CN112876460B - 7-diethylamino-3-acetyl coumarin derivative and synthetic method and application thereof - Google Patents
7-diethylamino-3-acetyl coumarin derivative and synthetic method and application thereof Download PDFInfo
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- 238000010189 synthetic method Methods 0.000 title description 3
- 239000000523 sample Substances 0.000 claims abstract description 43
- 238000001514 detection method Methods 0.000 claims abstract description 19
- 238000000034 method Methods 0.000 claims abstract description 6
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 25
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 24
- 239000007850 fluorescent dye Substances 0.000 claims description 20
- JQYDECSZBZCKFQ-UHFFFAOYSA-N 3-acetyl-7-(diethylamino)chromen-2-one Chemical class C1=C(C(C)=O)C(=O)OC2=CC(N(CC)CC)=CC=C21 JQYDECSZBZCKFQ-UHFFFAOYSA-N 0.000 claims description 10
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- 239000000203 mixture Substances 0.000 claims description 10
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- 150000001875 compounds Chemical class 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 6
- 239000001431 2-methylbenzaldehyde Substances 0.000 claims description 5
- PCNFLKVWBDNNOW-UHFFFAOYSA-N 4-hydrazinylbenzoic acid Chemical compound NNC1=CC=C(C(O)=O)C=C1 PCNFLKVWBDNNOW-UHFFFAOYSA-N 0.000 claims description 5
- 229960000583 acetic acid Drugs 0.000 claims description 5
- 201000010099 disease Diseases 0.000 claims description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 5
- 239000012362 glacial acetic acid Substances 0.000 claims description 5
- 239000002994 raw material Substances 0.000 claims description 5
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- FESPWYJNJWOYTL-UHFFFAOYSA-N C(C)N(C1=CC=C2C=C(C(OC2=C1)=O)C1=NN(C(C1)C1=CC=C(C=C1)C)C1=CC=C(C(=O)O)C=C1)CC Chemical group C(C)N(C1=CC=C2C=C(C(OC2=C1)=O)C1=NN(C(C1)C1=CC=C(C=C1)C)C1=CC=C(C(=O)O)C=C1)CC FESPWYJNJWOYTL-UHFFFAOYSA-N 0.000 abstract description 4
- 238000001308 synthesis method Methods 0.000 abstract description 3
- QXAMGWKESXGGNV-UHFFFAOYSA-N 7-(diethylamino)-1-benzopyran-2-one Chemical compound C1=CC(=O)OC2=CC(N(CC)CC)=CC=C21 QXAMGWKESXGGNV-UHFFFAOYSA-N 0.000 abstract 1
- WPYMKLBDIGXBTP-UHFFFAOYSA-N Benzoic acid Natural products OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 abstract 1
- 239000005711 Benzoic acid Substances 0.000 abstract 1
- 235000010233 benzoic acid Nutrition 0.000 abstract 1
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 12
- 239000000243 solution Substances 0.000 description 10
- ZYGHJZDHTFUPRJ-UHFFFAOYSA-N coumarin Chemical compound C1=CC=C2OC(=O)C=CC2=C1 ZYGHJZDHTFUPRJ-UHFFFAOYSA-N 0.000 description 9
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 4
- 238000005160 1H NMR spectroscopy Methods 0.000 description 4
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- 238000001228 spectrum Methods 0.000 description 4
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- 239000000975 dye Substances 0.000 description 3
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- 230000035945 sensitivity Effects 0.000 description 3
- SOJLFNLKNQHPJS-UHFFFAOYSA-N 3-acetyl-7-(diethylamino)-4-hydroxychromen-2-one Chemical compound C(C)N(CC)C1=CC=C2C(=C(C(OC2=C1)=O)C(C)=O)O SOJLFNLKNQHPJS-UHFFFAOYSA-N 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 241000252212 Danio rerio Species 0.000 description 2
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- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 2
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- 125000000218 acetic acid group Chemical group C(C)(=O)* 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- OSVXSBDYLRYLIG-UHFFFAOYSA-N chlorine dioxide Inorganic materials O=Cl=O OSVXSBDYLRYLIG-UHFFFAOYSA-N 0.000 description 1
- 229910001914 chlorine tetroxide Inorganic materials 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 125000001664 diethylamino group Chemical group [H]C([H])([H])C([H])([H])N(*)C([H])([H])C([H])([H])[H] 0.000 description 1
- 238000001917 fluorescence detection Methods 0.000 description 1
- QWPPOHNGKGFGJK-UHFFFAOYSA-N hypochlorous acid Chemical compound ClO QWPPOHNGKGFGJK-UHFFFAOYSA-N 0.000 description 1
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- 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
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K49/0039—Coumarin dyes
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Abstract
The invention provides a 7-diethylamino-3-acetyl coumarin derivative, a synthesis method and an application thereof, wherein the derivative is 4- (3- (7- (diethylamino) -2-oxo-2H-chromium-3-yl) -5- (p-tolyl) -4, 5-dihydro-1H-pyrazole-1-yl) benzoic acid, the English name is 4- (3- (7- (diethyl amino) -2-oxo-2H-chromen-3-yl) -5- (p-tolyl) -4, 5-dihydro-1H-pyrazole-1-yl) benzoic acid, and the derivative is named as Probe 1; or 4- (3- (7- (diethylamino) -4-hydroxy-2-oxo-2H-chromen-3-yl) -5- (p-tolyl) -4,5-dihydro-1H-pyrazol-1-yl) benzoic acid, having the english name 4- (3- (7- (diethylamino) -4-hydroxy-2-oxo-2H-chromen-3-yl) -5- (p-tolyl) -4,5-dihydro-1H-pyrazol-1-y l) benzoic acid and named Probe 2. Probe 1 and ClO in PBS (pH 7.4) buffer‑Non-reactive, Probe 2 can realize "turn-off" type detection of ClO‑. The detection process is simple, sensitive and quick, and the detection result is accurate.
Description
Technical Field
The invention relates to a 7-diethylamino-3-acetyl coumarin derivative, in particular to a 7-diethylamino-3-acetyl coumarin derivative and a synthesis method thereof, and the derivative is used as a detection reagent for detecting ClO-The use of (1).
Background
The coumarin has the advantages of easy modification of the structure, various photophysical properties and the like, and has been developed in the field of chemical biological probes. Coumarin is a dye with a benzopyrone structure, which is not fluorescent by itself. The introduction of electron donating groups at the 3, 4 positions or electron withdrawing groups at the 6, 7 positions produces intense fluorescence. The coumarin dye has larger Stokes shift, good water solubility and higher quantum yield. Due to the small size and the ability to penetrate cell membranes, more and more researchers in recent years have studied coumarin dyes as small-molecule fluorescent probes for detecting relevant disease markers. Wherein, 7-diethylamino-3-acetylcoumarin, diethylamino are used as strong electron-donating groups, acetyl is used as strong electron-withdrawing groups, so that intramolecular charge transfer effect is formed, and the whole molecule emits strong fluorescence. The work reported at present is all design and optimization at 3-bit. However, relatively few studies have been conducted on the 4-position of 7-diethylamino-3-acetylcoumarin.
In view of the above problems, it has become one of the leading challenges in the current biomedical development to study the 4-position of 7-diethylamino-3-acetylcoumarin and design a fluorescent probe with good selectivity, high sensitivity and low cytotoxicity for detecting disease markers.
In the invention, a compound based on 7-diethylamino-3-acetyl coumarin is synthesized, and a probe and ClO are used-The fluorescence changes before and after the reaction to realize the ClO reaction-Detection of (3).
Disclosure of Invention
The invention aims to provide a 7-diethylamino-3-acetyl coumarin derivative, a synthetic method thereof and application of the derivative in ClO-The detection method is simple, convenient to operate, good in selectivity and high in sensitivity.
The invention provides a 7-diethylamino-3-acetyl coumarin derivative: (1) the name of 4- (3- (7- (diethylamino) -2-oxo-2H-chromen-3-yl) -5- (p-tolyl) -4,5-dihydro-1H-pyrazol-1-yl) benzoic acid in the Chinese, the name of 4- (3- (7- (diethylamino) -2-oxo-2H-chromen-3-yl) -5- (p-tolyl) -4,5-dihydro-1H-pyrazol-1-yl) benzoic acid in the English, and the name of Probe 1; (2) the Chinese name is 4- (3- (7- (diethylamino) -4-hydroxy-2-oxo-2H-chromen-3-yl) -5- (p-tolyl) -4,5-dihydro-1H-pyrazol-1-yl) benzoic acid, the English name is 4- (3- (7- (dimethylamino) -4-hydroxy-2-oxo-2H-chromen-3-yl) -5- (p-tolyl) -4,5-dihydro-1H-pyrazol-1-yl) benzoic acid, and the name is Probe 2; the structural formula is as follows:
the invention provides a synthesis method of a 7-diethylamino-3-acetyl coumarin derivative Probe 1, which comprises the following steps:
(1) the molar ratio of the components is 1: 1 mixing 7-diethylamino-3-acetylcoumarin and 4-methylbenzaldehyde in absolute ethanol, adding a small amount of piperidine with stirring, and refluxing the mixture for 48 hours. Cooling to room temperature, filtering the precipitate and washing with ethanol, then drying in vacuo to obtain the compound (E) -7- (diethylamino) -3- (3- (p-tolyl) acryloyl) -2H-chromen-2-one as an orange solid;
(2) the molar ratio of the raw materials is 1: 3 (E) -7- (diethylamino) -3- (3- (p-tolyl) acryloyl) -2H-chromen-2-one and 4-hydrazinobenzoic acid were mixed in absolute ethanol, a small amount of glacial acetic acid was added, and the mixture was stirred under reflux for 4H. After cooling to room temperature, the precipitate was filtered and washed with anhydrous ethanol, then dried in vacuo to obtain a red solid, i.e., 4- (3- (7- (diethylamino) -2-oxo-2H-chromen-3-yl) -5- (p-tolyl) -4,5-dihydro-1H-pyrazol-1-yl) benzoic acid (Probe 1).
The invention provides a method for synthesizing a 7-diethylamino-3-acetylcoumarin derivative Probe 2, which comprises the following steps:
(1) the molar ratio of the raw materials is 1: 1 mixing 7-diethylamino-4-hydroxy-3-acetylcoumarin and 4-methylbenzaldehyde in absolute ethanol, adding a small amount of piperidine with stirring, and refluxing the mixture for 48 hours. Cooling to room temperature, filtering the precipitate and washing with ethanol, then drying in vacuo to obtain the compound as an orange solid, i.e. (E) -7- (diethylamino) -4-hydroxy-3- (3- (p-tolyl) acryloyl) -2H-chromen-2-one;
(2) the molar ratio of the raw materials is 1: 3 (E) -7- (diethylamino) -4-hydroxy-3- (3- (p-tolyl) acryloyl) -2H-chromen-2-one and 4-hydrazinobenzoic acid were mixed in absolute ethanol, a small amount of glacial acetic acid was added, and the mixture was stirred under reflux for 4H. After cooling to room temperature, the precipitate was filtered and washed with anhydrous ethanol, then dried in vacuo to obtain a red solid, i.e., 4- (3- (7- (diethylamino) -4-hydroxy-2-oxo-2H-chromen-3-yl) -5- (p-tolyl) -4,5-dihydro-1H-pyrazol-1-yl) benzoic acid (Probe 2).
The invention provides a ClO detection method for non-disease diagnosis or treatment-The method comprises the following steps:
(1) preparing a fluorescent Probe stock solution of 2mM Probe 2 by using dimethyl sulfoxide (DMSO);
(2) 2mL of PBS (pH 7.4) buffer and 10. mu.L of fluorescent probe stock were added to a fluorescence cuvette and detected on a spectrofluorometer with ClO-The fluorescence intensity at 520nm is gradually weakened, and the fluorescence changes in a turn-off type;
(3) with ClO-Concentration as abscissa, in fluorescence intensity F520nmPlotting the plot for ordinate to obtain ClO-The working curve of (c); the linear regression equation is: y is-303.7 x +3578.6, x has the unit of 10-6mol/L。
Compared with the prior art, the invention has the following advantages and effects:
1. the 7-diethylamino-3-acetyl coumarin derivative is simple to synthesize and low in cost;
2. the 7-diethylamino-3-acetyl coumarin derivative Probe 2 can realize the ClO pair-The detection has high sensitivity and good selectivity of the detection result; can also be used for ClO in cells-Detection, and can also be used for animal living body such as zebra fish ClO-Detecting;
3. the detection method is simple and can be realized only by means of a fluorescence spectrometer;
4. the invention adopts turn-off fluorescence detection, and the detection signal is obvious.
Drawings
FIG. 1 nuclear magnetic hydrogen spectrum of Probe 1 as a fluorescent Probe prepared in example 1
FIG. 2 nuclear magnetic carbon spectrum of Probe 1, a fluorescent Probe prepared in example 1
FIG. 3 Mass Spectrum of Probe 1, a fluorescent Probe prepared in example 1
FIG. 4 nuclear magnetic hydrogen spectrum of Probe 2, a fluorescent Probe prepared in example 1
FIG. 5 nuclear magnetic carbon spectrum of Probe 2, a fluorescent Probe prepared in example 1
FIG. 6 Mass Spectrum of Probe 2, a fluorescent Probe prepared in example 1
FIG. 7 fluorescent probes Probe 2 and ClO-Fluorescence emission map of action
FIG. 8 fluorescent histogram of fluorescent Probe Probe 2 and various analytes
FIG. 9 fluorescent Probe 2 measurement of ClO-Working curve of
FIG. 10 fluorescent Probe 2 measurement of ClO-Imaging of cells
FIG. 11 fluorescent Probe 2 measurement of ClO-Zebra fish imaging map
Detailed Description
The present invention will be further described with reference to the following examples and drawings, but the present invention is not limited to the following examples.
Example 1
Preparation and characterization of Probe 1
7-diethylamino-3-acetylcoumarin (1.55g,6mmol) and 4-methylbenzaldehyde (0.72g,6mmol) were mixed in 30mL of anhydrous ethanol, 200. mu.L of piperidine was added with stirring, and the mixture was refluxed for 48 hours. Cooled to room temperature, the precipitate was filtered and washed with ethanol, and then dried in vacuo to obtain an orange solid compound (1.25g, yield: 57.6%) which was (E) -7- (diethylamino) -3- (3- (p-tolyl) acryloyl) -2H-chromen-2-one.1H NMR(600MHz,Chloroform-d)δ8.57(s,1H),8.13(d,J＝15.7Hz,1H),7.84(d,J＝15.7Hz,1H),7.61(d,J＝7.9Hz,2H),7.45(d,J＝8.9Hz,1H),7.22(d,J＝7.8Hz,2H),6.65(d,J＝8.9Hz,1H),6.52(s,1H),3.49(q,J＝7.1Hz,4H),2.40(s,3H),1.27(t,J＝7.0Hz,6H).13C NMR(150MHz,Chloroform-d)δ186.62,160.89,158.61,152.72,148.67,143.58,140.74,132.63,131.81,129.55,128.83,123.86,117.03,110.04,108.87,96.91,45.36,45.29,21.59,12.46,12.44.
(E) -7- (diethylamino) -3- (3- (p-tolyl) acryloyl) -2H-chromen-2-one (0.36g,1mmol) and 4-hydrazinobenzoic acid (0.46g,3mmol) were mixed in anhydrous ethanol (25mL), 1mL of glacial acetic acid was added, and the mixture was stirred at reflux for 4H. After cooling to room temperature, the precipitate was filtered and washed with anhydrous ethanol, and then dried in vacuo to obtain Probe 1(0.20g, yield: 40.4%) as a red solid.1H NMR(600MHz,DMSO-d6) δ 8.41(s,1H),7.72(d, J-9.0 Hz,2H),7.60(d, J-9.0 Hz,1H),7.14(q, J-8.3 Hz,4H),7.02(d, J-8.7 Hz,2H),6.77(dd, J-9.0, 2.3Hz,1H),6.57(d, J-2.1 Hz,1H),5.53(dd, J-12.1, 4.9Hz,1H),3.96(dd, J-18.0, 12.1Hz,1H),3.47(q, J-6.9 Hz,4H),3.22(dd, J-18.0, 4.9, 1H),2.25(s,3H),1.14(t, 7.0Hz, 7H), 7.7H (H, 7H)13C NMR(150MHz,DMSO-d6) Delta 167.68,159.56,156.77,151.57,147.50,147.24,141.23,139.32,137.17,131.23,130.77,130.08,126.00,120.09,112.36,111.43,110.08,108.53,96.58,62.24,45.56,44.66,21.12,12.85. (FIG. 2) ESI-MS M/z [ M + H]+calcd for 496.2236；Found496.2224；[M+Na]+calcd for 518.2056; found 518.2046 (fig. 3)
Preparation and characterization of Probe 2
7-diethylamino-4-hydroxy-3-acetylcoumarin (1.65g,6mmol) and 4-methylbenzaldehyde (0.72g,6mmol) were mixed in 30mL of anhydrous ethanol, 200. mu.L of piperidine was added with stirring, and the mixture was refluxed for 48 hours. Cooled to room temperature, the precipitate was filtered and washed with ethanol, and then dried in vacuo to obtain an orange solid compound (1.40g, yield: 61.9%) which was (E) -7- (diethylamino) -4-hydroxy-3- (3- (p-tolyl) acryloyl) -2H-chromen-2-one.1H NMR(600MHz,DMSO-d6)δ8.27(d,J＝15.8Hz,1H),7.91(d,J＝15.8Hz,1H),7.75(d,J＝9.1Hz,1H),7.64(d,J＝7.3Hz,2H),7.31(d,J＝7.4Hz,2H),6.79(d,J＝9.1Hz,1H),6.51(s,1H),3.49(dd,J＝10.6,7.1Hz,4H),2.36(s,3H),1.14(t,J＝6.7Hz,6H).13C NMR(150MHz,DMSO-d6)δ190.89,179.62,160.72,157.42,154.42,145.29,141.94,132.29,130.33,129.35,127.35,122.30,110.14,103.04,98.10,96.36,44.89,21.64,12.80.
Reacting (E) -7- (diethylamino) -4-hydroxy-3- (3- (p-tolyl) acryloyl) -2H-chromen-2-one (0.37g,1mmol) and 4-hydrazinobenzoic acid (0.46g,3mmol) were combined in anhydrous ethanol (25mL) and 1mL glacial acetic acid was added and the mixture was stirred at reflux for 4H. After cooling to room temperature, the precipitate was filtered and washed with anhydrous ethanol, and then dried in vacuo to obtain Probe 2(0.25g, yield: 48.9%) as a red solid.1H NMR(600MHz,DMSO-d6) δ 13.48(s,1H), 7.78-7.74 (m,3H), 7.18-7.15 (m,4H),6.88(d, J ═ 8.6Hz,2H),6.81(d, J ═ 9.1Hz,1H),6.53(s,1H),5.49(dd, J ═ 12.0,5.2Hz,1H),4.13(dd, J ═ 18.7,12.1Hz,1H), 3.48-3.46 (m,4H),3.45(s,1H),2.26(s,3H),1.14(t, J ═ 7.0Hz,6H) (fig. 4)13C NMR(150MHz,DMSO-d6) Delta 167.56,166.72,160.95,155.82,153.10,152.42,146.38,138.94,137.34,131.50,130.13,126.07,125.55,120.64,112.06,109.92,102.50,96.52,91.88,60.76,46.79,44.65,21.13,12.79. (FIG. 5) ESI-MS M/z: [ M + H]+calcd for 512.2185; found 512.2175 (fig. 6)
Example 2
Preparing a fluorescent Probe stock solution of 2mM Probe 2 by using dimethyl sulfoxide (DMSO); 2mL of PBS (pH 7.4) buffer and 10. mu.L of stock solution of fluorescent probe were added to a fluorescent cuvette, and ClO was taken-The solution was gradually added to the cuvette by a microsyringe and simultaneously detected on a spectrofluorometer. With ClO-With the addition of (2), the fluorescence intensity at 520nm gradually decreased in a "turn-off" type change. (FIG. 7)
Example 3
Preparing a fluorescent Probe stock solution of 2mM Probe 2 by using dimethyl sulfoxide (DMSO); to a fluorescence cuvette, 2mL of PBS (pH 7.4) and 10. mu.L of stock solutions of fluorescent probes were added, and then 10-fold equivalents of the other analytes and ClO were added, respectively-(1-Blank；2-Na+；3-K+；4-Mg2+；5-Cu2+；6-Zn2+；7-Fe2+；8-Fe3+；9-Al3+；10-Ca2+；11-NO2 -；12-NO3 -；13-ClO2 -；14-ClO3 -；15-ClO4 -；16-MnO4 -；17-Cl-；18-H2O2；19-ClO-) And detected on a fluorescence spectrophotometer (fig. 8). ClO-So that the fluorescence intensity of the detection system is obviously reduced at 520nm, and other analytes basically do not cause the change of the fluorescence intensity of the detection system.
Example 4
With ClO-Concentration as abscissa, in fluorescence intensity F520nmPlotting the plot for ordinate to obtain ClO-Working curve of (fig. 9); the linear regression equation is: y is-303.7 x +3578.6, and x has a unit of 10-6mol/L。
Example 5
Preparing a PBS buffer solution with the pH value of 7.4 and the concentration of 10mM, and preparing a DMSO solution of 2mM Probe 2; add 10. mu.L of LProbe 2 in DMSO to 2mL of PBS; adding the probe solution into a HeLa cell culture solution to enable the concentration of the probe solution to be 10 mu M, and reacting the probe solution with HeLa cells at 37 ℃ for 10min to enable the system to have obvious fluorescence in a green channel; then adding exogenous ClO-And reacting for 15min at 37 ℃, and observing the disappearance of green channel fluorescence of the system under a fluorescence imager. When the HeLa cells were incubated with LPS first and the probe solution was added, the fluorescence decreased in the green channel. See fig. 10.
Example 6
Preparing a PBS buffer solution with the pH value of 7.4 and the concentration of 10mM, and preparing a DMSO solution of 2mM Probe 2; add 10. mu.L of SSN in DMSO to 2mL of PBS; culturing the probe solution and the zebra fish together to ensure that the concentration of the probe solution is 10 mu M, and reacting for 10min at 37 ℃ so that the system has obvious fluorescence in a green channel; then adding exogenous hypochlorous acid, reacting for 15min at 37 deg.C, and observing the disappearance of green channel fluorescence by the system under fluorescence imager, as shown in FIG. 11.
Claims (6)
1. A7-diethylamino-3-acetyl coumarin derivative is characterized in that the structural formula is represented by Probe 2:
2. the method for synthesizing 7-diethylamino-3-acetylcoumarin derivative Probe 2 according to claim 1, comprising the steps of:
(1) the molar ratio of the raw materials is 1: 1 mixing 7-diethylamino-4-hydroxy-3-acetyl coumarin and 4-methylbenzaldehyde in absolute ethanol, adding a small amount of piperidine while stirring, and refluxing the mixture for 48 hours; cooling to room temperature, filtering the precipitate and washing with ethanol, then drying in vacuo to obtain the compound as an orange solid, i.e. (E) -7- (diethylamino) -4-hydroxy-3- (3- (p-tolyl) acryloyl) -2H-chromen-2-one;
(2) the molar ratio of the raw materials is 1: 3 mixing (E) -7- (diethylamino) -4-hydroxy-3- (3- (p-tolyl) acryloyl) -2H-chromen-2-one and 4-hydrazinobenzoic acid in absolute ethanol, adding a small amount of glacial acetic acid, and stirring and refluxing the mixture for 4H; after cooling to room temperature, the precipitate was filtered and washed with anhydrous ethanol, then dried in vacuo to obtain 4- (3- (7- (diethylamino) -4-hydroxy-2-oxo-2H-chromen-3-yl) -5- (p-tolyl) -4,5-dihydro-1H-pyrazol-1-yl) benzoic acid as a yellow solid.
3. ClO for non-disease diagnosis or treatment of 7-diethylamino-3-acetylcoumarin derivative Probe 2 according to claim 1-Application in detection.
4. ClO detection for non-disease diagnosis or treatment-The method is characterized by comprising the following steps:
(1) preparing 2mM of a stock solution of the Probe 2 of claim 1 using dimethyl sulfoxide (DMSO);
(2) 2mL of a pH 7.4 PBS buffer and 10. mu.L of a stock solution of fluorescent probe were added to a fluorescence cuvette and detected on a spectrofluorometer with ClO-The fluorescence intensity at 520nm is gradually weakened, and the fluorescence changes in a turn-off type;
(3) with ClO-Concentration as abscissa, in fluorescence intensity F520nmPlotting a chart for the ordinate to obtainClO-The working curve of (a); the linear regression equation is: y is-303.7 x +3578.6, and x has a unit of 10-6mol/L。
5. ClO in a cell-producing process using the 7-diethylamino-3-acetylcoumarin derivative Probe 2 as defined in claim 1-The application of the detection reagent.
6. The use of the 7-diethylamino-3-acetylcoumarin derivative Probe 2 as defined in claim 1 for preparing ClO in living animal body-The application of the detection reagent.
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015177839A1 (en) * | 2014-05-19 | 2015-11-26 | 株式会社日本化学工業所 | Novel pyrazoline compound and photoelectric conversion element using same |
| WO2016132802A1 (en) * | 2015-02-20 | 2016-08-25 | 国立大学法人 東京大学 | Sulfane sulfur-selective fluorescent probe |
| CN107235946A (en) * | 2017-06-20 | 2017-10-10 | 济南大学 | A kind of glutathione fluorescence probe and its preparation method and application |
| CN111217799A (en) * | 2020-02-27 | 2020-06-02 | 山西大学 | Indole salt-coumarin derivative and synthesis method and application thereof |
| CN111233804A (en) * | 2020-03-03 | 2020-06-05 | 山西大学 | Benzopyranoylium ion-coumarin derivative and synthetic method and application thereof |
| CN111848661A (en) * | 2020-08-03 | 2020-10-30 | 南通大学 | Coumarin fluorescent probe and preparation method and application thereof |
| CN112159378A (en) * | 2020-08-27 | 2021-01-01 | 中山大学 | Gilmane type sesquiterpene lactone compound and preparation method and application thereof |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| NZ529221A (en) * | 2001-04-25 | 2005-04-29 | Altana Pharma Ag | Phthalazinones with PDE-inhibiting properties for treatment of airway disorders and other inflammatory conditions |
| US7176242B2 (en) * | 2001-11-08 | 2007-02-13 | Elan Pharmaceuticals, Inc. | N,N′-substituted-1,3-diamino-2-hydroxypropane derivatives |
| CN111253356A (en) * | 2020-03-03 | 2020-06-09 | 山西大学 | Coumarin-benzopyrylium salt derivative and synthesis method and application thereof |
-
2021
- 2021-02-05 CN CN202110160194.1A patent/CN112876460B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015177839A1 (en) * | 2014-05-19 | 2015-11-26 | 株式会社日本化学工業所 | Novel pyrazoline compound and photoelectric conversion element using same |
| WO2016132802A1 (en) * | 2015-02-20 | 2016-08-25 | 国立大学法人 東京大学 | Sulfane sulfur-selective fluorescent probe |
| CN107235946A (en) * | 2017-06-20 | 2017-10-10 | 济南大学 | A kind of glutathione fluorescence probe and its preparation method and application |
| CN111217799A (en) * | 2020-02-27 | 2020-06-02 | 山西大学 | Indole salt-coumarin derivative and synthesis method and application thereof |
| CN111233804A (en) * | 2020-03-03 | 2020-06-05 | 山西大学 | Benzopyranoylium ion-coumarin derivative and synthetic method and application thereof |
| CN111848661A (en) * | 2020-08-03 | 2020-10-30 | 南通大学 | Coumarin fluorescent probe and preparation method and application thereof |
| CN112159378A (en) * | 2020-08-27 | 2021-01-01 | 中山大学 | Gilmane type sesquiterpene lactone compound and preparation method and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| An innovative hypochlorite-sensing scaffold and its imaging application;Yongfei Huang等;《Dyes and Pigments 》;20210420;第191卷;全文 * |
| Discovery of a novel fluorescent HSP90 inhibitor and its anti-lung cancer effect;Su-Yun Bai等;《RSC Advances》;20140604;第38卷;第19887页 * |
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