CN110386898A - A kind of quinoline ring analog derivative fluorescence probe and its preparation method and application - Google Patents
A kind of quinoline ring analog derivative fluorescence probe and its preparation method and application Download PDFInfo
- Publication number
- CN110386898A CN110386898A CN201810364213.0A CN201810364213A CN110386898A CN 110386898 A CN110386898 A CN 110386898A CN 201810364213 A CN201810364213 A CN 201810364213A CN 110386898 A CN110386898 A CN 110386898A
- Authority
- CN
- China
- Prior art keywords
- solution
- fluorescent molecular
- fluorescence
- molecular probe
- cell
- 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
- 239000000523 sample Substances 0.000 title claims abstract description 13
- 125000002943 quinolinyl group Chemical group N1=C(C=CC2=CC=CC=C12)* 0.000 title claims abstract description 9
- 238000002360 preparation method Methods 0.000 title abstract description 6
- 238000001514 detection method Methods 0.000 claims abstract description 13
- 238000006243 chemical reaction Methods 0.000 claims abstract description 12
- 150000001875 compounds Chemical class 0.000 claims abstract description 9
- -1 sulfhydryl compound Chemical class 0.000 claims abstract description 3
- 239000000243 solution Substances 0.000 claims description 38
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 28
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 18
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 18
- 235000019441 ethanol Nutrition 0.000 claims description 15
- 239000000047 product Substances 0.000 claims description 13
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 12
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 9
- JPJALAQPGMAKDF-UHFFFAOYSA-N selenium dioxide Chemical compound O=[Se]=O JPJALAQPGMAKDF-UHFFFAOYSA-N 0.000 claims description 8
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 claims description 6
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 6
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 6
- CMEWLCATCRTSGF-UHFFFAOYSA-N N,N-dimethyl-4-nitrosoaniline Chemical compound CN(C)C1=CC=C(N=O)C=C1 CMEWLCATCRTSGF-UHFFFAOYSA-N 0.000 claims description 6
- MLUCVPSAIODCQM-NSCUHMNNSA-N crotonaldehyde Chemical compound C\C=C\C=O MLUCVPSAIODCQM-NSCUHMNNSA-N 0.000 claims description 6
- MLUCVPSAIODCQM-UHFFFAOYSA-N crotonaldehyde Natural products CC=CC=O MLUCVPSAIODCQM-UHFFFAOYSA-N 0.000 claims description 6
- 238000010898 silica gel chromatography Methods 0.000 claims description 6
- 239000007787 solid Substances 0.000 claims description 5
- 239000000126 substance Substances 0.000 claims description 5
- 238000003760 magnetic stirring Methods 0.000 claims description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 3
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 claims description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical class [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 3
- SMWDFEZZVXVKRB-UHFFFAOYSA-N anhydrous quinoline Natural products N1=CC=CC2=CC=CC=C21 SMWDFEZZVXVKRB-UHFFFAOYSA-N 0.000 claims description 3
- 239000000706 filtrate Substances 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 238000010438 heat treatment Methods 0.000 claims description 3
- 239000011259 mixed solution Substances 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- 239000003208 petroleum Substances 0.000 claims description 3
- 238000007259 addition reaction Methods 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- 230000003595 spectral effect Effects 0.000 claims description 2
- 238000003756 stirring Methods 0.000 claims description 2
- 241001448862 Croton Species 0.000 claims 1
- 150000001299 aldehydes Chemical class 0.000 claims 1
- 239000003795 chemical substances by application Substances 0.000 claims 1
- 238000001035 drying Methods 0.000 claims 1
- 238000011503 in vivo imaging Methods 0.000 claims 1
- 229910052938 sodium sulfate Inorganic materials 0.000 claims 1
- 235000011152 sodium sulphate Nutrition 0.000 claims 1
- 239000003068 molecular probe Substances 0.000 abstract description 42
- LSNNMFCWUKXFEE-UHFFFAOYSA-L sulfite Chemical compound [O-]S([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-L 0.000 abstract description 24
- 238000000034 method Methods 0.000 abstract description 7
- 238000012360 testing method Methods 0.000 abstract description 6
- 230000000694 effects Effects 0.000 abstract description 4
- 239000012528 membrane Substances 0.000 abstract description 3
- 230000035699 permeability Effects 0.000 abstract description 3
- NIXOWILDQLNWCW-UHFFFAOYSA-M Acrylate Chemical compound [O-]C(=O)C=C NIXOWILDQLNWCW-UHFFFAOYSA-M 0.000 abstract description 2
- UCKMPCXJQFINFW-UHFFFAOYSA-N Sulphide Chemical compound [S-2] UCKMPCXJQFINFW-UHFFFAOYSA-N 0.000 abstract description 2
- 230000007246 mechanism Effects 0.000 abstract description 2
- BUIHQKXJJJBLGS-UHFFFAOYSA-N n,n-dimethylquinolin-6-amine Chemical compound N1=CC=CC2=CC(N(C)C)=CC=C21 BUIHQKXJJJBLGS-UHFFFAOYSA-N 0.000 abstract description 2
- 231100000252 nontoxic Toxicity 0.000 abstract description 2
