CN104262373B - A kind of targeting detection the fluorescence probe of inhibition cancer cell, preparation method and application - Google Patents
A kind of targeting detection the fluorescence probe of inhibition cancer cell, preparation method and application Download PDFInfo
- Publication number
- CN104262373B CN104262373B CN201410412644.1A CN201410412644A CN104262373B CN 104262373 B CN104262373 B CN 104262373B CN 201410412644 A CN201410412644 A CN 201410412644A CN 104262373 B CN104262373 B CN 104262373B
- Authority
- CN
- China
- Prior art keywords
- add
- dried
- good solvent
- dichloromethane
- solid
- 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.)
- Expired - Fee Related
Links
- 239000000523 sample Substances 0.000 title claims abstract description 36
- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 23
- 201000011510 cancer Diseases 0.000 title claims abstract description 23
- 230000008685 targeting Effects 0.000 title claims abstract description 22
- 230000005764 inhibitory process Effects 0.000 title claims abstract description 19
- 238000001514 detection method Methods 0.000 title claims abstract description 16
- 238000002360 preparation method Methods 0.000 title claims abstract description 15
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium Chemical compound [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 claims abstract description 42
- 238000006243 chemical reaction Methods 0.000 claims abstract description 23
- 229910052763 palladium Inorganic materials 0.000 claims abstract description 21
- RIOQSEWOXXDEQQ-UHFFFAOYSA-N triphenylphosphine Chemical compound C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1 RIOQSEWOXXDEQQ-UHFFFAOYSA-N 0.000 claims abstract description 14
- -1 aryl butadiene derivatives Chemical class 0.000 claims abstract description 12
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 95
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 57
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 52
- 239000000047 product Substances 0.000 claims description 34
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 32
- 239000007787 solid Substances 0.000 claims description 32
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 30
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 30
- 239000002904 solvent Substances 0.000 claims description 29
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 claims description 26
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 24
- 239000003208 petroleum Substances 0.000 claims description 22
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 claims description 20
- 239000000706 filtrate Substances 0.000 claims description 20
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 claims description 20
- 238000000926 separation method Methods 0.000 claims description 17
- 230000001681 protective effect Effects 0.000 claims description 16
- 238000003756 stirring Methods 0.000 claims description 16
- 239000003480 eluent Substances 0.000 claims description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- DMBMXJJGPXADPO-UHFFFAOYSA-N 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzaldehyde Chemical compound O1C(C)(C)C(C)(C)OB1C1=CC=C(C=O)C=C1 DMBMXJJGPXADPO-UHFFFAOYSA-N 0.000 claims description 12
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 12
- 239000003054 catalyst Substances 0.000 claims description 12
- 239000012074 organic phase Substances 0.000 claims description 12
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 12
- ODWXUNBKCRECNW-UHFFFAOYSA-M bromocopper(1+) Chemical compound Br[Cu+] ODWXUNBKCRECNW-UHFFFAOYSA-M 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 10
- 239000012046 mixed solvent Substances 0.000 claims description 10
- 229910000027 potassium carbonate Inorganic materials 0.000 claims description 10
- 238000013019 agitation Methods 0.000 claims description 8
- 239000000376 reactant Substances 0.000 claims description 8
- 238000000967 suction filtration Methods 0.000 claims description 8
- BGMHQBQFJYJLBP-UHFFFAOYSA-N 4-ethynylbenzaldehyde Chemical compound O=CC1=CC=C(C#C)C=C1 BGMHQBQFJYJLBP-UHFFFAOYSA-N 0.000 claims description 7
- 229920001184 polypeptide Polymers 0.000 claims description 7
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 7
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 claims description 6
- 238000005859 coupling reaction Methods 0.000 claims description 6
- 239000012153 distilled water Substances 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 6
- 239000000725 suspension Substances 0.000 claims description 6
- 238000004440 column chromatography Methods 0.000 claims description 5
- 239000012043 crude product Substances 0.000 claims description 5
- 238000001914 filtration Methods 0.000 claims description 4
- 239000000463 material Substances 0.000 claims description 4
- 150000003233 pyrroles Chemical class 0.000 claims description 4
- 125000001140 1,4-phenylene group Chemical group [H]C1=C([H])C([*:2])=C([H])C([H])=C1[*:1] 0.000 claims description 3
- UQIDNSKBUXCODH-UHFFFAOYSA-N diazaborine Chemical compound C1=CC(C)=CC=C1S(=O)(=O)N1B(O)C2=CC=CC=C2C=N1 UQIDNSKBUXCODH-UHFFFAOYSA-N 0.000 claims description 3
- 238000005406 washing Methods 0.000 claims description 3
- JXYITCJMBRETQX-UHFFFAOYSA-N 4-ethynylaniline Chemical compound NC1=CC=C(C#C)C=C1 JXYITCJMBRETQX-UHFFFAOYSA-N 0.000 claims description 2
- SJXHLZCPDZPBPW-UHFFFAOYSA-M 4-ethynylbenzoate Chemical compound [O-]C(=O)C1=CC=C(C#C)C=C1 SJXHLZCPDZPBPW-UHFFFAOYSA-M 0.000 claims description 2
- ZOXJGFHDIHLPTG-UHFFFAOYSA-N Boron Chemical compound [B] ZOXJGFHDIHLPTG-UHFFFAOYSA-N 0.000 claims description 2
- UAKJHPKLCNNVDO-UHFFFAOYSA-N C#N.C(#C)C1=CC=CC=C1 Chemical compound C#N.C(#C)C1=CC=CC=C1 UAKJHPKLCNNVDO-UHFFFAOYSA-N 0.000 claims description 2
- 206010011224 Cough Diseases 0.000 claims description 2
- 229910052796 boron Inorganic materials 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 claims description 2
- 238000000502 dialysis Methods 0.000 claims description 2
- 229910052731 fluorine Inorganic materials 0.000 claims description 2
- 239000011737 fluorine Substances 0.000 claims description 2
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 claims 3
- CTWQGTOWGFCWNW-UHFFFAOYSA-N 1,3-dimethylpyrrole Chemical compound CC=1C=CN(C)C=1 CTWQGTOWGFCWNW-UHFFFAOYSA-N 0.000 claims 2
- AQZMINLSVARCSL-UHFFFAOYSA-N 4-chloro-3,6-dioxocyclohexa-1,4-diene-1,2-dicarbonitrile Chemical class ClC1=CC(=O)C(C#N)=C(C#N)C1=O AQZMINLSVARCSL-UHFFFAOYSA-N 0.000 claims 2
- 150000001335 aliphatic alkanes Chemical class 0.000 claims 1
- RPGWZZNNEUHDAQ-UHFFFAOYSA-N phenylphosphine Chemical compound PC1=CC=CC=C1 RPGWZZNNEUHDAQ-UHFFFAOYSA-N 0.000 claims 1
- BMIBJCFFZPYJHF-UHFFFAOYSA-N 2-methoxy-5-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine Chemical compound COC1=NC=C(C)C=C1B1OC(C)(C)C(C)(C)O1 BMIBJCFFZPYJHF-UHFFFAOYSA-N 0.000 abstract description 6
- LIQLLTGUOSHGKY-UHFFFAOYSA-N [B].[F] Chemical compound [B].[F] LIQLLTGUOSHGKY-UHFFFAOYSA-N 0.000 abstract description 3
- 239000002253 acid Substances 0.000 abstract description 3
- 150000003935 benzaldehydes Chemical class 0.000 abstract description 3
- 239000003814 drug Substances 0.000 abstract description 2
- 229940079593 drug Drugs 0.000 abstract description 2
- 150000004702 methyl esters Chemical class 0.000 abstract description 2
- 238000011275 oncology therapy Methods 0.000 abstract description 2
- KAKZBPTYRLMSJV-UHFFFAOYSA-N Butadiene Chemical compound C=CC=C KAKZBPTYRLMSJV-UHFFFAOYSA-N 0.000 abstract 4
- 238000006069 Suzuki reaction reaction Methods 0.000 abstract 2
- 125000000950 dibromo group Chemical group Br* 0.000 abstract 2
- 239000002262 Schiff base Substances 0.000 abstract 1
- 150000004753 Schiff bases Chemical class 0.000 abstract 1
- 238000006555 catalytic reaction Methods 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 91
- 239000000243 solution Substances 0.000 description 31
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 30
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 29
- 239000008103 glucose Substances 0.000 description 29
- 239000001963 growth medium Substances 0.000 description 24
- 239000006285 cell suspension Substances 0.000 description 15
- 238000010186 staining Methods 0.000 description 9
- 210000000170 cell membrane Anatomy 0.000 description 8
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 8
