, by measuring the content of halfcystine in organism, it may be achieved to the diagnosis of some metabolic trouble. Therefore, it is achieved quick, sensitive detection halfcystine tool is of great significance.
At present, picture colorimetry, electrochemical analysis, the methods such as stratographic analysis all can be used to the semicystinol concentration measured in blood plasma and homogenised tissue, but these methods often need sample pretreatment, fluorescent probe detection method has highly sensitive because of it and the advantage such as direct-detection can become one of important detection means of analyzing halfcystine in life entity in viable cell or tissue. A fluorescent probe with application prospect should have before and after effect that change in fluorescence is obviously, fast to target molecule response, selectivity is good, can be used for the advantages such as real-time reversible detection. MengZhang etc. disclose a class in order to detect the fluorescent probe (M.Zhanget.al, J.Am.Chem.Soc., 2007,129,10322) of halfcystine, with Fluorescence Increasing after halfcystine effect thus detect the existence of halfcystine. But this kind of fluorescent probe is slow to halfcystine response, and detection limit for height, can not be used for detecting halfcystine fast. And the excitation-emission wavelength of this probe is positioned at ultraviolet region, can not effectively avoiding the interference of organism itself fluorescence, UV-light is very big to organism photobleaching effect simultaneously, is easy to damage biological sample. Fully it is penetrated into organization internal for reaching and avoids cell autofluorescence interference object, greatly reduce the interference of outside atmosphere, it is achieved detection by quantitative, still need to develop the operability fluorescent probe with longer excitation-emission wavelength. Therefore, exploitation has good selectivity, and the fluorescent probe that can carry out detecting in near-infrared region halfcystine in living things system is significant.
Summary of the invention
It is an object of the invention to provide a kind of organic compound based on cyanine and application thereof.
For achieving the above object, the technical solution used in the present invention is:
Based on an organic compound for cyanine, based on the organic compound of cyanine, structural formula such as formula shown in I,
Based on an application for the organic compound of cyanine, the organic compound based on cyanine shown in described formula I is as the fluorescent probe detecting halfcystine.
Halfcystine under the organic compound based on cyanine shown in described formula I is used for the detection physiological environment of qualitative/quantitative as probe, inside and outside cell or organism.
The useful effect of the present invention:
The present invention is used for the compound as halfcystine fluorescent probe, its fluorescence intensity corresponding under halfcystine exists and emission wavelength change, simultaneously uv-absorbing also correspondence change, and then can be used for the detection of aqueous systems, simulation physiological environment and intracellular cysteine level, and can greatly reduce the interference of external detection condition, it is to increase accuracy of detection. The compounds of this invention is used as fluorescent probe, can be used for the detection of intracellular cysteine, but also the plastosome in cell can be positioned, this is to the kinetics mechanism of the processes such as the further investigation generation of halfcystine in organism, conveying and accumulation, the physiological action of understanding halfcystine further, especially research halfcystine resist oxidative stress environment institute role at plastosome and have important biomedical meaning.
The present invention is used for the compound as halfcystine fluorescent probe, and fluorescence intensity generation considerable change corresponding under halfcystine exists, can be used for the detection of halfcystine, and can greatly reduce the interference of external detection condition, it is to increase accuracy of detection. This compounds of this invention halfcystine fluorescent probe, when halfcystine exists also there is considerable change in uv-absorbing, can take into account naked eyes with ultraviolet scene luminosity simultaneously detect. This compounds can be used for the detection of intraor extracellular cysteine levels as fluorescent probe, this, to the kinetics mechanism of the processes such as the further investigation generation of halfcystine in organism, conveying and accumulation, especially studies halfcystine and has important biomedical meaning in plastosome physiological action.
Accompanying drawing explanation
The fluorescence intensity change curve of the fluorescent probe adopted under the different pH that Fig. 1 provides for the embodiment of the present invention.
At 560nm and 720nm place fluorescence intensity ratio value change curve after the fluorescent probe adopted under the different pH that Fig. 2 provides for the embodiment of the present invention and halfcystine effect.
The fluorescent probe adopted that Fig. 3 provides for the embodiment of the present invention is to the selectivity schematic diagram of halfcystine; Wherein, X-coordinate is followed successively by blank, halfcystine, homocysteine from left to right, gsh, tryptophane, glycine, Threonine, proline(Pro), arginine, Serine, methionine(Met), ��-amino-isovaleric acid.
The linear fit curve of the fluorescent probe 560nm adopted that Fig. 4 provides for the embodiment of the present invention and 720nm place fluorescence intensity ratio value and semicystinol concentration change.