- 230000003000 nontoxic effect Effects 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 36
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 15
- RAHZWNYVWXNFOC-UHFFFAOYSA-N sulfur dioxide Inorganic materials O=S=O RAHZWNYVWXNFOC-UHFFFAOYSA-N 0.000 description 11
- 238000011534 incubation Methods 0.000 description 8
- 238000002189 fluorescence spectrum Methods 0.000 description 6
- LSNNMFCWUKXFEE-UHFFFAOYSA-M Bisulfite Chemical compound OS([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-M 0.000 description 5
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 4
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 4
- UCTWMZQNUQWSLP-UHFFFAOYSA-N adrenaline Chemical compound CNCC(O)C1=CC=C(O)C(O)=C1 UCTWMZQNUQWSLP-UHFFFAOYSA-N 0.000 description 4
- 230000001413 cellular effect Effects 0.000 description 4
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 4
- 235000018417 cysteine Nutrition 0.000 description 4
- 230000009977 dual effect Effects 0.000 description 4
- 238000002474 experimental method 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
- 239000002609 medium Substances 0.000 description 4
- 238000000862 absorption spectrum Methods 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 238000004113 cell culture Methods 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 231100000135 cytotoxicity Toxicity 0.000 description 3
- 230000003013 cytotoxicity Effects 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 230000005284 excitation Effects 0.000 description 3
- RMVRSNDYEFQCLF-UHFFFAOYSA-N thiophenol Chemical compound SC1=CC=CC=C1 RMVRSNDYEFQCLF-UHFFFAOYSA-N 0.000 description 3
- 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
- 108010024636 Glutathione Proteins 0.000 description 2
- HDFGOPSGAURCEO-UHFFFAOYSA-N N-ethylmaleimide Chemical compound CCN1C(=O)C=CC1=O HDFGOPSGAURCEO-UHFFFAOYSA-N 0.000 description 2
- LSNNMFCWUKXFEE-UHFFFAOYSA-N Sulfurous acid Chemical compound OS(O)=O LSNNMFCWUKXFEE-UHFFFAOYSA-N 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- MLIREBYILWEBDM-UHFFFAOYSA-N cyanoacetic acid Chemical compound OC(=O)CC#N MLIREBYILWEBDM-UHFFFAOYSA-N 0.000 description 2
- 239000003480 eluent Substances 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 229960003180 glutathione Drugs 0.000 description 2
- 235000003969 glutathione Nutrition 0.000 description 2
- 229910000037 hydrogen sulfide Inorganic materials 0.000 description 2
- 231100001231 less toxic Toxicity 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- QSOTZUMQTPFZAX-UHFFFAOYSA-N C1(=CC=CC=C1)O.[S].C1=CC=CC=C1 Chemical compound C1(=CC=CC=C1)O.[S].C1=CC=CC=C1 QSOTZUMQTPFZAX-UHFFFAOYSA-N 0.000 description 1
- UGFAIRIUMAVXCW-UHFFFAOYSA-N Carbon monoxide Chemical compound [O+]#[C-] UGFAIRIUMAVXCW-UHFFFAOYSA-N 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- GHAZCVNUKKZTLG-UHFFFAOYSA-N N-ethyl-succinimide Natural products CCN1C(=O)CCC1=O GHAZCVNUKKZTLG-UHFFFAOYSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 208000012902 Nervous system disease Diseases 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 210000002821 alveolar epithelial cell Anatomy 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 239000012736 aqueous medium Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 229910002091 carbon monoxide Inorganic materials 0.000 description 1
- 238000007334 copolymerization reaction Methods 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 239000012043 crude product Substances 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000005611 electricity Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 125000005909 ethyl alcohol group Chemical group 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 230000001605 fetal effect Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 210000003494 hepatocyte Anatomy 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 1
- 210000003292 kidney cell Anatomy 0.000 description 1
- 150000002611 lead compounds Chemical class 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 210000003470 mitochondria Anatomy 0.000 description 1
- 201000011682 nervous system cancer Diseases 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 230000005622 photoelectricity Effects 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000002516 radical scavenger Substances 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 239000012265 solid product Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 150000003573 thiols Chemical class 0.000 description 1
- 231100001234 toxic pollutant Toxicity 0.000 description 1
- 231100000820 toxicity test Toxicity 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 description 1
- 238000005406 washing Methods 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
- A61K49/0032—Methine dyes, e.g. cyanine dyes
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
- C07D215/02—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom
- C07D215/16—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D215/38—Nitrogen atoms
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pathology (AREA)