- 239000007789 gas Substances 0.000 description 7
- 238000003384 imaging method Methods 0.000 description 7
- 230000002401 inhibitory effect Effects 0.000 description 7
- 239000000843 powder Substances 0.000 description 7
- 230000004663 cell proliferation Effects 0.000 description 6
- 229940024606 amino acid Drugs 0.000 description 5
- 150000001413 amino acids Chemical class 0.000 description 5
- 210000003855 cell nucleus Anatomy 0.000 description 4
- QTMDXZNDVAMKGV-UHFFFAOYSA-L copper(ii) bromide Chemical compound [Cu+2].[Br-].[Br-] QTMDXZNDVAMKGV-UHFFFAOYSA-L 0.000 description 4
- 125000003963 dichloro group Chemical group Cl* 0.000 description 4
- 239000000975 dye Substances 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 210000000633 nuclear envelope Anatomy 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 3
- 102000004142 Trypsin Human genes 0.000 description 3
- 108090000631 Trypsin Proteins 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 230000029087 digestion Effects 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 230000035755 proliferation Effects 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 231100000419 toxicity Toxicity 0.000 description 3
- 239000012588 trypsin Substances 0.000 description 3
- XPNGNIFUDRPBFJ-UHFFFAOYSA-N (2-methylphenyl)methanol Chemical compound CC1=CC=CC=C1CO XPNGNIFUDRPBFJ-UHFFFAOYSA-N 0.000 description 2
- LKNKAEWGISYACD-UHFFFAOYSA-N 1-bromobuta-1,3-diene Chemical class BrC=CC=C LKNKAEWGISYACD-UHFFFAOYSA-N 0.000 description 2
- 238000005160 1H NMR spectroscopy Methods 0.000 description 2
- HZNVUJQVZSTENZ-UHFFFAOYSA-N 2,3-dichloro-5,6-dicyano-1,4-benzoquinone Chemical class ClC1=C(Cl)C(=O)C(C#N)=C(C#N)C1=O HZNVUJQVZSTENZ-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 229910015900 BF3 Inorganic materials 0.000 description 2
- 229910021590 Copper(II) bromide Inorganic materials 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 description 2
- 238000005481 NMR spectroscopy Methods 0.000 description 2
- 101150003085 Pdcl gene Proteins 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical class [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 230000007541 cellular toxicity Effects 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 239000007795 chemical reaction product Substances 0.000 description 2
- 210000000805 cytoplasm Anatomy 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 230000031700 light absorption Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 125000001160 methoxycarbonyl group Chemical group [H]C([H])([H])OC(*)=O 0.000 description 2
- QPJVMBTYPHYUOC-UHFFFAOYSA-N methyl benzoate Chemical compound COC(=O)C1=CC=CC=C1 QPJVMBTYPHYUOC-UHFFFAOYSA-N 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 210000004940 nucleus Anatomy 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 238000012549 training Methods 0.000 description 2
- 238000007039 two-step reaction Methods 0.000 description 2
- 238000001291 vacuum drying Methods 0.000 description 2
- DWNBOPVKNPVNQG-LURJTMIESA-N (2s)-4-hydroxy-2-(propylamino)butanoic acid Chemical compound CCCN[C@H](C(O)=O)CCO DWNBOPVKNPVNQG-LURJTMIESA-N 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 206010070834 Sensitisation Diseases 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 238000005415 bioluminescence Methods 0.000 description 1
- 230000029918 bioluminescence Effects 0.000 description 1
- 239000000090 biomarker Substances 0.000 description 1
- 208000019065 cervical carcinoma Diseases 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 230000001934 delay Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000003255 drug test Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 229960000310 isoleucine Drugs 0.000 description 1
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 description 1
- 210000001161 mammalian embryo Anatomy 0.000 description 1
- 238000001840 matrix-assisted laser desorption--ionisation time-of-flight mass spectrometry Methods 0.000 description 1
- 229940095102 methyl benzoate Drugs 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 238000006862 quantum yield reaction Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 238000009777 vacuum freeze-drying Methods 0.000 description 1
- 239000004474 valine Substances 0.000 description 1
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
-
- 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"
-
- 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
- C09K2211/1022—Heterocyclic compounds bridged by heteroatoms, e.g. N, P, Si or B
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Health & Medical Sciences (AREA)
- Optics & Photonics (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Materials Engineering (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Heterocyclic Carbon Compounds Containing A Hetero Ring Having Oxygen Or Sulfur (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention discloses a kind of targeting detection the fluorescence probe of inhibition cancer cell, preparation method and application, belong to biological sensor field.Described fluorescence probe is containing 4,4', 4 " 1,4 4 aryl butadiene derivatives of double fluorine boron dipyrryls group；By 4,4'((1E, 3E) 1,4 dibromos 1,3 butadiene 1,4 diyls) two benzaldehydes (or 4,4'((1E, 3E) 1,4 dibromos 1,3 butadiene 1,4 diyls) dibenzoic acid methyl esters) and BODIPY pinacol ester four (triphenylphosphine) close palladium catalysis under obtain by suzuki reaction (Suzuki reaction) or obtained by a step schiff base reaction by DIBODIPY DICHO and biomolecule；Described preparation method is simple, easy to operate；Described fluorescence probe has the function of targeting mark and inhibition cancer cell simultaneously, can be used as potential cancer therapy drug.
Description
Technical field
The present invention relates to a kind of targeting detection the fluorescence probe of inhibition cancer cell, preparation method and use, specifically
Relate to a kind of containing 4,4', 4 "-bis-fluorine boron dipyrryls group 1,4-tetra-aryl butadiene derivatives fluorescence probe and
Its preparation method, described fluorescence probe can be used for carrying out cancer cell the bioluminescence imaging technique of targeting detection,
Belong to fluorescent optical sensor field.
Background technology
Imaging-PAM is indispensable one for biomedical research, clinical diagnosis, drug test
Technology.Preferably fluorescence probe should possess and suitably excites and launch wavelength, higher brightness, the most raw
The advantages such as thing stability, relatively low bio-toxicity, preferable photostability.Especially for living cells and body
Interior imaging, water-soluble and fat-soluble balance is to reach the key condition of higher film infiltration.But, traditional is glimmering
Light probe due to higher photobleaching, poor living cells targeting, the intrinsic shortcoming such as fat-soluble without
Method meets the requirement of people.Therefore, the development and application of new preferable fluorescence probe has become present stage section
Grind a popular research of worker.
At present, most of documents are it has been reported that double fluorine boron two pyrroles (BODIPYs) class dyestuff is glimmering as one
Light probe is widely used in the fields such as biomarker, fluorescent device, chemical sensitisation, artificial light assembly system.BODIPYs
Dyestuff has higher molar extinction coefficient, biocompatibility, preferable chemical and photochemical stability, relatively
The advantages such as high fluorescence quantum yield.But, the BODIPYs dyestuff that existing document was reported is for living cells
During major part is distributed in cytoplasm during imaging, thus do not possesses targeting.(Karolin,J.；Johansson,L.
B.-A.；Strandberg,L.J.Am.Chem.Soc.,1994,116,7801-7806.Loudet,A.；Burgess,
K.Chem.Rev.,2007,107,4891-4932.Hu,R.；Lager,E.；Aguilar-Aguilar,A.J.Phys.
Chem.C,2009,113,15845-15853.Whited,M.T.；Djurovich,P.I.；Roberts,S.T.J.
Am.Chem.Soc.,2011,133,88-96.Zhu,S.L.；Zhang,J.T.；Vegesna,G.Org.Lett.,
2011,13,438-441.Kamkaew,A.；Lim,S.H.；Lee,H.B.Chem.Soc.Rev.2013,42,
77-78.Lin,Y.；Lin,W.Y.；Zheng,K.B.Chem.Soc.Rev.2013,42,622-661.Fu,Y.Y.；
He,Q.G.；Zhu,D.F.Chem.Comm.2013,49,11266-11268.Ye,J.H.；Chen,H.C.；
Bai,Y.RSC Adv.,2014,4,6691-6695.Saha,T.；Kand,D.；Talukdar,P.Org.Biomol.
