CN112939950A - Carbonyl azetidine substituted coumarin fluorescent dye and synthetic method and application thereof - Google Patents
Carbonyl azetidine substituted coumarin fluorescent dye and synthetic method and application thereof Download PDFInfo
- Publication number
- CN112939950A CN112939950A CN201911258845.XA CN201911258845A CN112939950A CN 112939950 A CN112939950 A CN 112939950A CN 201911258845 A CN201911258845 A CN 201911258845A CN 112939950 A CN112939950 A CN 112939950A
- Authority
- CN
- China
- Prior art keywords
- methyl
- benzopyran
- azetidine
- bromo
- mass ratio
- 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.)
- Pending
Links
- ZYGHJZDHTFUPRJ-UHFFFAOYSA-N benzo-alpha-pyrone Natural products C1=CC=C2OC(=O)C=CC2=C1 ZYGHJZDHTFUPRJ-UHFFFAOYSA-N 0.000 title claims abstract description 35
- 239000007850 fluorescent dye Substances 0.000 title claims abstract description 34
- 229960000956 coumarin Drugs 0.000 title claims abstract description 27
- 235000001671 coumarin Nutrition 0.000 title claims abstract description 27
- ZLAFIVONWPDODG-UHFFFAOYSA-N O=C=C1CCN1 Chemical group O=C=C1CCN1 ZLAFIVONWPDODG-UHFFFAOYSA-N 0.000 title claims abstract description 24
- 125000000332 coumarinyl group Chemical class O1C(=O)C(=CC2=CC=CC=C12)* 0.000 title claims 2
- 238000010189 synthetic method Methods 0.000 title description 3
- 239000000975 dye Substances 0.000 claims abstract description 43
- 230000015572 biosynthetic process Effects 0.000 claims abstract description 18
- 150000004775 coumarins Chemical class 0.000 claims abstract description 18
- 238000003786 synthesis reaction Methods 0.000 claims abstract description 18
- 238000000799 fluorescence microscopy Methods 0.000 claims abstract description 6
- 125000000217 alkyl group Chemical group 0.000 claims abstract 8
- 125000004178 (C1-C4) alkyl group Chemical group 0.000 claims abstract 2
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 51
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 33
- VRLDNEKULNVMFD-UHFFFAOYSA-N 7-bromo-4-methylchromen-2-one Chemical compound C1=C(Br)C=CC2=C1OC(=O)C=C2C VRLDNEKULNVMFD-UHFFFAOYSA-N 0.000 claims description 31
- 239000002904 solvent Substances 0.000 claims description 26
- 238000003756 stirring Methods 0.000 claims description 17
- CYPYTURSJDMMMP-WVCUSYJESA-N (1e,4e)-1,5-diphenylpenta-1,4-dien-3-one;palladium Chemical compound [Pd].[Pd].C=1C=CC=CC=1\C=C\C(=O)\C=C\C1=CC=CC=C1.C=1C=CC=CC=1\C=C\C(=O)\C=C\C1=CC=CC=C1.C=1C=CC=CC=1\C=C\C(=O)\C=C\C1=CC=CC=C1 CYPYTURSJDMMMP-WVCUSYJESA-N 0.000 claims description 15
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 claims description 15
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 15
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 15
- FJDQFPXHSGXQBY-UHFFFAOYSA-L caesium carbonate Chemical compound [Cs+].[Cs+].[O-]C([O-])=O FJDQFPXHSGXQBY-UHFFFAOYSA-L 0.000 claims description 15
- 229910000024 caesium carbonate Inorganic materials 0.000 claims description 15
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 14
- 238000006243 chemical reaction Methods 0.000 claims description 14
- 239000000741 silica gel Substances 0.000 claims description 14
- 229910002027 silica gel Inorganic materials 0.000 claims description 14
- 239000003480 eluent Substances 0.000 claims description 12
- 238000001816 cooling Methods 0.000 claims description 11
- 238000010438 heat treatment Methods 0.000 claims description 11
- 239000000243 solution Substances 0.000 claims description 7
- PSGQCCSGKGJLRL-UHFFFAOYSA-N 4-methyl-2h-chromen-2-one Chemical compound C1=CC=CC2=C1OC(=O)C=C2C PSGQCCSGKGJLRL-UHFFFAOYSA-N 0.000 claims description 6
- XDLOYRPBXLOSTK-UHFFFAOYSA-N C(C)(C)C1=C(C(=CC(=C1)C(C)C)C(C)C)C1=CC=CC=C1.P Chemical group C(C)(C)C1=C(C(=CC(=C1)C(C)C)C(C)C)C1=CC=CC=C1.P XDLOYRPBXLOSTK-UHFFFAOYSA-N 0.000 claims description 6
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 claims description 6
- ASOKPJOREAFHNY-UHFFFAOYSA-N 1-Hydroxybenzotriazole Chemical compound C1=CC=C2N(O)N=NC2=C1 ASOKPJOREAFHNY-UHFFFAOYSA-N 0.000 claims description 5
- GIAFURWZWWWBQT-UHFFFAOYSA-N 2-(2-aminoethoxy)ethanol Chemical compound NCCOCCO GIAFURWZWWWBQT-UHFFFAOYSA-N 0.000 claims description 5
- 239000012295 chemical reaction liquid Substances 0.000 claims description 5
- 239000012299 nitrogen atmosphere Substances 0.000 claims description 5
- -1 1-ethyl- (3-dimethylaminopropyl) Chemical group 0.000 claims description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide Substances CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 claims description 2
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 claims description 2
- CHNOQXRIMCFHKZ-UHFFFAOYSA-N methyl azetidine-2-carboxylate Chemical compound COC(=O)C1CCN1 CHNOQXRIMCFHKZ-UHFFFAOYSA-N 0.000 claims description 2
- WGEYNHXHVVYOSK-UHFFFAOYSA-N methyl azetidine-3-carboxylate Chemical compound COC(=O)C1CNC1 WGEYNHXHVVYOSK-UHFFFAOYSA-N 0.000 claims description 2
- 239000002243 precursor Substances 0.000 claims description 2