- Biomedical Technology (AREA)
- General Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Molecular Biology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Optics & Photonics (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The present invention relates to a kind of quinoline ring analog derivative fluorescence probe and its preparation method and application, compound name is 2- cyano -3- (6- (dimethylamino) quinoline -2- base) acrylate.Compound molecule quality of the present invention is small, and structure is simple, can be used as a molecule sensor, the sulphite in sensitive, selective detection living cells, and open fluorescence reaction.The testing mechanism is not influenced by sulfide and sulfhydryl compound, have the characteristics that selectivity is high, more outstanding to be, the fluorescent molecular probe is almost non-toxic to cell and has very excellent membrane permeability effect, 2min just can penetrate living cells film, open fluorescence reaction.It provides a kind of method of new detection endogenous sulphite of living cells, is of great practical significance.
Description
The present invention relates to a kind of quinoline ring analog derivative fluorescent molecular probe and its preparation method and in detection sulphite
Using.
Background technique
SO2Gas is listed in toxic pollutant since the last century, recent studies have shown that the molecule may be after an oxygen
Change nitrogen, another after carbon monoxide and hydrogen sulfide participates in the important gas signaling molecule of various pathology and physiology course, high
Exogenous SO2Level can cause cardiovascular disease, the diseases such as the nervous system disease and cancer.United States Food and Drag Administration rule
Fixed, the sulfite content that product contains such as meets or exceeds 10 mcg/mls needs and is especially marked.On the other hand, In
In cytoplasm and mitochondria also SO can be generated by the approach such as hydrogen sulfide and biological thiol oxidation2, can also generate in an aqueous medium
SO3 2-And HSO3 -.Normal endogenous SO2Level has the effects that adjust blood pressure, but abnormal endogenous SO2It is horizontal also with mind
It is closely bound up through the diseases such as systemic disease and cancer.Therefore, develop quick, sensitive, selective sulphite detection probe or
Method is most important.
We obtain a kind of effectively sensitive and less toxic fluorescent molecular probe, same normalizing by preparing quinoline ring analog derivative
Serial experiment shows that the fluorescent molecule is with good performance, and potential practical application value is very high.
Summary of the invention
The purpose of the present invention is to provide a kind of novel quinoline ring analog derivative fluorescent molecular probe and its preparation method with
Application in practice.
Technical scheme is as follows:
One kind is by quinoline cyclammonium analog derivative fluorescent molecular probe, it is characterized in that it is just like flowering structure:
A method of above-mentioned quinoline ring analog derivative being prepared, it includes the following steps:
4-N, accelerine are added in hydrochloric acid solution by step 1., dissolve it sufficiently, add crotonaldehyde,
Magnetic stirring makes to be uniformly mixed, normal-temperature reaction 1h, TLC detection reaction carry out degree, the ratio between amount of substance 4-N, accelerine:
Crotonaldehyde=1: 2, toluene addition reaction solution is further flowed back 115 DEG C overnight, after being cooled to room temperature, removes toluene layer, water layer
It is neutralized with saturation sodium hydroxide solution, solution is extracted with dichloromethane again, is washed twice with saturated sodium chloride solution, uses after dry
Anhydrous sodium sulfate filtering, is concentrated under reduced pressure, which is purified by silica gel column chromatography, and obtains yellow-brown solid, is first step product；
Selenium dioxide is added to dioxane/water by step 2., and volume ratio is dioxane: water=10: 1 solution
In, first step product is added after 80 DEG C of heating 30min, the ratio between amount of substance first step product: selenium dioxide=1: 2, magnetic stirs
Mixing makes to be uniformly mixed, and reacts 4h at 80 DEG C, after being cooled to room temperature, is filtered by diatomite, then is rinsed with a small amount of methylene chloride
Filter residue, filtrate are concentrated under reduced pressure, and are separated with silica gel column chromatography, and eluant, eluent is petroleum ether and ethyl acetate volume ratio is 6: 1
Mixed solution obtains second step product；
Step 3. by step 2 in product and cyanoacetic acid second rouge be dissolved in ethanol solution, 1h is stirred at room temperature,
Obtained alcohol mixture is washed 3 times with cold ethyl alcohol, and obtained solid recrystallizes in the mixed liquor of ethyl alcohol and acetone, volume ratio
It is ethyl alcohol: acetone=9: 1, obtain target compound.