Chem.,2013,11,8166-8170.Collado,D.；Vida,Y.；Najera,F.RSC Adv.,2014,4,
2306-2309.Zhang,H.X.；Chen,J.B.；Guo,X.F.Anal.Chem.2014,86,
3115-3123.)。
Summary of the invention
When being used for living cells imaging for current BODIPYs dyestuff, major part is distributed in cytoplasm, does not possesses
The problem of targeting, an object of the present invention is to provide a kind of for targeting detection inhibition cancer cell glimmering
Light probe, described fluorescence probe has the function of targeting mark and inhibition cancer cell simultaneously；The two of purpose are to carry
For a kind of preparation method for the targeting detection also fluorescence probe of inhibition cancer cell, described preparation method is simple,
Easy to operate；The three of purpose are to provide a kind of application for the targeting detection also fluorescence probe of inhibition cancer cell.
The purpose of the present invention is realized by techniques below scheme:
It is a kind of that for targeting detection the fluorescence probe of inhibition cancer cell, described fluorescence probe is containing 4,4', 4 "-
Isosorbide-5-Nitrae-four aryl butadiene derivatives of double fluorine boron dipyrryls group, its structural formula is as follows:
R preferably-CHO ,-COOCH3、-NH3,-CN, amino acid, one in small peptide and polypeptide；Its
In, R ' is identical with R；
A kind of preparation method for the targeting detection also fluorescence probe of inhibition cancer cell, described method step is such as
Under:
As R preferably-CHO ,-COOCH3、-NH3Or the preparation of fluorescence probe during-CN:
(1) catalyst is added in reactor 1, be sequentially added into good solvent 1 and reactant 1, under agitation
Carry out coupling reaction, after reaction 11h, filter, take filtrate and be spin-dried for, obtain reacting coarse product 1；Will reaction
Crude product 1 is dissolved in chloroform, utilizes pillar layer separation to purify, and is dried, obtains solid a；
Wherein, the preferred palladium bichloride of described catalyst and the mixture of copper bromide, palladium bichloride and the material of copper bromide
The preferred 0.04:3 of ratio of amount；
The preferred 1:50 of volume ratio of the preferred acetonitrile of good solvent 1 and the mixed solvent of toluene, acetonitrile and toluene；
The preferred 4-ethynylbenzaldehyde of reactant 1,4-ethynyl-benzoate, 4-acetylenylbenzene formonitrile HCN and
One in 4-acetylenylaniline；
Described reactant 1, catalyst and the preferred 1:5:28 of mass ratio of good solvent 1；
Described pillar layer separation purifies with petroleum ether and dichloromethane as eluant, eluent, wherein, and petroleum ether and dichloro
The preferred 1:1 of mass ratio of methane；
(2) 4-formylphenylboronic acid pinacol ester is added in reactor 2, add dichloromethane, obtain dense
Degree is the 4-formylphenylboronic acid pinacol ester solution of 5.73g/L, is passed through protective gas in described solution,
Add Isosorbide-5-Nitrae-dimethyl pyrrole, be under agitation added dropwise over trifluoroacetic acid, react 12h, add 2,3-dichloro
-5,6-dicyano-Isosorbide-5-Nitrae-benzoquinones, reacts 4h, adds DIPEA, treats N, N-diisopropyl second
After amine is completely dissolved, reactor 2 is put in ice bath, adds BFEE, react 12h, wash,
Separatory, takes organic phase and adds anhydrous sodium sulfate in organic phase, filtering, take filtrate and be spin-dried for, reacted
Crude product 2；Reacting coarse product 2 is dissolved in dichloromethane, utilizes pillar layer separation to purify, be dried,
To solid b；
Wherein, 4-formylphenylboronic acid pinacol ester, Isosorbide-5-Nitrae-dimethyl pyrrole, trifluoroacetic acid, 2,3-bis-chloro-5,6-
Dicyano-1,4-benzoquinones, N, N-diisopropylethylamine are preferred with the mass ratio of BFEE: 1:1:0.25:
1:6.5:9.8；
Described pillar layer separation purifies with petroleum ether and dichloromethane as eluant, eluent, wherein, and petroleum ether and dichloro
The preferred 1:4 of mass ratio of methane；
The preferred N of described protective gas2With the one in Ar；
(3) solid a, solid b, potassium carbonate and four (triphenylphosphines) are closed palladium and add in reactor 3, repeat
Vacuumize, be passed through protective gas, add good solvent 2, at 70 DEG C, react 12h, suction filtration, take filtrate rotation
Dry, obtain reacting coarse product 3, reacting coarse product 3 is dissolved in chloroform, purify with pillar layer separation,
It is dried, obtains fluorescence probe of the present invention；
Wherein, the preferred N of described protective gas2With the one in Ar；
The preferred 3:1 of volume ratio of the described preferred toluene of good solvent 2 and the mixed solvent of methyl alcohol, toluene and methyl alcohol
Solid a, solid b, potassium carbonate, four (triphenylphosphine) close palladium, good solvent 2 mass ratio preferred: 1:2.5:
2:0.18:210；
Described pillar layer separation purifies with petroleum ether and dichloromethane as eluant, eluent, wherein, and petroleum ether and dichloro
The preferred 1:1 of mass ratio of methane；
The preparation of fluorescence probe when R preferred amino acid, small peptide or polypeptide:
(1) catalyst is added in reactor 4, be sequentially added into good solvent 4 and 4-ethynylbenzaldehyde,
Carry out coupling reaction under stirring, after reaction 11h, filter, take filtrate and be spin-dried for, obtain reacting coarse product 4；
Reacting coarse product 4 is dissolved in chloroform, with petroleum ether and dichloromethane as eluant, eluent, utilizes column chromatography
Separating-purifying, is dried, obtains solid 1；
Wherein, the preferred palladium bichloride of described catalyst and the mixture of copper bromide, palladium bichloride and the material of copper bromide
The preferred 0.04:3 of ratio of amount；
The preferred 1:50 of volume ratio of the preferred acetonitrile of good solvent 4 and the mixed solvent of toluene, acetonitrile and toluene；
Described 4-ethynylbenzaldehyde, catalyst and the preferred 1:5:28 of mass ratio of good solvent 1；
Described petroleum ether and the preferred 1:1 of mass ratio of dichloromethane；
(2) 4-formylphenylboronic acid pinacol ester is added in reactor 5, add dichloromethane, obtain dense
Degree is the 4-formylphenylboronic acid pinacol ester solution of 5.73g/L, is passed through protective gas in described solution,
Add Isosorbide-5-Nitrae-dimethyl pyrrole, be under agitation added dropwise over trifluoroacetic acid, react 12h, add 2,3-bis-chloro-5,6-
Dicyano-Isosorbide-5-Nitrae-benzoquinones, reacts 4h, adds DIPEA, treats that DIPEA is complete
After dissolving, reactor 5 is put in ice bath, adds BFEE, react 12h, wash, separatory,
Take organic phase and in organic phase, add anhydrous sodium sulfate, filtering, take filtrate and be spin-dried for, obtain reacting coarse product 5；
Reacting coarse product 5 is dissolved in dichloromethane, with petroleum ether and dichloromethane as eluant, eluent, utilizes column chromatography
Separating-purifying, is dried, obtains solid 2；
Wherein, 4-formylphenylboronic acid pinacol ester, Isosorbide-5-Nitrae-dimethyl pyrrole, trifluoroacetic acid, 2,3-bis-chloro-5,6-
Dicyano-1,4-benzoquinones, N, N-diisopropylethylamine, BFEE mass ratio preferred: 1:1:0.25:
1:6.5:9.8；
Described petroleum ether and the preferred 1:4 of mass ratio of dichloromethane；
(3) solid 1, solid 2, potassium carbonate and four (triphenylphosphines) are closed palladium to add in reactor 6, take out true
Sky, is passed through protective gas, adds good solvent 5, reacts 12h, suction filtration, take filtrate and be spin-dried at 70 DEG C,
Obtain reacting coarse product 6, reacting coarse product 6 be dissolved in chloroform, purify with pillar layer separation, be dried,
Obtain DIBODIPY-DICHO；
Wherein, the preferred N of described protective gas2With the one in Ar；
The preferred 3:1 of volume ratio of the described preferred toluene of good solvent 5 and the mixed solvent of methyl alcohol, toluene and methyl alcohol
Solid a, solid b, potassium carbonate, four (triphenylphosphine) close palladium, good solvent 2 mass ratio preferred: 1:2.5:
2:0.18:210；
Described pillar layer separation purifies with petroleum ether and dichloromethane as eluant, eluent, wherein, and petroleum ether and dichloro
The preferred 1:1 of mass ratio of methane；
(4) DIBODIPY-DICHO is added in reactor 7, add good solvent 6, fully dissolve,
To the DIBODIPY-DICHO solution that concentration is 0.53g/L；Biomolecule is added in described solution,
30 DEG C of stirring reaction 40h, are dried, add dimethyl sulfoxide (DMSO), obtain suspension a；Add in suspension a
Distilled water, stirs, dialysis, is dried, obtains fluorescence probe of the present invention；
Wherein, DIBODIPY-DICHO is 10,10'-(((1Z, 3Z)-Isosorbide-5-Nitrae-two (4-Fonnylphenyl)-1,3-fourth two
Alkene-1,4-diyl) two (1,4-phenylenes)) two (5,5-bis-fluoro-1,3,7,9-tetramethyl-5H-two pyrroles
Cough up) abbreviation of [1,2-c:2', 1'-f] [1,3,2] diaza borine.