The employing fluorescent probe halfcystine that Fig. 5 provides for the embodiment of the present invention is for detecting the Laser Scanning Confocal Microscope imaging of intracellular hydrogen sulfide.
Embodiment
Embodiment is used for the present invention is described further, but the invention is not restricted to embodiment.
Embodiment 1
Organic compound structure formula based on cyanine is:
Halfcystine during type I compound is inside and outside with water body to be determined, simulation physiological environment or organism is combined, obtain the compound of formula II structure thus cause the fluorescence intensity of type I compound and the change of wavelength, and the change of uv-absorbing, and then utilize type I compound halfcystine can be carried out qualitative, quantitative detection.
Preparation based on formula I organic compound of cyanine:
(1) preparation of compound one
Graduated cylinder measures 20-50mLDMF and is dissolved in 20-40mL methylene dichloride, pours in 250mL there-necked flask and (puts magneton to stir), cools in ice-water bath. Measure 20-40mL phosphorus oxychloride and dissolve in 20-40mL methylene dichloride, pour into and constant pressure funnel is added dropwise to after above-mentioned cooling in solution, and stir instrument with magnetic and constantly stir.
Dropwise, take 5-10g pimelinketone and pour above-mentioned constant pressure funnel into and drip and add. Now solution colour turns into yellow gradually from colourless. Then ice-water bath is removed, reflux 3 hours. Reaction solution is poured into and is filled in trash ice after terminating by reaction. It is placed in stink cupboard hold over night. In tank, upper strata is be yellow flocks and have a little red oil bottom yellow liquid. Using B��chner funnel decompress filter, mother liquor washs, and obtains yellow solid compound one. By solid transfer in small beaker, dry in vacuum drying oven. Fusing point: 130��131 DEG C, results of elemental analyses (theoretical value): C, 55.4 (55.7); H5.4 (5.3); Cl, 20.4 (20.5). LC-MS (API-ES): m/zC8H9ClO2Calcd172,found[M+]173.
(2) preparation of compound two
By 100-150mL iodoethane, 20-30mL2,3,3-tri-methyl indole quinoline, 10-20mL toluene three adds in 250mL round-bottomed flask, and reflux 10-15h reacts. On a rotary evaporator solvent is spin-dried for after complete, after being spin-dried for, obtains amaranth solid. Add after methyl alcohol heating makes dissolution of solid in flask, transfer to beaker drips and add ether solid precipitation is gone out. Take out filter, by washed with diethylether precipitation, obtain baby pink pulverulent solids compound two. Stand-by by transferring to after solid drying in wide-necked bottle. Results of elemental analyses (theoretical value): C, 82.9 (82.8); H9.7 (9.8); N, 7.4 (7.5). LC-MS (API-ES): m/zC13H18N+Calcd188,found[M+]189.
(3) preparation of compound three
0.5-1g compound one and 0.2-0.5g compound two are dissolved in the mixed solvent of 100-150mL propyl carbinol and benzene (7:3v/v), fill a water separator on reactor, separate the water that reaction generates, reaction is carried out to positive reaction direction. Mixture reflux, and constantly stir. After reaction 2-4h, cool to room temperature, underpressure distillation is except desolventizing. Precipitation washed with diethylether, obtains amaranth solid crude product, column chromatography separating purification, and eluent is methyl alcohol-ethyl acetate (7:12V/V), obtains green product compound three. Results of elemental analyses (theoretical value): C, 79.7 (79.8); H, 7.9 (8.0); Cl, 6.9 (6.8); N, 5.5 (5.6). LC-MS (API-ES): m/zC34H40ClN2 +Calcd511,found[M+]512.
(4) preparation of compound four
Under nitrogen protection, 0.3-0.5g compound three and 0.1-0.3g sodium-acetate, at the anhydrous N of 10-20ml, react 5-8 hour in dinethylformamide at 80-90 DEG C. After cooling, by quartz sand termination reaction and filter. Silica gel column chromatogram separating purification, eluent is methyl alcohol-ethyl acetate (7:12V/V), receives red component, obtains red product, be compound four.
Results of elemental analyses (theoretical value): C, 88.9 (88.8); H, 8.2 (8.3); N, 5.7 (5.8); O, 3.2 (3.3). LC-MS (API-ES): m/zC34H40N2OCalcd492,found[M+]493.
