CN114835698B - (diphenylamino) phenyl flavonoid fluorescent probe for detecting cysteine and preparation method thereof - Google Patents
(diphenylamino) phenyl flavonoid fluorescent probe for detecting cysteine and preparation method thereof Download PDFInfo
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- -1 (diphenylamino) phenyl flavonoid Chemical class 0.000 title claims abstract description 49
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 title claims abstract description 33
- 235000018417 cysteine Nutrition 0.000 title claims abstract description 33
- 239000007850 fluorescent dye Substances 0.000 title claims abstract description 25
- 229930003935 flavonoid Natural products 0.000 title claims abstract description 20
- 235000017173 flavonoids Nutrition 0.000 title claims abstract description 20
- 238000002360 preparation method Methods 0.000 title claims abstract description 16
- FPLSTPZGCDCWID-UHFFFAOYSA-N OC(C(C1=CC=CC=C1)=O)C(C=C1)=CC=C1N(C1=CC=CC=C1)C1=CC=CC=C1 Chemical compound OC(C(C1=CC=CC=C1)=O)C(C=C1)=CC=C1N(C1=CC=CC=C1)C1=CC=CC=C1 FPLSTPZGCDCWID-UHFFFAOYSA-N 0.000 claims abstract description 20
- HYBBIBNJHNGZAN-UHFFFAOYSA-N furfural Chemical compound O=CC1=CC=CO1 HYBBIBNJHNGZAN-UHFFFAOYSA-N 0.000 claims abstract description 16
- XVWSAMCMMBIZQS-UHFFFAOYSA-N OC(C(C1=C2)=O)=C(C3=CC=CO3)OC1=CC=C2C(C=C1)=CC=C1N(C1=CC=CC=C1)C1=CC=CC=C1 Chemical compound OC(C(C1=C2)=O)=C(C3=CC=CO3)OC1=CC=C2C(C=C1)=CC=C1N(C1=CC=CC=C1)C1=CC=CC=C1 XVWSAMCMMBIZQS-UHFFFAOYSA-N 0.000 claims abstract description 11
- LQCMMXGKEGWUIM-UHFFFAOYSA-N 1-(4-bromo-2-hydroxyphenyl)ethanone Chemical compound CC(=O)C1=CC=C(Br)C=C1O LQCMMXGKEGWUIM-UHFFFAOYSA-N 0.000 claims abstract description 7
- HFBMWMNUJJDEQZ-UHFFFAOYSA-N acryloyl chloride Chemical compound ClC(=O)C=C HFBMWMNUJJDEQZ-UHFFFAOYSA-N 0.000 claims abstract description 7
- TWWQCBRELPOMER-UHFFFAOYSA-N [4-(n-phenylanilino)phenyl]boronic acid Chemical compound C1=CC(B(O)O)=CC=C1N(C=1C=CC=CC=1)C1=CC=CC=C1 TWWQCBRELPOMER-UHFFFAOYSA-N 0.000 claims abstract description 5
- 239000002994 raw material Substances 0.000 claims abstract description 5
- 238000005859 coupling reaction Methods 0.000 claims abstract description 4
- 238000005886 esterification reaction Methods 0.000 claims abstract description 4
- 238000007363 ring formation reaction Methods 0.000 claims abstract description 4
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 27
- 238000006243 chemical reaction Methods 0.000 claims description 20
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 18
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 18
- 239000000243 solution Substances 0.000 claims description 18
- 238000000034 method Methods 0.000 claims description 14
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 12
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 9
- 239000012043 crude product Substances 0.000 claims description 9
- 238000001914 filtration Methods 0.000 claims description 8
- 239000007787 solid Substances 0.000 claims description 8
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 claims description 6
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 6
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 6
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 6
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 claims description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical class [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 6
- 239000005457 ice water Substances 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 6
- 230000007935 neutral effect Effects 0.000 claims description 6
- 239000012074 organic phase Substances 0.000 claims description 6
- NFHFRUOZVGFOOS-UHFFFAOYSA-N palladium;triphenylphosphane Chemical compound [Pd].C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1 NFHFRUOZVGFOOS-UHFFFAOYSA-N 0.000 claims description 6
- 239000003208 petroleum Substances 0.000 claims description 6
- 239000000741 silica gel Substances 0.000 claims description 6
- 229910002027 silica gel Inorganic materials 0.000 claims description 6
- 238000005406 washing Methods 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 238000012544 monitoring process Methods 0.000 claims description 4
- 239000007864 aqueous solution Substances 0.000 claims description 3
- 238000009833 condensation Methods 0.000 claims description 3
- 230000005494 condensation Effects 0.000 claims description 3
- 238000004821 distillation Methods 0.000 claims description 3
- 229910052757 nitrogen Inorganic materials 0.000 claims description 3
- 239000002244 precipitate Substances 0.000 claims description 3
- 239000000047 product Substances 0.000 claims description 3