The invention has the advantages that compound of the present invention has very sensitive effect to detection inferior sulfate radical, and
And checkout procedure is quick, performance stablize, it is less toxic the advantages that.Experiment shows that the fluorescent molecule can fast and effectively detect interior external source
Sulfur dioxide molecule, it is most important that the fluorescent molecule very short membrane permeability time, it is only necessary to 2min；Benefit of the invention
It is the probe molecule that lead compound synthesis has fluorescent characteristic with crotonaldehyde, can effectively detects intracellular sulfur dioxide molecule
Content, be with a wide range of applications.
Specific embodiment
By following embodiment, present invention be described in more detail, but should be noted that the scope of the present invention is not implemented by these
Any restrictions of example.
The preparation of one: 2- cyano -3- of embodiment (6- (dimethylamino) quinoline -2- base) acrylate
By 36.7mmol, the 4-N of 5g, accelerine is added in 6mol, the hydrochloric acid solution of 66mL, fills it
Divide dissolution, add 73.5mmol, the crotonaldehyde of 6mL, magnetic stirring makes to be uniformly mixed, normal-temperature reaction 1h, TLC detection reaction progress
Degree, the ratio between amount of substance 4-N, accelerine: crotonaldehyde=1: 2, it is further that reaction solution is added in the toluene of 35mL
115 DEG C of back flow reactions are stayed overnight, and after being cooled to room temperature, remove toluene layer, water layer is neutralized with saturation sodium hydroxide solution, acquired solution
It is extracted with dichloromethane, is washed twice with saturated sodium chloride solution again, is concentrated under reduced pressure after being filtered after dry with anhydrous sodium sulfate, it should
Crude product is purified by silica gel column chromatography, and obtains yellow-brown solid product；It is two that selenium dioxide, which is added to dioxane/water volume ratio,
Six ring of oxygen: water=10: 1, i.e. dioxane 140mL in the solution of water 40mL, are added after 80 DEG C of heating 30min
18.8mmol, 3.5g products therefrom, magnetic stirring make to be uniformly mixed, react 4h at 80 DEG C, after being cooled to room temperature, pass through diatomite
Filtering, then filter residue is rinsed with a small amount of methylene chloride, filtrate is being concentrated under reduced pressure, is being separated with silica gel column chromatography, and eluant, eluent is petroleum ether
The mixed solution for being 6: 1 with ethyl acetate volume ratio, obtains product；3.0mmol, the resulting product of 0.6g and 3.3mmol are taken,
The cyan-acetic ester of 0.37g is dissolved in ethanol solution, and 1h is stirred at room temperature, and obtained alcohol mixture is clear with cold ethyl alcohol
The solid obtained after washing 3 times recrystallizes in the mixed liquor of ethyl alcohol and acetone, volume ratio ethyl alcohol: acetone=9: 1, obtain red powder
The target compound 0.73g at end, yield 83%.1H NMR (600MHz, DMSO-d6) δ 8.37 (s, 1H), 8.16 (d, J=
8.5Hz, 1H), 7.89 (d, J=8.6Hz, 1H), 7.57 (dd, J=9.4,2.8Hz, 1H), 6.93 (d, J=2.8Hz, 1H),
4.34 (q, J=7.1Hz, 2H), 3.12 (s, 6H), 4.34 (t, J=7.1Hz, 3H) .13C NMR (150MHz, DMSO-d6)δ
(ppm) 162.80,153.71,150.44,144.67,141.85,134.14,131.23,130.97,124.77,121.04,
115.94,103.88,103.14,62.76,40.43,14.50.HRMS (ESI-TOF) m/z:[M+H]+Calcd for
C17H18N3O2296.13 Found 296.1410.
The property of the fluorescent molecule compound and active application experiment, by the fluorescent molecular probe prepared in embodiment one into
The test of row embodiment two to ten, specific data and is analyzed as follows:
Embodiment two:
Fig. 1: in PBS solution, the uv absorption spectra of the fluorescent molecular probe
10 μM of fluorescent molecular probes are dissolved in PBS (pH7.4,10mM, 5%DMSO) solution, after 37 DEG C are incubated for, In
It is detected on Shimadzu UV-2550 instrument, the uv-visible absorption spectra of the fluorescent molecular probe is as shown in Figure 1.
Embodiment three:
Fig. 2: in PBS solution, the fluorescent molecular probe is with SO3 2-The fluorescence spectrum and change in fluorescence curve of concentration variation
Figure
10 μM of fluorescent molecular probes are dissolved in PBS (pH7.4,10mM, 5%DMSO) solution, after 37 DEG C of incubation 1h,
Respectively in different SO3 2-Its fluorescence spectral characteristic, SO are detected under concentration3 2-Concentration range is 0-1000 μM, in Hitachi F-7000
It is detected on instrument, excitation wavelength 364nm, slit width 5nm, Photomultiplier tube voltage 500V.
The results show that by SO at 483nm3 2-Concentration increase to 1000 μM (100 equivalents for being equivalent to probe) from 0
To a kind of standard curve；SO at 483nm3 2-Concentration shows stronger linear relationship at 0-15 μM, and related coefficient is
0.9862.It follows that with SO3 2-The increase of concentration, fluorescence intensity constantly become by force, SO3 2-When concentration reaches 15 equivalents or so,
Fluorescence intensity reaches maximum and keeps stable.
Example IV:
Fig. 3: in PBS solution, the selective lab diagram of the fluorescent molecular probe
10 μM of fluorescent molecular probes are dissolved in PBS (pH7.4,10mM, 5%DMSO) solution, after 37 DEG C of incubation 1h,
Respectively using different tested its selectivity of analyte detection, SO3 2-Concentration and HSO3 2-Concentration be 100 μM, other are tested
Material concentration is 1mM, is detected on Hitachi's F-7000 instrument, excitation wavelength 364nm, slit width 5nm, photoelectricity times
Increasing tube voltage is 500V.
The fluorescent molecular probe be capable of specificity to SO3 2-And HSO3 2-The substance of response, other tests has cysteine,
Glutathione, cysteine, benzenethiol and other anion.Compared to cysteine, glutathione, cysteine and benzene sulphur
Phenol, the fluorescent molecular probe is to SO3 2-And HSO3 2-With good selectivity.The fluorescent molecule can be used as SO3 2-And HSO3 2-'s
Probe.
Embodiment five:
Fig. 4: in PBS solution, fluorescence spectra which responds with pH
10 μM of fluorescent molecular probes are dissolved in PBS (pH7.4,10mM, 5%DMSO) solution, after 37 DEG C of incubation 1h,
Its performance, the range of pH value are as follows: 3-12 are detected under different pH value respectively.It is detected, is swashed on Hitachi's F-7000 instrument
Hair wavelength is 364nm, slit width 5nm, Photomultiplier tube voltage 500V.
From the figure, it can be seen that the fluorescent molecular probe itself is hardly influenced by pH.The fluorescent molecular probe and 100
μM SO3 2-When being incubated for altogether, it can be seen that when pH value is 1-5 and 8-12, the fluorescence intensity of the molecule is constantly declining, however pH
In 5-8, which stablizes value, and stable part is sufficient for experiment in vivo.
Embodiment six:
Fig. 5: in PBS solution, fluorescence spectra which responds at any time
10 μM of fluorescent molecular probes are dissolved in PBS (pH7.4,10mM, 5%DMSO) solution, add 100 μM
SO3 2-It is incubated at 37 DEG C, detects its performance, incubation time range are as follows: 0-12h under different incubation times respectively.In F-
It is detected on 7000 instruments, excitation wavelength 364nm, launch wavelength 483nm, slit width 5nm, photomultiplier tube electricity
Pressure is 500V.
The fluorescent molecular probe shows that very strong fluorescence increases (fluorescence intensity more than half increases) within 1min,
After time reaches 1h, fluorescence property is continual and steady.By the accurate control response time, can shorter incubation time (such as
The detection system is used in 10min), while guaranteeing that it continues 12h or more, this illustrates the steady of the fluorescent molecular probe detection system
It is qualitative.