(5) preparation of formula one
By 0.2-0.5g compound four, 0.1-0.3g paranitrobenzoyl chloride and the anhydrous quadrol of 150-200 �� L at the anhydrous N of 20-30ml, dinethylformamide reacts 0.5-2h under condition of ice bath. Silica gel column chromatogram separating purification, eluent is methyl alcohol-ethyl acetate (1:3V/V), receives green color component, obtains green product formula one compound. Results of elemental analyses (theoretical value): C, 76.6 (76.7); H, 6.9 (6.8); N, 6.5 (6.6); O, 9.9 (10.0). LC-MS (API-ES): m/zC41H44N3O4 +Calcd642,found[M+]643.
Embodiment 2
Using prepare gained formula one compound as probe application in aqueous systems, simulation physiological environment and cell in carry out the detection to halfcystine, simulation physiological condition, the following experiment all carries out (HEPES buffered soln when pH7-8, concentration is 1-30mM), concentration and probe concentration adopts 0.5-10 ��M.
The above-mentioned pH effect preparing the double pipe ammonia acid response of gained probe:
PH gradient adopts the control of HEPES buffered soln. In 1.5ml colorimetric cylinder, add the 1-10mMHEPES of the different pH gradient of 0.8-1mL, then add 1-10 �� L100 ��M of probe, shake even solution, after balancing 10-30min at 25-40 DEG C, above-mentioned mixed working fluid is added in fluorescence ware and measures fluorescence spectrum. Fluorescence intensity with pH change as shown in Figure 1. In the scope of pH4.0��8.0, fluorescence intensity does not have considerable change as shown in Figure 1, and namely in the system of pH6.0��8.0, probe is all stable, may be used to detection halfcystine. 0.8-1mL1-10mMHEPES is added in 1.5ml colorimetric cylinder, add 1-10 �� L100 ��M of probe again, then add 50-100 �� L200 ��M of halfcystine, shake even solution, after balancing 10-30min at 25-40 DEG C, above-mentioned working fluid is added in fluorescence ware and measures fluorescence spectrum. Ratio fluorescent with pH change as shown in Figure 2. The system in pH6.0��8.0 can be found, probe halfcystine has corresponding, and the rising with pH, corresponding reinforcement, due to probe in the basic conditions, relatively unstable, so, when pH is 7.40, probe was both stablized, again can the sensitive response of acid of double pipe ammonia, can be used for the detection realizing in organism.
Embodiment 3
Probe is to the selectivity of halfcystine
PH adopts the control of HEPES buffered soln. Get multiple 1.5ml colorimetric cylinder, in colorimetric cylinder, add 0.9-1mL1-10mMHEPES, pH7-8, adding 1-10 �� L100 ��M of probe again, then add determinand respectively as shown in Figure 2, determinand working fluid is followed successively by: blank (without amino acid), halfcystine, homocysteine, gsh, tryptophane, glycine, Threonine, proline(Pro), arginine, Serine, methionine(Met), ��-amino-isovaleric acid, wherein the final concentration of halfcystine and homocysteine is 20 ��Ms, and other final concentrations are 2mM. Shake even solution, after balancing 10-30min at 25-40 DEG C, working fluid in each colorimetric cylinder is poured in fluorescence ware respectively and measures fluorescence spectrum. Probe is to the selectivity of halfcystine as shown in Figure 2. And halfcystine is had good selectivity by probe as seen from the figure, after halfcystine effect, the fluorescence that 720nM place excites obviously weakens, and the fluorescence that 560nM place excites obviously strengthens. Homocysteine under condition determination, gsh, tryptophane, glycine, Threonine, proline(Pro), arginine, Serine, methionine(Met), ��-amino-isovaleric acid etc. can not make fluorescence probe change.
Embodiment 4
Probe is to the detection by quantitative of halfcystine
Get multiple 1.5ml colorimetric cylinder, 0.9-1mL1-10mMHEPES is added in each colorimetric cylinder, pH7-8, add 1-10 �� L100 ��M of probe again, then the halfcystine of different final concentration is added respectively, final concentration is respectively 2, 4, 6, 8, 10, 12 ��Ms, shake even solution, after balancing 10-30min at 25-40 DEG C, working fluid in each colorimetric cylinder is poured into respectively fluorescence ware and measures fluorescence spectrum, get each 560nm and 720nm place fluorescence intensity ratio value, Input Software OriginPro8.0, obtain linear work curve as shown in Figure 4, wherein the linear regression constant of linear fit curve is 0.996, show the concentration of the mensuration halfcystine that probe can be quantitative.
Representing the change of the change system fluorescence intensity with semicystinol concentration by Fig. 3, show the increase with semicystinol concentration, system 560nm absorption intensity is obviously weakening in obvious enhancing, 720nm absorption intensity.