DIBODIPY-DICHO, biomolecule, the preferred 1:1:688 of mass ratio of dimethyl sulfoxide (DMSO)；
The described preferred N,N-dimethylformamide of good solvent 6；
One in described biomolecule preferred amino acid, small peptide and polypeptide；
Described dry preferred vacuum freezedrying；Suspension a and the preferred 1:200 of volume ratio of distilled water.
The fluorescence probe of a kind of targeting detection inhibition cancer cell is applied to targeting detection cancer cell and suppresses simultaneously
The propagation of cancer cell.
Beneficial effect:
(1) fluorescence probe of the present invention can carry out targeting mark to cancer cell, and fluorescence probe becomes at cell
For different cell-targeting site differences during picture；For human cervical carcinoma cell (Hela cell), fluorescence is visited
The target site of pin is cell membrane and nucleus；For differing bigger mouse embryo fibroblast with Hela cell
For cell (3T3swiss cell is called for short 3T3cell), the target site of fluorescence probe is the nuclear membrane of cell；
And human breast cancer cell (MCF-7cell), the target site of fluorescence probe is cell membrane；
(2) fluorescence probe of the present invention has inhibitory action to the propagation of Hela cell, MCF-7cell, but
Being for non-cancerous cells, the propagation of 3T3cell is having no significant effect；
(3) fluorescence probe of the present invention has the function of targeting mark and inhibition cancer cell simultaneously, can be used as
Potential cancer therapy drug；
(4) fluorescence probe preparation method of the present invention is simple, easy to operate.
Accompanying drawing explanation
Fig. 1 is that in embodiment 1, DIBODIPY-DICHO concentration is 5 × 10-6Mol/L and 1 × 10-5mol/L
Time respectively to Hela cell, MCF-7cell, 3T3cell at the concentration-survival rate figure of 24h, 48h, 72h；
Fig. 2 is that in embodiment 1, DIBODIPY-DICHO concentration is 2.5 × 10-5mol/L、5×10-5mol/L
With 1 × 10-4During mol/L respectively to Hela cell, MCF-7cell, 3T3cell at 24h, 48h, 72h
Concentration-survival rate figure；
Fig. 3 is DIBODIPY-DICOOCH in embodiment 23Concentration is 5 × 10-6Mol/L and 1 × 10-5
During mol/L respectively to Hela cell, MCF-7cell, 3T3cell in the concentration-survival of 24h, 48h, 72h
Rate figure；
Fig. 4 is DIBODIPY-DICOOCH in embodiment 23Concentration is 2.5 × 10-5mol/L、5×10-5
Mol/L and 1 × 10-4During mol/L respectively to Hela cell, MCF-7cell, 3T3cell 24h, 48h,
The concentration of 72h-survival rate figure；
Fig. 5 is that in embodiment 3, DIBOODIPY-PEPTIDE concentration is 5 × 10-6Mol/L and 1 × 10-5
During mol/L respectively to Hela cell, MCF-7cell, 3T3cell in the concentration-survival of 24h, 48h, 72h
Rate figure；
Fig. 6 is that in embodiment 3, DIBOODIPY-PEPTIDE concentration is 2.5 × 10-5mol/L、5×10-5
Mol/L and 1 × 10-4During mol/L respectively to Hela cell, MCF-7cell, 3T3cell 24h, 48h,
The concentration of 72h-survival rate figure.
Detailed description of the invention
The present invention is described in detail in detail with specific embodiment below in conjunction with the accompanying drawings, but is not limited to this.
The main agents information mentioned in following example is shown in Table 1；Key instrument and facility information are shown in Table 2.
Table 1
Embodiment 1
(1) palladium bichloride (PdCl is weighed2, 0.1383g), copper bromide (CuBr2, 10.4105g) join
In 250mL round-bottomed flask, add acetonitrile (1.26mL) and toluene (63mL), be slowly added to 4-acetenyl
Benzaldehyde (2.0g), at 25 DEG C, stirring carries out coupling reaction.Reaction is stopped, by flask after reaction 11h
Middle mixture filtered on buchner funnel, filtrate is spin-dried for, and obtains reacting coarse product 1, by reacting coarse product 1 with 5
After mL chloroform dissolves, with petroleum ether: dichloromethane=1:1, as eluant, eluent, utilizes column chromatography analysis to divide
From, vacuum drying obtains 4,4'-(the bromo-1,3-butadiene-Isosorbide-5-Nitrae-diyl of (1E, 3E)-Isosorbide-5-Nitrae-two) two benzaldehydes；
(2) weigh in the there-necked flask that 573mg4-formylphenylboronic acid pinacol ester joins 250mL,
Add 100mL CaH2Dried dichloromethane, plug is airtight to its logical N230min, by 565mg1,4-
Dimethyl pyrrole adds, agitation and dropping 0.1mL trifluoroacetic acid, and stirring reaction 12h at 25 DEG C, reactant is complete
Entirely run out of, disposably add 560mg2,3-bis-chloro-5,6-dicyano-Isosorbide-5-Nitrae-benzoquinones, N2The lower stirring of protection is anti-
Add 5mL DIPEA after answering 4h and continue stirring 15min, in ice bath, then add 5mL tri-
Boron fluoride ether reaction 12h.Crude product is moved on in 500mL separatory funnel with distilling washing three times, every time
100mL, then wash three times with saturated nacl aqueous solution, each 100mL, separate to obtain organic phase.By organic phase
Being dried except water with anhydrous sodium sulfate, suction filtration obtains filtrate.Filtrate is spin-dried for, and obtains reacting coarse product 2, by thick for reaction
Product 2 is dissolved in 5mL dichloromethane, purifies with pillar layer separation, and eluant, eluent is petroleum ether: dichloromethane
Alkane=1:4, obtains red solid BODIPY pinacol ester.
(3) in two mouthfuls of flasks of 100mL, product prepared by above-mentioned two-step reaction is added
4,4'-((1E, 3E)-1,4-two bromo-1,3-butadiene-1,4-diyl) two benzaldehydes (122.2mg) and BODIPY frequency
Any alcohol ester (302.1mg), and add 250.9mg potassium carbonate and 21.5mg tetra-(triphenylphosphine) conjunction palladium, weight
Vacuumize again, fill N2Three times, (toluene with methyl alcohol volume ratio is to inject 30mL toluene-methanol mixed solvent
3:1), at N2Under protective condition, back flow reaction 12h at 70 DEG C, treat (1E, 3E)-Isosorbide-5-Nitrae-diphenyl-Isosorbide-5-Nitrae-
Two bromo-1,3-butadienes stop reaction, suction filtration after ruing out of completely, filtrate is spin-dried for, and obtains reacting coarse product 3, will
Reacting coarse product 3 is dissolved in 5mL chloroform, purifies with pillar layer separation, and eluant, eluent is oil
Ether: dichloromethane=1:1, obtains red solid powder.By nuclear magnetic resonance chemical analyser and mass-spectroscopic characterization
The nucleus magnetic hydrogen spectrum and the mass spectrometric data that obtain red solid powder are as follows:
1H-NMR(400MHz,CDCl2) δ (ppm): 9.97 (s, 2H), 7.83-7.75 (d, 5H), 7.50 (d, 7H),
7.35-7.29 (d, 4H), 6.97 (d, 2H), 6.12 (d, 2H), 1.54 (d, 24H) .MS (MALDI-TOF): calcd.for
C56H48B2F4N4O2, 906.4；Found, 906.5.
Illustrate that red solid powder is DIBODIPY-DICHO of the present invention.
(4) DIBODIPY-DICHO prepared by step (3) is dissolved in 0.01mL dimethyl sulfoxide (DMSO)
(DMSO), in, it is added thereto to 9mL DMEM in high glucose culture medium and is configured to 10-4The solution of mol/L.
10 will prepared again-4Mol/L solution DMEM in high glucose culture medium is diluted to 5 × 10 respectively-5mol/L、
2.5×10-5mol/L、1×10-5mol/L、0.5×10-5The solution of mol/L.
Respectively by bottom confluent cultures ware 80～90% Hela cell, MCF-7cell, 3T3cell with 0.05%
Trypsin Induced become round to 50～60% cell detachment culture dish bottom, in the culture dish of three kinds of cells
Each 3mL DMEM in high glucose culture medium that adds terminates digestion, more respectively by three kinds of cell suspensions from culture dish
Suck in three 15mL centrifuge tubes, respectively add DMEM in high glucose culture medium to 10mL, by three centrifuge tubes
It is centrifuged 5min as 1000rpm in centrifuge, removes supernatant；1mL is respectively added again in three centrifuge tubes
DMEM in high glucose culture medium, and the mixing with cells bottom centrifuge tube is uniform, by cell suspension with high sugared
About 2500000 (cell blood counting chambers in cell in DMEM culture medium dilution 50mL cell suspension
Counting).Three kinds of cell suspension liquid-transfering guns are injected separately in 96 porocyte culture plates (every kind of cell is each again
With 3 96 porocyte culture plates), every hole 100 μ L cell suspension (about 5000 cells), separately 96
Orifice plate adds a white control group of emptying, i.e. 100 μ L acellular DMEM in high glucose culture medium.By 96 holes
Tissue Culture Plate is placed in 37 DEG C, CO2The incubator of content 5% is cultivated 24h.
After cell in 96 porocyte culture plates cultivates 24h, each addition 100 μ L in each hole having cell
The DIBODIPY-DICHO solution of the above-mentioned variable concentrations prepared, every kind of concentration has 4 parallel holes.Separately
100 μ L DMEM in high glucose culture mediums are respectively added in the hole of outer blank control group.96 porocyte culture plates are continued
Continue and be placed in 37 DEG C, CO2Volume content be 5% incubator in cultivate 24h.
After adding DIBODIPY-DICHO solution 24h, three kinds of cells respectively take a plate 96 porocyte culture plate,
(MTS is 3-(4,5-with toxicity detection reagent MTS to give addition 20 μ L cell proliferations in every hole
dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazoliu
The abbreviation of m) continue to put into 37 DEG C, CO2Exist with enzyme connection detector after 4h cultivated by the incubator of content 5%
Detect the absorbance value (OD) in each hole at 492nm, calculate 24h DIBODIPY-DICHO according to OD value
The solution inhibiting rate to three kinds of cell proliferations；According to said method, detect DIBODIPY-DICHO respectively
Respectively to three kinds of cells at the proliferation inhibition rate of 48h and 72h.Draw concentration-survival rate figure such as Fig. 1 and Fig. 2
Shown in, it can be seen that DIBODIPY-DICHO is all to cancer cell Hela when 24h, 48h, 72h
The propagation of cell, MCF-7cell has inhibitory action, and has no significant effect the propagation of 3T3cell.
(5) respectively Hela cell, MCF-7cell, 3T3cell are passaged to confocal (laser co-focusing
Microscope) special culture dish, at 37 DEG C, CO2The incubator of content 5% is cultivated 24h.After 24h, use
PH7.4PBS (phosphate buffer) washes three times, in each culture dish in each addition 0.5mL step (4)
The 10 of preparation-5The DIBODIPY-DICHO solution of mol/L, then add the cultivation of 0.5mL DMEM in high glucose
Base.Continue to be put in 37 DEG C, CO2The incubator of content 5% is cultivated 4h.Sucking-off solution after 4h, uses pH7.4
PBS washes three times, and every ware adds 1mL DMEM in high glucose culture medium.Observe with confocal
The DIBODIPY-DICHO imaging contexts in three kinds of cells respectively.It is observed that for Hela cell,
The staining foci of DIBODIPY-DICHO is cell membrane and nucleus；For differing bigger with Hela cell
For 3T3cell, the staining foci of DIBODIPY-DICHO is the nuclear membrane of cell；And MCF-7cell,
The staining foci of DIBODIPY-DICHO is cell membrane.
Embodiment 2
(1) palladium bichloride (PdCl is weighed2, 0.1383g), copper bromide (CuBr2, 10.4105g) join
In 250mL round-bottomed flask, add acetonitrile (1.26mL) and toluene (63mL), be slowly added to 4-acetenyl
Methyl benzoate (2.0g), at 25 DEG C, stirring carries out coupling reaction.Reaction is stopped after reaction 11h, will
Mixture filtered on buchner funnel in flask, filtrate is spin-dried for, and obtains reacting coarse product 1, by reacting coarse product 1
After dissolving with 5mL chloroform, with petroleum ether: dichloromethane=1:1, as eluant, eluent, utilizes column chromatography to divide
Analysis separates, and vacuum drying obtains 4,4'-(the bromo-1,3-butadiene-Isosorbide-5-Nitrae-diyl of (1E, 3E)-Isosorbide-5-Nitrae-two) dibenzoic acid first
Ester.
(2) weigh in the there-necked flask that 573mg4-formylphenylboronic acid pinacol ester joins 250mL,
Add 100mL CaH2Dried dichloromethane, plug is airtight to its logical N230min, by 565mg1,4-
Dimethyl pyrrole adds, agitation and dropping 0.1mL trifluoroacetic acid, and stirring reaction 12h at 25 DEG C, reactant is complete
Entirely run out of, disposably add 560mg2,3-bis-chloro-5,6-dicyano-Isosorbide-5-Nitrae-benzoquinones, N2The lower stirring of protection is anti-
Add 5mL DIPEA after answering 4h and continue stirring 15min, in ice bath, then add 5mL tri-
Boron fluoride ether reaction 12h.Crude product is moved on in 500mL separatory funnel with distilling washing three times, every time
100mL, then wash three times with saturated nacl aqueous solution, each 100mL, separate to obtain organic phase.By organic phase
Being dried except water with anhydrous sodium sulfate, suction filtration obtains filtrate.Filtrate is spin-dried for, and obtains reacting coarse product 2, by thick for reaction
Product 2 is dissolved in 5mL dichloromethane, purifies with pillar layer separation, and eluant, eluent is petroleum ether: dichloromethane=
1:4, obtains red solid BODIPY pinacol ester.
(3) in two mouthfuls of flasks of 100mL, product prepared by above-mentioned two-step reaction is added
4,4'-((1E, 3E)-1,4-two bromo-1,3-butadiene-1,4-diyl) dibenzoic acid methyl esters (122.2mg) and BODIPY
Pinacol ester (302.1mg), and add 250.9mg potassium carbonate and 21.5mg tetra-(triphenylphosphine) conjunction palladium,
Repeat to vacuumize, fill N2Three times, inject 30mL toluene-methanol mixed solvent (toluene and methyl alcohol volume ratio
For 3:1), at N2Under protective condition at 70 DEG C back flow reaction 12h, treat (1E, 3E)-Isosorbide-5-Nitrae-diphenyl-Isosorbide-5-Nitrae-
Two bromo-1,3-butadienes stop reaction, suction filtration after ruing out of completely, filtrate is spin-dried for, and obtains reacting coarse product 3, will
Reacting coarse product 3 is dissolved in 5mL chloroform, purifies with pillar layer separation, and eluant, eluent is oil
Ether: dichloromethane=1:1, obtains red solid powder.By nuclear magnetic resonance chemical analyser and mass-spectroscopic characterization
The nucleus magnetic hydrogen spectrum and the mass spectrometric data that obtain red solid powder are as follows:
1H-NMR(400MHz,CDCl2) δ (ppm): 8.08-8.05 (d, 4H), 7.92-7.90 (d, 3H),
7.79-7.76 (d, 4H), 7.47 (d, 2H), 7.31-7.29 (d, 5H), 5.97 (d, 4H), 3.95 (s, 6H),
1.41-1.35(d,24H).
Illustrate that red solid powder is DIBODIPY-DICOOCH of the present invention3(10,10'-(((1Z,3Z)
-1,4-two (4-(methoxycarbonyl group) phenyl)-1,3-butadiene-1,4-diyl) two (1,4-phenylenes)) two (5,5-difluoros
-1,3,7,9-tetramethyl-5H-two pyrroles) abbreviation of [1,2-c:2', 1'-f] [1,3,2] diaza borine).
(4) DIBODIPY-DICOOCH prepared by step (3)3It is dissolved in 0.01mL DMSO, to
Wherein add 9mL DMEM in high glucose culture medium and be configured to 10-4The solution of mol/L.10 will prepared again-4
Mol/L solution DMEM in high glucose culture medium is diluted to 5 × 10 respectively-5mol/L、2.5×10-5mol/L、1×10-5
mol/L、0.5×10-5The solution of mol/L.
Respectively by bottom confluent cultures ware 80～90% Hela cell, MCF-7cell, 3T3cell with 0.05%
Trypsin Induced become round to 50～60% cell detachment culture dish bottom, in the culture dish of three kinds of cells
Each 3mL DMEM in high glucose culture medium that adds terminates digestion, more respectively by three kinds of cell suspensions from culture dish
Suck in three 15mL centrifuge tubes, respectively add DMEM in high glucose culture medium to 10mL, by three centrifuge tubes
It is centrifuged 5min as 1000rpm in centrifuge, takes out and pour out supernatant, three centrifuge tubes respectively add 1mL
Cell suspension DMEM in high glucose uniformly, is trained by DMEM in high glucose culture medium by cell piping and druming bottom centrifuge tube
Support in base dilution 50mL cell suspension containing about 2500000, cell (cell blood counting chamber counts).Again
Three kinds of cell suspension liquid-transfering guns are injected separately in 96 porocyte culture plates that (every kind of cell is respectively with 3 96
Porocyte culture plate), every hole 100 μ L cell suspension (about 5000 cells), separately add in 96 orifice plates
One empties white control group, i.e. 100 μ L acellular DMEM in high glucose culture medium.By 96 porocyte culture plates
It is placed in 37 DEG C, CO2The incubator of content 5% is cultivated 24h.
After cell in 96 porocyte culture plates cultivates 24h, each addition 100 μ L in each hole having cell
The DIBODIPY-DICOOCH of the above-mentioned variable concentrations prepared3Solution, every kind of concentration has 4 parallel holes.
Additionally respectively add 100 μ L DMEM in high glucose culture mediums in the hole of blank group.By 96 porocyte culture plates
Continue to be placed in 37 DEG C, CO2The incubator of content 5% is cultivated 24h.
Add DIBODIPY-DICOOCH3After solution 24h, three kinds of cells respectively take a plate 96 porocyte training
Support plate, give in every hole and add 20 μ L cell proliferations and toxicity detection reagent MTS, continue to put into 37 DEG C,
CO2At 492nm, detect the light absorption in each hole with enzyme connection detector after 4h cultivated by the incubator of content 5%
Value (OD), calculates 24h DIBODIPY-DICOOCH according to OD value3Solution is to three kinds of cell proliferations
Inhibiting rate, according to said method, detects DIBODIPY-DICOOCH respectively3Respectively to three kinds of cells 48
The proliferation inhibition rate of h and 72h.Draw concentration-survival rate figure as shown in Figure 3 and Figure 4, it can be seen that
The DIBODIPY-DICOOCH when 24h, 48h, 72h3All to cancer cell Hela cell, MCF-7cell
Propagation have inhibitory action, and the propagation of 3T3cell is had no significant effect.
(5) respectively Hela cell, MCF-7cell, 3T3cell are passaged to confocal special culture dish, 37 DEG C,
CO2The incubator of content 5% is cultivated 24h.After 24h, wash with pH 7.4PBS (phosphate buffer)
Three times, 10 prepared in each addition 0.5mL step (4) in each culture dish-5Mol/L's
DIBODIPY-DICOOCH3Solution, then add 0.5mL DMEM in high glucose culture medium.Continue to be put in 37 DEG C,
CO2The incubator of content 5% is cultivated 4h.Sucking-off solution after 4h, washes three times with pH7.4PBS, every ware
Each addition 1mL DMEM in high glucose culture medium.DIBODIPY-DICOOCH is observed with confocal3Respectively
Imaging contexts in three kinds of cells.It is observed that for Hela cell, DIBODIPY-DICOOCH3
Staining foci be cell membrane and nucleus；For differ bigger 3T3cell with Hela cell,
DIBODIPY-DICOOCH3The nuclear membrane that staining foci is cell；And MCF-7cell,
DIBODIPY-DICOOCH3Staining foci be cell membrane.
Embodiment 3
(1) DIBODIPY-DICHO of preparation in embodiment 1 being weighed 8mg, to be dissolved in 15mL the driest
The DMF (DMF) crossed, is added thereto to 8mg polypeptide and (is called for short cc-259, its structure
It is made up of 11 amino acid, including glycine (Gly), histidine (His), isoleucine (Ile), silk
Propylhomoserin (Ser), valine (Val)), oil bath 30 DEG C stirring reaction 40h, solvent is directly dried up with air pump
DMF.Product after drying up again is scattered in the dimethyl sulfoxide (DMSO) (DMSO) of 5mL, to remove not
The raw material DIBODIPY-DICHO of reaction.Then 200mL distilled water it is added thereto to, after stirring
Mixed solution is loaded in the bag filter of molecular cut off 1000, bag filter is immersed in distilled water and stand thoroughly
After analysis 48h, take out the solution in bag filter after dialysing, after liquid nitrogen frozen, be put in freeze drier cold
Freeze to vacuumize to be dried and i.e. can get pressed powder DIBOODIPY-PEPTIDE.
Wherein, end has 11 amino acid whose peptide sequences with amino:
(2) DIBOODIPY-PEPTIDE prepared by step (1) is dissolved in 0.01mL DMSO, to
Wherein add 9mL DMEM in high glucose culture medium and be configured to 10-4The solution of mol/L.10 will prepared again-4
Mol/L solution DMEM in high glucose culture medium is diluted to 5 × 10 respectively-5mol/L、2.5×10-5mol/L、1×10-5
mol/L、0.5×10-5The solution of mol/L.
Respectively by bottom confluent cultures ware 80～90% Hela cell, MCF-7cell, 3T3cell with 0.05%
Trypsin Induced become round to 50～60% cell detachment culture dish bottom, in the culture dish of three kinds of cells
Each 3mL DMEM in high glucose culture medium that adds terminates digestion, more respectively by three kinds of cell suspensions from culture dish
Suck in three 15mL centrifuge tubes, respectively add DMEM in high glucose culture medium to 10mL, by three centrifuge tubes
It is centrifuged 5min as 1000rpm in centrifuge, takes out and pour out supernatant, three centrifuge tubes respectively add 1mL
Cell suspension DMEM in high glucose uniformly, is trained by DMEM in high glucose culture medium by cell piping and druming bottom centrifuge tube
Support in base dilution 50mL cell suspension containing about 2500000, cell (cell blood counting chamber counts).Again
Three kinds of cell suspension liquid-transfering guns are injected separately in 96 porocyte culture plates that (every kind of cell is respectively with 3 96
Porocyte culture plate), every hole 100 μ L cell suspension (about 5000 cells), separately add in 96 orifice plates
One empties white control group, i.e. 100 μ L acellular DMEM in high glucose culture medium.By 96 porocyte culture plates
It is placed in 37 DEG C, CO2The incubator of content 5% is cultivated 24h.
After cell in 96 porocyte culture plates cultivates 24h, each addition 100 μ L in each hole having cell
The DIBOODIPY-PEPTIDE solution of the above-mentioned variable concentrations prepared, every kind of concentration has 4 parallel holes.
Additionally respectively add 100 μ L DMEM in high glucose culture mediums in the hole of blank group.By 96 porocyte culture plates
Continue to be placed in 37 DEG C, CO2The incubator of content 5% is cultivated 24h.
After adding DIBOODIPY-PEPTIDE solution 24h, three kinds of cells respectively take a plate 96 porocyte training
Support plate, give in every hole and add 20 μ L cell proliferations and toxicity detection reagent MTS, continue to put into 37 DEG C,
CO2At 492nm, detect the light absorption in each hole with enzyme connection detector after 4h cultivated by the incubator of content 5%
Value (OD), calculates 24h DIBOODIPY-PEPTIDE solution to three kinds of cell proliferations according to OD value
Inhibiting rate；According to said method, detect that three kinds of cells are existed by DIBOODIPY-PEPTIDE respectively respectively
The proliferation inhibition rate of 48h and 72h.Draw concentration-survival rate figure as shown in Figure 5 and Figure 6, it can be seen that
When 24h, 48h, 72h, DIBOODIPY-PEPTIDE is all to cancer cell Hela cell, MCF-7cell
Propagation have inhibitory action, and the propagation of 3T3cell is had no significant effect.And with after peptide modified with modify before
Comparing, the impact that 3T3cell is bred by DIBODIPY-DICHO is without significant change.
(5) respectively Hela cell, MCF-7cell, 3T3cell are passaged to confocal special culture dish,
37℃、CO2The incubator of content 5% is cultivated 24h.After 24h, with pH7.4PBS, (phosphate delays
Rush liquid) wash three times, 10 prepared in each addition 0.5mL step (2) in each culture dish-5Mol/L's
DIBOODIPY-PEPTIDE solution, then add 0.5mL DMEM in high glucose culture medium.Continue to be put in
37℃、CO2The incubator of content 5% is cultivated 4h.Sucking-off solution after 4h, washes three with pH7.4PBS
Secondary, every ware adds 1mL DMEM in high glucose culture medium.Observe with confocal
The DIBOODIPY-PEPTIDE imaging contexts in three kinds of cells respectively.It is observed that for Hela
The staining foci of cell, DIBOODIPY-PEPTIDE is cell membrane and nucleus；For with Hela cell
For differing bigger 3T3cell, the staining foci of DIBOODIPY-PEPTIDE is the nuclear membrane of cell；
And the staining foci of MCF-7cell, DIBOODIPY-PEPTIDE is cell membrane.
The present invention includes but not limited to above example, every carry out under the principle of spirit of the present invention appoint
What equivalent or local improvement, all will be regarded as within protection scope of the present invention.
Claims (3)
1. the fluorescence probe for targeting detection also inhibition cancer cell, it is characterised in that: described fluorescence is visited
Pin is containing 4,4', 4 " Isosorbide-5-Nitrae-four aryl butadiene derivatives of-bis-fluorine boron dipyrryls group, its structural formula is as follows:
Wherein, R is-CHO ,-COOCH3With the one in polypeptide；R ' is identical with R.
2. the preparation detecting the also fluorescence probe of inhibition cancer cell as claimed in claim 1 for targeting
Method, it is characterised in that: when R is-CHO or-COOCH3Time, described method step is as follows:
(1) catalyst is added in reactor 1, be sequentially added into good solvent 1 and reactant 1, under agitation
Carry out coupling reaction, after reaction 11h, filter, take filtrate and be spin-dried for, obtain reacting coarse product 1；Will reaction
Crude product 1 is dissolved in chloroform, with petroleum ether and dichloromethane as eluant, eluent, utilizes pillar layer separation to carry
Pure, it is dried, obtains solid a；
(2) 4-formylphenylboronic acid pinacol ester is added in reactor 2, add dichloromethane, obtain dense
Degree is the 4-formylphenylboronic acid pinacol ester solution of 5.73g/L, is passed through protective gas in described solution,
Add Isosorbide-5-Nitrae-dimethyl pyrrole, be under agitation added dropwise over trifluoroacetic acid, react 12h, add 2,3-bis-chloro-5,6-
Dicyano-Isosorbide-5-Nitrae-benzoquinones, reacts 4h, adds DIPEA, treats that DIPEA is complete
After dissolving, reactor 2 is put in ice bath, adds BFEE, react 12h, wash, separatory,
Take organic phase and in organic phase, add anhydrous sodium sulfate, filtering, take filtrate and be spin-dried for, obtain reacting coarse product 2；
Reacting coarse product 2 is dissolved in dichloromethane, with petroleum ether and dichloromethane as eluant, eluent, utilizes column chromatography
Separating-purifying, is dried, obtains solid b；
(3) solid a, solid b, potassium carbonate and four (triphenylphosphines) are closed palladium to add in reactor 3, take out true
Sky, is passed through protective gas, adds good solvent 2, reacts 12h, suction filtration, take filtrate and be spin-dried at 70 DEG C,
Obtain reacting coarse product 3, reacting coarse product 3 be dissolved in chloroform, with petroleum ether with dichloromethane for washing
De-agent, purifies with pillar layer separation, is dried, obtains described fluorescence probe；
Wherein, step (1) described catalyst is the mixture of palladium bichloride and copper bromide, palladium bichloride and copper bromide
The ratio of amount of material be 0.04:3；Described good solvent 1 is the mixed solvent of acetonitrile and toluene, acetonitrile and first
The volume ratio of benzene is 1:50；Described reactant 1 be 4-ethynylbenzaldehyde, 4-ethynyl-benzoate,
One in 4-acetylenylbenzene formonitrile HCN and 4-acetylenylaniline；Described reactant 1, catalyst and good solvent 1
Mass ratio is 1:5:28；Described petroleum ether is 1:1 with the mass ratio of dichloromethane.
Step (2) described 4-formylphenylboronic acid pinacol ester, 1,4-dimethyl pyrrole, trifluoroacetic acid, 2,3-
Two chloro-5,6-dicyano-1,4-benzoquinones, N, N-diisopropylethylamine are 1:1 with the mass ratio of BFEE:
0.25:1:6.5:9.8；Described petroleum ether is 1:4 with the mass ratio of dichloromethane；Described protective gas is N2
With the one in Ar.
Step (3) described protective gas is N2With the one in Ar；Described good solvent 2 is toluene and methyl alcohol
Mixed solvent, the volume ratio of toluene and methyl alcohol is 3:1；Described solid a, solid b, potassium carbonate, four (three
Phenylphosphine) to close palladium, the mass ratio of good solvent 2 be 1:2.5:2:0.18:210；Described petroleum ether and dichloromethane
The mass ratio of alkane is 1:1.
3. the preparation detecting the also fluorescence probe of inhibition cancer cell as claimed in claim 1 for targeting
Method, it is characterised in that: when R is polypeptide, described method step is as follows:
(1) catalyst is added in reactor 4, be sequentially added into good solvent 4 and 4-ethynylbenzaldehyde,
Carry out coupling reaction under stirring, after reaction 11h, filter, take filtrate and be spin-dried for, obtain reacting coarse product 4；
Reacting coarse product 4 is dissolved in chloroform, utilizes pillar layer separation to purify, be dried, obtain solid 1；
(2) 4-formylphenylboronic acid pinacol ester is added in reactor 5, add dichloromethane, obtain dense
Degree is the 4-formylphenylboronic acid pinacol ester solution of 5.73g/L, is passed through protective gas in described solution,
Add Isosorbide-5-Nitrae-dimethyl pyrrole, be under agitation added dropwise over trifluoroacetic acid, react 12h, add 2,3-bis-chloro-5,6-
Dicyano-Isosorbide-5-Nitrae-benzoquinones, reacts 4h, adds DIPEA, treats that DIPEA is complete
After dissolving, reactor 5 is put in ice bath, adds BFEE, react 12h, wash, separatory,
Take organic phase and in organic phase, add anhydrous sodium sulfate, filtering, take filtrate and be spin-dried for, obtain reacting coarse product 5；
Reacting coarse product 5 is dissolved in dichloromethane, utilizes pillar layer separation to purify, be dried, obtain solid 2；
(3) solid 1, solid 2, potassium carbonate and four (triphenylphosphines) are closed palladium to add in reactor 6, take out true
Sky, is passed through protective gas, adds good solvent 5, reacts 12h, suction filtration, take filtrate and be spin-dried at 70 DEG C,
Obtain reacting coarse product 6, reacting coarse product 6 be dissolved in chloroform, purify with pillar layer separation, be dried,
Obtain DIBODIPY-DICHO；
(4) DIBODIPY-DICHO is added in reactor 7, add good solvent 6, fully dissolve,
To the DIBODIPY-DICHO solution that concentration is 0.53g/L；Biomolecule is added in described solution,
30 DEG C of stirring reaction 40h, are dried, add dimethyl sulfoxide (DMSO), obtain suspension a；Add in suspension a
Distilled water, stirs, dialysis, is dried, obtains described fluorescence probe；
Wherein, DIBODIPY-DICHO is 10,10'-(((1Z, 3Z)-Isosorbide-5-Nitrae-two (4-Fonnylphenyl)-1,3-fourth two
Alkene-1,4-diyl) two (1,4-phenylenes)) two (5,5-bis-fluoro-1,3,7,9-tetramethyl-5H-two pyrroles
Cough up) abbreviation of [1,2-c:2', 1'-f] [1,3,2] diaza borine；
Step (1) described catalyst is the mixture of palladium bichloride and copper bromide, palladium bichloride and the material of copper bromide
The ratio of amount be 0.04:3；Described good solvent 4 is the mixed solvent of acetonitrile and toluene, acetonitrile and the body of toluene
Long-pending ratio is 1:50；Described 4-ethynylbenzaldehyde, catalyst are 1:5:28 with the mass ratio of good solvent 4；
Step (2) described 4-formylphenylboronic acid pinacol ester, 1,4-dimethyl pyrrole, trifluoroacetic acid, 2,3-
Two chloro-5,6-dicyano-1,4-benzoquinones, N, N-diisopropylethylamine with the mass ratio of BFEE be: 1:1:
0.25:1:6.5:9.8；
Step (2) and step (3) described protective gas are N2With the one in Ar；
Step (3) described good solvent 5 is the mixed solvent of toluene and methyl alcohol, and toluene with the volume ratio of methyl alcohol is
3:1；Described solid 1, solid 2, potassium carbonate, four (triphenylphosphine) conjunction palladium, the mass ratio of good solvent 5 are 1:
2.5:2:0.18:210；
Step (4) described good solvent 6 is N,N-dimethylformamide；Described biomolecule is polypeptide；Described
DIBODIPY-DICHO, biomolecule, the mass ratio of dimethyl sulfoxide (DMSO) are 1:1:688；Described suspension
A is 1:200 with the volume ratio of distilled water.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410412644.1A CN104262373B (en) | 2014-08-20 | 2014-08-20 | A kind of targeting detection the fluorescence probe of inhibition cancer cell, preparation method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410412644.1A CN104262373B (en) | 2014-08-20 | 2014-08-20 | A kind of targeting detection the fluorescence probe of inhibition cancer cell, preparation method and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104262373A CN104262373A (en) | 2015-01-07 |
| CN104262373B true CN104262373B (en) | 2016-08-17 |
Family
ID=52153984
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410412644.1A Expired - Fee Related CN104262373B (en) | 2014-08-20 | 2014-08-20 | A kind of targeting detection the fluorescence probe of inhibition cancer cell, preparation method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104262373B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108444959B (en) * | 2018-02-08 | 2020-09-11 | 北京理工大学 | Lighting type fluorescent probe for identifying and detecting liver cancer cells |
| CN108578714A (en) * | 2018-04-26 | 2018-09-28 | 天津大学 | The method that group and fluorophor utilization rate are targeted in BODIPY fluorescence probes is improved using phosphatide |
| CN108865119B (en) * | 2018-08-10 | 2019-08-27 | 北京理工大学 | For marking cancer cell to light type fluorescent chemicals and its preparation method and application under freezing conditions |
| CN110776434B (en) * | 2019-09-29 | 2021-01-15 | 中山大学 | Hole transport material based on tetraaryl butadiene and preparation method and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101108860A (en) * | 2007-08-06 | 2008-01-23 | 浙江理工大学 | Beta-enol imine structure boron fluoride complex and method of synthesizing the same |
-
2014
- 2014-08-20 CN CN201410412644.1A patent/CN104262373B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101108860A (en) * | 2007-08-06 | 2008-01-23 | 浙江理工大学 | Beta-enol imine structure boron fluoride complex and method of synthesizing the same |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104262373A (en) | 2015-01-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104262373B (en) | A kind of targeting detection the fluorescence probe of inhibition cancer cell, preparation method and application | |
| CN104945322B (en) | Detect the compound and preparation method thereof of tumor hypoxia | |
| JP5517624B2 (en) | Antioxidant indicator molecule | |
| Liang et al. | AIEgen based light-up probes for live cell imaging | |
| Xu et al. | Fluorescent boronic acid polymer grafted on silica particles for affinity separation of saccharides | |
| Anorma et al. | Surveillance of cancer stem cell plasticity using an isoform-selective fluorescent probe for aldehyde dehydrogenase 1A1 | |
| WO2021103700A1 (en) | Nitroreductase responsive hypoxia probe compound, and preparation and application thereof | |
| Kang et al. | Multimodal imaging probe development for pancreatic β cells: from fluorescence to PET | |
| JP6463005B2 (en) | Macrophage discrimination agent, discrimination method using the macrophage discrimination agent, sorting method, evaluation method, screening method, and kit | |
| Li et al. | Hypoxia-activated probe for NIR fluorescence and photoacoustic dual-mode tumor imaging | |
| Li et al. | Detection of Aβ oligomers in early Alzheimer’s disease diagnose by in vivo NIR-II fluorescence imaging | |
| WO2022218362A1 (en) | Application of nitrofuran small-molecule compound in preparation of drug for inducing ferroptosis and/or slowing chemotherapy resistance in gastric cancer | |
| CN110862819B (en) | PH fluorescent probe based on near-infrared fluorescent dye and preparation method and application thereof | |
| Zhang et al. | A near-infrared ratiometric fluorescent probe for highly selective recognition and bioimaging of cysteine | |
| Qin et al. | Identification of a glypican‐3‐binding peptide for in vivo non‐invasive human hepatocellular carcinoma detection | |
| CN113004306A (en) | Near-infrared two-region fluorescent molecule containing benzodithiadiazole, preparation method thereof, fluorescent nanoparticles, and preparation method and application thereof | |
| Cheng et al. | Synthesis of a novel HER2 targeted aza-BODIPY–antibody conjugate: synthesis, photophysical characterisation and in vitro evaluation | |
| Bao et al. | NIR absorbing DICPO derivatives applied to wide range of pH and detection of glutathione in tumor | |
| Zhang et al. | Glycine-conjugated porphyrin fluorescent probe with iRGD for live cell imaging | |
| Gurram et al. | Near-infrared fluorescent probe for fast track of cyclooxygenase-2 in Golgi apparatus in cancer cells | |
| Wang et al. | A photostable cationic fluorophore for long-term bioimaging | |
| JPWO2018174253A1 (en) | Nitrobenzene derivatives or their salts and their uses | |
| Zhong et al. | In vivo study of a novel, safe, rapid, and targeted red carbon dot probe for recognition of tumors with high expression of folate enzyme | |
| Li et al. | Biocompatible Porphyrin‐Peptide Conjugates as Theranostic Agents Targeting the Epstein‐Barr Virus | |
| Wang et al. | Design, synthesis and application of near-infrared fluorescence probe IR-780-Crizotinib in detection of ALK positive tumors |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20160817 Termination date: 20210820 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |