CN114702518A - 一种酪氨酸酶和过氧化氢双响应型的近红外荧光探针及其制备与应用 - Google Patents
一种酪氨酸酶和过氧化氢双响应型的近红外荧光探针及其制备与应用 Download PDFInfo
- Publication number
- CN114702518A CN114702518A CN202210485902.3A CN202210485902A CN114702518A CN 114702518 A CN114702518 A CN 114702518A CN 202210485902 A CN202210485902 A CN 202210485902A CN 114702518 A CN114702518 A CN 114702518A
- Authority
- CN
- China
- Prior art keywords
- tyrosinase
- fluorescent probe
- hydrogen peroxide
- infrared fluorescent
- ion
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000007850 fluorescent dye Substances 0.000 title claims abstract description 51
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 title claims abstract description 47
- 102000003425 Tyrosinase Human genes 0.000 title claims abstract description 39
- 108060008724 Tyrosinase Proteins 0.000 title claims abstract description 39
- 230000004044 response Effects 0.000 title claims abstract description 22
- 238000002360 preparation method Methods 0.000 title claims description 14
- 150000001875 compounds Chemical class 0.000 claims abstract description 21
- -1 borate compound Chemical class 0.000 claims abstract description 17
- 238000000034 method Methods 0.000 claims abstract description 9
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 12
- 239000003513 alkali Substances 0.000 claims description 10
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 9
- 125000003172 aldehyde group Chemical group 0.000 claims description 8
- 125000000304 alkynyl group Chemical group 0.000 claims description 8
- 125000003368 amide group Chemical group 0.000 claims description 8
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 8
- 125000004093 cyano group Chemical group *C#N 0.000 claims description 8
- 125000004185 ester group Chemical group 0.000 claims description 8
- 229910052736 halogen Inorganic materials 0.000 claims description 8
- 150000002367 halogens Chemical class 0.000 claims description 8
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 8
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 8
- 125000001424 substituent group Chemical group 0.000 claims description 8
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 claims description 7
- 230000009977 dual effect Effects 0.000 claims description 7
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 claims description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 6
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 6
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical group CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 6
