CN114773381B - Pyridine ring-containing aromatic acrylonitrile carbazole reaction type fluoride ion fluorescent probe and preparation method and application thereof - Google Patents
Pyridine ring-containing aromatic acrylonitrile carbazole reaction type fluoride ion fluorescent probe and preparation method and application thereof Download PDFInfo
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- -1 acrylonitrile carbazole Chemical compound 0.000 title claims abstract description 81
- 238000002360 preparation method Methods 0.000 title claims abstract description 56
- KRHYYFGTRYWZRS-UHFFFAOYSA-M Fluoride anion Chemical compound [F-] KRHYYFGTRYWZRS-UHFFFAOYSA-M 0.000 title claims abstract description 45
- 239000007850 fluorescent dye Substances 0.000 title claims abstract description 29
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical group C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 title claims abstract description 24
- 125000003118 aryl group Chemical group 0.000 title claims abstract description 16
- 238000006757 chemical reactions by type Methods 0.000 title claims abstract description 16
- UJOBWOGCFQCDNV-UHFFFAOYSA-N 9H-carbazole Chemical compound C1=CC=C2C3=CC=CC=C3NC2=C1 UJOBWOGCFQCDNV-UHFFFAOYSA-N 0.000 claims abstract description 22
- 238000001514 detection method Methods 0.000 claims abstract description 18
- 239000000523 sample Substances 0.000 claims abstract description 13
- 239000012472 biological sample Substances 0.000 claims abstract description 9
- 150000001875 compounds Chemical class 0.000 claims abstract description 6
- 150000002825 nitriles Chemical class 0.000 claims abstract description 6
- 210000004369 blood Anatomy 0.000 claims abstract description 4
- 239000008280 blood Substances 0.000 claims abstract description 4
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 203
- 238000006243 chemical reaction Methods 0.000 claims description 85
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 40
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 38
- FNJVVKBJFIJRCZ-UHFFFAOYSA-N CCCN1C(C=C(C(C=O)=C2)O)=C2C2=CC=CC=C12 Chemical compound CCCN1C(C=C(C(C=O)=C2)O)=C2C2=CC=CC=C12 FNJVVKBJFIJRCZ-UHFFFAOYSA-N 0.000 claims description 33
- XLJIGLNQSNXOQD-UHFFFAOYSA-N CCCN1C(C=C(C(C=O)=C2)OC)=C2C2=CC=CC=C12 Chemical compound CCCN1C(C=C(C(C=O)=C2)OC)=C2C2=CC=CC=C12 XLJIGLNQSNXOQD-UHFFFAOYSA-N 0.000 claims description 31
- AJHYASMURHSAHE-UHFFFAOYSA-N CCCN1C(C=C(C(C=O)=C2)O[Si](C)(C)C(C)(C)C)=C2C2=CC=CC=C12 Chemical compound CCCN1C(C=C(C(C=O)=C2)O[Si](C)(C)C(C)(C)C)=C2C2=CC=CC=C12 AJHYASMURHSAHE-UHFFFAOYSA-N 0.000 claims description 31
- RPDJLFIOPNAQHK-UHFFFAOYSA-N CCCN1C(C=C(C=C2)OC)=C2C2=CC=CC=C12 Chemical compound CCCN1C(C=C(C=C2)OC)=C2C2=CC=CC=C12 RPDJLFIOPNAQHK-UHFFFAOYSA-N 0.000 claims description 31
- RWRDLPDLKQPQOW-UHFFFAOYSA-N tetrahydropyrrole Natural products C1CCNC1 RWRDLPDLKQPQOW-UHFFFAOYSA-N 0.000 claims description 28
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical group [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 27
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical group CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 24
- XHXFXVLFKHQFAL-UHFFFAOYSA-N phosphoryl trichloride Chemical compound ClP(Cl)(Cl)=O XHXFXVLFKHQFAL-UHFFFAOYSA-N 0.000 claims description 24
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical group CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 claims description 20
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 20
- WSLDOOZREJYCGB-UHFFFAOYSA-N 1,2-Dichloroethane Chemical compound ClCCCl WSLDOOZREJYCGB-UHFFFAOYSA-N 0.000 claims description 19
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 19
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 claims description 15
- MDKVPSJRUXOFFV-UHFFFAOYSA-N 2-methoxy-9h-carbazole Chemical compound C1=CC=C2C3=CC=C(OC)C=C3NC2=C1 MDKVPSJRUXOFFV-UHFFFAOYSA-N 0.000 claims description 14
- 239000003153 chemical reaction reagent Substances 0.000 claims description 14
- SCYULBFZEHDVBN-UHFFFAOYSA-N 1,1-Dichloroethane Chemical compound CC(Cl)Cl SCYULBFZEHDVBN-UHFFFAOYSA-N 0.000 claims description 12
- FFNVQNRYTPFDDP-UHFFFAOYSA-N 2-cyanopyridine Chemical compound N#CC1=CC=CC=N1 FFNVQNRYTPFDDP-UHFFFAOYSA-N 0.000 claims description 12
- PVWOIHVRPOBWPI-UHFFFAOYSA-N n-propyl iodide Chemical compound CCCI PVWOIHVRPOBWPI-UHFFFAOYSA-N 0.000 claims description 12
- BCNZYOJHNLTNEZ-UHFFFAOYSA-N tert-butyldimethylsilyl chloride Chemical compound CC(C)(C)[Si](C)(C)Cl BCNZYOJHNLTNEZ-UHFFFAOYSA-N 0.000 claims description 12
- 239000002904 solvent Substances 0.000 claims description 11
- VSCWAEJMTAWNJL-UHFFFAOYSA-K aluminium trichloride Chemical group Cl[Al](Cl)Cl VSCWAEJMTAWNJL-UHFFFAOYSA-K 0.000 claims description 10
- 239000003054 catalyst Substances 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 8
- 238000006467 substitution reaction Methods 0.000 claims description 8
- 239000000376 reactant Substances 0.000 claims description 7
- 239000003513 alkali Substances 0.000 claims description 4
- 239000002585 base Substances 0.000 claims description 4
- HPNMFZURTQLUMO-UHFFFAOYSA-N diethylamine Chemical compound CCNCC HPNMFZURTQLUMO-UHFFFAOYSA-N 0.000 claims description 4
- WSFSSNUMVMOOMR-UHFFFAOYSA-N formaldehyde Substances O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims description 2
- 238000006170 formylation reaction Methods 0.000 claims description 2
- 230000003301 hydrolyzing effect Effects 0.000 claims description 2
- ILMRJRBKQSSXGY-UHFFFAOYSA-N tert-butyl(dimethyl)silicon Chemical compound C[Si](C)C(C)(C)C ILMRJRBKQSSXGY-UHFFFAOYSA-N 0.000 claims description 2
- 125000003554 tetrahydropyrrolyl group Chemical group 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 claims 1
- 239000011737 fluorine Substances 0.000 abstract description 15
- 229910052731 fluorine Inorganic materials 0.000 abstract description 15
- 230000035945 sensitivity Effects 0.000 abstract description 7
- 238000006862 quantum yield reaction Methods 0.000 abstract description 4
- 230000004044 response Effects 0.000 abstract description 4
- 238000007363 ring formation reaction Methods 0.000 abstract description 4
- 125000000391 vinyl group Chemical group [H]C([*])=C([H])[H] 0.000 abstract description 3
- 229920002554 vinyl polymer Polymers 0.000 abstract description 3
- 238000006000 Knoevenagel condensation reaction Methods 0.000 abstract description 2
- 239000002841 Lewis acid Substances 0.000 abstract description 2
- 229910018557 Si O Inorganic materials 0.000 abstract description 2
- 238000010670 acid alkali reaction Methods 0.000 abstract description 2
- 238000010504 bond cleavage reaction Methods 0.000 abstract description 2
- 230000021615 conjugation Effects 0.000 abstract description 2
- 150000007517 lewis acids Chemical class 0.000 abstract description 2
- LIVNPJMFVYWSIS-UHFFFAOYSA-N silicon monoxide Inorganic materials [Si-]#[O+] LIVNPJMFVYWSIS-UHFFFAOYSA-N 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 83
- 239000012043 crude product Substances 0.000 description 72
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 48
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 48
- 238000004440 column chromatography Methods 0.000 description 40
- 239000007787 solid Substances 0.000 description 40
- 238000003756 stirring Methods 0.000 description 37
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 32
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical class [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 32
- 239000012074 organic phase Substances 0.000 description 32
- 239000012071 phase Substances 0.000 description 32
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 25
- 229910052786 argon Inorganic materials 0.000 description 24
- 238000000605 extraction Methods 0.000 description 16
- 239000000203 mixture Substances 0.000 description 14
- 210000004027 cell Anatomy 0.000 description 9
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical class [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 8
- 239000005457 ice water Substances 0.000 description 8
- 239000002994 raw material Substances 0.000 description 8
- 230000000704 physical effect Effects 0.000 description 5
- 230000005284 excitation Effects 0.000 description 4
- 238000003384 imaging method Methods 0.000 description 4
- 239000008213 purified water Substances 0.000 description 4
- 239000011541 reaction mixture Substances 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000002189 fluorescence spectrum Methods 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- YCKRFDGAMUMZLT-UHFFFAOYSA-N Fluorine atom Chemical compound [F] YCKRFDGAMUMZLT-UHFFFAOYSA-N 0.000 description 2
- 239000007995 HEPES buffer Substances 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000000295 emission spectrum Methods 0.000 description 2
- 238000000695 excitation spectrum Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000004448 titration Methods 0.000 description 2
- AKYHKWQPZHDOBW-UHFFFAOYSA-N (5-ethenyl-1-azabicyclo[2.2.2]octan-7-yl)-(6-methoxyquinolin-4-yl)methanol Chemical compound OS(O)(=O)=O.C1C(C(C2)C=C)CCN2C1C(O)C1=CC=NC2=CC=C(OC)C=C21 AKYHKWQPZHDOBW-UHFFFAOYSA-N 0.000 description 1
- 125000004105 2-pyridyl group Chemical group N1=C([*])C([H])=C([H])C([H])=C1[H] 0.000 description 1
- 239000001576 FEMA 2977 Substances 0.000 description 1
- 208000001132 Osteoporosis Diseases 0.000 description 1
- 208000037273 Pathologic Processes Diseases 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000000601 blood cell Anatomy 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- LOUPRKONTZGTKE-UHFFFAOYSA-N cinchonine Natural products C1C(C(C2)C=C)CCN2C1C(O)C1=CC=NC2=CC=C(OC)C=C21 LOUPRKONTZGTKE-UHFFFAOYSA-N 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 208000002925 dental caries Diseases 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000022244 formylation Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000009054 pathological process Effects 0.000 description 1
