CN116425820A - Preparation and application of targeted nitroreductase fluorescent probe - Google Patents
Preparation and application of targeted nitroreductase fluorescent probe Download PDFInfo
- Publication number
- CN116425820A CN116425820A CN202310401806.0A CN202310401806A CN116425820A CN 116425820 A CN116425820 A CN 116425820A CN 202310401806 A CN202310401806 A CN 202310401806A CN 116425820 A CN116425820 A CN 116425820A
- Authority
- CN
- China
- Prior art keywords
- fluorescent probe
- ntr
- probe
- nitroreductase
- targeted
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 102000004459 Nitroreductase Human genes 0.000 title claims abstract description 88
- 108020001162 nitroreductase Proteins 0.000 title claims abstract description 88
- 239000007850 fluorescent dye Substances 0.000 title claims abstract description 74
- 238000002360 preparation method Methods 0.000 title abstract description 10
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims abstract description 16
- BHQCQFFYRZLCQQ-UHFFFAOYSA-N (3alpha,5alpha,7alpha,12alpha)-3,7,12-trihydroxy-cholan-24-oic acid Natural products OC1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 BHQCQFFYRZLCQQ-UHFFFAOYSA-N 0.000 claims abstract description 8
- 239000004380 Cholic acid Substances 0.000 claims abstract description 8
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 claims abstract description 8
- BHQCQFFYRZLCQQ-OELDTZBJSA-N cholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-OELDTZBJSA-N 0.000 claims abstract description 8
- 229960002471 cholic acid Drugs 0.000 claims abstract description 8
- 235000019416 cholic acid Nutrition 0.000 claims abstract description 8
- KXGVEGMKQFWNSR-UHFFFAOYSA-N deoxycholic acid Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 KXGVEGMKQFWNSR-UHFFFAOYSA-N 0.000 claims abstract description 8
- 229910052757 nitrogen Inorganic materials 0.000 claims abstract description 8
- 238000003384 imaging method Methods 0.000 claims abstract description 4
- 238000000034 method Methods 0.000 claims abstract 2
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 11
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 9
- 230000008685 targeting Effects 0.000 claims description 6
- 238000006243 chemical reaction Methods 0.000 claims description 4
- 238000004440 column chromatography Methods 0.000 claims description 3
- 239000012043 crude product Substances 0.000 claims description 3
- 239000003480 eluent Substances 0.000 claims description 3
- 239000012074 organic phase Substances 0.000 claims description 3
- JHJLBTNAGRQEKS-UHFFFAOYSA-M sodium bromide Chemical class [Na+].[Br-] JHJLBTNAGRQEKS-UHFFFAOYSA-M 0.000 claims description 3
- 239000007787 solid Substances 0.000 claims description 3
- 239000002904 solvent Substances 0.000 claims description 3
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 claims description 2
- 238000001035 drying Methods 0.000 claims 1
- 230000004044 response Effects 0.000 abstract description 9
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 abstract description 5
- 229910052760 oxygen Inorganic materials 0.000 abstract description 5
- 239000001301 oxygen Substances 0.000 abstract description 5
- 230000035945 sensitivity Effects 0.000 abstract description 5
- 102000004190 Enzymes Human genes 0.000 abstract description 4
- 108090000790 Enzymes Proteins 0.000 abstract description 4
- 230000003834 intracellular effect Effects 0.000 abstract description 4
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 abstract description 3
- 150000001413 amino acids Chemical class 0.000 abstract description 3
- 229910021645 metal ion Inorganic materials 0.000 abstract description 3
- 229910052717 sulfur Inorganic materials 0.000 abstract description 3
- 239000011593 sulfur Substances 0.000 abstract description 3
- XTVRPSBYSGATQQ-UHFFFAOYSA-N 1,2-dinitrosodioxidane Chemical compound O=NOON=O XTVRPSBYSGATQQ-UHFFFAOYSA-N 0.000 abstract 1
- RZVAJINKPMORJF-UHFFFAOYSA-N Acetaminophen Chemical compound CC(=O)NC1=CC=C(O)C=C1 RZVAJINKPMORJF-UHFFFAOYSA-N 0.000 abstract 1
- 239000003054 catalyst Substances 0.000 abstract 1
- 239000002994 raw material Substances 0.000 abstract 1
- 239000000523 sample Substances 0.000 description 39
- 210000004027 cell Anatomy 0.000 description 23
- 239000000243 solution Substances 0.000 description 11
- 230000000694 effects Effects 0.000 description 9
- 238000001514 detection method Methods 0.000 description 8
- 206010021143 Hypoxia Diseases 0.000 description 7
- 238000002189 fluorescence spectrum Methods 0.000 description 7
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 description 7