- GRVDJDISBSALJP-UHFFFAOYSA-N methyloxidanyl Chemical group [O]C GRVDJDISBSALJP-UHFFFAOYSA-N 0.000 claims description 6
- 125000005245 nitryl group Chemical group [N+](=O)([O-])* 0.000 claims description 6
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 claims description 6
- 125000003396 thiol group Chemical group [H]S* 0.000 claims description 6
- UCPYLLCMEDAXFR-UHFFFAOYSA-N triphosgene Chemical compound ClC(Cl)(Cl)OC(=O)OC(Cl)(Cl)Cl UCPYLLCMEDAXFR-UHFFFAOYSA-N 0.000 claims description 5
- LSNNMFCWUKXFEE-UHFFFAOYSA-M Bisulfite Chemical compound OS([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-M 0.000 claims description 4
- CPELXLSAUQHCOX-UHFFFAOYSA-M Bromide Chemical compound [Br-] CPELXLSAUQHCOX-UHFFFAOYSA-M 0.000 claims description 4
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 claims description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 claims description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 4
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 claims description 4
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims description 4
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical group [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 4
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 claims description 4
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 4
- AYJRCSIUFZENHW-UHFFFAOYSA-L barium carbonate Chemical compound [Ba+2].[O-]C([O-])=O AYJRCSIUFZENHW-UHFFFAOYSA-L 0.000 claims description 4
- 229940006460 bromide ion Drugs 0.000 claims description 4
- MVPPADPHJFYWMZ-UHFFFAOYSA-N chlorobenzene Chemical compound ClC1=CC=CC=C1 MVPPADPHJFYWMZ-UHFFFAOYSA-N 0.000 claims description 4
- 238000004440 column chromatography Methods 0.000 claims description 4
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 claims description 4
- XMBWDFGMSWQBCA-UHFFFAOYSA-M iodide Chemical compound [I-] XMBWDFGMSWQBCA-UHFFFAOYSA-M 0.000 claims description 4
- 229940006461 iodide ion Drugs 0.000 claims description 4
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 4
- 239000003960 organic solvent Substances 0.000 claims description 4
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 claims description 4
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 3
- 229910052760 oxygen Inorganic materials 0.000 claims description 3
- 239000001301 oxygen Substances 0.000 claims description 3
- 229910000027 potassium carbonate Inorganic materials 0.000 claims description 3
- 235000011181 potassium carbonates Nutrition 0.000 claims description 3
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 2
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 claims description 2