- 238000005375 photometry Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 229960003110 quinine sulfate Drugs 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 229910052938 sodium sulfate Inorganic materials 0.000 description 1
- 235000011152 sodium sulphate Nutrition 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F7/00—Compounds containing elements of Groups 4 or 14 of the Periodic Table
- C07F7/02—Silicon compounds
- C07F7/08—Compounds having one or more C—Si linkages
- C07F7/18—Compounds having one or more C—Si linkages as well as one or more C—O—Si linkages
- C07F7/1804—Compounds having Si-O-C linkages
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F7/00—Compounds containing elements of Groups 4 or 14 of the Periodic Table
- C07F7/02—Silicon compounds
- C07F7/08—Compounds having one or more C—Si linkages
- C07F7/18—Compounds having one or more C—Si linkages as well as one or more C—O—Si linkages
- C07F7/1804—Compounds having Si-O-C linkages
- C07F7/1872—Preparation; Treatments not provided for in C07F7/20
- C07F7/1892—Preparation; Treatments not provided for in C07F7/20 by reactions not provided for in C07F7/1876 - C07F7/1888
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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"
- G01N21/643—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" non-biological material
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- 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
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
- G01N21/6458—Fluorescence microscopy
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- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1029—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
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Abstract
The invention relates to the technical field of detection, in particular to an aromatic vinyl nitrile carbazole reaction type fluoride ion fluorescent probe containing pyridine rings, and a preparation method and application thereof. According to the invention, carbazole is taken as a fluorophore, a silicon-oxygen bond is taken as an identification group, a pyridine group is introduced through Knoevenagel condensation reaction, and a synthetic compound 3- ((2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-yl) 2- (pyridine-2-yl) acrylonitrile fluoride ion is subjected to Lewis acid alkali reaction to induce Si-O bond cleavage, so that O is formed ‑ The ortho-aryl allyldinitrile group is attacked to undergo cyclization, so that the conjugation area of the probe is greatly increased, the quantum yield is greatly improved, and the fluorescence intensity is remarkably changed to realize the detection of fluoride ions in a biological sample. The ratio type fluorescent probe has the characteristics of wide range and high sensitivity in response to fluorine ions, and can realize quantitative detection of trace fluorine ions in blood samples.
Description
Technical Field
The invention relates to the technical field of detection, in particular to an aromatic vinyl nitrile carbazole reaction type fluoride ion fluorescent probe containing pyridine rings, and a preparation method and application thereof.
Background
Fluoride ion is one of trace elements necessary for human body, and proper amount of fluorine can play a role in strengthening bones and teeth, so that caries can be effectively prevented and cured, osteoporosis of human body can be treated, and the like. Excessive fluoride ions can inhibit synthesis of DNA and protein in human body, cause metabolic disorder of immune system, and reduce immunity of human body. In addition, the blood pressure is reduced, and the growth and development of human bodies are affected. Therefore, the determination of the concentration of fluoride ions in biological samples is of great importance for further understanding of the pathological processes of various diseases associated with fluoride ions.
The method for detecting the systematic fluoride ions by using the fluorine reagent colorimetry, the absorbance photometry, the ion chromatography, the ion selective electrode method and the like has the defects of high instrument price, complicated sample treatment, easy interference, inapplicability to detection of the fluoride ions in organisms and the like. The molecular fluorescent probe can convert the environmental information of the molecules into fluorescent signals, has the advantages of high sensitivity, high selectivity, convenient use, low cost, no need of pretreatment, capability of carrying out in-situ detection in real time and the like, and is considered as the most efficient fluoride ion detection method. Although the research of the fluoride ion fluorescent probes is remarkably advanced at present, most probes have low sensitivity and poor hydrophilicity, and are difficult to be used for quantitatively detecting fluoride ions in biological samples (cells, blood and urine) of an aqueous-phase-like system. For example, CN111039972A, the probe has poor water solubility, is only suitable for quantitatively detecting fluorine ions in pure acetonitrile, is easy to interfere by Ar-OH formed after responding to the fluorine ions, has a smaller conjugated system and lower sensitivity, and is not suitable for quantitatively detecting trace fluorine ions.
Therefore, the fluoride ion fluorescent probe with high sensitivity, high selectivity and strong anti-interference performance, which is suitable for detecting fluoride ions in biological samples and imaging living cells, is an urgent problem to be solved at present.
Disclosure of Invention
In order to solve the technical problems in the prior art, the invention provides an aromatic vinyl nitrile carbazole reaction type fluoride ion fluorescent probe containing a pyridine ring, and a preparation method and application thereof.
An arylenenitrile carbazole-based reactive fluoride ion fluorescent probe (named 3- ((2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazol-3-yl) 2- (pyridin-2-yl) acrylonitrile with the English name 3- (2- ((tert-butylsilyl) oxy) -9-propyl-9H-carbazol-3-yl) -2- (pyridin-2-yl) acylonitrile) is abbreviated as SCP, and has the following specific structural formula:
the preparation method of the pyridine ring-containing aromatic acrylonitrile carbazole reaction type fluoride ion fluorescent probe comprises the following steps:
(a) 2-methoxy-9H-carbazole and iodopropane are subjected to substitution reaction to generate 2-methoxy-9-propyl-9H-carbazole:
the molar ratio of the substitution reagent to the 2-methoxy-9H-carbazole is 2-5:1, the molar ratio of the alkali to the 2-methoxy-9H-carbazole is 2-5:1, and the reaction temperature is 60-70 ℃;
(b) Formylation of 2-methoxy-9-propyl-9H-carbazole with DMF and phosphorus oxychloride to produce 2-methoxy-9-propyl-9H-carbazole-3-carbaldehyde:
the molar ratio of the reactant to the 2-methoxy-9-propyl-9H-carbazole is 2-5:1, and the reaction temperature is 55-65 ℃;
(c) Hydrolyzing the 2-methoxy-9-propyl-9H-carbazole-3-formaldehyde to generate 2-hydroxy-9-propyl-9H-carbazole-3-formaldehyde:
the feeding mole ratio of the reaction reagent to the 2-methoxy-9-propyl-9H-carbazole-3-formaldehyde is 2-5:1, and the reaction temperature is room temperature;
(d) 2-hydroxy-9-propyl-9H-carbazole-3-formaldehyde and tert-butyl dimethyl chlorosilane undergo substitution reaction to obtain 2- ((tert-butyl dimethyl silicon based) oxy) -9-propyl-9H-carbazole-3-formaldehyde:
the molar ratio of the reaction reagent to the 2-hydroxy-9-propyl-9H-carbazole-3-formaldehyde is 2-5:1, the molar ratio of the catalyst to the 2-hydroxy-9-propyl-9H-carbazole-3-formaldehyde is 1-2:1, the molar ratio of the alkali to the compound 4 is 2-5:1, and the reaction temperature is-10 ℃;
(e) 2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde was reacted with 2-cyanopyridine to give the target compound 3- ((2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazol-3-yl) 2- (pyridin-2-yl) acrylonitrile:
The feeding molar ratio of the reactant to the 2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-formaldehyde is 2-3:1, the ratio of the catalyst to the 2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-formaldehyde is 0.04-0.06:1, and the reaction temperature is room temperature.
Further, in the step (a), the solvent is dimethyl sulfoxide (DMSO) or N, N-Dimethylformamide (DMF), the substitution reagent is iodopropane, and the base is sodium hydroxide or potassium hydroxide.
Further, in the step (b), the solvent is anhydrous N, N-Dimethylformamide (DMF), and the reaction reagent is phosphorus oxychloride.
Further, in the step (c), the solvent is anhydrous dichloromethane, anhydrous 1, 2-dichloroethane or anhydrous 1, 1-dichloroethane, and the reaction reagent is aluminum trichloride.
Further, in the step (d), the solvent is dichloromethane, 1, 2-dichloroethane or 1, 1-dichloroethane, the reaction reagent is tert-butyldimethyl chlorosilane, the catalyst is 4-dimethylaminopyridine, and the base is triethylamine or diethylamine.
Further, in the step (e), the solvent is anhydrous Tetrahydrofuran (THF), the reaction reagent is 2-cyanopyridine, and the catalyst is tetrahydropyrrole.
The pyridine ring-containing aromatic acrylonitrile carbazole reaction type fluoride ion fluorescent probe is applied to fluoride ion detection.
Further, the fluoride ion detection aspect is a fluoride ion detection aspect in a biological sample. Further, the aspect of detecting fluoride ions in the biological sample is an aspect of detecting fluoride ions in a blood sample.