- BOPGDPNILDQYTO-NNYOXOHSSA-N nicotinamide-adenine dinucleotide Chemical compound C1=CCC(C(=O)N)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]2[C@H]([C@@H](O)[C@@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 BOPGDPNILDQYTO-NNYOXOHSSA-N 0.000 description 7
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- 238000010521 absorption reaction Methods 0.000 description 6
- 238000000862 absorption spectrum Methods 0.000 description 4
- 230000009471 action Effects 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 230000005284 excitation Effects 0.000 description 4
- 230000001146 hypoxic effect Effects 0.000 description 4
- 238000012544 monitoring process Methods 0.000 description 4
- 238000003556 assay Methods 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 239000000975 dye Substances 0.000 description 3
- 230000007954 hypoxia Effects 0.000 description 3
- 230000003993 interaction Effects 0.000 description 3
- 210000004185 liver Anatomy 0.000 description 3
- 239000011550 stock solution Substances 0.000 description 3
- 150000003573 thiols Chemical class 0.000 description 3
- 108090000371 Esterases Proteins 0.000 description 2
- 239000007821 HATU Substances 0.000 description 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 2
- 102000004142 Trypsin Human genes 0.000 description 2
- 108090000631 Trypsin Proteins 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 231100000263 cytotoxicity test Toxicity 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 210000000936 intestine Anatomy 0.000 description 2
- 238000011835 investigation Methods 0.000 description 2
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000012588 trypsin Substances 0.000 description 2
- 229910021642 ultra pure water Inorganic materials 0.000 description 2
- 239000012498 ultrapure water Substances 0.000 description 2
- HSINOMROUCMIEA-FGVHQWLLSA-N (2s,4r)-4-[(3r,5s,6r,7r,8s,9s,10s,13r,14s,17r)-6-ethyl-3,7-dihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1h-cyclopenta[a]phenanthren-17-yl]-2-methylpentanoic acid Chemical compound C([C@@]12C)C[C@@H](O)C[C@H]1[C@@H](CC)[C@@H](O)[C@@H]1[C@@H]2CC[C@]2(C)[C@@H]([C@H](C)C[C@H](C)C(O)=O)CC[C@H]21 HSINOMROUCMIEA-FGVHQWLLSA-N 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 108020003264 Cotransporters Proteins 0.000 description 1
- 102000034534 Cotransporters Human genes 0.000 description 1
- AVXURJPOCDRRFD-UHFFFAOYSA-N Hydroxylamine Chemical compound ON AVXURJPOCDRRFD-UHFFFAOYSA-N 0.000 description 1
- 102100021711 Ileal sodium/bile acid cotransporter Human genes 0.000 description 1
- BKAYIFDRRZZKNF-VIFPVBQESA-N N-acetylcarnosine Chemical compound CC(=O)NCCC(=O)N[C@H](C(O)=O)CC1=CN=CN1 BKAYIFDRRZZKNF-VIFPVBQESA-N 0.000 description 1
- 238000005481 NMR spectroscopy Methods 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 108091006611 SLC10A1 Proteins 0.000 description 1
- 108091006614 SLC10A2 Proteins 0.000 description 1
- 102100021988 Sodium/bile acid cotransporter Human genes 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 238000004847 absorption spectroscopy Methods 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- XJHABGPPCLHLLV-UHFFFAOYSA-N benzo[de]isoquinoline-1,3-dione Chemical compound C1=CC(C(=O)NC2=O)=C3C2=CC=CC3=C1 XJHABGPPCLHLLV-UHFFFAOYSA-N 0.000 description 1
- 210000000941 bile Anatomy 0.000 description 1
- 239000003613 bile acid Substances 0.000 description 1
- 108091022863 bile acid binding Proteins 0.000 description 1
- 102000030904 bile acid binding Human genes 0.000 description 1
- 238000012984 biological imaging Methods 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 239000000090 biomarker Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 125000003716 cholic acid group Chemical group 0.000 description 1
- 239000005515 coenzyme Substances 0.000 description 1
- 238000004624 confocal microscopy Methods 0.000 description 1
- 238000002784 cytotoxicity assay Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- DOBMPNYZJYQDGZ-UHFFFAOYSA-N dicoumarol Chemical compound C1=CC=CC2=C1OC(=O)C(CC=1C(OC3=CC=CC=C3C=1O)=O)=C2O DOBMPNYZJYQDGZ-UHFFFAOYSA-N 0.000 description 1
- 229960001912 dicoumarol Drugs 0.000 description 1
- HFCSXCKLARAMIQ-UHFFFAOYSA-L disodium;sulfate;hydrate Chemical compound O.[Na+].[Na+].[O-]S([O-])(=O)=O HFCSXCKLARAMIQ-UHFFFAOYSA-L 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- 210000003494 hepatocyte Anatomy 0.000 description 1
- 208000027866 inflammatory disease Diseases 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 208000028774 intestinal disease Diseases 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 208000028867 ischemia Diseases 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 208000019423 liver disease Diseases 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 229950006238 nadide Drugs 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 238000006862 quantum yield reaction Methods 0.000 description 1