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 claims description 2
- FJDQFPXHSGXQBY-UHFFFAOYSA-L caesium carbonate Chemical compound [Cs+].[Cs+].[O-]C([O-])=O FJDQFPXHSGXQBY-UHFFFAOYSA-L 0.000 claims description 2
- 229910000024 caesium carbonate Inorganic materials 0.000 claims description 2
- 239000003153 chemical reaction reagent Substances 0.000 claims description 2
- 125000001841 imino group Chemical group [H]N=* 0.000 claims description 2
- 150000007529 inorganic bases Chemical class 0.000 claims description 2
- 150000007530 organic bases Chemical class 0.000 claims description 2
- 229910000028 potassium bicarbonate Inorganic materials 0.000 claims description 2
- 235000015497 potassium bicarbonate Nutrition 0.000 claims description 2
- 239000011736 potassium bicarbonate Substances 0.000 claims description 2
- TYJJADVDDVDEDZ-UHFFFAOYSA-M potassium hydrogencarbonate Chemical compound [K+].OC([O-])=O TYJJADVDDVDEDZ-UHFFFAOYSA-M 0.000 claims description 2
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims description 2
- 238000000926 separation method Methods 0.000 claims description 2
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 2
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 2
- 239000008096 xylene Substances 0.000 claims description 2
- 229960001701 chloroform Drugs 0.000 claims 1
- 238000012632 fluorescent imaging Methods 0.000 claims 1
- 201000001441 melanoma Diseases 0.000 abstract description 24
- 238000003384 imaging method Methods 0.000 abstract description 11
- 206010028980 Neoplasm Diseases 0.000 abstract description 10
- 238000000799 fluorescence microscopy Methods 0.000 abstract description 10
- 238000004458 analytical method Methods 0.000 abstract description 6
- 238000003745 diagnosis Methods 0.000 abstract description 4
- 230000015572 biosynthetic process Effects 0.000 abstract description 3
- 230000008569 process Effects 0.000 abstract description 3
- 239000000523 sample Substances 0.000 abstract description 3
- 230000035945 sensitivity Effects 0.000 abstract description 3
- 230000001737 promoting effect Effects 0.000 abstract description 2
- 238000003786 synthesis reaction Methods 0.000 abstract description 2
- 230000008685 targeting Effects 0.000 abstract description 2
- 230000009471 action Effects 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 12
- 238000005481 NMR spectroscopy Methods 0.000 description 10
- 238000001514 detection method Methods 0.000 description 7
- 241000699666 Mus <mouse, genus> Species 0.000 description 6
- 229940125898 compound 5 Drugs 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 238000001228 spectrum Methods 0.000 description 6