Compared with the prior art, the invention has the technical effects that:
(1) According to the invention, carbazole is taken as a fluorophore, a silicon-oxygen bond is taken as an identification group, a pyridine group is introduced through Knoevenagel condensation reaction, and a synthetic compound 3- ((2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-yl) 2- (pyridine-2-yl) acrylonitrile fluoride ion is subjected to Lewis acid alkali reaction to induce Si-O bond cleavage, so that O is formed - The ortho-aryl allyldinitrile group is attacked to undergo cyclization, so that the conjugation area of the probe is greatly increased, the quantum yield is greatly improved, and the fluorescence intensity is remarkably changed to realize the detection of fluoride ions in a biological sample. Experiments prove that: in ethanol (EtOH), the recognition of fluoride ions by probe SCP produces a significant change in fluorescence signal. SCP has weak fluorescence intensity before reacting with fluoride ions in EtOH; the solution after the addition of fluoride ions gave a bright blue color. [ F ]]In the interval of=0-5.6 mu M, SCP fluorescence intensity and fluoride ion concentration at λEm=485 nm are in linear relation, and the detection limit is 4.88 And nM. With quinine sulfate (Φ350 nm=0.456) as a reference, the quantum yield of SCP after reaction with fluoride ions in EtOH was tested to be 0.857; the quantum yield after reaction of SCP with fluoride ions in EtOH-HEPES/NaOH (pH=7.4, 9:1, v/v) was 0.628. Cell imaging showed that under the Cyan channel, the fluorescence of probe SCP was significantly enhanced after reaction with fluoride ions. And (3) performing a standard adding recovery experiment on the fluoride ions in serum, wherein the recovery rate of the fluoride ions reaches 104.63%. In conclusion, SCP is a ratio type fluorescent probe with the characteristics of wide range and high sensitivity to fluoride ion response, and can realize quantitative detection of trace fluoride ions in blood samples.
Drawings
FIG. 1 is a flow chart of the preparation of pyridine ring-containing aralkenyl nitrile carbazole-based reactive fluoride ion fluorescent probe in the present application.
FIG. 2 is a fluorescence spectrum of the pyridine ring-containing aralkenylcarbazole-based reactive fluoride ion fluorescent probe of the present application in EtOH, wherein SCP and fluoride ion coexist (a) and SCP-F (b).
FIG. 3 is a graph showing fluorescence spectra of pyridine ring-containing aralkenyl carbazole-based reactive fluoride ion fluorescent probe SCP (0.2. Mu.M) titrated with fluoride ions (0-5.6. Mu.M) in the present application.
FIG. 4 is a fluorescent image of pyridine ring-containing aralkenyl carbazole-based reactive fluorogenic probe SCP (1. Mu.M) and fluoride ion (1. Mu.M) -treated HepG2 cells of the present application.
Detailed Description
The technical scheme of the present invention is further defined below in conjunction with the specific embodiments, but the scope of the claimed invention is not limited to the description.
Example 1
The preparation method of the pyridine ring-containing aromatic acrylonitrile carbazole type reaction type fluoride ion fluorescent probe comprises the following steps:
(a) Preparation of 2-methoxy-9-propyl-9H-carbazole
In a 100mL round bottom flask, 2-methoxy-9H-carbazole (5.91 g,30 mmol) and sodium hydroxide (6 g,150 mmol) were added and dissolved in DMSO (60 mL) and iodopropane (10.2 g,60 mmol) was added with stirring. After all the raw materials are added, a condensing tube and an Ar balloon are placed above the round-bottom flask, the vacuum is pumped for three times, and the temperature is raised to 60 ℃ for reaction for 16 hours. After the reaction was completed, cooled to room temperature and diluted with 60mL of water, dichloromethane (60 ml×3) was added for extraction, and after the organic phase was combined and washed with saturated NaCl solution (100 ml×2), anhydrous sodium sulfate was added for standing for 15min, and the dichloromethane was dried under reduced pressure to give crude product, which was subjected to column chromatography (PE: ea=10:1) to give 5.87g of 2-methoxy-9-propyl-9H-carbazole as a white solid, yield 81.8%.
(b) Preparation of 2-methoxy-9-propyl-9H-carbazole-3-carbaldehyde
In a 250mL three-necked flask, anhydrous DMF (9 mL) was added first, phosphorus oxychloride (4.59 g,30 mmol) was added dropwise to the three-necked flask under the protection of argon at 0℃and stirred for 15min, 2-methoxy-9-propyl-9H-carbazole (3.6 g,15 mmol) was dissolved in anhydrous DMF (26 mL) and then added dropwise to the three-necked flask, the temperature was raised to 55℃and stirred for 5H, ice water (35 mL) was added after the reaction solution cooled to room temperature and stirring was continued for 0.5H, extraction was performed with dichloromethane (35 mL. Times.3), the organic phases were combined and washed with saturated NaCl solution, anhydrous sodium sulfate was added and allowed to stand for 15min, and the dichloromethane was evaporated under reduced pressure to obtain a crude product. EA: pe=1: 5 as mobile phase, the crude product was purified by column chromatography to give 3.02g of 2-methoxy-9-propyl-9H-carbazole-3-carbaldehyde as yellow solid with a yield of 75.6%.
(c) Preparation of 2-hydroxy-9-propyl-9H-carbazole-3-carbaldehyde
Anhydrous aluminum trichloride (3.19 g,24 mmol) and anhydrous 1, 2-dichloroethane (16 mL) are added into a 100mL round bottom flask, anhydrous 1, 2-dichloroethane (16 mL) is dissolved 2-methoxy-9-propyl-9H-carbazole-3-formaldehyde (3.2 g,12 mmol) and then added dropwise into the round bottom flask, stirring is carried out for 10H at room temperature, after the reaction is finished, 2M HCl solution (12 mL) is added dropwise and stirring is continued for 0.5H, and 1, 2-dichloroethane in the reaction solution is evaporated under reduced pressure. Ethyl acetate (30 ml×3) was extracted, the organic phases were combined and washed with saturated NaCl solution (60 ml×2), anhydrous sodium sulfate was added and left to stand for 15min, and ethyl acetate was evaporated under reduced pressure to give a crude product. EA: pe=1: 5 as mobile phase, the crude product was purified by column chromatography to give 0.97g of 2-hydroxy-9-propyl-9H-carbazole-3-carbaldehyde as yellow solid with a yield of 31.9%.
(d) Preparation of 2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde
In a 100mL two-necked flask, 2-hydroxy-9-propyl-9H-carbazole-3-carbaldehyde (1.77 g,7 mmol), 4-dimethylaminopyridine (0.85 g,7 mmol), triethylamine (1.41 g,14 mmol) and 1, 2-dichloroethane (10 mL) were added, stirred and protected with argon, t-butyldimethylchlorosilane (2.11 g,14 mmol) was dissolved in 1, 2-dichloroethane (10 mL), then the mixture was added dropwise to the two-necked flask at-10℃and stirred for 9 hours, saturated sodium carbonate solution (20 mL, purified water) was added to the reaction mixture, extracted with dichloromethane (20 mL. Times.3), the organic phase was combined and washed with saturated NaCl solution (40 mL. Times.2, purified water), anhydrous sodium sulfate was added and allowed to stand for 15min, and dichloromethane was evaporated under reduced pressure to give a crude product. EA: pe=1: 20 as mobile phase, the crude product was purified by column chromatography to give 0.79g of 2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde as yellow solid in 30.8% yield.
(e) Preparation of 3- ((2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazol-3-yl) 2- (pyridin-2-yl) acrylonitrile
2- ((tert-Butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde (0.74 g,2 mmol), 2-cyanopyridine (0.29 g,4 mmol), tetrahydropyrrole (0.0071 g,0.1 mmol) and anhydrous THF (8 mL) were added to a 100mL round bottom flask, and the mixture was stirred at room temperature under argon for 10H, and the anhydrous THF in the reaction solution was evaporated under reduced pressure to give a crude product. EA: pe=1: 10 as mobile phase, the crude product was purified by column chromatography to give 0.2g of 3- ((2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazol-3-yl) 2- (pyridin-2-yl) acrylonitrile as a yellow-green solid in 20.9% yield.
Example 2
The preparation method of the pyridine ring-containing aromatic acrylonitrile carbazole type reaction type fluoride ion fluorescent probe comprises the following steps:
(a) Preparation of 2-methoxy-9-propyl-9H-carbazole
In a 100mL round bottom flask, 2-methoxy-9H-carbazole (4.93 g,25 mmol) and sodium hydroxide (5 g,125 mmol) were added and dissolved in DMSO (50 mL) and iodopropane (10.6 g,62.5 mmol) was added with stirring. After all the raw materials are added, a condensing tube and an Ar balloon are placed above the round-bottom flask, the vacuum is pumped for three times, and the temperature is raised to 60 ℃ for reaction for 16 hours. After the reaction was completed, cooled to room temperature and diluted with 50mL of water, dichloromethane (50 ml×3) was added for extraction, and after the organic phase was combined and washed with saturated NaCl solution (80 ml×2), anhydrous sodium sulfate was added for standing for 15min, and the dichloromethane was dried under reduced pressure to give crude product, which was subjected to column chromatography (PE: ea=10:1) to give 5.11g of 2-methoxy-9-propyl-9H-carbazole as a white solid, yield 85.4%.
(b) Preparation of 2-methoxy-9-propyl-9H-carbazole-3-carbaldehyde
In a 250mL three-necked flask, anhydrous DMF (8 mL) was added first, phosphorus oxychloride (3.83 g,25 mmol) was added dropwise to the three-necked flask under the protection of argon at 0℃and stirred for 15min, 2-methoxy-9-propyl-9H-carbazole (2.4 g,10 mmol) was dissolved in anhydrous DMF (16 mL) and then added dropwise to the three-necked flask, the temperature was raised to 60℃and stirred for 4H, ice water (25 mL) was added after the reaction solution cooled to room temperature and stirring was continued for 0.5H, extraction was performed with dichloromethane (25 mL. Times.3 mL), the organic phases were combined and washed with saturated NaCl solution, anhydrous sodium sulfate was added and allowed to stand for 15min, and the dichloromethane was evaporated under reduced pressure to obtain a crude product. EA: pe=1: 5 as mobile phase, the crude product was purified by column chromatography to give 2.06g of 2-methoxy-9-propyl-9H-carbazole-3-carbaldehyde as yellow solid in 77.4% yield.