- 238000000034 method Methods 0.000 claims 6
- 230000002194 synthesizing effect Effects 0.000 claims 6
- 238000001514 detection method Methods 0.000 abstract 1
- 238000001917 fluorescence detection Methods 0.000 abstract 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- 238000000695 excitation spectrum Methods 0.000 description 6
- 239000012452 mother liquor Substances 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 5
- 238000005286 illumination Methods 0.000 description 5
- WFKWXMTUELFFGS-UHFFFAOYSA-N tungsten Chemical compound [W] WFKWXMTUELFFGS-UHFFFAOYSA-N 0.000 description 5
- 229910052721 tungsten Inorganic materials 0.000 description 5
- 239000010937 tungsten Substances 0.000 description 5
- 238000000295 emission spectrum Methods 0.000 description 4
- 230000005284 excitation Effects 0.000 description 4
- 239000005457 ice water Substances 0.000 description 4
- 238000001228 spectrum Methods 0.000 description 4
- FYSNRJHAOHDILO-UHFFFAOYSA-N thionyl chloride Chemical compound ClS(Cl)=O FYSNRJHAOHDILO-UHFFFAOYSA-N 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 3
- 238000010226 confocal imaging Methods 0.000 description 3
- 238000002189 fluorescence spectrum Methods 0.000 description 3
- 238000001819 mass spectrum Methods 0.000 description 3
- FKHBGZKNRAUKEF-UHFFFAOYSA-N methyl azetidine-2-carboxylate;hydrochloride Chemical compound Cl.COC(=O)C1CCN1 FKHBGZKNRAUKEF-UHFFFAOYSA-N 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000010992 reflux Methods 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 238000005160 1H NMR spectroscopy Methods 0.000 description 2
- GOLORTLGFDVFDW-UHFFFAOYSA-N 3-(1h-benzimidazol-2-yl)-7-(diethylamino)chromen-2-one Chemical group C1=CC=C2NC(C3=CC4=CC=C(C=C4OC3=O)N(CC)CC)=NC2=C1 GOLORTLGFDVFDW-UHFFFAOYSA-N 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- IADUEWIQBXOCDZ-UHFFFAOYSA-N azetidine-2-carboxylic acid Chemical compound OC(=O)C1CCN1 IADUEWIQBXOCDZ-UHFFFAOYSA-N 0.000 description 2
- 238000009795 derivation Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000012265 solid product Substances 0.000 description 2
- 239000012085 test solution Substances 0.000 description 2
- IADUEWIQBXOCDZ-VKHMYHEASA-N Azetidine-2-carboxylic acid Natural products OC(=O)[C@@H]1CCN1 IADUEWIQBXOCDZ-VKHMYHEASA-N 0.000 description 1
- UXKCLTPQRBKROC-UHFFFAOYSA-N C(C)(C)C1=C(C(=CC(=C1)C(C)C)C(C)C)C1=CC=CC=C1.[P] Chemical group C(C)(C)C1=C(C(=CC(=C1)C(C)C)C(C)C)C1=CC=CC=C1.[P] UXKCLTPQRBKROC-UHFFFAOYSA-N 0.000 description 1
- 210000001789 adipocyte Anatomy 0.000 description 1
- GFZWHAAOIVMHOI-UHFFFAOYSA-N azetidine-3-carboxylic acid Chemical compound OC(=O)C1CNC1 GFZWHAAOIVMHOI-UHFFFAOYSA-N 0.000 description 1
- 150000001539 azetidines Chemical group 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- UOCWTLBPYROHEF-UHFFFAOYSA-N methyl azetidine-3-carboxylate;hydrochloride Chemical compound Cl.COC(=O)C1CNC1 UOCWTLBPYROHEF-UHFFFAOYSA-N 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
- C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
- C07D405/04—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
-
- 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
- C09B57/00—Other synthetic dyes of known constitution
- C09B57/02—Coumarine dyes
-
- 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
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1029—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1088—Heterocyclic compounds characterised by ligands containing oxygen as the only heteroatom
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
- G01N2001/302—Stain compositions
Abstract
The invention provides a novel carbonyl azetidine substituted coumarin fluorescent dye and synthesis and application thereof, and the structural innovation of the dye is that carbonyl azetidine is covalently connected to the 7 th position of a coumarin parent. The novel fluorescent dye has excellent fluorescence intensity and light stability, so that the novel fluorescent dye has wide application prospect in the fields of fluorescence detection, fluorescence imaging and the like, particularly emerging fields of monomolecular detection, super-resolution fluorescence imaging and the like.
R1Is C1-4 alkyl, R2Is composed of
Wherein X1Is H, C1-2 alkyl, X2Is a C1-2 alkyl group,
X3is a C1-2 alkyl group,
X4is C1-2 alkyl.
Description
Technical Field
The invention belongs to the field of fluorescent dyes, and particularly relates to a novel carbonyl azetidine substituted coumarin fluorescent dye and synthesis and application thereof.
Background
Coumarin is a very large family of compounds, and coumarin derivatives are widely found in nature, particularly in plants. The chemical history of coumarin dates back to nearly 200 years, and in recent years, the coumarin parent has been widely applied to the design of small-molecule fluorescent dyes due to the advantages of excellent biocompatibility, strong and stable fluorescence emission, good structural flexibility and the like.