- 241000699670 Mus sp. Species 0.000 description 5
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 4
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 4
- 229910052799 carbon Inorganic materials 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 3
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 3
- 229910052739 hydrogen Inorganic materials 0.000 description 3
- 239000001257 hydrogen Substances 0.000 description 3
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 2
- 238000005160 1H NMR spectroscopy Methods 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 2
- XUMBMVFBXHLACL-UHFFFAOYSA-N Melanin Chemical compound O=C1C(=O)C(C2=CNC3=C(C(C(=O)C4=C32)=O)C)=C2C4=CNC2=C1C XUMBMVFBXHLACL-UHFFFAOYSA-N 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000000090 biomarker Substances 0.000 description 2
- 239000001569 carbon dioxide Substances 0.000 description 2
- 229910002092 carbon dioxide Inorganic materials 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 2
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 2
- 239000002953 phosphate buffered saline Substances 0.000 description 2
- 230000035790 physiological processes and functions Effects 0.000 description 2
- 239000003642 reactive oxygen metabolite Substances 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 2
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 108010019160 Pancreatin Proteins 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 125000005621 boronate group Chemical group 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 230000012292 cell migration Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 229940126214 compound 3 Drugs 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000001647 drug administration Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 239000013067 intermediate product Substances 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 210000002752 melanocyte Anatomy 0.000 description 1
- 210000002780 melanosome Anatomy 0.000 description 1
- 230000009401 metastasis Effects 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000036542 oxidative stress Effects 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 238000010791 quenching Methods 0.000 description 1
- 230000000171 quenching effect Effects 0.000 description 1
- 150000004053 quinones Chemical class 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 201000000849 skin cancer Diseases 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Images
Classifications
-
- 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
- C07F5/00—Compounds containing elements of Groups 3 or 13 of the Periodic Table
- C07F5/02—Boron compounds
- C07F5/025—Boronic and borinic acid compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
- A61K49/0032—Methine dyes, e.g. cyanine dyes
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- 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
- G01N21/6458—Fluorescence microscopy
-
- 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
-
- 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
- C09K2211/1096—Heterocyclic compounds characterised by ligands containing other heteroatoms
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- General Health & Medical Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Engineering & Computer Science (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Optics & Photonics (AREA)
- Biomedical Technology (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
本发明公开了一种酪氨酸酶和过氧化氢双响应型的近红外荧光探针，是将半花菁类化合物与硼酸酯类化合物连接制备而得，该探针在酪氨酸酶和过氧化氢共同作用下，高灵敏、高选择性地点亮近红外荧光信号，同时响应黑色素瘤中的酪氨酸酶和H2O2这两种特征因子，避免产生“假阳性”信号的干扰，可实现黑色素瘤的高选择性荧光成像分析。本发明的一种酪氨酸酶和过氧化氢双响应型的近红外荧光探针具有结构新颖、合成工艺简单快捷、靶向性高和成像性能好等优点，对于推动肿瘤的精准诊断具有重要意义。
Description
技术领域
本发明属于荧光探针检测技术领域，具体涉及一种酪氨酸酶和过氧化氢双响应型的近红外荧光探针及其制备方法，同时涉及该荧光探针在荧光成像中的应用应用。
背景技术
黑色素瘤是一种侵袭性很强的皮肤恶性肿瘤，具有快速生长和早期转移的特点。统计显示，黑色素瘤晚期时5年相对生存率为23%。目前黑素瘤的临床治疗仍以手术切除为主，而对黑素瘤精准诊断是临床黑素瘤治疗的关键。生物标志物激活的荧光探针，特别是近红外(NIR)荧光探针为黑素瘤的诊断提供了强有力的手段。近年来，国内外研究者开发了多种黑素瘤特征因子激活的荧光探针用于黑素瘤特异性荧光成像检测（王晨羽等，中华皮肤科杂志，2021, 54, 60-63；Li, Z. et al., Anal. Chem.2018, 90, 3666–3669; Park,S. Y. et al., Sensors and Actuators B: Chemical, 2020, 319, 128306）。然而，目前已开发的荧光探针通常仅对黑素瘤中单一的特征因子进行响应，易产生“假阳性”信号的干扰（Wu, L. et al.,Nat. Rev. Chem.2021, 5, 406-421）。因此，开发对黑素瘤中多种特征因子响应的近红外荧光探针对于黑素瘤的精准成像检测具有重大意义。
酪氨酸酶是一种多功能、糖基化的黑色素细胞特异性含铜氧化酶，主要存在于细胞器黑素体中，通过介导酪氨酸氧化成一些醌类的而引发黑色素的形成。研究表明，酪氨酸酶在与酪氨酸相关的生理过程中发挥着重要作用，黑色素瘤中酪氨酸酶含量远高于正常细胞和组织。由于酪氨酸酶在黑色素瘤细胞中过量表达，酪氨酸酶被作为黑色素瘤诊断和预后的独立生物标志物。此外，过氧化氢（H2O2）是生物体内最稳定的活性氧（ROS）。作为信号分子和氧化还原稳态的维持者，低水平的H2O2调控细胞的生长、增殖、迁移等多种生理过程。然而，高浓度的H2O2会促进氧化应激导致癌症等各种疾病的发展。黑色素瘤中H2O2的含量是正常细胞的10倍。因此，构建黑色素瘤中的酪氨酸酶和H2O2共同激活的荧光探针，可有效地区分黑色素瘤和正常组织，对黑色素瘤的成像检测具有重要意义。
发明内容：
本发明的目的在于提供一种酪氨酸酶和过氧化氢双响应型的近红外荧光探针；本发明的另一目的是提供该荧光探针的制备方法及其应用。
一、酪氨酸酶和过氧化氢双响应型的近红外荧光探针及其制备
本发明酪氨酸酶和过氧化氢双响应型的近红外荧光探针，结构式为：
其中：取代基R1，R2，R3为H、甲基、乙基、丙基、甲氧基、羟基、硝基、氨基、卤素、醛基、羧基、磺酸基、巯基、酯基、氰基、酰胺基或炔基；
取代基R4为H、甲氧基、羟基、硝基、氨基、卤素、醛基、羧基、巯基、磺酸基、巯基、酯基、氰基、酰胺基或炔基；
X-为碘离子、溴离子、氯离子、硝酸根离子、硫酸根离子、碳酸根离子或亚硫酸根离子；n为1~20的整数；Y为氧醚基、巯基或亚氨基。
本发明酪氨酸酶和过氧化氢双响应型的近红外荧光探针的制备方法，是将半花菁类化合物、碱和三光气加入到有机溶剂中，0~25℃下反应0.01~1小时后，再加入硼酸酯类化合物，在0~80℃下反应0.1~20小时后减压浓缩，柱层析分离即得。
所述半花菁类化合物的结构式如下：
其中，取代基R4为H、甲氧基、羟基、硝基、氨基、卤素、醛基、羧基、巯基、磺酸基、巯基、酯基、氰基、酰胺基或炔基；
X-为碘离子、溴离子、氯离子、硝酸根离子、硫酸根离子、碳酸根离子或亚硫酸根离子；n为1~20的整；Y为羟基、巯基或氨基。半花菁类化合物的制备方法见文献（Anal. Chem.,2021, 93, 15080−15087）。
所述硼酸酯类化合物的结构式如下：
其中，取代基R1，R2，R3为H、甲基、乙基、丙基、甲氧基、羟基、硝基、氨基、卤素、醛基、羧基、磺酸基、巯基、酯基、氰基、酰胺基或炔基。
所述半花菁类化合物和硼酸酯类化合物的摩尔比为1:0.2~1:5；所述半花菁类化合物与三光气的摩尔比为1:0.1~1:0.5；所述半花菁类化合物与碱的摩尔比为1:1~1:5。
所述有机溶剂为四氢呋喃、乙腈、二甲基亚砜、N, N-二甲基甲酰胺、二氯甲烷、苯、甲苯、二甲苯、氯苯、甲醇、乙醇、三氯甲烷中至少一种，优选二氯甲烷。
所述碱为有机碱或无机碱；其中有机碱为三乙胺、吡啶或4-二甲胺基吡啶；无机碱为碳酸钠、碳酸钾、氢氧化钠、碳酸铯、碳酸氢钠、碳酸氢钾或碳酸钡；优选4-二甲胺基吡啶。
二、荧光探针在荧光成像中的应用
本发明中的近红外荧光探针，可同时响应酪氨酸酶和H2O2两种黑色素瘤特征因子，利用近红外荧光信号穿透性强、背景干扰小等优势有利于细胞或体内黑色素瘤的成像分析。下面以制备的荧光探针7为例说明本发明荧光探针在荧光成像中的应用。
1、荧光探针的肿瘤细胞成像分析中的应用
首先将A549细胞在37℃，21 %氧气，5％二氧化碳环境的细胞培养箱中培养24小时后，用胰酶消化细胞转移到细胞成像皿中，分为实验组和对照组。实验组加入本发明实施例1制备的近红外荧光探针7溶液、酪氨酸酶和H2O2；对照组a仅加入本发明实施例1制备的近红外荧光探针7溶液；对照组b加入本发明实施例1制备的近红外荧光探针7溶液和H2O2；对照组c加入本发明实施例1制备的近红外荧光探针7溶液和酪氨酸酶。将上述四组细胞继续在37℃，5%二氧化碳环境的细胞培养箱中培养4小时后，用PBS缓冲液洗去成像皿里的培养基，用激光扫描共聚焦显微镜进行荧光成像。从图5中可以看出，实验组细胞表现出较强荧光，表明探针能进入细胞并被酪氨酸酶和H2O2激活产生显著荧光信号。与之相比，对照组细胞中未见明显的荧光信号，说明机制干扰较小。
2、近红外荧光探针的肿瘤活体荧光成像分析中的应用
通过给两只荷瘤小鼠瘤内注射0.1mL浓度为100 μM的近红外荧光探针7后，对照组小鼠注射0.1 mL 生理盐水，给药组小鼠注射0.1 mL酪氨酸酶和H2O2的混合溶液，将上述小鼠给药后8 h后，使用小动物活体荧光成像仪进行荧光成像，结果如图6所示。图6中1号小鼠为对照组小鼠，2号小鼠为给药组小鼠。由图可以看出，给药组小鼠肿瘤部位荧光信号较为显著，与之相比，对照组小鼠肿瘤部位未见荧光信号。这是由于给药组小鼠中探针和酪氨酸酶和H2O2相互作用导致近红外荧光信号点亮，实现动物体内肿瘤的成像分析。
半花菁类化合物具有近红外荧光发射性能，本发明将荧光基团半花菁类化合物与猝灭基团硼酸酯类化合物连接，由于硼酸酯类化合物猝灭了荧光基团半花菁类化合物的荧光，使得半花菁类化合物无荧光发射性能。在硼酸酯基团被肿瘤中高浓度的H2O2激活后，中间产物进一步被酪氨酸酶活化，释放荧光基团半花菁类化合物。通过这种级联激活，荧光基团半花菁类化合物点亮近红外荧光信号，实现对酪氨酸酶和H2O2两种黑色素瘤特征因子的同时检测。相较与目前已开发的仅对肿瘤中单一的特征因子（酪氨酸酶、H2O2、微酸和缺氧等）激活型的荧光探针，本发明所提供的双响应型近红外荧光探针，可同时响应肿瘤中的酪氨酸酶和H2O2这两种特征因子，对肿瘤的选择性高、灵敏性强，对于黑素瘤的精准检测方面具有重要意义。
本发明的有益效果为：
1、本发明所提供的一种酪氨酸酶和过氧化氢双响应型的近红外荧光探针，具有结构新颖、靶向性高、灵敏性强和成像性能好等优点，可同时响应黑色素瘤中的酪氨酸酶和H2O2这两种特征因子，避免产生“假阳性”信号的干扰，可实现黑色素瘤的高选择性荧光成像分析。
2、本发明所提供的一种酪氨酸酶和过氧化氢双响应型的近红外荧光探针的制备方法，合成工艺简单有效，可以高产率、高纯度的得到荧光探针。
3、鉴于良好的成像性能，本申请所提供的一种酪氨酸酶和过氧化氢双响应型的近红外荧光探针，对于推动黑色素瘤的成像检测具有重要意义，在可视化检测领域有潜在的应用价值。
附图说明
图1是本发明制备的化合物5的核磁共振氢谱图；
图2是本发明制备的化合物5的核磁共振碳谱图；
图3是本发明制备的荧光探针7的核磁共振氢谱图；
图4是本发明制备的荧光探针7的核磁共振碳谱图；
图5是本发明近红外荧光探针7在A549细胞的成像图；
图6是本发明近红外荧光探针7在荷瘤小鼠体内成像图。
具体实施方式
下面结合若干优选实施例及附图对本发明的技术方案做进一步详细说明，本实施例在以发明技术方案为前提下进行实施，给出了详细的实施方式和具体的操作过程，但本发明的保护范围不限于下述的实施例。
下面所用的实施例中所采用的实验材料，如无特殊说明，均可由常规的生化试剂公司购买得到。
实施例
（1）化合物5的制备：
在圆底烧瓶中，加入化合物3（1.0 mmol）和化合物4（1.1 mmol），加入干燥的乙腈（10 mL）后，再加入碳酸钾（3 mmol）。在氮气保护下室温搅拌30分钟后，在60℃下继续反应5小时后，过滤、干燥、柱层析分离得到化合物5；反应式如下：
制备的化合物5核磁共振氢谱图如图1所示，核磁共振碳谱图如图2所示，1H NMR和13C NMR数据如下：1H NMR (400 MHz, CDCl3): δ 7.82 (d, J=7.6 Hz, 2H), 7.43 (d,J=8.0 Hz, 2H), 7.24-7.28 (m, 1H), 7.00 (s, 1H), 6.94 (d, J=7.6 Hz, 1H), 6.88-6.90 (m, 1H), 5.10 (s, 2H), 4.66 (d, J=5.2 Hz, 2H), 1.35 (s, 12H). 13C NMR(101 MHz, CDCl3): δ 159.0, 142.6, 140.2, 135.1, 129.6, 126.5, 119.4, 114.1,113.3, 83.9, 69.8, 65.2, 24.9。
（2）荧光探针7的制备：
在圆底烧瓶中，化合物6（1mmol）和干燥的二氯甲烷（50mL），加入对二甲氨基吡啶（4mmol），在0℃下加入三光气（0.4 mmol），反应在0℃搅拌30分钟后，加入化合物5（2mmol），在室温条件下继续反应8小时后，减压浓缩，柱层析分离得到荧光探针7。其中，化合物6的制备方法见文献（Anal. Chem.,2021, 93, 15080−15087）。其反应式如下：
本实施例制备的荧光探针7核磁共振氢谱图如图3所示，核磁共振碳谱图如图4所示，1H NMR和13C NMR数据如下：1H NMR (400 MHz, DMSO-d 6): δ10.44 (s, 1H), 8.54 (d,J=15.2 Hz, 1H), 7.79-7.80 (m, 2H),7.67-7.69 (m, 2H), 7.44-7.57 (m, 6H), 7.31-7.33 (d, J=8.0 Hz, 2H), 7.11 (s, 1H), 6.99-7.03 (m, 2H), 6.55 (d, J=15.2 Hz,1H), 5.17 (d, J=5.6 Hz, 4H), 3.88 (s, 3H), 2.67-2.72 (m, 4H), 1.83 (m, 2H),1.75 (s, 6H), 1.258 (s, 12H). 13C NMR (101 MHz, DMSO-d 6): δ 178.2, 160.5,158.8, 153.6, 142.8, 142.4, 140.8, 138.3, 135.0, 130.2, 128.1, 127.6, 127.3,120.7, 117.0, 114.9, 114.7, 114.2, 104.3, 84.1, 69.4, 50.8, 33.1, 30.1, 27.6,25.1。荧光成像结果如图5、图6。
Claims (8)
1.一种酪氨酸酶和过氧化氢双响应型的近红外荧光探针，结构式为：
其中：取代基R1，R2，R3为H、甲基、乙基、丙基、甲氧基、羟基、硝基、氨基、卤素、醛基、羧基、磺酸基、巯基、酯基、氰基、酰胺基或炔基；
取代基R4为H、甲氧基、羟基、硝基、氨基、卤素、醛基、羧基、巯基、磺酸基、巯基、酯基、氰基、酰胺基或炔基；
X-为碘离子、溴离子、氯离子、硝酸根离子、硫酸根离子、碳酸根离子或亚硫酸根离子；n为1~20的整数；Y为氧醚基、巯基或亚氨基。
2.如权利要求1所述的一种酪氨酸酶和过氧化氢双响应型的近红外荧光探针的制备方法，是将半花菁类化合物、碱和三光气加入到有机溶剂中，0~25℃下反应0.01~1小时后，再加入硼酸酯类化合物，在0~80℃下反应0.1~20小时后减压浓缩，柱层析分离即得；所述半花菁类化合物的结构式如下：
其中，取代基R4为H、甲氧基、羟基、硝基、氨基、卤素、醛基、羧基、巯基、磺酸基、巯基、酯基、氰基、酰胺基或炔基；
X-为碘离子、溴离子、氯离子、硝酸根离子、硫酸根离子、碳酸根离子或亚硫酸根离子；n为1~20的整；Y为羟基、巯基或氨基；
所述硼酸酯类化合物的结构式如下：
其中，取代基R1，R2，R3为H、甲基、乙基、丙基、甲氧基、羟基、硝基、氨基、卤素、醛基、羧基、磺酸基、巯基、酯基、氰基、酰胺基或炔基。
3.如权利要求2所述的一种酪氨酸酶和过氧化氢双响应型的近红外荧光探针的制备方法，其特征在于：所述半花菁类化合物和硼酸酯类化合物的摩尔比为1:0.2~1:5。
4.如权利要求2所述的一种酪氨酸酶和过氧化氢双响应型的近红外荧光探针的制备方法，其特征在于：所述有机溶剂为四氢呋喃、乙腈、二甲基亚砜、N, N-二甲基甲酰胺、二氯甲烷、苯、甲苯、二甲苯、氯苯、甲醇、乙醇、三氯甲烷中至少一种。
5.如权利要求2所述的一种酪氨酸酶和过氧化氢双响应型的近红外荧光探针的制备方法，其特征在于：所述碱为有机碱或无机碱；其中有机碱为三乙胺、吡啶或4-二甲胺基吡啶；无机碱为碳酸钠、碳酸钾、氢氧化钠、碳酸铯、碳酸氢钠、碳酸氢钾或碳酸钡。
6.如权利要求2所述的一种酪氨酸酶和过氧化氢双响应型的近红外荧光探针的制备方法，其特征在于：所述半花菁类化合物与三光气的摩尔比为1:0.1~1:0.5。
7.如权利要求2所述的一种酪氨酸酶和过氧化氢双响应型的近红外荧光探针的制备方法，其特征在于：所述半花菁类化合物与碱的摩尔比为1:1~1:5。
8.如权利要求1所述的一种酪氨酸酶和过氧化氢双响应型的近红外荧光探针在制备荧光成像试剂中的应用。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210485902.3A CN114702518A (zh) | 2022-05-06 | 2022-05-06 | 一种酪氨酸酶和过氧化氢双响应型的近红外荧光探针及其制备与应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210485902.3A CN114702518A (zh) | 2022-05-06 | 2022-05-06 | 一种酪氨酸酶和过氧化氢双响应型的近红外荧光探针及其制备与应用 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114702518A true CN114702518A (zh) | 2022-07-05 |
Family
ID=82176999
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210485902.3A Pending CN114702518A (zh) | 2022-05-06 | 2022-05-06 | 一种酪氨酸酶和过氧化氢双响应型的近红外荧光探针及其制备与应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114702518A (zh) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109810101A (zh) * | 2017-11-21 | 2019-05-28 | 中国科学院宁波材料技术与工程研究所 | 化合物、其制备方法及含有其的荧光探针和应用 |