- 238000006462 rearrangement reaction Methods 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 150000003431 steroids Chemical group 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J43/00—Normal steroids having a nitrogen-containing hetero ring spiro-condensed or not condensed with the cyclopenta(a)hydrophenanthrene skeleton
- C07J43/003—Normal steroids having a nitrogen-containing hetero ring spiro-condensed or not condensed with the cyclopenta(a)hydrophenanthrene skeleton not condensed
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1003—Carbocyclic compounds
- C09K2211/1011—Condensed systems
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1029—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1088—Heterocyclic compounds characterised by ligands containing oxygen as the only heteroatom
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- General Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Immunology (AREA)
- Analytical Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Materials Engineering (AREA)
- Optics & Photonics (AREA)
- Engineering & Computer Science (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to preparation and application of a targeted nitroso peroxide fluorescent probe. The structural formula of the fluorescent probe is as follows:
the invention provides a method for preparing the catalyst by Cy-NH 2 The preparation method of the fluorescent probe is synthesized by using cholic acid, 2- (7-aza-benzotriazol) -N, N, N ', N' -tetramethyl urea hexafluorophosphate, N-ethyldiisopropylamine and the like as raw materials. The fluorescent probe is a near infrared and targeted nitroreductase fluorescent probe. First, the fluorescent probe exhibits very high sensitivity to NTR, and the fluorescence intensity is enhanced by 41 times. Secondly, the fluorescent probe shows high selectivity to NTR and is not influenced by other metal ions, active oxygen, active nitrogen, active sulfur and biological sulfurAlcohol, common amino acids, and interference of several other enzymes. The fluorescent probe has rapid action with NTR, and the response time is about 20 minutes. In addition, the fluorescent probe has been applied to cell imaging studies, and can detect changes in the intracellular NTR content.
Description
Technical Field
The invention belongs to the technical field of fluorescent probes, and particularly relates to preparation and application of a targeted nitroreductase fluorescent probe.
Background
Nitroreductase (NTR) is a reductase that reduces aromatic nitro groups (L.F.Xu, L.H.Sun, F.Zeng, S.Z.Wu, anal.Chim.Acta,2020,1125,152-161) in the presence of Nicotinamide Adenine Dinucleotide (NADH). NTR is overexpressed under hypoxic conditions and can be an effective biomarker for hypoxia (Y.J.Liu, W.Liu, H.J.Li, W.X.Yan, X.J Yang, D.D.Liu, anal.Chim.Acta,2018,1024,177-186). Hypoxia caused by inadequate oxygen supply is closely associated with many diseases, such as cardiovascular disease, inflammatory disease, cancer and acute ischemia (H.K.Eltzschig, D.L.Bratton, S.P.Colgan.Nat.Rev.Drug discover, 2014,13,852-869; R.A. Edwards, M.Witherspoon, K.Wang, K.Afrasiani, T.Pham, L.Birnbaumer, canc. Res.,2009,69,6423-6429). The NTR levels can be used to assess the degree of hypoxia, and thus determine the disease state. Therefore, it is of great importance to develop a strategy for high-sensitivity detection of NTR.
The fluorescent probe has the advantages of high sensitivity, real-time monitoring, high-resolution imaging and the like. To date, many fluorescent probes for detecting NTR have been reported (Z.Li, X.Y.He, Z.Wang, R.H.Yang, W.Shi, H.M.Ma, biosens.Bioelectron.,2015,63,112-116; R.Peng, J.Yuan, D.Cheng, T.B.ren, F.P.jin, R.H.Yang, L.Yuan, X.B.Zhang, anal.chem.,2019,91,15974-15981; S.xu, Q.Wang, Q.Zhang, L.Zhang, L.Zuo, J.D.Jiang, H.Y.Hu, chem.Commun.,2017,53,11177-11180; X.F.Zhang, X.H.Li, W.Shi, H.M.Ma, chem.Commun, 2021,57,8174-8177; H.S.Wang, X.F.Zhang, H.Dong, Q.n, X.Q.Cao, S.L.Chien, anal.202 m, 2, 107). However, these NTR probes suffer from a number of disadvantages: (1) The emission wavelength of fluorophores (such as fluorescein, naphthalimide, borofluoride and the like) is short, so that the application of the probes in organisms is limited; (2) Without active targeting of cells and tissues, it is difficult to aggregate in specific tissues, and the scattered probes reduce detection efficiency and further enhance background fluorescence.