Organic fluorescent dyes are widely applied to biomolecule labeling, and the position and the effect of functional biomolecules are researched by tracking the biomolecules in a WYSIWYG (what you see is what you get) mode. The light intensity and light stability of fluorescent dyes are the primary factors affecting the imaging quality, and although a large number of commercial dyes have been developed and widely used, with the rapid development of single-molecule fluorescence imaging supported by ultra-high resolution, higher requirements are made on the light intensity and light stability of fluorescent dyes.
Quaternary azetidines have been shown to improve both the brightness and photostability of dyes, and newly developed carbonyl azetidines have been shown to further improve the brightness and photostability of coumarin fluorophore precursors.
Disclosure of Invention
One of the purposes of the invention is to provide a novel carbonyl azetidine substituted coumarin fluorescent dye.
The invention also aims to provide a synthesis method of the novel carbonyl azetidine substituted coumarin fluorescent dye, which has the advantages of simple and convenient operation, convenient purification, easy derivation and the like.
The novel carbonyl azetidine substituted coumarin fluorescent dye takes coumarin as a fluorophore matrix, and carbonyl azetidine is covalently connected to the 7 th position of coumarin, so that the brightness and the light stability of the dye are improved.
The carbonyl azetidine substituted coumarin fluorescent dye has the following structural formula:
the novel carbonyl azetidine substituted coumarin fluorescent dye comprises the following specific synthesis steps:
(1) synthesis of dye 1- (4-methyl-2H-1-benzopyran-2-one) azetidine-2-carboxylic acid methyl ester:
adding 4-methyl-7-bromo-2H-1-benzopyran-2-one, azetidine-2-methyl formate hydrochloride, cesium carbonate, tris (dibenzylideneacetone) dipalladium and 2-dicyclohexyl-phosphorus-2, 4, 6-triisopropylbiphenyl into a reactor, adding 1, 4-dioxane under anhydrous and oxygen-free conditions, slowly heating the reaction solution to 80-120 deg.C under stirring, reacting for 2-10 hr, cooling to room temperature, removing solvent under reduced pressure, separating with silica gel column, and removing the solvent by using dichloromethane and methanol in a volume ratio of 200-5:1 as an eluent to obtain the dye 1- (4-methyl-2H-1-benzopyran-2-ketone) azetidine-2-carboxylic acid methyl ester.
(2) Synthesis of dye 1- (4-methyl-2H-benzopyran-2-one) azetidine-3-carboxylic acid methyl ester:
adding 4-methyl-7-bromo-2H-1-benzopyran-2-one, azetidine-3-methyl formate hydrochloride, cesium carbonate, tris (dibenzylideneacetone) dipalladium and 2-dicyclohexylphosphonium-2, 4, 6-triisopropylbiphenyl into a reactor, adding 1, 4-dioxane under anhydrous and oxygen-free conditions, slowly heating the reaction solution to 80-120 deg.C under stirring, reacting for 2-10 hr, cooling to room temperature, removing solvent under reduced pressure, separating with silica gel column, and removing the solvent by using dichloromethane and methanol in a volume ratio of 200-5:1 as an eluent to obtain the dye 1- (4-methyl-2H-1-benzopyran-2-ketone) azetidine-3-carboxylic acid methyl ester.
(3) Synthesis of the dye 1- (4-methyl-2H-1-benzopyran-2-one) azetidine-2- (2- (2-hydroxyethoxy) ethyl) carboxamide:
1-Boc-L-acridine-2-carboxylic acid is dissolved in dry N, N-dimethylformamide and diglycolamine, 1-hydroxybenzotriazole, 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride and triethylamine are added thereto in this order and stirred at room temperature for 12 to 36 hours. Stopping the reaction, removing the solvent under reduced pressure, separating by a silica gel column, adding dichloromethane into the obtained product, adding trifluoroacetic acid into the dichloromethane, continuing to stir overnight, stopping the reaction, separating by the silica gel column, and removing the solvent by taking dichloromethane as an eluent to obtain a white viscous liquid intermediate azetidine-2- (2- (2-hydroxyethoxy) ethyl) formamide.
Adding 4-methyl-7-bromo-2H-1-benzopyran-2-one, azetidine-2- (2- (2-hydroxyethoxy) ethyl) formamide, cesium carbonate, tris (dibenzylideneacetone) dipalladium and 2-dicyclohexylphosphorus-2, 4, 6-triisopropylbiphenyl into a reactor, adding 1, 4-dioxane under anhydrous and oxygen-free (nitrogen atmosphere), slowly heating the reaction liquid to 80-120 ℃ under stirring, continuing to react for 2-10 hours, cooling to room temperature, removing the solvent under reduced pressure, separating by using a silica gel column, removing the solvent by using dichloromethane and methanol with the volume ratio of 200-5:1 as eluent to obtain the dye 1- (4-methyl-2H-1-benzopyran-2-one) azetidine-2- (2- (2-hydroxyethoxy) ethyl) formamide.
Wherein in (1), 4-methyl-7-bromo-2H-1-benzopyran-2-ketone: the mass ratio of the azetidine-2-methyl formate hydrochloride is 1: 0.5-2;
4-methyl-7-bromo-2H-1-benzopyran-2-one: the mass ratio of the cesium carbonate is 1: 3-6;
4-methyl-7-bromo-2H-1-benzopyran-2-one: the mass ratio of the tris (dibenzylideneacetone) dipalladium is 10-2: 1;
4-methyl-7-bromo-2H-1-benzopyran-2-one: the mass ratio of the 2-dicyclohexyl phosphonium-2, 4, 6-triisopropyl biphenyl is 12-1: 1;
the volume ratio of the mass of the 4-methyl-7-bromo-2H-1-benzopyran-2-ketone to the volume of the 1, 4-dioxane is 1:40-100 g/mL.