| CN110540837A (zh) * | 2019-09-23 | 2019-12-06 | 湘潭大学 | 一种过氧化氢近红外荧光探针的制备和应用 |
| CN112574243A (zh) * | 2020-12-21 | 2021-03-30 | 大连理工大学 | 一种快速响应的过氧化氢长波长荧光探针的合成及应用 |
-
2022
- 2022-05-06 CN CN202210485902.3A patent/CN114702518A/zh active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109810101A (zh) * | 2017-11-21 | 2019-05-28 | 中国科学院宁波材料技术与工程研究所 | 化合物、其制备方法及含有其的荧光探针和应用 |
| CN110540837A (zh) * | 2019-09-23 | 2019-12-06 | 湘潭大学 | 一种过氧化氢近红外荧光探针的制备和应用 |
| CN112574243A (zh) * | 2020-12-21 | 2021-03-30 | 大连理工大学 | 一种快速响应的过氧化氢长波长荧光探针的合成及应用 |
Non-Patent Citations (3)
| Title |
|---|
| ALI HARIRI 等: "Molecular imaging of oxidative stress using an LED-based photoacoustic imaging system", SCIENTIFIC REPORTS, 6 August 2019 (2019-08-06), pages 9 * |
| JIAHANG ZHANG等: "Near-infrared fluorescence probe for hydrogen peroxide detection: design, synthesis, and application in living systems", ANALYST, vol. 144, no. 11, 25 April 2019 (2019-04-25), pages 3643 - 3648 * |
| NING DING等: "Detection of Tyrosinase in Real Food Samples and Living Cells by a Novel Near-Infrared Fluorescence Probe", JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, vol. 69, no. 6, 2 February 2021 (2021-02-02), pages 1994 - 2000 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110746410B (zh) | 一种亮氨酸氨肽酶和单胺氧化酶激活的近红外荧光探针、合成方法及生物应用 | |
| CN112409322B (zh) | Ggt激活型化学发光探针及其合成方法和应用 | |
| CN110283583B (zh) | γ-谷氨酰转肽酶响应型分子探针及其应用 | |
| CN114380808B (zh) | 用于中性粒细胞弹性蛋白酶双模态成像检测的分子探针及制备方法与应用 | |
| CN109336835B (zh) | 用于检测髓过氧化物酶活性荧光探针及其制备方法和应用 | |
| CN108117547B (zh) | 基于喹喔啉酮芳基硫醚类的荧光探针及其制备方法和应用 | |
| CN113461609B (zh) | 一种硫酸酯酶响应的aie纳米探针及其制备方法与应用 | |
| CN110357865A (zh) | 一种用于检测hNQO1酶的近红外荧光探针及其合成方法和应用 | |
| CN111518083A (zh) | 一种检测一氧化碳的打开型荧光探针的制备和应用 | |
| CN111410652B (zh) | 一种具有聚集诱导发光效应的线粒体靶向型近红外荧光探针的制备 | |
| CN109180716B (zh) | 一种多信号比率型区分检测h2o2和h2s的荧光探针的设计、合成及应用 | |
| CN114478473A (zh) | 一种亮氨酸氨基肽酶化学发光检测试剂的合成及应用 | |
| CN114702518A (zh) | 一种酪氨酸酶和过氧化氢双响应型的近红外荧光探针及其制备与应用 | |
| CN109485627B (zh) | 一种香豆素衍生物及其合成方法和在检测硫化氢中的应用 | |
| CN113278413B (zh) | 一种基于萘醌触发基团的nqo1近红外荧光探针、制备方法及应用 | |
| CN109776379A (zh) | 一种可用于响应活细胞内和慢性伤口发展过程中pH变化的近红外荧光探针及其制备方法 | |
| CN111689937B (zh) | 一种过氧化氢激活阿司匹林可视化前药及其制法和应用 | |
| CN110229203B (zh) | 一种氨基己糖酶荧光探针及其制备方法和应用 | |
| CN113527389A (zh) | 快速检测β-半乳糖苷酶的荧光探针及其制备方法和应用 | |
| CN110452187B (zh) | 一种光控酪氨酸酶荧光分子探针及其制备方法与应用 | |
| CN112457360A (zh) | 一种肝靶向的过氧亚硝酸根荧光探针及制备方法和应用 | |
| CN113135904B (zh) | 羟自由基近红外荧光分子探针及其制备方法与应用 | |
| CN110003291B (zh) | 一种氟代糖基修饰的紫杉醇类化合物及其合成方法和应用 | |
| CN112142766A (zh) | 一种苯并吲哚类过氧化氢荧光探针及其制备方法 | |
| CN116396283B (zh) | 一种具有大斯托克斯位移特征的羧酸酯酶2识别近红外荧光探针及其制备方法与应用 |
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 |