The hemicyanine dye is a dye which is widely applied in the field of fluorescent probes at present, and has the advantages of stable chemical property, high fluorescence quantum yield and the like. In particular, the probe based on the hemicyanine has near infrared fluorescence emission, so that the tissue penetrating power is strong, the probe is not easy to be interfered by biological autofluorescence, and the probe is more beneficial to biological imaging. The literature reports that hemicyanine fluorescent probes have been used to detect pH, reactive oxygen, biological thiols and various enzymes (L.L.Wu, Y.Wang, T.D.James, N.Q.Jia, C.S.Huang, chem.Commun.,2018,54,5518-5521; X.Xie, X.Yang, T.Wu, Y.Li, M.Li, Q.Tan, X.Wang, B.Tang, anal.chem.,2016,88,8019-8025; C.S.park, T.H.Ha, S.A.Choi, D.N.Nguyen, S.Noh, O.S.Kwon, C.S.Lee, H.Yoon, biosens.bioelectron, 2017,89,919-926; S.J.Li, C.Y.Li, Y.F.Li, J.J.Fei, P.Wu, B.Yang, J.Ou-Yang, S.X.Nie, anal.Chem.,2017,89,6854-6860). Cholic acid is an organic acid having a steroid structure present in bile, and is one of the components of bile acid. Cholic acid can be taken in by hepatocyte sodium ion-taurocholate cotransporter (NTCP) and then reabsorbed by intestinal cell apical membrane sodium-dependent bile acid transporter (ASBT), so as to improve liver and intestine targeting of the medicine. In addition, the advantage of cholic acid amphoteric molecules is utilized, so that the permeability of the medicine to cell membranes can be improved. The introduction of cholic acid structure into the probe is an effective strategy for realizing targeting of liver and intestine related diseases, and the specific distribution of the probe in cells and tissues is expected to improve the sensitivity and the signal-to-noise ratio of NTR detection in the cells. However, there is no fluorescent probe that can simultaneously achieve targeting of liver and intestinal diseases and detection of NTR. Therefore, it is very necessary to design and synthesize a targeted nitroreductase fluorescent probe based on hemicyanine dye as an effective tool for detecting NTR in cells.
Disclosure of Invention
In light of the requirements set forth, the inventors have conducted intensive studies on this, and after a great deal of creative effort, provided a targeted nitroreductase fluorescent probe.
The technical scheme of the invention is that the targeted nitroreductase fluorescent probe has the following structural formula:
a preparation method of a targeted nitroreductase fluorescent probe comprises the following steps:
at room temperature, 1 equivalent of Cy-NH 2 1 to 1.5 equivalents of Cholic Acid (CA), 1 to 3 equivalents of 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea Hexafluorophosphate (HATU), 2 to 5 equivalents of N-ethyldiisopropylamine (DIPEA) were dissolved in 10 to 20mL of N, N-Dimethylformamide (DMF), then added to a 50mL round bottom flask and stirred under nitrogen for 2 hours; after the reaction was completed, the product was extracted with saturated sodium bromide solution and dichloromethane, and the organic phase was taken after 3 times of extraction with noDrying with sodium sulfate hydrate, removing solvent under reduced pressure, and using CH with volume ratio of 10:1-9:1 as the obtained crude product 2 Cl 2 /CH 3 And (3) performing column chromatography separation by using an OH eluent to obtain blue solid Cy-CA, namely the fluorescent probe.
The invention has the beneficial effects that the targeting nitroreductase fluorescent probe has good performance. First, the fluorescence spectrum properties of the probe were studied. The fluorescent probe has no obvious near infrared emission peak, and after NTR is added, the obvious near infrared emission peak appears at 720 nm. And as the concentration of NTR increases, the near infrared fluorescence intensity of the probe is continuously enhanced. The detection range of the probe is 0.01-5.0 mug/mL, and the detection limit is 20ng/mL, which shows that the probe has high sensitivity to NTR. Next, the ultraviolet absorption spectrum of the probe was studied. The probe has absorption bands at 572nm and 668 nm; after addition of NTR, the absorption at 572nm was significantly reduced, the 668nm absorption was enhanced and red shifted to 688nm. Then, the selectivity of the probe was investigated. Investigation of the probe and the Metal ion (K + ,Ca 2+ ) Active oxygen (H) 2 O 2 ,ClO - ) Active Nitrogen (NO) 2 - ,ONOO - ) Active Sulfur (S) 2 O 3 2- ,H 2 S), biological thiols (Cys, hcy, GSH), common amino acids (Met, lys, trp, phe, thr, ile, leu, val), and the fluorescent response of several enzymes (Esterase, trypsin). As a result, it was found that only NTR could cause a change in fluorescence spectrum, and that other detection substances had no significant effect on the fluorescence spectrum of the probe. Finally, the effect of pH on the determination of NTR by a fluorescent probe was studied, and the fluorescence at 720nm was gradually increased with increasing pH, without affecting the determination of NTR by a fluorescent probe when the pH was between 7.0 and 10.0. In addition, the fluorescent probe responds faster to NTR, and the response time is within 20 min.
An application of a targeted nitroreductase fluorescent probe. The cells were added with fluorescent probes under normoxic conditions with only weak fluorescence, indicating a low NTR content in the cells. The fluorescent probe is added into the cells under the anoxic condition, and the fluorescence is obviously enhanced; treatment of cells with biscoumarin under hypoxic conditions inhibited the production of intracellular NTR, and the fluorescence of the red channel was found to decrease. These results demonstrate that fluorescent probe Cy-CA can monitor changes in intracellular NTR content, which provides a reliable means for monitoring nitroreductase-related lesions in humans.
Drawings
FIG. 1 shows the synthetic route of fluorescent probes.
FIG. 2 is a graph showing fluorescence spectra of fluorescent probes after interaction with NTR at different concentrations.
The abscissa is wavelength and the ordinate is fluorescence intensity. The concentrations of the fluorescent probes are 10 mu M, and the NTR concentrations are respectively as follows: 0,0.10,0.2,0.3,0.4,0.5,0.6,0.7,0.8,0.9,1.0,1.2,1.4,1.6,1.8,2.0,2.5,3.0,4.0,5.0. Mu.g/mL. The fluorescence excitation wavelength was 688nm.
FIG. 3 is a graph showing the linear response of fluorescent probes to NTR fluorescence at different concentrations.
The abscissa indicates the NTR concentration, and the ordinate indicates the fluorescence intensity.
FIG. 4 is a graph showing the UV-visible absorption spectra of a fluorescent probe before and after interaction with NTR.
The abscissa is wavelength and the ordinate is absorbance. The concentration of the fluorescent probe was 10. Mu.M, and the NTR concentration was 5.0. Mu.g/mL.
FIG. 5 is a selective diagram of fluorescent probes.
The concentration of the fluorescent probe was 10. Mu.M, and the NTR concentration was 5.0. Mu.g/mL.
FIG. 6 is a graph showing the effect of pH on fluorescent probes.
The abscissa indicates pH value, and the ordinate indicates fluorescence intensity. The concentration of the fluorescent probe was 10. Mu.M, and the NTR concentration was 5.0. Mu.g/mL.
FIG. 7 is a graph showing the relationship between fluorescence intensity and time after the action of the fluorescent probe and NTR.
The abscissa is time, and the ordinate is fluorescence intensity. The concentration of the fluorescent probe was 10. Mu.M, and the NTR concentration was 0,1.0,3.0,5.0. Mu.g/mL.
FIG. 8 is a cytotoxicity test. The abscissa indicates the concentration of the fluorescent probe, and the ordinate indicates the viability of the cells.
FIG. 9 is a cell image of the effect of fluorescent probes on NTR. A (a) staining cells with probes under normoxic conditions for 0.5h. (b) staining the cells with the probe under anaerobic conditions for 0.5h. (c) Cells were treated with dicoumarol for 0.5h and then stained with probes for 0.5h. B: relative fluorescence intensity plot. Excitation wavelength is 640nm, and emission wavelength is 663-738 nm.
Detailed Description
The invention is described in detail below with reference to the drawings and the specific examples, but is not limited thereto.
Example 1:
synthesis of fluorescent probes
The synthetic route is shown in FIG. 1. At 25 ℃,1 equivalent of Cy-NH 2 1 equivalent of cholic acid, 1.5 equivalent of 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethyluronium Hexafluorophosphate (HATU), 2 equivalent of N-ethyldiisopropylamine (DIPEA) was dissolved in 15mL of N, N-Dimethylformamide (DMF) and then added to a 50mL round bottom flask and stirred under nitrogen for 2 hours; after the reaction is completed, the product is extracted by saturated sodium bromide solution and dichloromethane, the organic phase is taken after 3 times of extraction and dried by anhydrous sodium sulfate, the solvent is removed under the condition of reduced pressure, and the obtained crude product is subjected to CH with the volume ratio of 10:1 2 Cl 2 /CH 3 Separating by OH eluent column chromatography to obtain blue solid Cy-CA (yield 60%), namely fluorescent probe. 1 H NMR(400MHz,DMSO)δ8.56(d,J＝14.8Hz,1H),8.31(dd,J＝8.7,2.1Hz,2H),8.08(s,1H),7.78(t,J＝8.8Hz,3H),7.69(d,J＝8.0Hz,1H),7.61–7.43(m,3H),7.19(s,1H),7.11(dd,J＝8.6,2.3Hz,1H),6.54(d,J＝14.9Hz,1H),5.75(d,J＝1.9Hz,1H),5.46(s,1H),4.45–4.34(m,2H),3.72(s,1H),3.50(s,1H),2.72(s,1H),2.21–2.04(m,3H),1.94(dt,J＝16.4,7.7Hz,4H),1.83(t,J＝5.9Hz,2H),1.75(s,6H),1.68(d,J＝17.3Hz,3H),1.63–1.53(m,4H),1.41(d,J＝12.8Hz,4H),1.35–1.13(m,15H),0.92–0.84(m,5H),0.82(s,1H),0.79(s,1H),0.73(s,3H),0.49(s,2H).