Embodiment 5
Probe is to the detection by quantitative of halfcystine in artificial cerebrospinal fluid
Get multiple 1.5ml colorimetric cylinder, 0.9-1mL1-10mM artificial cerebrospinal fluid is added in each colorimetric cylinder, pH7-8, add 1-10 �� L100 ��M of probe again, then adding final concentration respectively is 2, 4, 6, 8, 10 ��Ms of halfcystines, shake even solution, after balancing 10-30min at 25-40 DEG C, working fluid in each colorimetric cylinder is poured into respectively fluorescence ware and measures fluorescence spectrum, obtain each 560nm and 720nm place fluorescence intensity ratio value, and by institute's value with embodiment 7 is obtained in linear work curve compared with numerical value, find that coincidence is better (see table 1), show the concentration of the mensuration halfcystine that probe can be quantitative in cerebrospinal fluid.
Table 1 is the result detecting halfcystine in cerebrospinal fluid with fluorescent probe
Embodiment 6
Probe is used for the detection of intracellular hydrogen sulfide
It is about 50% that rat liver cancer cell HepG2 carries out being cultured to cell density according to AmericantypeTissueCultureCollection regulation. Then by HepG2 cell in 1-10 ��M of probe 25-40 DEG C hatch 10-30 minute, wash 3 times with DMEM-1640 substratum, under being placed in confocal fluorescent microscope, under 560nM exciting light, take pictures (see Fig. 5). As shown in Figure 5, under 560nM exciting light, fluorescence intensity obviously strengthens; Cell mitochondrial is mainly dyeed by this probe.
Above content is in conjunction with concrete preferred implementation further description made for the present invention, can not assert that specific embodiment of the invention is confined to these explanations. For general technical staff of the technical field of the invention, without departing from the inventive concept of the premise, it is also possible to make some simple deduction or replace, all should be considered as belonging to protection scope of the present invention. As a kind of purposes that fluorescence dye is new compound of the present invention; can not assert that the compound of the present invention is only for fluorescence dye; for general technical staff of the technical field of the invention; under the consideration of the identical mechanism of action being used as fluorescence dye based on the compounds of this invention; some simple inferences can also be made; draw other application purpose of the compound of the present invention, all should be considered as belonging to protection scope of the present invention.
Claims (3)
1. the organic compound based on cyanine, it is characterised in that: based on the organic compound of cyanine, structural formula such as formula shown in I,
2. the application of the organic compound based on cyanine according to claim 1, it is characterised in that: the organic compound based on cyanine shown in described formula I is as the fluorescent probe of non-diseases diagnostic detection halfcystine.
3. by the application of the organic compound based on cyanine according to claim 2, it is characterised in that: the probe that the organic compound based on cyanine shown in described formula I diagnose as non-diseases is for the halfcystine under the detection physiological environment of qualitative/quantitative, inside and outside cell or organism.
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| CN107266347B (en) * | 2017-07-27 | 2020-03-31 | 中国科学院烟台海岸带研究所 | Organic compound based on cyanine and application thereof |
| CN107722058B (en) * | 2017-11-08 | 2019-07-05 | 中国科学院烟台海岸带研究所 | A kind of organic compound and its application |
| CN107903289B (en) * | 2017-11-08 | 2020-07-31 | 中国科学院烟台海岸带研究所 | Organic compound based on cyanine and application thereof |
| CN107722057B (en) * | 2017-11-08 | 2019-07-05 | 中国科学院烟台海岸带研究所 | Organic compound and its application based on flower cyanines |
| CN107954920A (en) * | 2017-11-28 | 2018-04-24 | 中国科学院烟台海岸带研究所 | A kind of organic compound and application based on flower cyanines |
| CN108129376A (en) * | 2017-11-28 | 2018-06-08 | 中国科学院烟台海岸带研究所 | Organic compound, intermediate and application based on flower cyanines |
| CN109651835A (en) * | 2018-12-26 | 2019-04-19 | 云南师范大学 | A kind of IR780 keto-acid Molecule of Cyanine Dyes and preparation method thereof and crystal structure |
| CN112694431B (en) * | 2019-10-23 | 2022-09-16 | 中国医学科学院药物研究所 | Nitroreductase in hypersensitive fluorescent probe detection bacteria and application in bacterial infection |
| CN112062708B (en) * | 2020-09-27 | 2021-09-24 | 中国科学院烟台海岸带研究所 | Fluorescent probe compound for detecting cysteine and application thereof |
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