(c) Preparation of 2-hydroxy-9-propyl-9H-carbazole-3-carbaldehyde
Anhydrous aluminum trichloride (3.99 g,30 mmol) and anhydrous 1, 2-dichloroethane (14 mL) are added into a 100mL round-bottomed flask, anhydrous 1, 2-dichloroethane (13 mL) is dissolved 2-methoxy-9-propyl-9H-carbazole-3-carbaldehyde (2.67 g,10 mmol) and then added dropwise into the round-bottomed flask, stirring is carried out at room temperature for 10H, after the reaction is completed, 2M HCl solution (15 mL) is added dropwise and stirring is continued for 0.5H, and 1, 2-dichloroethane in the reaction solution is evaporated under reduced pressure. Ethyl acetate (25 mL x 3) was extracted, the organic phases were combined and washed with saturated NaCl solution (50 mL x 2), anhydrous sodium sulfate was added and left to stand for 15min, and ethyl acetate was evaporated under reduced pressure to give a crude product. EA: pe=1: 5 as mobile phase, the crude product was purified by column chromatography to give 0.88g of 2-hydroxy-9-propyl-9H-carbazole-3-carbaldehyde as yellow solid with a yield of 34.7%.
(d) Preparation of 2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde
In a 100mL two-necked flask, 2-hydroxy-9-propyl-9H-carbazole-3-carbaldehyde (1.52 g,6 mmol), 4-dimethylaminopyridine (0.73 g,6 mmol), diethylamine (0.88 g,12 mmol) and 1, 2-dichloroethane (7 mL) were added, stirred and protected with argon, t-butyldimethylchlorosilane (1.81 g,12 mmol) was dissolved in 1, 2-dichloroethane (8 mL), then the mixture was added dropwise to the two-necked flask at-10℃and stirred for 9 hours, saturated sodium carbonate solution (15 mL, pure water) was added to the reaction mixture, extracted with dichloromethane (15 mL. Times.3), the organic phase was combined and washed with saturated NaCl solution (30 mL. Times.2, pure water), anhydrous sodium sulfate was added and allowed to stand for 15 minutes, and dichloromethane was evaporated under reduced pressure to give a crude product. EA: pe=1: 20 as mobile phase, the crude product was purified by column chromatography to give 0.71g of 2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde as a yellow solid in 32% yield.
(e) Preparation of 3- ((2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazol-3-yl) 2- (pyridin-2-yl) acrylonitrile
2- ((tert-Butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde (1.1 g,3 mmol), 2-cyanopyridine (0.53 g,7.5 mmol), tetrahydropyrrole (0.0107 g,0.15 mmol) and anhydrous THF (11 mL) were added to a 100mL round bottom flask, and the reaction was stirred at room temperature under argon for 12H, and the anhydrous THF in the reaction solution was evaporated under reduced pressure to give the crude product. EA: pe=1: 10 as mobile phase, the crude product was purified by column chromatography to give 0.3g of 3- ((2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazol-3-yl) 2- (pyridin-2-yl) acrylonitrile as a yellow-green solid in 21.3% yield.
Example 3
The preparation method of the pyridine ring-containing aromatic acrylonitrile carbazole type reaction type fluoride ion fluorescent probe comprises the following steps:
(a) Preparation of 2-methoxy-9-propyl-9H-carbazole
In a 100mL round bottom flask, 2-methoxy-9H-carbazole (5.91 g,30 mmol) and potassium hydroxide (6.72 g,120 mmol) were added and dissolved in DMSO (60 mL) and iodopropane (15.3 g,90 mmol) was added with stirring. After all the raw materials are added, a condensing tube and an Ar balloon are placed above the round-bottom flask, the vacuum is pumped for three times, and the temperature is raised to 65 ℃ for reaction for 14 hours. After the reaction was completed, cooled to room temperature and diluted with 60mL of water, dichloromethane (60 ml×3) was added for extraction, and after the organic phase was combined and washed with saturated NaCl solution (100 ml×2), anhydrous sodium sulfate was added for standing for 15min, and the dichloromethane was dried under reduced pressure to give crude product, which was subjected to column chromatography (PE: ea=10:1) to give 6.43g of 2-methoxy-9-propyl-9H-carbazole as a white solid, yield 89.6%.
(b) Preparation of 2-methoxy-9-propyl-9H-carbazole-3-carbaldehyde
In a 250mL three-necked flask, anhydrous DMF (9 mL) was added first, phosphorus oxychloride (6.89 g,45 mmol) was added dropwise to the three-necked flask under the protection of argon at 0℃and stirred for 15min, 2-methoxy-9-propyl-9H-carbazole (3.59 g,15 mmol) was dissolved in anhydrous DMF (26 mL) and then added dropwise to the three-necked flask, the temperature was raised to 65℃and stirred for 3H, ice water (60 mL) was added after the reaction solution cooled to room temperature and stirring was continued for 0.5H, extraction was performed with dichloromethane (60 mL. Times.3 mL), the organic phases were combined and washed with saturated NaCl solution, anhydrous sodium sulfate was added and allowed to stand for 15min, and dichloromethane was evaporated under reduced pressure to obtain a crude product. EA: pe=1: 5 as mobile phase, the crude product was purified by column chromatography to give 3.28g of 2-methoxy-9-propyl-9H-carbazole-3-carbaldehyde as yellow solid with a yield of 82.1%.
(c) Preparation of 2-hydroxy-9-propyl-9H-carbazole-3-carbaldehyde
Anhydrous aluminum trichloride (4.79 g,36 mmol) and anhydrous dichloromethane (16 mL) are added into a 100mL round-bottomed flask, anhydrous dichloromethane (16 mL) is added dropwise into the round-bottomed flask after dissolving 2-methoxy-9-propyl-9H-carbazole-3-formaldehyde (3.2 g,12 mmol), stirring is carried out at room temperature for 12H, after the reaction is finished, 2M HCl solution (18 mL) is added dropwise for continuous stirring for 0.5H, and dichloromethane in the reaction solution is evaporated under reduced pressure. Ethyl acetate (30 ml×3) was extracted, the organic phases were combined and washed with saturated NaCl solution (60 ml×2), anhydrous sodium sulfate was added and left to stand for 15min, and ethyl acetate was evaporated under reduced pressure to give a crude product. EA: pe=1: 5 as mobile phase, the crude product was purified by column chromatography to give 1.38g of 2-hydroxy-9-propyl-9H-carbazole-3-carbaldehyde as yellow solid with a yield of 45.6%.
(d) Preparation of 2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde
In a 100mL two-necked flask, 2-hydroxy-9-propyl-9H-carbazole-3-carbaldehyde (1.77 g,7 mmol), 4-dimethylaminopyridine (0.85 g,7 mmol), triethylamine (1.41 g,14 mmol) and methylene chloride (10 mL) were added, stirred and protected with argon, t-butyldimethylchlorosilane (3.16 g,21 mmol) was dissolved in methylene chloride (10 mL), then the mixture was added dropwise to the two-necked flask at-10℃with stirring, the reaction was stirred for 9 hours, a saturated sodium carbonate solution (20 mL, prepared with pure water) was added to the reaction solution, extracted with methylene chloride (20 mL. Times.3), the organic phase was combined and washed with a saturated NaCl solution (40 mL. Times.2, prepared with pure water), and the mixture was allowed to stand for 15min with anhydrous sodium sulfate, and the methylene chloride was evaporated under reduced pressure to give a crude product. EA: pe=1: 20 as mobile phase, the crude product was purified by column chromatography to give 0.88g of 2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde as yellow solid in 34.2% yield.
(e) Preparation of 3- ((2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazol-3-yl) 2- (pyridin-2-yl) acrylonitrile
2- ((tert-Butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde (0.73 g,2 mmol), 2-cyanopyridine (0.43 g,6 mmol), tetrahydropyrrole (0.0071 g,0.1 mmol) and anhydrous THF (8 mL) were added to a 100mL round bottom flask, and the mixture was stirred at room temperature under argon for 14H, and the anhydrous THF in the reaction solution was evaporated under reduced pressure to give a crude product. EA: pe=1: 10 as mobile phase, the crude product was purified by column chromatography to give 0.22g of 3- ((2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazol-3-yl) 2- (pyridin-2-yl) acrylonitrile as a yellow-green solid in 23.7% yield.
Example 4
The preparation method of the pyridine ring-containing aromatic acrylonitrile carbazole type reaction type fluoride ion fluorescent probe comprises the following steps:
(a) Preparation of 2-methoxy-9-propyl-9H-carbazole
In a 100mL round bottom flask, 2-methoxy-9H-carbazole (4.93 g,25 mmol) and potassium hydroxide (2.8 g,50 mmol) were added and dissolved in DMSO (50 mL) and iodopropane (14.9 g,87.5 mmol) was added with stirring. After all the raw materials are added, a condensing tube and an Ar balloon are placed above the round-bottom flask, the temperature is raised to 70 ℃ for reaction for 10 hours after three times of vacuumizing. After the reaction was completed, cooled to room temperature and diluted with 50mL of water, dichloromethane (50 ml×3) was added for extraction, and after the organic phase was combined and washed with saturated NaCl solution (80 ml×2), anhydrous sodium sulfate was added for standing for 15min, and the dichloromethane was dried under reduced pressure to give crude product, which was subjected to column chromatography (PE: ea=10:1) to give 5.46g of 2-methoxy-9-propyl-9H-carbazole as a white solid, yield 91.3%.