(2) And (3) 4-methyl-7-bromo-2H-1-benzopyran-2-one: the mass ratio of the azetidine-3-methyl formate hydrochloride is 1: 0.5-2;
4-methyl-7-bromo-2H-1-benzopyran-2-one: the mass ratio of the cesium carbonate is 1: 3-6;
4-methyl-7-bromo-2H-1-benzopyran-2-one: the mass ratio of the tris (dibenzylideneacetone) dipalladium is 10-2: 1;
4-methyl-7-bromo-2H-1-benzopyran-2-one: the mass ratio of the 2-dicyclohexyl phosphonium-2, 4, 6-triisopropyl biphenyl is 12-1: 1;
the volume ratio of the mass of the 4-methyl-7-bromo-2H-1-benzopyran-2-ketone to the volume of the 1, 4-dioxane is 1:40-100 g/mL.
(3) In (1), 1-Boc-L-acridine-2-carboxylic acid: the mass ratio of diglycolamine is 1: 1-3;
1-Boc-L-acridine-2-carboxylic acid: the mass ratio of the 1-hydroxybenzotriazole is 1: 2-4;
1-Boc-L-acridine-2-carboxylic acid: the mass ratio of the 1-ethyl- (3-dimethylaminopropyl) carbonyldiimine hydrochloride is 1: 3-6;
the volume ratio of the mass of the 1-Boc-L-acridine-2-carboxylic acid to the volume of triethylamine is 1:4-10 g/mL;
the volume ratio of the mass of the 1-Boc-L-acridine-2-carboxylic acid to the volume of the N, N-dimethylformamide is 1:20-80 g/mL;
(3) and (3) 4-methyl-7-bromo-2H-1-benzopyran-2-one: the mass ratio of the azetidine-2- (2- (2-hydroxyethoxy) ethyl) formamide is 1: 1-4;
4-methyl-7-bromo-2H-1-benzopyran-2-one: the mass ratio of the cesium carbonate is 1: 3-6;
4-methyl-7-bromo-2H-1-benzopyran-2-one: the mass ratio of the tris (dibenzylideneacetone) dipalladium is 10-2: 1;
4-methyl-7-bromo-2H-1-benzopyran-2-one: the mass ratio of the 2-dicyclohexyl phosphonium-2, 4, 6-triisopropyl biphenyl is 12-1: 1;
the volume ratio of the mass of the 4-methyl-7-bromo-2H-1-benzopyran-2-ketone to the volume of the 1, 4-dioxane is 1:40-100 g/mL.
The application of the novel carbonyl azetidine substituted coumarin fluorescent dye in the field of cell and slice fluorescence imaging.
The invention has the advantages and beneficial effects that:
the fluorescent dye provided by the invention has the advantages of simple synthetic method, cheap and easily available raw materials, convenience in derivation and the like.
The dye structure provided by the invention is carbonyl azetidine 4-substituted coumarin, and the brightness and light stability of the dye are effectively improved.
The fluorescent dye provided by the invention has the advantages of high brightness, good light stability and good confocal imaging effect.
Drawings
FIG. 1 is a NMR spectrum of dye Cou-Aze-1 prepared in example 1.
FIG. 2 is a NMR spectrum of dye Cou-Aze-2 prepared in example 2.
FIG. 3 is a high resolution mass spectrum of dye Cou-Aze-3 prepared in example 3.
FIG. 4 is a fluorescence excitation spectrum and a fluorescence emission spectrum of dye Cou-Aze-1 prepared in example 1 normalized to water, with wavelength on the abscissa, normalized fluorescence intensity on the ordinate, and fluorescent dye concentration of 5. mu.M.
FIG. 5 is a fluorescence excitation spectrum and a fluorescence emission spectrum of dye Cou-Aze-2 prepared in example 2 normalized to water, with wavelength on the abscissa, normalized fluorescence intensity on the ordinate, and fluorescent dye concentration of 5. mu.M.
FIG. 6 is a graph of the change in fluorescence emission intensity of dye Cou-Aze-1 prepared in example 1 under 500W tungsten lamp illumination, wavelength on the abscissa and fluorescence intensity on the ordinate.
FIG. 7 is a graph of the change in fluorescence emission intensity of dye Cou-Aze-2 prepared in example 2 under 500W tungsten lamp illumination, wavelength on the abscissa and fluorescence intensity on the ordinate.
FIG. 8 is a photograph of a fluorescent confocal image of HeLa cells of dye Cou-Aze-1 prepared in example 1.
Detailed Description
Example 1
Synthesis of dye Cou-Aze-1:
(1) synthesis of intermediate azetidine-2-carboxylic acid methyl ester hydrochloride:
adding 10mL of methanol into a 50mL single-neck bottle, cooling to 0 ℃ in an ice water bath, slowly dropwise adding 0.19mL of thionyl chloride into the single-neck bottle, keeping the temperature at 0 ℃ and continuing stirring for 30 minutes, adding 150mg of azetidine-2-carboxylic acid into the reaction bottle, removing the ice water bath, slowly heating to room temperature, and stirring overnight. The reaction was stopped, the solvent was removed and the resulting solid was washed twice with anhydrous ether to give the pale yellow solid product azetidine-2-carboxylic acid methyl ester hydrochloride in 86% yield.