Embodiment seven:
Fig. 6: the fluorescent molecular probe is to four kinds of differential cytotoxicity lab diagrams
The cytotoxicity of the fluorescent molecular probe is evaluated using mtt assay.Test cell be HeLa cell (tumour cell),
HEK293T cell (human embryonic kidney cell line), A549 (people alveolar epithelium AL cell line) and LO2 cell (Fetal hepatocyte strain).Carefully
Born of the same parents' culture solution is comprising 10%FBS, 0.1% dual anti-complete medium (DMEM), and cell kind is cultivated in 96 orifice plates 37
DEG C, 5%CO2In incubator, cell density is 5 × 105A/mL.
It may be concluded that the fluorescent molecular probe has lesser cytotoxicity, even if in higher probe from figure
Under concentration, this proves that the fluorescent molecular probe can be with further progress clinical test.
Embodiment eight:
Fig. 7: in living cells, which is total to focused view to the time dependence experimental fluorescence for entering cell
By 10 μM of fluorescent molecular probes and HeLa cell at 37 DEG C, 5%CO2It is incubated for altogether in incubator, cell culture exists
Comprising 10% fetal calf serum FBS, in 0.1% dual anti-complete medium, each group and probe are total to incubation time and are set to 1,1,
2,10,30 and 30min, then by 150 μM of SO3 2-It is incubated for 30min altogether with cell.It analyzes HeLa cell fluorescence and (λ is imagedex=
364nm, λem=425-475nm), scale bar: 25 μM.
It can be seen from the figure that the fluorescent molecular probe is completely into HeLa cell when incubation time is 2min
In, this quick Premeabilisation of cells process be very remarkably with it is important, it can shorten entire detection cycle.
Embodiment nine:
Fig. 8: in living cells, the fluorescent molecular probe is detected with cell external source SO3 2-The fluorescence co-focusing figure of concentration variation
HeLa cell culture is comprising 10%FBS, and in 0.1% dual anti-complete medium (DMEM, high sugar), cell exists
37 DEG C, 5%CO2After cultivating 12h in incubator, it is added the 10 μM fluorescent molecular probe into cell, after 2min into cell
The SO of various concentration is added3 2-Solution (50 μM, 100 μM, 150 μM), then fluorescence co-focusing picture is shot after being incubated for 30min.
It is available by the analysis to image, the fluorescence intensity of the fluorescent molecular probe and the increase of sulfite concentration
It is positively correlated, and does not observe apparent fluorescence intensity after cell and the fluorescent molecule are incubated for altogether.This shows the fluorescent molecule
Probe can be used as the fluorescent optical sensor of a unlatching, detect the sulphite of exceeded external source, and it is in terms of Premeabilisation of cells
Being fine into image space face with work has preferable performance.
Embodiment ten:
Fig. 9: in living cells, the fluorescent molecular probe is detected at different conditions to cellular endogenous SO3 2-Concentration variation
Fluorescence co-focusing figure
Then further detect the cellular endogenous imaging performance of the fluorescent molecular probe.HeLa cell culture is including 10%
FBS, in 0.1% dual anti-complete medium (DMEM, high sugar), cell is at 37 DEG C, 5% CO2After cultivating 12h in incubator, to
It is added the 10 μM fluorescent molecular probe in cell, after 2min, 200 μM of SO is added into cell2(liquor epinephrinae bitartratis ophthalmicus goes first to donor
Adrenaline) be incubated for 30min again after, at this time under confocal fluorescent microscopic it can be seen that there is apparent fluorescence enhancement, the knot
Fruit may also indicate that the fluorescent molecular probe can effectively detect the SO of cellular endogenous3 2-.It is incubated for after probe and adds into cell
Enter a kind of 2mM NEM (n-ethylmaleimide, sulfhydryl compound scavenger) and 200 μM of SO2(liquor epinephrinae bitartratis ophthalmicus goes first to donor
Adrenaline) be incubated for 30min altogether, then without apparent fluorescence enhancement, therefore, the enhancing of above-mentioned fluorescence, be in living cells in
Source SO3 2-Generation it is related.Then, the SO of 150 μM of external sources is added into cell3 2-, it is incubated for 30min, at this point, can then see again
To apparent fluorescence enhancement.These the experimental results showed that, the fluorescent molecular probe can detect it is exogenous also can detecte it is interior
SO in source property biological cell3 2-.The result shows that compound of the present invention can be used as a molecule sensor, Neng Gouling
It is quick.The selectively sulphite in detection living cells, and fluorescence reaction is opened, the testing mechanism is not by sulfide and sulfydryl
Compounds affect has the characteristics that selectivity is high, and more outstanding to be, the fluorescent molecular probe is almost non-toxic to cell and has
Very excellent membrane permeability effect, 2min just can penetrate living cells film, open fluorescence reaction.It provides a kind of new inspection
The method for surveying the endogenous sulphite of living cells, is of great practical significance.