(b) Preparation of 2-methoxy-9-propyl-9H-carbazole-3-carbaldehyde
In a 250mL three-necked flask, anhydrous DMF (8 mL) was added first, phosphorus oxychloride (5.36 g,35 mmol) was added dropwise to the three-necked flask at 0℃under argon protection, stirred for 15min, 2-methoxy-9-propyl-9H-carbazole (2.39 g,10 mmol) was dissolved in anhydrous DMF (16 mL) and then added dropwise to the three-necked flask, the temperature was raised to 55℃and stirred for 5H, ice water (25 mL) was added after the reaction solution cooled to room temperature, stirring was continued for 0.5H, extraction was performed with dichloromethane (25 mL. Times.3 mL), the organic phase was combined and saturated NaCl solution (50 mL. Times.2 mL) was washed, anhydrous sodium sulfate was added and allowed to stand for 15min, and dichloromethane was evaporated under reduced pressure to give a crude product. EA: pe=1: 5 as mobile phase, the crude product was purified by column chromatography to give 2.14g of 2-methoxy-9-propyl-9H-carbazole-3-carbaldehyde as yellow solid with a yield of 80.1%.
(c) Preparation of 2-hydroxy-9-propyl-9H-carbazole-3-carbaldehyde
Anhydrous aluminum trichloride (3.99 g,30 mmol) and anhydrous 1, 1-dichloroethane (14 mL) are added into a 100mL round-bottomed flask, anhydrous 1, 1-dichloroethane (13 mL) is dissolved 2-methoxy-9-propyl-9H-carbazole-3-carbaldehyde (2.7 g,10 mmol) and then added dropwise into the round-bottomed flask, stirring is carried out at room temperature for 14H, after the reaction is completed, 2M HCl solution (15 mL) is added dropwise and stirring is continued for 0.5H, and 1, 1-dichloroethane in the reaction solution is evaporated under reduced pressure. Ethyl acetate (25 mL x 3) was extracted, the organic phases were combined and washed with saturated NaCl solution (50 mL x 2), anhydrous sodium sulfate was added and left to stand for 15min, and ethyl acetate was evaporated under reduced pressure to give a crude product. EA: pe=1: 5 as mobile phase, the crude product was purified by column chromatography to give 1.18g of 2-hydroxy-9-propyl-9H-carbazole-3-carbaldehyde as yellow solid with a yield of 46.8%.
(d) Preparation of 2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde
In a 100mL two-necked flask, 2-hydroxy-9-propyl-9H-carbazole-3-carbaldehyde (1.52 g,6 mmol), 4-dimethylaminopyridine (0.88 g,7.2 mmol), triethylamine (1.82 g,18 mmol) and 1, 1-dichloroethane (7 mL) were added, stirred and protected with argon, t-butyldimethylchlorosilane (2.71 g,18 mmol) was dissolved in 1, 1-dichloroethane (8 mL), then the mixture was added dropwise to the flask at-10℃and stirred for 12 hours, a saturated sodium carbonate solution (15 mL, prepared with pure water) was added to the reaction mixture, extracted with dichloromethane (15 mL. Times.3), the organic phases were combined and washed with a saturated NaCl solution (30 mL. Times.2, prepared with pure water), and anhydrous sodium sulfate was added and allowed to stand for 15 minutes, and the dichloromethane was evaporated under reduced pressure to give a crude product. EA: pe=1: 20 as mobile phase, the crude product was purified by column chromatography to give 0.8g of 2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde as yellow solid in 36.4% yield.
(e) Preparation of 3- ((2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazol-3-yl) 2- (pyridin-2-yl) acrylonitrile
2- ((tert-Butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde (1.1 g,3 mmol), 2-cyanopyridine (0.43 g,6 mmol), tetrahydropyrrole (0.0107 g,0.15 mmol) and anhydrous THF (11 mL) were added to a 100mL round bottom flask, and the mixture was stirred at room temperature under argon for 12H, and the anhydrous THF in the reaction solution was evaporated under reduced pressure to give a crude product. EA: pe=1: 10 as mobile phase, the crude product was purified by column chromatography to give 0.28g of 3- ((2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazol-3-yl) 2- (pyridin-2-yl) acrylonitrile as a yellow-green solid in 20.1% yield.
Example 5
The preparation method of the pyridine ring-containing aromatic acrylonitrile carbazole type reaction type fluoride ion fluorescent probe comprises the following steps:
(a) Preparation of 2-methoxy-9-propyl-9H-carbazole
In a 100mL round bottom flask, 2-methoxy-9H-carbazole (5.91 g,30 mmol) and potassium hydroxide (8.4 g,150 mmol) were added and dissolved in DMF (60 mL) and iodopropane (20.4 g,120 mmol) was added with stirring. After all the raw materials are added, a condensing tube and an Ar balloon are placed above the round-bottom flask, the vacuum is pumped for three times, and the temperature is raised to 60 ℃ for reaction for 16 hours. After the reaction was completed, cooled to room temperature and diluted with 60mL of water, dichloromethane (60 ml×3) was added for extraction, and after the organic phase was combined and washed with saturated NaCl solution (100 ml×2), anhydrous sodium sulfate was added for standing for 15min, and the dichloromethane was dried under reduced pressure to give crude product, which was subjected to column chromatography (PE: ea=10:1) to give 6.6g of 2-methoxy-9-propyl-9H-carbazole as a white solid in 92% yield.
(b) Preparation of 2-methoxy-9-propyl-9H-carbazole-3-carbaldehyde
In a 250mL three-necked flask, anhydrous DMF (9 mL) was added first, phosphorus oxychloride (9.18 g,60 mmol) was added dropwise to the three-necked flask at 0℃under argon protection, stirred for 15min, 2-methoxy-9-propyl-9H-carbazole (3.6 g,15 mmol) was dissolved in anhydrous DMF (26 mL) and then added dropwise to the three-necked flask, the temperature was raised to 65℃and stirred for 3H, ice water (40 mL) was added after the reaction solution cooled to room temperature, stirring was continued for 0.5H, extraction was performed with dichloromethane (40 mL. Times.3 mL), the organic phase was combined and washed with saturated NaCl solution (60 mL. Times.2 mL), anhydrous sodium sulfate was added and allowed to stand for 15min, and dichloromethane was evaporated under reduced pressure to give a crude product. EA: pe=1: 5 as mobile phase, the crude product was purified by column chromatography to give 3.34g of 2-methoxy-9-propyl-9H-carbazole-3-carbaldehyde as yellow solid with a yield of 83.4%.
(c) Preparation of 2-hydroxy-9-propyl-9H-carbazole-3-carbaldehyde
Anhydrous aluminum trichloride (6.38 g,48 mmol) and anhydrous dichloromethane (16 mL) are added into a 100mL round-bottomed flask, 2-methoxy-9-propyl-9H-carbazole-3-formaldehyde (3.2 g,12 mmol) is dissolved in anhydrous dichloromethane (16 mL), then the mixture is dropwise added into the round-bottomed flask, stirring is carried out at room temperature for 10H, after the reaction is finished, 2M HCl solution (24 mL) is dropwise added, stirring is continued for 0.5H, and dichloromethane in the reaction solution is evaporated under reduced pressure. Ethyl acetate (30 ml×3) was extracted, the organic phases were combined and washed with saturated NaCl solution (60 ml×2), anhydrous sodium sulfate was added and left to stand for 15min, and ethyl acetate was evaporated under reduced pressure to give a crude product. EA: pe=1: 5 as mobile phase, the crude product was purified by column chromatography to give 1.2g of 2-hydroxy-9-propyl-9H-carbazole-3-carbaldehyde as yellow solid with a yield of 39.6%.
(d) Preparation of 2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde
In a 100mL two-necked flask, 2-hydroxy-9-propyl-9H-carbazole-3-carbaldehyde (1.77 g,7 mmol), 4-dimethylaminopyridine (1.37 g,11.2 mmol), diethylamine (2.3 g,32 mmol) and 1, 2-dichloroethane (10 mL) were added, stirred and protected with argon, t-butyldimethylchlorosilane (3.16 g,21 mmol) was dissolved in 1, 2-dichloroethane (10 mL), then added dropwise to the flask at-10℃under stirring, the reaction was stirred for 14 hours, saturated sodium carbonate solution (20 mL, purified water) was added to the reaction solution, extracted with dichloromethane (20 mL. Times.3), the organic phases were combined and washed with saturated NaCl solution (40 mL. Times.2, purified water), anhydrous sodium sulfate was added and allowed to stand for 15min, and the dichloromethane was evaporated under reduced pressure to give a crude product. EA: pe=1: 20 as mobile phase, the crude product was purified by column chromatography to give 0.99g of 2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde as yellow solid in 38.6% yield.
(e) Preparation of 3- ((2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazol-3-yl) 2- (pyridin-2-yl) acrylonitrile
2- ((tert-Butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde (0.73 g,2 mmol), 2-cyanopyridine (0.36 g,5 mmol), tetrahydropyrrole (0.0071 g,0.1 mmol) and anhydrous THF (8 mL) were added to a 100mL round bottom flask, and the mixture was stirred at room temperature under argon for 12H, and the anhydrous THF in the reaction solution was evaporated under reduced pressure to give a crude product. EA: pe=1: 10 as mobile phase, the crude product was purified by column chromatography to give 0.21g of 3- ((2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazol-3-yl) 2- (pyridin-2-yl) acrylonitrile as a yellow-green solid in 22.5% yield.