(2) Synthesis of dye 1- (4-methyl-2H-1-benzopyran-2-one) azetidine-2-carboxylic acid methyl ester:
sequentially adding 59.5mg of 4-methyl-7-bromo-2H-1-benzopyran-2-one, 45mg of azetidine-2-methyl formate hydrochloride, 196mg of cesium carbonate, 23mg of tris (dibenzylideneacetone) dipalladium and 36mg of 2-dicyclohexyl-phosphorus-2, 4, 6-triisopropylbiphenyl into a 25mL double-mouth bottle, assembling a reflux device, vacuumizing and replacing nitrogen for three times, adding 4mL of 1, 4-dioxane into a reactor by using an injector in a nitrogen atmosphere, slowly heating the reaction liquid to 105 ℃ under stirring, continuing to react for 4 hours, cooling to room temperature, removing the solvent under reduced pressure, separating by using a silica gel column, and removing the solvent by using dichloromethane and methanol with a volume ratio of 100-10:1 as an eluent to obtain the dye 1- (4-methyl-2H-1-benzopyran-2-one ) Azetidine-2-carboxylic acid methyl ester.
The nuclear magnetic spectrum hydrogen spectrum is shown in figure 1, and the specific data is as follows:
1H NMR(400MHz,Chloroform-d)δ7.41(d,J＝8.6Hz,1H),6.41(dd,J＝8.6,2.3Hz,1H),6.35(d,J＝2.3Hz,1H),6.02(q,J＝1.2Hz,1H),4.65(dd,J＝8.9,6.6Hz,1H),4.12(ddd,J＝8.7,7.1,4.9Hz,1H),3.91–3.84(m,1H),3.82(s,3H),2.73–2.58(m,2H),2.35(d,J＝1.2Hz,3H).
the specific data of the high-resolution mass spectrum are as follows:
C15H16NO4[M+H]+theoretical value: 274.1074, actual value: 274.1082.
the structure of the compound is shown as Cou-Aze-1. The fluorescence properties are as follows:
the dye is dissolved in DMSO solution to prepare mother liquor with the concentration of 2mM, 10 mu L of Cou-Aze-1 mother liquor is added into 4mL of water each time to prepare fluorescent dye test solution with the concentration of 5 mu M, and fluorescence excitation and emission spectrum test are carried out. The fluorescence excitation and emission spectra in water are shown in FIG. 4: the absorption wavelength was 350nm and the emission wavelength was 457 nm.
Example 2
Synthesis of dye Cou-Aze-2:
(1) synthesis of intermediate azetidine-3-carboxylic acid methyl ester hydrochloride:
adding 10mL of methanol into a 50mL single-neck bottle, cooling to 0 ℃ in an ice water bath, slowly dropwise adding 0.19mL of thionyl chloride into the single-neck bottle, keeping the temperature at 0 ℃ and continuing stirring for 30 minutes, adding 150mg of azetidine-3-carboxylic acid into the reaction bottle, removing the ice water bath, slowly heating to room temperature, and stirring overnight. The reaction was stopped, the solvent was removed and the resulting solid was washed twice with anhydrous ether to give the pale yellow solid product azetidine-2-carboxylic acid methyl ester hydrochloride in 89% yield.
(2) Synthesis of dye 1- (4-methyl-2H-benzopyran-2-one) azetidine-3-carboxylic acid methyl ester:
sequentially adding 59.5mg of 4-methyl-7-bromo-2H-1-benzopyran-2-one, 45mg of azetidine-3-methyl formate hydrochloride, 196mg of cesium carbonate, 23mg of tris (dibenzylideneacetone) dipalladium and 36mg of 2-dicyclohexyl phosphorus-2, 4, 6-triisopropylbiphenyl into a 25mL double-mouth bottle, assembling a reflux device, vacuumizing and replacing nitrogen for three times, adding 4mL of 1, 4-dioxane into a reactor by using an injector in a nitrogen atmosphere, slowly heating the reaction liquid to 105 ℃ under stirring, continuing to react for 4 hours, cooling to room temperature, removing the solvent under reduced pressure, separating by using a silica gel column, and removing the solvent by using dichloromethane and methanol with a volume ratio of 100-10:1 as an eluent to obtain the dye 1- (4-methyl-2H-1-benzopyran-2-one ) Azetidine-3-carboxylic acid methyl ester.
The nuclear magnetic spectrum hydrogen spectrum is shown in figure 2, and the specific data is as follows:
1H NMR(400MHz,Chloroform-d)δ7.41(d,J＝8.6Hz,1H),6.34(dd,J＝8.6,2.3Hz,1H),6.26(d,J＝2.3Hz,1H),6.01(d,J＝1.2Hz,1H),4.22–4.12(m,4H),3.78(s,3H),3.61(tt,J＝8.5,6.0Hz,1H),2.35(d,J＝1.1Hz,3H).
the structure of the compound is shown as Cou-Aze-2. The fluorescence properties are as follows:
the dye is dissolved in DMSO solution to prepare mother liquor with the concentration of 2mM, 10 mu L of Cou-Aze-2 mother liquor is added into 4mL of water each time to prepare fluorescent dye test solution with the concentration of 5 mu M, and fluorescence excitation and emission spectrum test are carried out. The fluorescence excitation and emission spectra in water are shown in FIG. 5: the absorption wavelength was 352nm and the emission wavelength was 464 nm.