Example 2:
fluorescent probe and NTR solution preparation
Preparation of probe solution: weighing a certain amount of probe and dissolving in DMSO to prepare 4×10 probe -4 M probe solution. Preparation of NTR stock solution: a certain amount of NTR was dissolved in ultrapure water to prepare a NTR solution of 20.0. Mu.g/mL, and the solution was stored in a frozen environment at-80℃and thawed before use. Preparation of NADH stock solution: will be a certain amountDissolving NADH in ultrapure water to prepare 10mM NADH solution, and storing in frozen environment at-80deg.C for thawing before use. mu.L of NADH stock solution, 250. Mu.L of probe solution and different volumes of nitroreductase sample solution were added to a 10mL volumetric flask, and the volume was fixed to 10mL with PBS buffer (10 mM, pH 7.4) and shaken well.
Example 3:
determination of fluorescence spectra of fluorescent probes and NTR interactions
FIG. 2 shows a fluorescence spectrum of the effect of a fluorescent probe at a concentration of 10. Mu.M and NTR at a concentration of 0,0.10,0.2,0.3,0.4,0.5,0.6,0.7,0.8,0.9,1.0,1.2,1.4,1.6,1.8,2.0,2.5,3.0,4.0,5.0. Mu.g/mL in that order. The excitation wavelength is 688nm, and the emission wavelength range is 700-800 nm. The excitation and emission slit widths were 5nm, and the fluorometric instrument used was a Hitachi F4600 fluorescence spectrophotometer. As can be seen from fig. 2, the fluorescent probe has little fluorescence emission before the NTR is added; after addition of NTR, a near infrared emission peak appears at 720 nm. This is because in the presence of the coenzyme NADH, the probe molecule is reduced by NTR, its nitro group is reduced to hydroxylamine or amino group, and 1, 6-rearrangement and elimination reaction occurs, thereby releasing the fluorophore, and near infrared fluorescence is generated. And, as the concentration of NTR increases, the near infrared fluorescence intensity of the probe molecules is continuously enhanced. When NTR of 5. Mu.g/mL was added, the fluorescence intensity was enhanced by 41-fold, so that NTR could be detected. FIG. 3 is a graph of the linear response of the probe to NTR at different concentrations. The fluorescence intensity and the concentration of NTR show a linear relation, the linear range is 0.1-5 mug/mL, and the detection limit is 20ng/mL. This indicates that the probe can detect NTR with high sensitivity.
Example 4:
determination of ultraviolet visible absorption Spectrometry by fluorescent Probe and NTR action
FIG. 4 is a graph showing the ultraviolet-visible absorption spectrum of a fluorescent probe after the action of NTR, wherein the concentration of the fluorescent probe is 10. Mu.M, and the concentration of NTR is 5. Mu.g/mL. The ultraviolet-visible absorption spectrum measuring instrument is an Agilent Cary60 ultraviolet-visible spectrophotometer. As can be seen from FIG. 4, in the absence of NTR, the probe has an absorption band at 572nm and 668 nm; after addition of NTR, the absorption at 572nm decreased, the absorption at 668nm increased and red shifted to 688nm.
Example 5:
selectivity of fluorescent probe for NTR assay
FIG. 5 is a plot of selectivity of fluorescent probe versus NTR assay. Investigation of the addition of NTR (5. Mu.g/mL) and its association with Metal ion (K) to a fluorescent Probe solution at a concentration of 10. Mu.M + ,Ca 2+ ) Active oxygen (H) 2 O 2 ,ClO - ) Active Nitrogen (NO) 2 - ,ONOO - ) Active Sulfur (S) 2 O 3 2- ,H 2 S), biological thiols (Cys, hcy, GSH), common amino acids (Met, lys, trp, phe, thr, ile, leu, val) (200 μm), and fluorescent responses of several enzymes (Esterase, trypsin) (10 μg/mL). As can be seen from fig. 5, only NTR causes a change in fluorescence intensity, and other detectors have no significant effect on the fluorescence intensity of the probe. These results indicate that the fluorescent probe has better selectivity to NTR.
Example 6:
effect of solution pH on fluorescence properties of fluorescent Probe assay NTR
The effect of pH on the fluorescence spectrum of NTR measured by a fluorescent probe was examined, and the results are shown in FIG. 6. The pH range studied was 4.0-10.0, the concentration of fluorescent probe was 10. Mu.M, and the concentration of NTR was 5. Mu.g/mL. As can be seen from the figure, the fluorescence intensity is substantially unchanged with changes in pH, indicating that pH has no significant effect on the probe itself. However, after addition of NTR, the change in fluorescence intensity with pH is not significant when the pH is in the range of 4 to 6; when the pH is in the range of 7.0 to 10.0, the fluorescence intensity is remarkably enhanced and reaches a higher level. In summary, when the pH is between 7.0 and 10.0, the determination of NTR by the fluorescent probe is not affected, and the pH range is suitable, which is very favorable for the determination of NTR in biological samples by the probe.