- 239000011541 reaction mixture Substances 0.000 claims description 3
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 3
- CFKJKWKYGPECGI-UHFFFAOYSA-N OB(O)Oc1ccc(cc1)N(c1ccccc1)c1ccccc1 Chemical compound OB(O)Oc1ccc(cc1)N(c1ccccc1)c1ccccc1 CFKJKWKYGPECGI-UHFFFAOYSA-N 0.000 claims description 2
- 238000001514 detection method Methods 0.000 abstract description 5
- 150000001875 compounds Chemical class 0.000 abstract description 3
- 239000000523 sample Substances 0.000 abstract description 3
- 238000005882 aldol condensation reaction Methods 0.000 abstract 1
- 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 10
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 9
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 6
- FFFHZYDWPBMWHY-VKHMYHEASA-N L-homocysteine Chemical compound OC(=O)[C@@H](N)CCS FFFHZYDWPBMWHY-VKHMYHEASA-N 0.000 description 5
- 229960003180 glutathione Drugs 0.000 description 5
- 238000002189 fluorescence spectrum Methods 0.000 description 4
- 239000007853 buffer solution Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- 239000011734 sodium Substances 0.000 description 3
- 230000007547 defect Effects 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- RBTBFTRPCNLSDE-UHFFFAOYSA-N 3,7-bis(dimethylamino)phenothiazin-5-ium Chemical compound C1=CC(N(C)C)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 RBTBFTRPCNLSDE-UHFFFAOYSA-N 0.000 description 1
- 208000024827 Alzheimer disease Diseases 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 108010024636 Glutathione Proteins 0.000 description 1
- 206010067125 Liver injury Diseases 0.000 description 1
- 206010029350 Neurotoxicity Diseases 0.000 description 1
- 208000018737 Parkinson disease Diseases 0.000 description 1
- 102000007562 Serum Albumin Human genes 0.000 description 1
- 108010071390 Serum Albumin Proteins 0.000 description 1
- 206010044221 Toxic encephalopathy Diseases 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 230000006851 antioxidant defense Effects 0.000 description 1
- 238000001479 atomic absorption spectroscopy Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000031018 biological processes and functions Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 238000004042 decolorization Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000001784 detoxification Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000000840 electrochemical analysis Methods 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 230000011132 hemopoiesis Effects 0.000 description 1
- 231100000234 hepatic damage Toxicity 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 201000002364 leukopenia Diseases 0.000 description 1
- 231100001022 leukopenia Toxicity 0.000 description 1
- 230000008818 liver damage Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229960000907 methylthioninium chloride Drugs 0.000 description 1
- 230000007135 neurotoxicity Effects 0.000 description 1
- 231100000228 neurotoxicity Toxicity 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 238000011897 real-time detection Methods 0.000 description 1
- 231100000272 reduced body weight Toxicity 0.000 description 1
- 230000037380 skin damage Effects 0.000 description 1
- 150000003573 thiols Chemical class 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
Classifications
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- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D407/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00
- C07D407/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00 containing two hetero rings
- C07D407/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
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- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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Abstract
The invention discloses a (diphenylamino) phenyl flavonoid fluorescent probe for detecting cysteine and a preparation method thereof. The invention uses 4-bromo-2-hydroxy acetophenone as raw material, and carries out coupling reaction with 4- (diphenylamino) phenylboronic acid to obtain 4- (diphenylamino) phenyl-2-hydroxy acetophenone, and then carries out aldol condensation and cyclization reaction with furfural to obtain 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxy chromone; then carrying out esterification reaction with acryloyl chloride to obtain 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxy chromone acrylate. The compound can react with cysteine rapidly and exclusively at room temperature to change the fluorescent color of the solution from colorless to orange red, and the minimum detection limit is 9.76X10 ‑8 M. The probe can be used as a fluorescent probe for detecting cysteine and has good application prospect.