Example 3
Synthesis of dye Cou-Aze-3:
(1) synthesis of intermediate azetidine-2- (2- (2-hydroxyethoxy) ethyl) carboxamide:
100mg of 1-Boc-L-acridine-2-carboxylic acid, 5mL of dried N, N-dimethylformamide, 210mg of diglycolamine, 270mg of 1-hydroxybenzotriazole, 380mg of 1-ethyl- (3-dimethylaminopropyl) carbonyldiimine hydrochloride and 0.55mL of triethylamine were sequentially added to a 25mL single-neck flask, and the mixture was stirred at room temperature for 24 hours. Stopping the reaction, removing the solvent under reduced pressure, separating with silica gel column, adding 10mL of dichloromethane into the obtained product, adding 0.1mL of trifluoroacetic acid into the obtained product, continuing to stir overnight, stopping the reaction, separating with silica gel column, and removing the solvent by using dichloromethane as an eluent to obtain white viscous liquid azetidine-2- (2- (2-hydroxyethoxy) ethyl) formamide.
(4) Synthesis of the dye 1- (4-methyl-2H-1-benzopyran-2-one) azetidine-2- (2- (2-hydroxyethoxy) ethyl) carboxamide:
adding 95mg of 4-methyl-7-bromo-2H-1-benzopyran-2-ketone, 113mg of azetidine-2- (2- (2-hydroxyethoxy) ethyl) formamide, 312mg of cesium carbonate, 18mg of tris (dibenzylideneacetone) dipalladium and 28mg of 2-dicyclohexyl-phosphorus-2, 4, 6-triisopropylbiphenyl into a 25mL double-mouth bottle in sequence, assembling a reflux device, vacuumizing and replacing nitrogen for three times, adding 4mL of 1, 4-dioxane into the reactor by using an injector under the nitrogen atmosphere, slowly heating the reaction liquid to 105 ℃ under stirring, continuing to react for 4 hours, cooling to room temperature, removing the solvent under reduced pressure, separating by using dichloromethane and methanol with the volume ratio of 100-10:1 as an eluent, the solvent was removed to give the dye 1- (4-methyl-2H-1-benzopyran-2-one) azetidine-2- (2- (2-hydroxyethoxy) ethyl) carboxamide.
The high resolution mass spectrum is shown in fig. 3, and the specific data are as follows:
C18H23N2O5[M+H]+theoretical value: 347.1601, actual value: 347.1599.
example 4
Examples 1 and 2 relate to the measurement of the change in fluorescence emission intensity of the fluorescent dyes Cou-Aze-1 and Cou-Aze-2 under 500W tungsten lamp illumination. And adding 10 mu L of the dye mother liquor into 4mL of deionized water, then adding the mixture into a threaded cuvette, placing the front side of the cuvette at a position 50cm away from a tungsten lamp, and performing fluorescence emission spectrum test by respectively adopting time nodes of 0 hour, 0.5 hour, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours and 10 hours.
The plots of the change in fluorescence emission intensity of Cou-Aze-1 and Cou-Aze-2 fluorescent dyes of examples 1 and 2 under 500W tungsten lamp illumination are shown in FIGS. 6 and 7: the fluorescence intensity of the dye can still keep higher intensity after 10 hours of illumination.
Example 5
The invention relates to fluorescence confocal imaging of partial dyes in living cells (RWPE, HeLa, fat cells and the like). Dissolving more than 0.5 mu L of dye mother liquor into 1mL of cell culture solution, and then placing the cell at 37 ℃ for incubation for 10-30 minutes and then carrying out fluorescence imaging.
Cou-Aze-1 fluorescence confocal imaging in HeLa cells as shown in FIG. 8: the dye Cou-Aze-1 is capable of staining the cytoplasm and has a high signal-to-noise ratio.
Claims (8)
1. A novel carbonyl azetidine substituted coumarin fluorescent dye is characterized in that: covalently linking the carbonyl azetidine to the 7-position of the coumarin fluorophore parent, the structure of this class of dyes is as follows:
R1is C1-4 alkyl, R2Is composed of
Wherein X1Is an H, C1-2 alkyl group,
X2is a C1-2 alkyl group,
X3is a C1-2 alkyl group,
X4is C1-2 alkyl.
2. A method for synthesizing novel carbonyl azetidine substituted coumarin fluorescent dye according to claim 1, wherein coumarin is used as the fluorophore precursor, and carbonyl azetidine is covalently linked to position 7 of coumarin to obtain the fluorescent dye.
3. The method for synthesizing the novel carbonyl azetidine substituted coumarin fluorescent dye according to claim 2, comprising the following steps:
(1) synthesis of dye 1- (4-methyl-2H-1-benzopyran-2-one) azetidine-2-carboxylic acid methyl ester:
adding 4-methyl-7-bromo-2H-1-benzopyran-2-one, azetidine-2-methyl formate hydrochloride, cesium carbonate, tris (dibenzylideneacetone) dipalladium and 2-dicyclohexyl-phosphorus-2, 4, 6-triisopropylbiphenyl into a reactor, adding 1, 4-dioxane under anhydrous and oxygen-free conditions, slowly heating the reaction solution to 80-120 deg.C under stirring, reacting for 2-10 hr, cooling to room temperature, removing solvent under reduced pressure, separating with silica gel column, removing the solvent by using dichloromethane and methanol as eluent in a volume ratio of 200-5:1 to obtain dye 1- (4-methyl-2H-1-benzopyran-2-ketone) azetidine-2-carboxylic acid methyl ester;
(2) synthesis of dye 1- (4-methyl-2H-benzopyran-2-one) azetidine-3-carboxylic acid methyl ester:
adding 4-methyl-7-bromo-2H-1-benzopyran-2-one, azetidine-3-methyl formate hydrochloride, cesium carbonate, tris (dibenzylideneacetone) dipalladium and 2-dicyclohexylphosphonium-2, 4, 6-triisopropylbiphenyl into a reactor, adding 1, 4-dioxane under anhydrous and oxygen-free conditions, slowly heating the reaction solution to 80-120 deg.C under stirring, reacting for 2-10 hr, cooling to room temperature, removing solvent under reduced pressure, separating with silica gel column, removing the solvent by using dichloromethane and methanol as eluent in a volume ratio of 200-5:1 to obtain dye 1- (4-methyl-2H-1-benzopyran-2-ketone) azetidine-3-carboxylic acid methyl ester;
(3) synthesis of the dye 1- (4-methyl-2H-1-benzopyran-2-one) azetidine-2- (2- (2-hydroxyethoxy) ethyl) carboxamide:
dissolving 1-Boc-L-acridine-2-carboxylic acid in dry N, N-dimethylformamide, and sequentially adding diglycolamine, 1-hydroxybenzotriazole, 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride and triethylamine thereto, stirring at room temperature for 12-36 hours; stopping the reaction, removing the solvent under reduced pressure, separating by using a silica gel column, adding dichloromethane into the obtained product, adding trifluoroacetic acid into the dichloromethane, continuously stirring the mixture overnight, stopping the reaction, separating by using the silica gel column, and removing the solvent by using dichloromethane as an eluent to obtain a white viscous liquid intermediate azetidine-2- (2- (2-hydroxyethoxy) ethyl) formamide;
adding 4-methyl-7-bromo-2H-1-benzopyran-2-one, the obtained azetidine-2- (2- (2-hydroxyethoxy) ethyl) formamide, cesium carbonate, tris (dibenzylideneacetone) dipalladium and 2-dicyclohexylphosphorus-2, 4, 6-triisopropylbiphenyl into a reactor, adding 1, 4-dioxane into the reactor under the anhydrous and oxygen-free (nitrogen atmosphere), slowly heating the reaction liquid to 80-120 ℃ under stirring, continuing to react for 2-10 hours, cooling to room temperature, removing the solvent under reduced pressure, separating by using a silica gel column, taking dichloromethane and methanol with the volume ratio of 200-5:1 as an eluent, removing the solvent to obtain the dye 1- (4-methyl-2H-1-benzopyran-2-one) azetidine-2- (2- (2-hydroxyethoxy) ethyl) formamide.
4. The method for synthesizing novel carbonyl azetidine substituted coumarin fluorescent dye according to claim 3, wherein in (1), 4-methyl-7-bromo-2H-1-benzopyran-2-one: the mass ratio of the azetidine-2-methyl formate hydrochloride is 1: 0.5-2;
4-methyl-7-bromo-2H-1-benzopyran-2-one: the mass ratio of the cesium carbonate is 1: 3-6;
4-methyl-7-bromo-2H-1-benzopyran-2-one: the mass ratio of the tris (dibenzylideneacetone) dipalladium is 10-2: 1;
4-methyl-7-bromo-2H-1-benzopyran-2-one: the mass ratio of the 2-dicyclohexyl phosphonium-2, 4, 6-triisopropyl biphenyl is 12-1: 1;
the volume ratio of the mass of the 4-methyl-7-bromo-2H-1-benzopyran-2-ketone to the volume of the 1, 4-dioxane is 1:40-100 g/mL.
5. The method for synthesizing novel carbonyl azetidine substituted coumarin fluorescent dye according to claim 3, wherein in (2), 4-methyl-7-bromo-2H-1-benzopyran-2-one: the mass ratio of the azetidine-3-methyl formate hydrochloride is 1: 0.5-2;
4-methyl-7-bromo-2H-1-benzopyran-2-one: the mass ratio of the cesium carbonate is 1: 3-6;
4-methyl-7-bromo-2H-1-benzopyran-2-one: the mass ratio of the tris (dibenzylideneacetone) dipalladium is 10-2: 1;
4-methyl-7-bromo-2H-1-benzopyran-2-one: the mass ratio of the 2-dicyclohexyl phosphonium-2, 4, 6-triisopropyl biphenyl is 12-1: 1;
the volume ratio of the mass of the 4-methyl-7-bromo-2H-1-benzopyran-2-ketone to the volume of the 1, 4-dioxane is 1:40-100 g/mL.
6. The method for synthesizing the novel carbonyl azetidine substituted coumarin fluorescent dye according to claim 3, wherein in (3), 1-Boc-L-acridine-2-carboxylic acid: the mass ratio of diglycolamine is 1: 1-3;
1-Boc-L-acridine-2-carboxylic acid: the mass ratio of the 1-hydroxybenzotriazole is 1: 2-4;
1-Boc-L-acridine-2-carboxylic acid: the mass ratio of the 1-ethyl- (3-dimethylaminopropyl) carbonyldiimine hydrochloride is 1: 3-6;
the volume ratio of the mass of the 1-Boc-L-acridine-2-carboxylic acid to the volume of triethylamine is 1:4-10 g/mL;
the volume ratio of the mass of the 1-Boc-L-acridine-2-carboxylic acid to the volume of the N, N-dimethylformamide is 1:20-80 g/mL.
7. The method for synthesizing the novel carbonyl azetidine substituted coumarin fluorescent dye according to claim 3, wherein in (3), the ratio of 4-methyl-7-bromo-2H-1-benzopyran-2-one: the mass ratio of the azetidine-2- (2- (2-hydroxyethoxy) ethyl) formamide is 1: 1-4;
4-methyl-7-bromo-2H-1-benzopyran-2-one: the mass ratio of the cesium carbonate is 1: 3-6;
4-methyl-7-bromo-2H-1-benzopyran-2-one: the mass ratio of the tris (dibenzylideneacetone) dipalladium is 10-2: 1;
4-methyl-7-bromo-2H-1-benzopyran-2-one: the mass ratio of the 2-dicyclohexyl phosphonium-2, 4, 6-triisopropyl biphenyl is 12-1: 1;
the volume ratio of the mass of the 4-methyl-7-bromo-2H-1-benzopyran-2-ketone to the volume of the 1, 4-dioxane is 1:40-100 g/mL.
8. The application of the novel carbonyl azetidine substituted coumarin fluorescent dye as claimed in claim 1 in the field of cell and slice fluorescence imaging.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911258845.XA CN112939950A (en) | 2019-12-10 | 2019-12-10 | Carbonyl azetidine substituted coumarin fluorescent dye and synthetic method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911258845.XA CN112939950A (en) | 2019-12-10 | 2019-12-10 | Carbonyl azetidine substituted coumarin fluorescent dye and synthetic method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112939950A true CN112939950A (en) | 2021-06-11 |
Family
ID=76225482
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911258845.XA Pending CN112939950A (en) | 2019-12-10 | 2019-12-10 | Carbonyl azetidine substituted coumarin fluorescent dye and synthetic method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112939950A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114459862A (en) * | 2022-01-13 | 2022-05-10 | 华腾实业（深圳）股份有限公司 | High-identification fungus fluorescent staining solution |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106471067A (en) * | 2014-04-01 | 2017-03-01 | 霍华德休斯医学研究所 | The fluorescent chemicalses that azetidine replaces |
-
2019
- 2019-12-10 CN CN201911258845.XA patent/CN112939950A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106471067A (en) * | 2014-04-01 | 2017-03-01 | 霍华德休斯医学研究所 | The fluorescent chemicalses that azetidine replaces |
Non-Patent Citations (4)
| Title |
|---|
| CONNER A. HOELZEL ET AL.: "A General Strategy to Enhance Donor-Acceptor Molecules Using Solvent-Excluding Substituents", 《ANGEW. CHEM. INT. ED.》 * |
| JOSHUA P. DELANEY ET AL.: "Synergistic effects within a C2-symmetric organocatalyst: the potential formation of a chiral catalytic pocket", 《ORG. BIOMOL. CHEM.,》 * |
| JUNLIANG ZHOU ET AL.: "Azetidine-Containing Heterospirocycles Enhance the Performance of Fluorophores", 《ORG. LETT.》 * |
| 乔庆龙: "萘酰亚胺类染料荧光构效关系研究及生物应用", 《中国优秀博硕士学位论文全文数据库(博士) 工程科技Ⅰ辑》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114459862A (en) * | 2022-01-13 | 2022-05-10 | 华腾实业（深圳）股份有限公司 | High-identification fungus fluorescent staining solution |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107603269B (en) | Fluorescent dye based on naphthalimide, preparation method and application thereof | |
| CN111333641B (en) | Enhanced fluorescent probe for tetrazine bio-orthogonal labeling and synthesis thereof | |
| CN112939950A (en) | Carbonyl azetidine substituted coumarin fluorescent dye and synthetic method and application thereof | |
| CN111793371B (en) | 3, 5-asymmetrically modified BODIPY near-infrared fluorescent dye and preparation method thereof | |
| CN111689950B (en) | Organic diheterocyclic near-infrared fluorescent probe based on diphenylamino and preparation method and application thereof | |
| CN112939960B (en) | Carbonyl azetidine substituted NBD fluorescent dye and synthetic method and application thereof | |
| CN111334070B (en) | 532nm excited rhodamine fluorescent dye and preparation method thereof | |
| CN112939936B (en) | Carbonyl azetidine substituted naphthalimide fluorescent dye and synthetic method and application thereof | |
| CN114773875B (en) | Azaindole-squaraine dye, and synthetic method and application thereof | |
| CN112940709B (en) | Self-flashing rhodamine spirothioester fluorescent dye, synthetic method thereof and application thereof in super-resolution imaging field | |
| CN112920195B (en) | Ratio type viscosity fluorescent probe and preparation method and application thereof | |
| CN111333624B (en) | High-stability wash-free SNAP-tag probe and preparation method and application thereof | |
| CN111333618B (en) | 488nm excited wash-free SNAP-tag probe and preparation method thereof | |
| CN114591632A (en) | Azaindole-hemicyanine dye, and synthesis method and application thereof | |
| CN111333619B (en) | 488nm excited high-stability super-resolution fluorescent dye and synthesis and application thereof | |
| CN111334076B (en) | High-brightness and high-light-stability cell nucleus fluorescent probe | |
| CN111333660B (en) | 550nm excited rhodamine dye and preparation method thereof | |
| CN112812088A (en) | Near-infrared luminescent triphenylamine derivative fluorescent molecule and preparation method and application thereof | |
| CN111961072A (en) | Lysosome-targeted infrared two-window emission fluorescent dye and preparation method and application thereof | |
| CN110804322B (en) | Fluorescent dye based on 1, 2-diaza-benzoperylene as well as preparation method and application thereof | |
| CN110790781A (en) | Asymmetric boron fluoride dye with red light emission and large Stokes shift | |
| CN111333623B (en) | Fluorescent dye for lysosome marking and synthetic method and application thereof | |
| CN111333621B (en) | 488nm excited washing-free Halo-tag probe and synthesis and biological application thereof | |
| CN115477654B (en) | Near-infrared rhodamine dye with large Stokes displacement and synthesis method thereof | |
| CN114262336B (en) | Autoflash fluorescent dye for lysosome super-resolution fluorescent imaging and synthesis method and application thereof |
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 | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20210611 |
|
| RJ01 | Rejection of invention patent application after publication |