Example 6
The preparation method of the pyridine ring-containing aromatic acrylonitrile carbazole type reaction type fluoride ion fluorescent probe comprises the following steps:
(a) Preparation of 2-methoxy-9-propyl-9H-carbazole
In a 100mL round bottom flask, 2-methoxy-9H-carbazole (4.93 g,25 mmol) and sodium hydroxide (2.5 g,62.5 mmol) were added and dissolved in DMSO (50 mL) and iodopropane (19.1 g,112.5 mmol) was added with stirring. After all the raw materials are added, a condensing tube and an Ar balloon are placed above the round-bottom flask, the temperature is raised to 70 ℃ for reaction for 10 hours after three times of vacuumizing. After the reaction was completed, cooled to room temperature and diluted with 60mL of water, dichloromethane (60 ml×3) was added for extraction, and after the organic phase was combined and washed with saturated NaCl solution (100 ml×2), anhydrous sodium sulfate was added for standing for 15min, and the dichloromethane was dried under reduced pressure to give crude product, which was subjected to column chromatography (PE: ea=10:1) to give 5.54g of 2-methoxy-9-propyl-9H-carbazole as a white solid, yield 92.7%.
(b) Preparation of 2-methoxy-9-propyl-9H-carbazole-3-carbaldehyde
In a 250mL three-necked flask, anhydrous DMF (9 mL) was added first, phosphorus oxychloride (6.9 g,45 mmol) was added dropwise to the three-necked flask under the protection of argon at 0℃and stirred for 15min, 2-methoxy-9-propyl-9H-carbazole (2.4 g,10 mmol) was dissolved in anhydrous DMF (16 mL) and then added dropwise to the three-necked flask, the temperature was raised to 55℃and stirred for 5H, ice water (25 mL) was added after the reaction solution cooled to room temperature and stirring was continued for 0.5H, extraction was performed with dichloromethane (25 mL. Times.3 mL), the organic phases were combined and saturated NaCl solution (50 mL. Times.2 mL) was washed, anhydrous sodium sulfate was added and allowed to stand for 15min, and dichloromethane was evaporated under reduced pressure to give a crude product. EA: pe=1: 5 as mobile phase, the crude product was purified by column chromatography to give 2.15g of 2-methoxy-9-propyl-9H-carbazole-3-carbaldehyde as yellow solid with a yield of 80.7%.
(c) Preparation of 2-hydroxy-9-propyl-9H-carbazole-3-carbaldehyde
Anhydrous aluminum trichloride (5.32 g,40 mmol) and anhydrous 1, 2-dichloroethane (14 mL) are added into a 100mL round-bottomed flask, anhydrous 1, 2-dichloroethane (13 mL) is dissolved 2-methoxy-9-propyl-9H-carbazole-3-carbaldehyde (2.67 g,10 mmol) and then added dropwise into the round-bottomed flask, stirring is carried out at room temperature for 12H, after the reaction is completed, 2M HCl solution (20 mL) is added dropwise and stirring is continued for 0.5H, and 1, 2-dichloroethane in the reaction solution is evaporated under reduced pressure. Ethyl acetate (25 mL x 3) was extracted, the organic phases were combined and washed with saturated NaCl solution (50 mL x 2), anhydrous sodium sulfate was added and left to stand for 15min, and ethyl acetate was evaporated under reduced pressure to give a crude product. EA: pe=1: 5 as mobile phase, the crude product was purified by column chromatography to give 1.05g of 2-hydroxy-9-propyl-9H-carbazole-3-carbaldehyde as yellow solid with a yield of 41.5%.
(d) Preparation of 2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde
In a 100mL two-necked flask, 2-hydroxy-9-propyl-9H-carbazole-3-carbaldehyde (1.52 g,6 mmol), 4-dimethylaminopyridine (0.73 g,6 mmol), triethylamine (1.21 g,12 mmol) and methylene chloride (7 mL) were added, stirred and protected with argon, tert-butyldimethylchlorosilane (4.52 g,30 mmol) was dissolved in methylene chloride (8 mL), then the mixture was added dropwise to the two-necked flask at-10℃with stirring, the reaction was stirred for 12 hours, a saturated sodium carbonate solution (15 mL, prepared with pure water) was added to the reaction solution, extracted with methylene chloride (15 mL. Times.3), the organic phase was combined and washed with a saturated NaCl solution (30 mL. Times.2, prepared with pure water), and anhydrous sodium sulfate was added to stand for 15min, and the methylene chloride was evaporated under reduced pressure to give a crude product. EA: pe=1: 20 as mobile phase, the crude product was purified by column chromatography to give 0.9g of 2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde as yellow solid in 40.8% yield.
(e) Preparation of 3- ((2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazol-3-yl) 2- (pyridin-2-yl) acrylonitrile
2- ((tert-Butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde (1.1 g,3 mmol), 2-cyanopyridine (0.64 g,9 mmol), tetrahydropyrrole (0.0107 g,0.15 mmol) and anhydrous THF (10 mL) were added to a 100mL round bottom flask, and the reaction was stirred at room temperature for 12H under argon, and THF in the reaction solution was evaporated under reduced pressure to give a crude product. EA: pe=1: 10 as mobile phase, the crude product was purified by column chromatography to give 0.33g of 3- ((2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazol-3-yl) 2- (pyridin-2-yl) acrylonitrile as a yellow-green solid in 23.8% yield.
Example 7
The preparation method of the pyridine ring-containing aromatic acrylonitrile carbazole type reaction type fluoride ion fluorescent probe comprises the following steps:
(a) Preparation of 2-methoxy-9-propyl-9H-carbazole
In a 100mL round bottom flask, 2-methoxy-9H-carbazole (5.91 g,30 mmol) and sodium hydroxide (4.8 g,120 mmol) were added and dissolved in DMF (60 mL) and iodopropane (25.5 g,150 mmol) was added with stirring. After all the raw materials are added, a condensing tube and an Ar balloon are placed above the round-bottom flask, the vacuum is pumped for three times, and the temperature is raised to 65 ℃ for reaction for 14 hours. After the reaction was completed, cooled to room temperature and diluted with 60mL of water, dichloromethane (60 ml×3) was added for extraction, and after the organic phase was combined and washed with saturated NaCl solution (100 ml×2), anhydrous sodium sulfate was added for standing for 15min, and the dichloromethane was dried under reduced pressure to give crude product, which was subjected to column chromatography (PE: ea=10:1) to give 6.71g of 2-methoxy-9-propyl-9H-carbazole as a white solid, yield 93.6%.
(b) Preparation of 2-methoxy-9-propyl-9H-carbazole-3-carbaldehyde
In a 250mL three-necked flask, anhydrous DMF (9 mL) was added first, phosphorus oxychloride (11.48 g,75 mmol) was added dropwise to the three-necked flask at 0℃under argon protection, stirred for 15min, 2-methoxy-9-propyl-9H-carbazole (3.59 g,15 mmol) was dissolved in anhydrous DMF (26 mL) and then added dropwise to the three-necked flask, the temperature was raised to 55℃and stirred for 5H, ice water (35 mL) was added after the reaction solution cooled to room temperature, stirring was continued for 0.5H, extraction was performed with dichloromethane (35 mL. Times.3 mL), the organic phase was combined and washed with saturated NaCl solution (70 mL. Times.2 mL), anhydrous sodium sulfate was added and allowed to stand for 15min, and dichloromethane was evaporated under reduced pressure to give a crude product. EA: pe=1: 5 as mobile phase, the crude product was purified by column chromatography to give 3.2g of 2-methoxy-9-propyl-9H-carbazole-3-carbaldehyde as yellow solid with 80% yield.
(c) Preparation of 2-hydroxy-9-propyl-9H-carbazole-3-carbaldehyde
Anhydrous aluminum trichloride (6.65 g,50 mmol) and anhydrous dichloromethane (14 mL) are added into a 100mL round-bottomed flask, anhydrous dichloromethane (13 mL) is added dropwise into the round-bottomed flask after 2-methoxy-9-propyl-9H-carbazole-3-formaldehyde (2.67 g,10 mmol) is dissolved, stirring is carried out at room temperature for 14H, after the reaction is finished, 2M HCl solution (25 mL) is added dropwise for continuous stirring for 0.5H, and dichloromethane in the reaction solution is evaporated under reduced pressure. Ethyl acetate (25 mL x 3) was extracted, the organic phases were combined and washed with saturated NaCl solution (60 mL x 2), anhydrous sodium sulfate was added and left to stand for 15min, and ethyl acetate was evaporated under reduced pressure to give a crude product. EA: pe=1: 5 as mobile phase, the crude product was purified by column chromatography to give 1.17g of 2-hydroxy-9-propyl-9H-carbazole-3-carbaldehyde as yellow solid with a yield of 46.3%.
(d) Preparation of 2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde
In a 100mL two-necked flask, 2-hydroxy-9-propyl-9H-carbazole-3-carbaldehyde (1.77 g,7 mmol), 4-dimethylaminopyridine (1.02 g,8.4 mmol), triethylamine (2.12 g,21 mmol) and methylene chloride (7 mL) were added, stirred and protected with argon, t-butyldimethylsilyl chloride (5.27 g,35 mmol) was dissolved in methylene chloride (8 mL), then the mixture was added dropwise to the two-necked flask at-10℃and stirred for 14 hours, a saturated sodium carbonate solution (20 mL, prepared with pure water) was added to the reaction mixture, extracted with methylene chloride (20 mL. Times.3), the organic phase was combined and washed with a saturated NaCl solution (40 mL. Times.2, prepared with pure water), and dried sodium sulfate was added and allowed to stand for 15 minutes, and the methylene chloride was evaporated under reduced pressure to give a crude product. EA: pe=1: 20 as mobile phase, the crude product was purified by column chromatography to give 1.11g of 2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde as yellow solid in 43% yield.