Description
Technical Field
The invention belongs to the technical field of fine organic synthesis, and relates to a (diphenylamino) phenyl flavonoid fluorescent probe for detecting cysteine and a preparation method thereof.
Background
Cysteine (Cys), homocysteine (Hcy) and Glutathione (GSH) are the main thiol-containing amino acids that play an important role in many biological processes. Among them, cys is directly related to a number of important cellular functions, such as antioxidant defense, cell growth, detoxification and metabolism, etc. Thus, abnormalities in Cys levels in cells are also associated with various diseases. Cys deficiency can lead to slow growth rate, reduced hematopoiesis, skin damage, hair decolorization, reduced body weight ratio, plasma albumin levels, leukopenia, liver damage, and Parkinson's disease; in contrast, excess Cys may lead to severe neurotoxicity, alzheimer's disease and cardiovascular disease. It is therefore of great importance to develop efficient methods for monitoring changes in cysteine concentration.
There are many reported methods for detecting cysteine, mainly including high performance liquid chromatography, atomic absorption spectrometry, gas chromatography, methylene blue method, electrochemical analysis and the like, but the methods have the defects of complex sample pretreatment, expensive equipment, complex operation, long detection time consumption and certain limitations. In contrast, the fluorescent probe can be used as a simple and effective Cys detection method, has the advantages of non-invasiveness, high sensitivity, high selectivity and the like, and can be imaged on living cells in real time. The organic micromolecular fluorescent probe has the characteristics of small volume, simple synthesis, quick reaction time, good selectivity, high sensitivity, convenience in real-time detection by taking fluorescence as an output signal and the like, and becomes an important method for detecting cysteine in recent years.
Disclosure of Invention
Aiming at the defects existing in the prior art, the technical problem to be solved by the invention is to provide the (diphenylamino) phenyl flavonoid fluorescent probe which meets the use requirement of cysteine detection. The invention aims to provide a preparation method of the (diphenylamino) phenyl flavonoid fluorescent probe. The invention also solves the technical problem of providing an application of the (diphenylamino) phenyl flavonoid fluorescent probe.
In order to solve the technical problems, the invention adopts the following technical scheme:
the (diphenylamino) phenyl flavonoid fluorescent probe for detecting cysteine is 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxy chromone acrylate, and has the structural formula:
the preparation method of the (diphenylamino) phenyl flavonoid fluorescent probe for detecting cysteine comprises the following process steps:
1) Taking 4-bromo-2-hydroxyacetophenone as a raw material, and carrying out coupling reaction with 4- (diphenylamino) phenylboronic acid to obtain 4- (diphenylamino) phenyl-2-hydroxyacetophenone;
2) 4- (diphenylamino) phenyl-2-hydroxy acetophenone and furfural undergo condensation cyclization reaction to obtain 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxy chromone;
3) Esterification reaction is carried out on 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxy chromanone and acryloyl chloride, so as to obtain 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxy chromanone acrylate.
In the step 1), 4-bromo-2-hydroxyacetophenone is used as a raw material and is subjected to a coupling reaction with 4- (diphenylamino) phenylboronic acid to obtain 4- (diphenylamino) phenyl-2-hydroxyacetophenone, and the specific preparation steps comprise:
(1) 10mmol of 4-bromo-2-hydroxyacetophenone, 12mmol of 4- (diphenylamino) phenylboronic acid, 40mmol of sodium carbonate and 1mmol of tetrakis (triphenylphosphine) palladium are sequentially added into a dry three-neck flask, 50mL of dioxane and 15mL of water are added to dissolve the solid, the mixture is heated to 90 ℃ under the condition of nitrogen to react overnight, and the reaction is stopped after the reaction is completed by tracking and monitoring by a TLC method;
(2) Removing dioxane and water from the reaction solution by reduced pressure distillation, adding ethyl acetate, washing with saturated saline solution to be neutral, drying an organic phase by anhydrous sodium sulfate, filtering and concentrating to obtain a crude product of 4- (diphenylamino) phenyl-2-hydroxyacetophenone;
(3) The crude 4- (diphenylamino) phenyl-2-hydroxyacetophenone product was separated by a silica gel column (ethyl acetate/petroleum ether=1/20, v/v) to give 4- (diphenylamino) phenyl-2-hydroxyacetophenone.