Fig. 1: in PBS solution, the uv absorption spectra of the fluorescent molecular probe
Fig. 2: in PBS solution, the fluorescent molecular probe is with SO2The fluorescence spectrum and change in fluorescence curve graph of concentration variation
Fig. 3: in PBS solution, the selective lab diagram of the fluorescent molecular probe
Fig. 4: in PBS solution, fluorescence spectra which responds with pH
Fig. 5: in PBS solution, fluorescence spectra which responds at any time
Fig. 6: toxicity test figure of the fluorescent molecular probe to four kinds of different cells
Fig. 7: in living cells, which is total to focused view to the time dependence experimental fluorescence for entering cell
Fig. 8: in living cells, the fluorescent molecular probe is detected with cell external source SO3 2-The fluorescence co-focusing figure of concentration variation
Fig. 9: in living cells, the fluorescent molecular probe is detected at different conditions to cellular endogenous SO3 2-Fluorescence copolymerization
Jiao Tu.
Claims (3)
1. a kind of quinoline ring class fluorescence probe, which is characterized in that the fluorescence probe is made of following structural formula:
2. quinoline ring analog derivative according to claim 1, which is characterized in that include the following steps to prepare:
4-N, accelerine are added in hydrochloric acid solution by step 1., dissolve it sufficiently, add crotonaldehyde, and magnetic stirs
Mixing makes to be uniformly mixed, normal-temperature reaction 1h, TLC detection reaction carry out degree, the ratio between amount of substance 4-N, accelerine: crotons
Aldehyde=1: 2, toluene addition reaction solution is further flowed back 115 DEG C overnight, after being cooled to room temperature, removes toluene layer, water layer is used full
It is neutralized with sodium hydroxide solution, solution is extracted with dichloromethane again, is washed twice with saturated sodium chloride solution, with anhydrous after drying
Filtered over sodium sulfate is concentrated under reduced pressure, which is purified by silica gel column chromatography, and obtains yellow-brown solid, is first step product；
Selenium dioxide is added to dioxane/water by step 2., and volume ratio is dioxane: water=10: in 1 solution, In
First step product is added after 80 DEG C of heating 30min, magnetic stirring makes to be uniformly mixed, and reacts 4h at 80 DEG C, after being cooled to room temperature, leads to
Diatomite filtering is crossed, then rinses filter residue with a small amount of methylene chloride, filtrate is concentrated under reduced pressure, is separated with silica gel column chromatography, elutes
The mixed solution that agent is petroleum ether and ethyl acetate volume ratio is 6: 1, obtains second step product；
Step 3. by step 2 in product and cyan-acetic ester be dissolved in ethanol solution, 1h is stirred at room temperature, obtains
Alcohol mixture washed 3 times with cold ethyl alcohol, obtained solid recrystallizes in the mixed liquor of ethyl alcohol and acetone, and volume ratio is second
Alcohol: acetone=9: 1, obtain target compound.
3. spectral signature, in-vivo imaging and the practical application of quinoline aldehydes fluorescent molecule according to claims 1 and 2.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810364213.0A CN110386898B (en) | 2018-04-20 | 2018-04-20 | Quinoline ring derivative fluorescent probe and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810364213.0A CN110386898B (en) | 2018-04-20 | 2018-04-20 | Quinoline ring derivative fluorescent probe and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110386898A true CN110386898A (en) | 2019-10-29 |
| CN110386898B CN110386898B (en) | 2022-07-29 |
Family
ID=68284330
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810364213.0A Active CN110386898B (en) | 2018-04-20 | 2018-04-20 | Quinoline ring derivative fluorescent probe and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110386898B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114436957A (en) * | 2022-02-25 | 2022-05-06 | 河南理工大学 | Ethylcyanoacetate-dihydroquinolinone derivative fluorescent probe and preparation method and application thereof |
| CN114621200A (en) * | 2022-03-15 | 2022-06-14 | 山西大学 | Near-infrared fluorescent probe and preparation method and application thereof |
-
2018
- 2018-04-20 CN CN201810364213.0A patent/CN110386898B/en active Active
Non-Patent Citations (4)
| Title |
|---|
| YUQING XU ET AL.: "A fluorescence probe acted on Site I binding for Human Serum Albumin", 《TALANTA》 * |
| ZHEN LI ET AL.: "Imaging of formaldehyde in plants with a ratiometric fluorescent probe", 《CHEM. SCI.》 * |
| 李东钰等.: "二氧化硫衍生物荧光探针的研究进展", 《中国科学：化学》 * |
| 柳志学.: "可逆荧光探针的设计、合成及其在药物研究中的应用", 《中国博士学位论文全文数据库·医药卫生科技辑》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114436957A (en) * | 2022-02-25 | 2022-05-06 | 河南理工大学 | Ethylcyanoacetate-dihydroquinolinone derivative fluorescent probe and preparation method and application thereof |
| CN114436957B (en) * | 2022-02-25 | 2023-04-25 | 河南理工大学 | Ethyl cyanoacetate-dihydroquinolinone derivative fluorescent probe and preparation method and application thereof |
| CN114621200A (en) * | 2022-03-15 | 2022-06-14 | 山西大学 | Near-infrared fluorescent probe and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110386898B (en) | 2022-07-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Ye et al. | A dual-channel responsive near-infrared fluorescent probe for multicolour imaging of cysteine in living cells | |
| Wei et al. | Simultaneous imaging of lysosomal and mitochondrial viscosity under different conditions using a NIR probe | |
| CN105924394B (en) | A kind of two-photon formaldehyde fluorescence probe and its preparation and application | |
| Lu et al. | A simple two-output near-infrared fluorescent probe for hydrazine detection in living cells and mice | |
| CN106518762A (en) | Fluorescent probe for detecting formaldehyde in cell endoplasmic reticulums | |
| CN105153214B (en) | A kind of silicon substrate rhodamine nitric oxide fluorescent probe and its preparation method and application | |
| Liu et al. | A dicyanoisophorone-quinolinium-based near-infrared-emission fluorescent probe for ratiometric sensing of bisulfite/sulfite in living cells | |
| Li et al. | A ratiometric fluorescent probe for fast detection of hydrogen sulfide and recognition of biological thiols | |
| Yang et al. | Thiol “click” chromene mediated cascade reaction Forming coumarin for in-situ imaging of thiol flux in drug-induced liver injury | |
| Xu et al. | The visualization of lysosomal and mitochondrial glutathione via near-infrared fluorophore and in vivo imaging application | |
| Yang et al. | A novel mitochondria-targeted ratiometric fluorescent probe for endogenous sulfur dioxide derivatives as a cancer-detecting tool | |
| Yu et al. | A near-infrared fluorogenic probe with fast response for detecting sodium dithionite in living cells | |
| Li et al. | A novel dual-channel fluorescent probe for nitroxyl detection and its application in HepG-2 cells | |
| Zheng et al. | A rhodol-derived probe for intracellular biothiols imaging and rapid labelling of sulfhydryl-containing proteins | |
| Kong et al. | A novel mitochondria-targeted fluorescent probe for imaging hydrazine in living cells, tissues and animals | |
| Li et al. | New carbazole fluorescent sensor for ultrasensitive and ratiometric sensing of SO2 derivatives and hydrazine | |
| CN106518855B (en) | It is a kind of using half river cyanines and flavonols as the Sulfur Dioxide-derivatives scale fluorescence probe of fluorogen and its application | |
| Du et al. | Rational design of a novel turn-on fluorescent probe for the detection and bioimaging of hydrazine with barbituric acid as a recognition group | |
| Shen et al. | A ratiometric and colorimetric fluorescent probe designed based on FRET for detecting SO32−/HSO3− in living cells and mice | |
| Yuan et al. | A novel formaldehyde fluorescent probe based on 1, 8-naphthalimide derivative and its application in living cell | |
| Zhou et al. | A highly sensitive fluorescent probe for selective detection of cysteine/homocysteine from glutathione and its application in living cells and tissues | |
| Qi et al. | Near-infrared turn-on fluorescent probe for discriminative detection of Cys and application in in vivo imaging | |
| KR101472318B1 (en) | WIDE pH RANGE RESPONSIBLE RATIOMETRIC pH PROBE | |
| CN110386898A (en) | A kind of quinoline ring analog derivative fluorescence probe and its preparation method and application | |
| CN109897625A (en) | Selective enumeration method cysteine fluorescence probe and its synthetic method and application |
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 |