Example 7:
determination of response time of fluorescent probe to NTR action
The response time of fluorescent probe to NTR was studied and the results are shown in fig. 7. From the graph, the response time of the probe to NTR is 20min, which can meet the requirement of real-time monitoring in actual samples. From FIG. 7, it can be seen that after the fluorescence intensity reaches the maximum value, the fluorescence intensity does not change any more in the later time, which indicates that the fluorescence probe has good light stability.
Example 8:
application of fluorescent probe in living cells
First, we performed cytotoxicity assays as shown in fig. 8. When 0-30 mu M NTR probe is added, the survival rate of cells is over 90%, so that the fluorescent probe has low toxicity and can be applied to detecting NTR in living cells. We then studied the use of fluorescent probes in living cells and selected HCT116 cells for confocal microscopy imaging, the results of which are shown in figure 9. FIG. 9A is a fluorescent image of cells under different conditions, stained with Cy-CA probe for 0.5h under normoxic conditions, with only weak fluorescence, indicating lower NTR concentrations in the cells (panel a). Cells were stained with probe for 0.5h under hypoxic conditions with a significant increase in fluorescence (panel b); cells were treated with the NTR inhibitor biscoumarin for 0.5h under hypoxic conditions, then stained with the probe for 0.5h, and fluorescence of the red channel was reduced (panel c). FIG. 9B is a graph of relative fluorescence intensity, which demonstrates that the fluorescent probe can monitor changes in intracellular NTR content, providing a reliable means for monitoring nitroreductase-related pathologies in humans.
Claims (3)
1. A targeting nitroreductase fluorescent probe, namely Cy-CA, is characterized by comprising the following structure:
2. the method for preparing the targeted nitroreductase fluorescent probe according to claim 1, wherein the reaction steps are as follows:
at room temperature, 1 equivalent of Cy-NH 2 0.8 to 1.2 equivalents of cholic acid, 1 to 2 equivalents of 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate1-3 equivalents of N-ethyldiisopropylamine is dissolved in 10-20 mL of N, N-dimethylformamide, added into a 50mL round bottom flask and stirred in nitrogen for 1-3 hours; after the reaction is completed, extracting for 3 times by using saturated sodium bromide solution and dichloromethane, drying an organic phase by using anhydrous sodium sulfate, removing the solvent under the condition of reduced pressure, and using CH with the volume ratio of 10:1-9:1 as the obtained crude product 2 Cl 2 /CH 3 Separating by OH eluent column chromatography to obtain blue solid Cy-CA, namely the fluorescent probe, wherein Cy-NH 2 The structure of (2) is as follows:
3. the use of a targeted nitroreductase fluorescent probe according to claim 1, wherein the fluorescent probe has been used in cell imaging studies to detect changes in nitroreductase content within cells.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310401806.0A CN116425820B (en) | 2023-04-14 | 2023-04-14 | Preparation and application of targeted nitroreductase fluorescent probe |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310401806.0A CN116425820B (en) | 2023-04-14 | 2023-04-14 | Preparation and application of targeted nitroreductase fluorescent probe |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN116425820A true CN116425820A (en) | 2023-07-14 |
| CN116425820B CN116425820B (en) | 2024-08-16 |
Family
ID=87088680
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310401806.0A Active CN116425820B (en) | 2023-04-14 | 2023-04-14 | Preparation and application of targeted nitroreductase fluorescent probe |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116425820B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106753341A (en) * | 2016-12-27 | 2017-05-31 | 湘潭大学 | A kind of preparation method and application of near-infrared alkaline phosphatase fluorescence probe |
| CN112457360A (en) * | 2020-11-26 | 2021-03-09 | 山西医科大学 | Liver-targeted peroxynitrite fluorescent probe, and preparation method and application thereof |
| CN113563279A (en) * | 2021-08-25 | 2021-10-29 | 安徽大学 | Two-photon fluorescent probe for detecting nitroreductase and preparation method and application thereof |
| CN115710299A (en) * | 2022-10-31 | 2023-02-24 | 北京工业大学 | Fluorescent/photoacoustic bimodal probes for liver-targeted early drug-induced hepatitis and autoimmune hepatitis |
-
2023
- 2023-04-14 CN CN202310401806.0A patent/CN116425820B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106753341A (en) * | 2016-12-27 | 2017-05-31 | 湘潭大学 | A kind of preparation method and application of near-infrared alkaline phosphatase fluorescence probe |
| CN112457360A (en) * | 2020-11-26 | 2021-03-09 | 山西医科大学 | Liver-targeted peroxynitrite fluorescent probe, and preparation method and application thereof |