(e) Preparation of 3- ((2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazol-3-yl) 2- (pyridin-2-yl) acrylonitrile
2- ((tert-Butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde (0.73 g,2 mmol), 2-cyanopyridine (0.36 g,5 mmol), tetrahydropyrrole (0.0057 g,0.08 mmol) and anhydrous THF (8 mL) were added to a 100mL round bottom flask, and the mixture was stirred at room temperature under argon for 12H, and the anhydrous THF in the reaction solution was evaporated under reduced pressure to give a crude product. EA: pe=1: 10 as mobile phase, the crude product was purified by column chromatography to give 0.2g of 3- ((2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazol-3-yl) 2- (pyridin-2-yl) acrylonitrile as a yellow-green solid in 21.3% yield.
Example 8
The preparation method of the pyridine ring-containing aromatic acrylonitrile carbazole type reaction type fluoride ion fluorescent probe comprises the following steps:
(a) Preparation of 2-methoxy-9-propyl-9H-carbazole
In a 100mL round bottom flask, 2-methoxy-9H-carbazole (4.93 g,25 mmol) and sodium hydroxide (4 g,100 mmol) were added and dissolved in DMSO (50 mL) and iodopropane (12.8 g,75 mmol) was added with stirring. After all the raw materials are added, a condensing tube and an Ar balloon are placed above the round-bottom flask, the vacuum is pumped for three times, and the temperature is raised to 60 ℃ for reaction for 16 hours. After the reaction was completed, cooled to room temperature and diluted with 50mL of water, dichloromethane (50 ml×3) was added for extraction, and after the organic phase was combined and washed with saturated NaCl solution (80 ml×2), anhydrous sodium sulfate was added for standing for 15min, and the dichloromethane was dried under reduced pressure to give crude product, which was subjected to column chromatography (PE: ea=10:1) to give 5.24g of 2-methoxy-9-propyl-9H-carbazole as a white solid, yield 87.7%.
(b) Preparation of 2-methoxy-9-propyl-9H-carbazole-3-carbaldehyde
In a 250mL three-necked flask, anhydrous DMF (8 mL) was added first, phosphorus oxychloride (6.12 g,40 mmol) was added dropwise to the three-necked flask at 0℃under argon protection, stirred for 15min, 2-methoxy-9-propyl-9H-carbazole (2.39 g,10 mmol) was dissolved in anhydrous DMF (16 mL) and then added dropwise to the three-necked flask, the temperature was raised to 60℃to stir the reaction for 4H, ice water (25 mL) was added after the reaction solution cooled to room temperature, stirring was continued for 0.5H, extraction was performed with dichloromethane (25 mL. Times.3 mL), the organic phase was combined and saturated NaCl solution (50 mL. Times.2 mL) was washed, anhydrous sodium sulfate was added and allowed to stand for 15min, and dichloromethane was evaporated under reduced pressure to give a crude product. EA: pe=1: 5 as mobile phase, the crude product was purified by column chromatography to give 2.22g of 2-methoxy-9-propyl-9H-carbazole-3-carbaldehyde as yellow solid with a yield of 83.3%.
(c) Preparation of 2-hydroxy-9-propyl-9H-carbazole-3-carbaldehyde
Anhydrous aluminum trichloride (7.98 g,60 mmol) and anhydrous 1, 1-dichloroethane (16 mL) are added into a 100mL round-bottomed flask, anhydrous 1, 1-dichloroethane (16 mL) is dissolved 2-methoxy-9-propyl-9H-carbazole-3-carbaldehyde (3.2 g,12 mmol) and then added dropwise into the round-bottomed flask, stirring is carried out at room temperature for 12H, after the reaction is completed, 2M HCl solution (30 mL) is added dropwise and stirring is continued for 0.5H, and 1, 1-dichloroethane in the reaction solution is evaporated under reduced pressure. Ethyl acetate (30 ml×3) was extracted, the organic phases were combined and washed with saturated NaCl solution (60 ml×2), anhydrous sodium sulfate was added and left to stand for 15min, and ethyl acetate was evaporated under reduced pressure to give a crude product. EA: pe=1: 5 as mobile phase, the crude product was purified by column chromatography to give 1.37g of 2-hydroxy-9-propyl-9H-carbazole-3-carbaldehyde as yellow solid with a yield of 45.2%.
(d) Preparation of 2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde
In a 100mL two-necked flask, 2-hydroxy-9-propyl-9H-carbazole-3-carbaldehyde (1.52 g,6 mmol), 4-dimethylaminopyridine (1.17 g,9.6 mmol), triethylamine (3.03 g,30 mmol) and methylene chloride (7 mL) were added, stirred and protected with argon, and tert-butyldimethylsilyl chloride (4.52 g,30 mmol) was dissolved in methylene chloride (8 mL), then the mixture was added dropwise to the two-necked flask at-10℃and reacted with stirring for 10 hours, a saturated sodium carbonate solution (15 mL, prepared with pure water) was added, extracted with methylene chloride (15 mL. Times.3), the organic phase was combined and washed with a saturated NaCl solution (30 mL. Times.2, prepared with pure water), and anhydrous sodium sulfate was added and allowed to stand for 15 minutes, and the methylene chloride was evaporated under reduced pressure to give a crude product. EA: pe=1: 20 as mobile phase, the crude product was purified by column chromatography to give 0.93g of 2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde as yellow solid in 42.2% yield.
(e) Preparation of 3- ((2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazol-3-yl) 2- (pyridin-2-yl) acrylonitrile
2- ((tert-Butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde (1.1 g,3 mmol), 2-cyanopyridine (0.53 g,7.5 mmol), tetrahydropyrrole (0.0128 g,0.18 mmol) and anhydrous THF (10 mL) were added to a 100mL round bottom flask, and the reaction was stirred at room temperature under argon for 12H, and the anhydrous THF in the reaction solution was evaporated under reduced pressure to give the crude product. EA: pe=1: 10 as mobile phase, the crude product was purified by column chromatography to give 0.3g of 3- ((2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazol-3-yl) 2- (pyridin-2-yl) acrylonitrile as a yellow-green solid in 21.7% yield.
The physical properties and the structure of the 2-methoxy-9-propyl-9H-carbazole obtained in the step (a) are as follows:
MP:100.3-101.7℃； 1 H NMR(400MHz,DMSO-d 6 )δ8.00(d,J＝
8.8Hz,2H),7.52(s,1H),7.34(d,J＝1.4Hz,1H),7.18–7.10(m,2H),6.80(dd,J＝8.4,2.4Hz,1H),4.34(s,2H),3.88(s,3H),1.79(d,J＝7.2Hz,2H),0.88(t,J＝7.6Hz,3H)； 13 C NMR(100MHz,CDCl 3 )δ159.0,141.9,140.7,124.3,123.0,121.1,119.5,118.9,116.8,108.5,106.9,93.3,55.7,44.6,22.2,11.9；ESI-HRMS C 16 H 17 NO([M+H] + ):calcd 240.1382,found 240.1385.
the physical properties and the structure of the 2-methoxy-9-propyl-9H-carbazole-3-formaldehyde obtained in the step (b) are as follows:
MP:104.5-105.9℃； 1 H NMR(400MHz,CDCl 3 )δ10.48(s,1H),8.57(s,1H),8.02(d,J＝7.6Hz,1H),7.49–7.39(m,1H),7.36(d,J＝8.0Hz,1H),7.26(s,1H),6.75(s,1H),4.22(t,J＝7.2Hz,2H),4.03(s,3H),1.93(d,J＝7.2Hz,2H),1.00(t,J＝7.6Hz,3H)； 13 C NMR(100MHz,CDCl 3 )δ189.4,161.6,145.8,141.3,125.6,123.5,121.7,120.4,120.2,118.4,116.9,109.0,90.4,55.9,44.8,22.1,11.8；ESI-HRMSC 17 H 17 NO 2 ([M+H] + ):calcd 268.1331,found 268.1331.
the physical properties and the structure of the 2-hydroxy-9-propyl-9H-carbazole-3-formaldehyde obtained in the step (c) are as follows:
MP:115.1-116.7℃； 1 H NMR(400MHz,CDCl 3 )δ11.52(s,1H),9.91(s,1H),8.14(s,1H),7.99(d,J＝8.8Hz,1H),7.49–7.40(m,1H),7.36(d,J＝8.4Hz,1H),7.30–7.25(m,1H),6.80(s,1H),4.17(t,J＝7.2Hz,2H),1.90(h,J＝7.6Hz,2H),0.98(t,J＝7.6Hz,3H)； 13 C NMR(101MHz,CDCl 3 )δ195.0,161.2,146.4,141.6,127.2,125.9,123.1,120.6,119.8,117.3,114.9,109.2,95.1,44.9,21.9,11.7；ESI-HRMS C 16 H 15 NO 2 ([M+H] + ):calcd 254.1175,found 245.1177.
the physical properties and the structure of the 2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-formaldehyde obtained in the step (d) are as follows:
MP:80.1-81.7℃； 1 H NMR(400MHz,CDCl 3 )δ10.48(s,1H),8.57(s,1H),8.03(d,J＝7.6Hz,1H),7.48–7.40(m,1H),7.36(d,J＝8.0Hz,1H),7.26(s,1H),6.72(s,1H),4.19(t,J＝6.8Hz,2H),1.91(h,J＝7.2Hz,2H),1.07(s,9H),0.98(t,J＝7.6Hz,3H),0.33(s,6H)； 13 CNMR(101MHz,CDCl 3 )δ189.77,158.05,145.77,141.51,125.82,123.46,121.16,120.56,120.35,120.34,118.17,108.99,98.71,44.86,25.80,22.12,18.47,11.91,-4.18；ESI-HRMS C 22 H 29 NO 2 Si([M+H] + ):calcd 368.2040,found 368.2035.
3- ((2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazol-3-yl) 2- (pyridin-2-yl) acrylonitrile obtained by step (e) has the following physical properties and structure:
MP:184.2-185.7℃；IR:υ2952.73,2925.95,2851.57,2202.98,1465.12,1453.22,1360.99,1250.91,1120.00,876.03,751.07cm -1 ； 1 HNMR(400MHz,CDCl 3 )δ9.19(s,1H),9.12(s,1H),8.63(d,J＝4.8Hz,1H),8.15(d,J＝7.6Hz,1H),7.80–7.74(m,2H),7.39(s,1H),7.23(d,J＝4.0Hz,2H),6.84(s,1H),4.22(t,J＝7.2Hz,2H),1.94(h,J＝7.2Hz,2H),1.13(s,9H),1.02(s,3H),0.32(s,6H)； 13 C NMR(101MHz,CDCl 3 )δ154.99,152.52,149.49,143.76,141.73,141.25,136.95,125.43,123.25,122.36,120.60,120.44,120.31,119.99,119.18,118.06,117.65,108.76,105.49,99.14,44.79,25.83,22.21,18.39,11.89,-4.26；ESI-HRMSC 29 H 33 N 3 OSi([M+H] + ):calcd 468.2473,found 468.2465.
fluoride fluorescent probe function test was performed on 3- ((2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazol-3-yl) 2- (pyridin-2-yl) acrylonitrile (SCP) prepared in examples 1-8 above:
test 3- ((2- ((tert-Butyldimethylsilyl) oxy) -9-propyl-9H-carbazol-3-yl) 2- (pyridin-2-yl) Acrylonitrile (SCP) with fluoride ions before and after reaction, changes in fluorescence of EtOH solution, see FIG. 2 in EtOH, spectrum of SCP with fluoride ions and SCP-F fluorescence Spectrum FIG. 2 (a) is excitation spectrum (Ex) and emission spectrum (Em) of SCP and SCP with fluoride ions, [ SCP ]]＝0.2μM，[F - ]The excitation spectrum is measured at a fixed emission wavelength (em=467nm) and the emission spectrum is measured at a fixed excitation wavelength (ex=384 nm), which is consistent with the excitation wavelength of the cyclization product after the reaction with fluorine ions measured in fig. 2 (b), and shows that the cyclization product is generated after the reaction of SCP and fluorine ions, and the reaction mechanism of SCP for recognizing fluorine ions is confirmed. The picture is a solution of SCP or SCP and fluoride ion together, which is photographed under 365nm ultraviolet lamp. It can be seen that at 384nm excitation wavelength, the fluorescence intensity of SCP solution without fluoride ion is weak, while the fluorescence intensity of SCP solution containing fluoride ion is remarkably enhanced, and the solution shows bright blue fluorescence. Experimental results show that in EtOH, the recognition of fluoride ions by probe SCP produces a significant change in fluorescence signal.
The effect of fluoride concentration on the fluorescence intensity of the solution after the reaction was determined by a fluorescence titration experiment, see FIG. 3 for spectra of fluoride (0-12. Mu.M) titrating SCP (0.2. Mu.M). The solvent was EtOH-HEPES/NaOH (50 mm, ph=7.4, 8:2, v/v) (emission wavelength ex=384 nm); FIG. 3a is a fluorescence titration spectrum of fluoride ions (0-12. Mu.M) of SCP (0.2. Mu.M). As can be seen from the graph, SCP (0.2. Mu.M) emits weak fluorescence at 467nm under 384nm monochromatic light excitation, and as fluorine ions are added, the fluorescence emission at 467nm gradually increases, and when the fluorine ion concentration reaches 5.6. Mu.M, the fluorescence intensity increases to saturation. The relationship between the fluorescence intensity of SCP at 467nm and the concentration of fluorine ions is fitted (see FIG. 3 b), and the fluorescence intensity and the concentration of fluorine ions have good linear relationship in the range of 0-5.6 mu M, so that the quantitative detection of fluorine ions can be realized.
The imaging ability of probe SCP on HepG2 cells was examined with an Olympus IX71 inverted fluorescence microscope. See FIG. 4 for fluorescent images of HepG2 cells treated by SCP (1. Mu.M) and fluoride ions (5. Mu.M). Under the Cyan channel, SCP (1 μm) incubated with cells produced a weaker blue fluorescence, which may result from the SCP response to endogenous fluoride ions. Subsequently, the addition of fluoride ions gives the fluorescence a brighter blue color. The experimental result shows that the probe SCP has good cell imaging capability and can detect fluorine ions in cells.
In conclusion, SCP is a ratio type fluorescent probe with wide range and high sensitivity to fluoride ion response, can be used for detecting fluoride ions in cells, and has good application prospect.
Finally, it should be noted that the above embodiments are merely representative examples of the present invention. Obviously, the technical solution of the invention is not limited to the above-described embodiments, but many variations are possible. All modifications directly derived or suggested to one skilled in the art from the present disclosure should be considered as being within the scope of the present invention.
Claims (10)
1. The pyridine ring-containing aromatic acrylonitrile carbazole type reaction type fluoride ion fluorescent probe is characterized by comprising the following specific structural formula:
2. the method for preparing pyridine ring-containing aralkenyl nitrile carbazole-based reactive fluoride ion fluorescent probe as claimed in claim 1, characterized by comprising the following steps:
(a) 2-methoxy-9H-carbazole and iodopropane are subjected to substitution reaction to generate 2-methoxy-9-propyl-9H-carbazole:
the molar ratio of the substitution reagent to the 2-methoxy-9H-carbazole is 2-5:1, the molar ratio of the alkali to the 2-methoxy-9H-carbazole is 2-5:1, and the reaction temperature is 60-70 ℃;
(b) Formylation reaction of 2-methoxy-9-propyl-9H-carbazole with N, N-dimethylformamide and phosphorus oxychloride to generate 2-methoxy-9-propyl-9H-carbazole-3-formaldehyde:
The molar ratio of the reactant to the 2-methoxy-9-propyl-9H-carbazole is 2-5:1, and the reaction temperature is 55-65 ℃;
(c) Hydrolyzing the 2-methoxy-9-propyl-9H-carbazole-3-formaldehyde to generate 2-hydroxy-9-propyl-9H-carbazole-3-formaldehyde:
the feeding mole ratio of the reaction reagent to the 2-methoxy-9-propyl-9H-carbazole-3-formaldehyde is 2-5:1, and the reaction temperature is room temperature;
(d) 2-hydroxy-9-propyl-9H-carbazole-3-formaldehyde and tert-butyl dimethyl chlorosilane undergo substitution reaction to obtain 2- ((tert-butyl dimethyl silicon based) oxy) -9-propyl-9H-carbazole-3-formaldehyde:
the molar ratio of the reaction reagent to the 2-hydroxy-9-propyl-9H-carbazole-3-formaldehyde is 2-5:1, the molar ratio of the catalyst to the 2-hydroxy-9-propyl-9H-carbazole-3-formaldehyde is 1-2:1, the molar ratio of the alkali to the compound 4 is 2-5:1, and the reaction temperature is-10 ℃;
(e) 2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-carbaldehyde was reacted with 2-cyanopyridine to give the target compound 3- ((2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazol-3-yl) 2- (pyridin-2-yl) acrylonitrile:
the feeding molar ratio of the reactant to the 2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-formaldehyde is 2-3:1, the ratio of the catalyst to the 2- ((tert-butyldimethylsilyl) oxy) -9-propyl-9H-carbazole-3-formaldehyde is 0.04-0.06:1, and the reaction temperature is room temperature.
3. The method according to claim 2, wherein the solvent in the step (a) is dimethyl sulfoxide (DMSO) or N, N-Dimethylformamide (DMF), the substitution reagent is iodopropane, and the base is sodium hydroxide or potassium hydroxide.
4. The method according to claim 2, wherein the solvent in the step (b) is anhydrous N, N-Dimethylformamide (DMF), and the reactant is phosphorus oxychloride.
5. The method according to claim 2, wherein the solvent in the step (c) is anhydrous dichloromethane, anhydrous 1, 2-dichloroethane or anhydrous 1, 1-dichloroethane, and the reaction agent is aluminum trichloride.
6. The process according to claim 2, wherein the solvent in step (d) is dichloromethane, 1, 2-dichloroethane or 1, 1-dichloroethane, the reactant is t-butyldimethylchlorosilane, the catalyst is 4-dimethylaminopyridine, and the base is triethylamine or diethylamine.
7. The process of claim 2 wherein in step (e) the solvent is anhydrous Tetrahydrofuran (THF), the reactant is 2-cyanopyridine and the catalyst is tetrahydropyrrole.
8. The use of the pyridine ring-containing aralkenyl nitrile carbazole-based reactive fluoride ion fluorescent probe in preparation of fluoride ion detection reagents.
9. The use according to claim 8, wherein the fluoride ion detection is of a biological sample.
10. The use according to claim 9, wherein the fluoride ion detection in the biological sample is a fluoride ion detection in a blood sample.
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| CN104418875A (en) * | 2013-08-28 | 2015-03-18 | 苏州罗兰生物科技有限公司 | Fluorescent molecular probe for detecting fluoride ions as well as synthesis method and application thereof |
| CN111039972A (en) * | 2019-11-29 | 2020-04-21 | 华南师范大学 | Application of 4-hydroxycarbazole compound as fluorine ion fluorescent probe |
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| CN104418875A (en) * | 2013-08-28 | 2015-03-18 | 苏州罗兰生物科技有限公司 | Fluorescent molecular probe for detecting fluoride ions as well as synthesis method and application thereof |
| CN111039972A (en) * | 2019-11-29 | 2020-04-21 | 华南师范大学 | Application of 4-hydroxycarbazole compound as fluorine ion fluorescent probe |
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| Development of carbazole-based fluorescent probe for highly sensitive application in fluoride ion detection;Dan Li et al.;《Molecular and Biomolecular Spectroscopy》;全文 * |
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