In the step 2), 4- (diphenylamino) phenyl-2-hydroxy acetophenone and furfural undergo condensation cyclization reaction to obtain 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxy chromone, wherein the specific preparation steps comprise:
(1) Sequentially adding 5mmol of 4- (diphenylamino) phenyl-2-hydroxyacetophenone, 5mmol of furfural, 10mmol of sodium hydroxide and 50mL of ethanol into a dry three-neck flask at the temperature of 0 ℃ and reacting for 8-10 h at room temperature;
(2) The reaction mixture was poured into ice water, acidified with dilute hydrochloric acid, filtered and washed to give a yellow solid precipitate. Then the yellow solid, 10mL of ethanol and 5mL of 20% NaOH aqueous solution are sequentially added into a dry three-neck flask, and 5mL of 30% H is added dropwise at 0 DEG C 2 O 2 Reacting for 5-6h at room temperature.
(3) The reaction solution is poured into ice water, neutralized by dilute hydrochloric acid, filtered to obtain a crude product of 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxy chromone, and then recrystallized by cold ethanol to obtain 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxy chromone.
In the step 3), the esterification reaction of the 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxy chromone and the acryloyl chloride is carried out to obtain the 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxy chromone acrylate, and the specific preparation steps comprise:
(1) 1mmol of 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxy chromone, 2mmol of acryloyl chloride and 2mmol of triethylamine are sequentially added into a dry three-neck flask, and the mixture is reacted for 10 to 12 hours at room temperature;
(2) Adding dichloromethane into the reaction solution again, washing the reaction solution to be neutral by using saturated saline solution, and drying, filtering and concentrating an organic phase by using anhydrous sodium sulfate to obtain a crude product of 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxy chromone acrylate;
(3) The crude 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxychromanone acrylate was isolated by column on silica gel (ethyl acetate/petroleum ether=1/5, v/v) to give 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxychromanone acrylate.
The (diphenylamino) phenyl flavonoid fluorescent probe for detecting cysteine is applied to cysteine detection.
The (diphenylamino) phenyl flavonoid fluorescent probe can react with cysteine rapidly and specifically at room temperature, and the solution changes from colorless to orange-red fluorescence.
The beneficial effects are that: compared with the prior art, the novel (diphenylamino) phenyl flavonoid fluorescent probe prepared by using the natural renewable resource furfural as a raw material can perform a specific reaction with cysteine, the fluorescent color of the solution is changed from colorless to orange red under 365nm ultraviolet light, and the fluorescent probe has the characteristics of good specificity and high sensitivity, and has a good application prospect as a fluorescent probe for detecting the cysteine.
Drawings
FIG. 1 is Cys, GSH, hcy, gly, arg, asp, ala, his, lys, glu, ser, val, tyr, F - ，C - ，Br - ，I - ，NO 3 - ，CO 3 2- ，SO 4 2- ，HSO 3 - ，SO 3 2- ，Mg 2+ ，Na + ，Cu 2+ ，Ca 2+ ，Ag + ，K + Equal to 28 different analytesA graph of the effect on the fluorescence spectrum of 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxychromanone acrylate;
FIG. 2 is a graph showing the effect of different concentrations of cysteine on the fluorescence spectrum of 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxychromanone acrylate.
Detailed Description
The invention will be further illustrated with reference to specific examples.
Example 1
The preparation of (diphenylamino) phenyl flavonoids has the following reaction formula:
the method comprises the following specific steps:
1) Preparation of 4- (diphenylamino) phenyl-2-hydroxyacetophenone:
10mmol of 4-bromo-2-hydroxyacetophenone, 12mmol of 4- (diphenylamino) phenylboric acid, 40mmol of sodium carbonate and 1mmol of tetrakis (triphenylphosphine) palladium are sequentially added into a dry three-neck flask, 50mL of dioxane and 15mL of water are added for dissolution, the mixture is heated to 90 ℃ under the protection of nitrogen for reaction overnight, and a TLC method is used for tracking and monitoring until the reaction is complete, and the reaction is stopped; removing dioxane and water from the reaction solution by reduced pressure distillation, adding ethyl acetate, washing with saturated saline solution to be neutral, drying an organic phase by anhydrous sodium sulfate, filtering and concentrating to obtain a crude product of 4- (diphenylamino) phenyl-2-hydroxyacetophenone; the crude 4- (diphenylamino) phenyl-2-hydroxyacetophenone product was separated by a silica gel column (ethyl acetate/petroleum ether=1/20, v/v) to give 4- (diphenylamino) phenyl-2-hydroxyacetophenone. Yield is as follows: 83.6% and purity of 99.2%. 1 H NMR(600MHz，CDCl 3 )δ：12.39(s，1H)，7.76(d，J＝8.4Hz，1H)，7.54-7.48(m，2H)，7.32-7.27(m，4H)，7.19(d，J＝1.8Hz，1H)，7.17-7.11(m，7H)，7.08(tt，J＝7.3，1.2Hz，2H)，2.65(s，3H). 13 C NMR(150MHz，CDCl 3 )δ：203.82，162.80，148.60，148.59，147.33，132.38，131.20，129.43，127.93，124.96，123.53，122.92，118.17，117.26，115.43，26.58.HRMS(m/z)：[M+H] + calcd for C 26 H 21 NO 2 +H + ，380.1651；found，380.1649。
2) Preparation of 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxychromanone:
sequentially adding 5mmol of 4- (diphenylamino) phenyl-2-hydroxyacetophenone, 5mmol of furfural, 10mmol of sodium hydroxide and 50mL of ethanol into a dry three-neck flask at the temperature of 0 ℃ and reacting for 8-10 h at room temperature; the reaction mixture was poured into ice water, acidified with dilute hydrochloric acid, and washed by filtration to give a yellow solid precipitate. Then the yellow solid, 10mL of ethanol and 5mL of 20% NaOH aqueous solution are sequentially added into a dry three-neck flask, and 5mL of 30% H is added dropwise at 0 DEG C 2 O 2 The reaction is carried out for 5 to 6 hours at room temperature. The reaction solution was poured into ice water and neutralized with dilute hydrochloric acid. Filtration gave a crude 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxychromanone which was then recrystallized from cold ethanol to give 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxychromanone. Yield is as follows: 60.6% and purity 98.5%. 1 H NMR(600MHz，DMSO-d 6 )δ：10.00(s，1H)，8.11(d，J＝8.3Hz，1H)，8.03(s，1H)，7.89(s，1H)，7.75(dd，J＝21.4，8.3Hz，3H)，7.34(t，J＝7.7Hz，4H)，7.28(d，J＝3.4Hz，1H)，7.09(dd，J＝17.7，7.7Hz，6H)，7.01(d，J＝8.2Hz，2H)，6.78(dd，J＝3.5，1.7Hz，1H). 13 C NMR(150MHz，DMSO-d 6 )δ：171.48，154.65，148.05，146.68，145.13，144.54，144.14，139.28，137.31，130.91，129.70，128.18，125.42，124.79，123.80，122.64，122.09，120.25，115.18，114.33，112.83.HRMS(m/z)：[M+H] + calcd for C 31 H 21 NO 4 +H + ，472.1549；found，472.1548。
3) Preparation of 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxychromanone acrylate:
1mmol of 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxychromanone, 2mmol of acryloyl chloride, 2mmol of triethylamine and 10 to 20mL of dichloromethane are added to the mixture in sequence to drynessIn a three-neck flask, reacting for 10-12 h at room temperature; washing the reaction solution to be neutral by using saturated saline, and drying, filtering and concentrating an organic phase by using anhydrous sodium sulfate to obtain a crude product of 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxy chromone acrylate; the crude 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxychromanone acrylate was isolated by column on silica gel (ethyl acetate/petroleum ether=1/5, v/v) to give 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxychromanone acrylate. Yield is as follows: 58.6% and purity 98.2%. 1 H NMR(600MHz，DMSO-d 6 )δ：8.14(d，J＝1.6Hz，1H)，8.09(d，J＝8.3Hz，1H)，8.02(d，J＝1.7Hz，1H)，7.85(dd，J＝8.4，1.7Hz，1H)，7.81(d，J＝8.3Hz，2H)，7.40-7.32(m，5H)，7.16-7.09(m，6H)，7.05(d，J＝8.3Hz，2H)，6.86(dd，J＝3.7，1.8Hz，1H)，6.69-6.63(m，1H)，6.59(dd，J＝17.3，10.2Hz，1H)，6.31-6.27(m，1H). 13 C NMR(150MHz，DMSO-d 6 )δ：163.18，155.64，148.79，148.15，147.76，147.12，146.19，143.33，135.53，131.00，130.65，130.22，128.78，127.20，126.13，125.34，124.39，124.14，122.51，121.63，117.37，115.11，113.69.HRMS(m/z)：[M+H] + calcd for C 34 H 23 NO 5 +H + ，526.1654；found：526.1716。
Example 2
Combining 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxychromanone acrylate with Cys, GSH, hcy, gly, arg, asp, ala, his, lys, glu, ser, val, tyr, F - ，Cl - ，Br - ，I - ，NO 3 - ，CO 3 2- ，SO 4 2- ，HSO 3 - ，SO 3 2- ，Mg 2+ ，Na + ，Cu 2+ ，Ca 2+ ，Ag + ，K + The 28 different analytes are respectively dissolved in DMSO/PBS (v/v=6/4) buffer solution to prepare the concentration of 1.0x10 -6 M probe solution and concentration of 1.0X10 -5 M28 different analyte solutions, fluorescence spectra of the solutions were measured using a fluorescence spectrometer, junctionThe result is shown in FIG. 1. FIG. 1 shows that the addition of cysteine changes the fluorescence color of the solution under 365nm ultraviolet light from colorless to orange-red, while by the addition of other interferents GSH, hcy, gly, arg, asp, ala, his, lys, glu, ser, val, tyr, F - ，Cl - ，Br - ，I - ，NO 3 - ，CO 3 2- ，SO 4 2- ，HSO 3 - ，SO 3 2- ，Mg 2+ ，Na + ，Cu 2+ ，Ca 2+ ，Ag + ，K + The fluorescence of the solution was not changed by equal comparison. The compound can be used as a fluorescent probe for specifically recognizing cysteine.
Example 3
6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxy chromanone acrylate was formulated to a concentration of 1.0X10 -5 M buffer solution of DMSO/PBS (v/v=6/4), cysteine was dissolved in DMSO/PBS (v/v=6/4) to prepare 0, 3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, 40. Mu.M buffer solution. The effect of different concentrations of cysteine on the fluorescence spectrum of 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxychromanone acrylate was measured by titration and the results are shown in figure 2. FIG. 2 shows that the fluorescence intensity at 575nm increases gradually with increasing cysteine concentration in the system, thus demonstrating that the compound can sensitively detect cysteine.
Claims (7)
1. A (diphenylamino) phenyl flavonoid fluorescent probe for detecting cysteine, which is characterized by having the structural formula:
2. the method for preparing a (diphenylamino) phenyl flavonoid fluorescent probe for detecting cysteine according to claim 1, which is characterized by comprising the following steps:
1) Taking 4-bromo-2-hydroxyacetophenone as a raw material, and carrying out coupling reaction with 4- (diphenylamino) phenylboronic acid to obtain 4- (diphenylamino) phenyl-2-hydroxyacetophenone;
2) 4- (diphenylamino) phenyl-2-hydroxy acetophenone and furfural undergo condensation cyclization reaction to obtain 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxy chromone;
3) Esterification reaction is carried out on 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxy chromanone and acryloyl chloride, so as to obtain 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxy chromanone acrylate.
3. The method for preparing a (diphenylamino) phenyl flavonoid fluorescent probe for detecting cysteine according to claim 2, wherein the preparation method of the step 1) is as follows:
(1) 10mmol of 4-bromo-2-hydroxyacetophenone, 12mmol of 4- (diphenylamino) phenylboric acid, 40mmol of sodium carbonate and 1mmol of tetrakis (triphenylphosphine) palladium are sequentially added into a dry three-neck flask, 50mL of dioxane and 15mL of water are added to dissolve the solid, the mixture is heated to 90 ℃ under the condition of nitrogen and reacted for 10 to 12 hours, a TLC method is used for tracking and monitoring, and the reaction is stopped after the reaction is completed;
(2) Removing dioxane and water from the reaction solution by reduced pressure distillation, adding ethyl acetate, washing with saturated saline solution to be neutral, drying an organic phase by anhydrous sodium sulfate, filtering and concentrating to obtain a crude product of 4- (diphenylamino) phenyl-2-hydroxyacetophenone;
(3) The crude 4- (diphenylamino) phenyl-2-hydroxyacetophenone product was separated by a silica gel column with ethyl acetate/petroleum ether=1/20, v/v to give 4- (diphenylamino) phenyl-2-hydroxyacetophenone.
4. The method for preparing a (diphenylamino) phenyl flavonoid fluorescent probe for detecting cysteine according to claim 2, wherein the specific preparation method of the step 2) is as follows:
(1) Sequentially adding 5mmol of 4- (diphenylamino) phenyl-2-hydroxyacetophenone, 5mmol of furfural, 10mmol of sodium hydroxide and 50mL of ethanol into a dry three-neck flask at the temperature of 0 ℃ and reacting for 8-10 h at room temperature;
(2) The reaction mixture was poured into ice water, acidified with dilute hydrochloric acid, filtered and washed to give a yellow solid precipitate. Then the yellow solid, 10mL of ethanol and 5mL of 20% NaOH aqueous solution are sequentially added into a dry three-neck flask, and 5mL of 30% H is added dropwise at 0 DEG C 2 O 2 Reacting for 5-6h at room temperature;
(3) The reaction solution is poured into ice water, neutralized by dilute hydrochloric acid, filtered to obtain a crude product of 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxy chromone, and then recrystallized by cold ethanol to obtain 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxy chromone.
5. The method for preparing a (diphenylamino) phenyl flavonoid fluorescent probe for detecting cysteine according to claim 2, wherein the preparation method of the step 3) specifically comprises the following steps:
(1) 1mmol of 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxy chromone, 2mmol of acryloyl chloride and 2mmol of triethylamine are sequentially added into a dry three-neck flask, and the mixture is reacted for 10 to 12 hours at room temperature;
(2) Washing the reaction solution to be neutral by using saturated saline, and drying, filtering and concentrating an organic phase by using anhydrous sodium sulfate to obtain a crude product of 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxy chromone acrylate;
(3) The crude product of 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxychromanone acrylate was separated by a silica gel column, ethyl acetate/petroleum ether=1/5, v/v, to give 6- (4- (diphenylamino) phenyl) -2- (furan-2-yl) -3-hydroxychromanone acrylate.
6. Use of the (diphenylamino) phenyl flavonoid fluorescent probe for detecting cysteine according to claim 1 for detecting cysteine.
7. The use according to claim 6, characterized in that: the (diphenylamino) phenyl flavonoid fluorescent probe can rapidly and specifically react with cysteine at room temperature, and the solution is changed from colorless to orange-red fluorescence.
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Non-Patent Citations (6)
| Title |
|---|
| Dinitrobenzene ether reactive turn-on fluorescence probes for the selective detection of H2S;Rajasekaran Dhivya, et al;《Analytical Method》;第14卷(第1期);58-66 * |
| Donor engineering on flavonoid-based probes to enhance the fluorescence brightness in water: Design, characterization, photophysical properties, and application for cysteine detection;Zhongyong Xu, et al.;《Sensors and Actuators B: Chemical》;第345卷;130367 * |
| Ratiometric fluorescent probe based on ESIPT for the highly selective detection of cysteine in living cells;Xinran Li, et al.;《Talanta》;第194卷;717-722 * |
| Two 3-hydroxyflavone derivatives as two-photon fluorescence turn-on chemosensors for cysteine and homocysteine in living cells;Qianqian Wu, et al.;《Talanta》;第181卷;118-124 * |
| 一种新型快速检测半胱氨酸的荧光探针;张晓辉;《分析试验室》;第41卷(第01期);55-58 * |
| 基于黄酮醇类半胱氨酸荧光探针的设计、合成及性能研究;曹小燕;《分析化学》;第48卷(第08期);1033-1049 * |
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