| CN113563279A (en) * | 2021-08-25 | 2021-10-29 | 安徽大学 | Two-photon fluorescent probe for detecting nitroreductase and preparation method and application thereof |
| CN115710299A (en) * | 2022-10-31 | 2023-02-24 | 北京工业大学 | Fluorescent/photoacoustic bimodal probes for liver-targeted early drug-induced hepatitis and autoimmune hepatitis |
Non-Patent Citations (5)
| Title |
|---|
| KEITH D. GREEN ET AL.: "Investigating the promiscuity of the chloramphenicol nitroreductase from Haemophilus influenzae towards the reduction of 4-nitrobenzene derivatives", 《BIOORGANIC & MEDICINAL CHEMISTRY LETTERS》, vol. 29, 21 February 2019 (2019-02-21), pages 1127 - 1132 * |
| SHUSHU WANG ET AL.: "Discovery of a highly efficient nitroaryl group for detection of nitroreductase and imaging of hypoxic tumor cells", 《ORG. BIOMOL. CHEM.》, vol. 19, 29 March 2021 (2021-03-29), pages 3469 - 3478 * |
| TING LI ET AL.: "An intestinal-targeting near-infrared probe for imaging nitroreductase in inflammatory bowel disease", 《SENSORS AND ACTUATORS: B. CHEMICAL》, vol. 403, 19 December 2023 (2023-12-19), pages 135181 * |
| YA-LIN QI ET AL.: "Recent progress in the design principles, sensing mechanisms, and applications of small-molecule probes for nitroreductases", 《COORDINATION CHEMISTRY REVIEWS》, vol. 421, 7 July 2020 (2020-07-07), pages 213460, XP086219728, DOI: 10.1016/j.ccr.2020.213460 * |
| ZHAO LI ET AL.: "in vivo imaging and detection of nitroreductase in zebrafish by a new near-infrared fluorescence off–on probe", 《BIOSENSORS AND BIOELECTRONICS》, vol. 63, 16 July 2014 (2014-07-16), pages 112 - 116, XP055487237, DOI: 10.1016/j.bios.2014.07.024 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN116425820B (en) | 2024-08-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110540837B (en) | Preparation and application of hydrogen peroxide near-infrared fluorescent probe | |
| CN111423423B (en) | Application of ratiometric fluorescent probe in detecting peroxynitrite anion | |
| Zhao et al. | Transforming the recognition site of 4-hydroxyaniline into 4-methoxyaniline grafted onto a BODIPY core switches the selective detection of peroxynitrite to hypochlorous acid | |
| CN111518083B (en) | Preparation and application of open type fluorescent probe for detecting carbon monoxide | |
| Wang et al. | Novel designed azo substituted semi-cyanine fluorescent probe for cytochrome P450 reductase detection and hypoxia imaging in cancer cells | |
| US8557811B2 (en) | Dual-color imaging method of sodium/calcium activities using two-photon fluorescent probes and preparation method of two-photon fluorescent probes | |
| CN109503515A (en) | A kind of methylene blue derivatives object and its synthetic method and application | |
| CN111253935A (en) | Two-photon fluorescent probe for detecting polarity and viscosity through two channels and preparation method and application thereof | |
| CN111116696A (en) | Preparation and application of adenosine triphosphate near-infrared nano fluorescent probe | |
| CN114672302B (en) | Preparation and application of near-infrared MOF fluorescent probe based on silicon rhodamine | |
| Liu et al. | A near-infrared fluorescence probe with a large Stokes shift for detecting carbon monoxide in living cells and mice | |
| CN113121580B (en) | Hydrogen peroxide fluorescent probe and preparation method and application thereof | |
| CN109762034A (en) | The preparation method of novel terephthalaldehyde contracting D- Glucosamine schiff bases | |
| CN111778014A (en) | Beta-galactosidase near-infrared fluorescent probe and preparation method and application thereof | |
| CN111825718B (en) | Preparation and application of alkaline phosphatase fluorescent probe based on quinoline-xanthene | |
| CN116425820B (en) | Preparation and application of targeted nitroreductase fluorescent probe | |
| CN110669503B (en) | Preparation and application of carbon monoxide near-infrared fluorescent probe | |
| CN114605432B (en) | Preparation and application of cyanine dye-based targeted cysteine fluorescent probe | |
| CN110229203B (en) | Hexosamine fluorescent probe and preparation method and application thereof | |
| CN113024445A (en) | Indocyanine-based fluorescent probe, preparation method and application | |
| CN115160391B (en) | Preparation and application of targeted nitrosoperoxide fluorescent probe | |
| CN115160338B (en) | Preparation and application of cyanine dye-based targeting ratio pH fluorescent probe | |
| He et al. | A novel fast and sensitive fluorescent hydrogen sulfide probe with large stokes shift for imaging living cells and zebrafish | |
| CN113429389B (en) | Preparation and application of solid-state fluorescent probe for detecting carbon monoxide | |
| CN115894293B (en) | Ratio type near infrared fluorescent probe for detecting hydrazine and synthetic method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |