CN104262378A - Silicon-based rhodamine derivative, preparation method and applications thereof - Google Patents
Silicon-based rhodamine derivative, preparation method and applications thereof Download PDFInfo
- Publication number
- CN104262378A CN104262378A CN201410428067.5A CN201410428067A CN104262378A CN 104262378 A CN104262378 A CN 104262378A CN 201410428067 A CN201410428067 A CN 201410428067A CN 104262378 A CN104262378 A CN 104262378A
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- Prior art keywords
- silica
- rhodamine derivatives
- sec
- hydrogen
- butyl
- Prior art date
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- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical class [Cl-].C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=CC=C1C(O)=O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 title claims abstract description 55
- 238000002360 preparation method Methods 0.000 title claims abstract description 21
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 title abstract description 6
- 229910052710 silicon Inorganic materials 0.000 title abstract description 6
- 239000010703 silicon Substances 0.000 title abstract description 6
- 238000006243 chemical reaction Methods 0.000 claims abstract description 48
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 129
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 93
- 239000000377 silicon dioxide Substances 0.000 claims description 64
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 45
- RTZKZFJDLAIYFH-UHFFFAOYSA-N ether Substances CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 44
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 42
- 238000000034 method Methods 0.000 claims description 36
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 31
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 31
- -1 isobutyl- Chemical group 0.000 claims description 31
- 150000001875 compounds Chemical class 0.000 claims description 30
- MZRVEZGGRBJDDB-UHFFFAOYSA-N N-Butyllithium Chemical compound [Li]CCCC MZRVEZGGRBJDDB-UHFFFAOYSA-N 0.000 claims description 26
- 229910052739 hydrogen Inorganic materials 0.000 claims description 24
- 239000001257 hydrogen Substances 0.000 claims description 24
- 239000003480 eluent Substances 0.000 claims description 19
- 125000004093 cyano group Chemical group *C#N 0.000 claims description 17
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 claims description 16
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 16
- 125000000217 alkyl group Chemical group 0.000 claims description 14
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims description 13
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 claims description 12
- 238000010898 silica gel chromatography Methods 0.000 claims description 12
- DHYHYLGCQVVLOQ-UHFFFAOYSA-N 3-bromoaniline Chemical class NC1=CC=CC(Br)=C1 DHYHYLGCQVVLOQ-UHFFFAOYSA-N 0.000 claims description 11
- 150000003935 benzaldehydes Chemical class 0.000 claims description 11
- 230000015572 biosynthetic process Effects 0.000 claims description 10
- 239000012046 mixed solvent Substances 0.000 claims description 10
- 238000003786 synthesis reaction Methods 0.000 claims description 9
- 239000003153 chemical reaction reagent Substances 0.000 claims description 8
- MROCJMGDEKINLD-UHFFFAOYSA-N dichlorosilane Chemical compound Cl[SiH2]Cl MROCJMGDEKINLD-UHFFFAOYSA-N 0.000 claims description 8
- 125000001424 substituent group Chemical group 0.000 claims description 8
- 125000003903 2-propenyl group Chemical group [H]C([*])([H])C([H])=C([H])[H] 0.000 claims description 7
- 150000002431 hydrogen Chemical class 0.000 claims description 7
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 7
- WHXSMMKQMYFTQS-UHFFFAOYSA-N Lithium Chemical compound [Li] WHXSMMKQMYFTQS-UHFFFAOYSA-N 0.000 claims description 6
- 125000000484 butyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 claims description 6
- 125000001972 isopentyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])C([H])([H])* 0.000 claims description 6
- 229910052744 lithium Inorganic materials 0.000 claims description 6
- 125000001280 n-hexyl group Chemical group C(CCCCC)* 0.000 claims description 6
- 125000000740 n-pentyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 claims description 6
- 125000004123 n-propyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])* 0.000 claims description 6
- 125000001971 neopentyl group Chemical group [H]C([*])([H])C(C([H])([H])[H])(C([H])([H])[H])C([H])([H])[H] 0.000 claims description 6
- 125000002914 sec-butyl group Chemical group [H]C([H])([H])C([H])([H])C([H])(*)C([H])([H])[H] 0.000 claims description 6
- 238000007789 sealing Methods 0.000 claims description 5
- KOPOQZFJUQMUML-UHFFFAOYSA-N chlorosilane Chemical compound Cl[SiH3] KOPOQZFJUQMUML-UHFFFAOYSA-N 0.000 claims description 4
- 238000004440 column chromatography Methods 0.000 claims description 4
- 125000004122 cyclic group Chemical group 0.000 claims description 4
- 125000004435 hydrogen atom Chemical group [H]* 0.000 claims description 4
- 238000006460 hydrolysis reaction Methods 0.000 claims description 4
- 229920006395 saturated elastomer Polymers 0.000 claims description 4
- 238000000926 separation method Methods 0.000 claims description 4
- UGNWTBMOAKPKBL-UHFFFAOYSA-N tetrachloro-1,4-benzoquinone Chemical compound ClC1=C(Cl)C(=O)C(Cl)=C(Cl)C1=O UGNWTBMOAKPKBL-UHFFFAOYSA-N 0.000 claims description 4
- 150000002596 lactones Chemical class 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 claims description 3
- 239000003208 petroleum Substances 0.000 claims description 3
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 claims description 2
- 229910021589 Copper(I) bromide Inorganic materials 0.000 claims description 2
- 125000003545 alkoxy group Chemical group 0.000 claims description 2
- 239000000460 chlorine Substances 0.000 claims description 2
- 229910052801 chlorine Inorganic materials 0.000 claims description 2
- 125000000753 cycloalkyl group Chemical group 0.000 claims description 2
- 125000001188 haloalkyl group Chemical group 0.000 claims description 2
- 229910052736 halogen Inorganic materials 0.000 claims description 2
- 150000002367 halogens Chemical class 0.000 claims description 2
- 239000000376 reactant Substances 0.000 claims 2
- 238000006722 reduction reaction Methods 0.000 claims 2
- 125000000852 azido group Chemical group *N=[N+]=[N-] 0.000 claims 1
- 230000003197 catalytic effect Effects 0.000 claims 1
- 125000001033 ether group Chemical group 0.000 claims 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims 1
- 239000002904 solvent Substances 0.000 claims 1
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- 230000005311 nuclear magnetism Effects 0.000 description 41
- 239000003921 oil Substances 0.000 description 26
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- 238000001819 mass spectrum Methods 0.000 description 20
- 239000002994 raw material Substances 0.000 description 16
- 238000003756 stirring Methods 0.000 description 14
- 238000000799 fluorescence microscopy Methods 0.000 description 13
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 11
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 10
- 210000004782 brain capillary endothelium Anatomy 0.000 description 9
- 238000013461 design Methods 0.000 description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 8
- 239000012472 biological sample Substances 0.000 description 8
- 239000000975 dye Substances 0.000 description 8
- 238000004043 dyeing Methods 0.000 description 8
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 8
- 239000012044 organic layer Substances 0.000 description 8
- 238000010025 steaming Methods 0.000 description 8
- HUMNYLRZRPPJDN-UHFFFAOYSA-N benzenecarboxaldehyde Natural products O=CC1=CC=CC=C1 HUMNYLRZRPPJDN-UHFFFAOYSA-N 0.000 description 7
- 238000001035 drying Methods 0.000 description 7
- QNGNSVIICDLXHT-UHFFFAOYSA-N para-ethylbenzaldehyde Natural products CCC1=CC=C(C=O)C=C1 QNGNSVIICDLXHT-UHFFFAOYSA-N 0.000 description 7
- 239000000523 sample Substances 0.000 description 7
- 239000011734 sodium Substances 0.000 description 7
- 239000005711 Benzoic acid Substances 0.000 description 6
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- 150000001299 aldehydes Chemical class 0.000 description 6
- 235000010233 benzoic acid Nutrition 0.000 description 6
- 238000000746 purification Methods 0.000 description 6
- 238000005406 washing Methods 0.000 description 6
- NCGICGYLBXGBGN-UHFFFAOYSA-N 3-morpholin-4-yl-1-oxa-3-azonia-2-azanidacyclopent-3-en-5-imine;hydrochloride Chemical compound Cl.[N-]1OC(=N)C=[N+]1N1CCOCC1 NCGICGYLBXGBGN-UHFFFAOYSA-N 0.000 description 5
- 0 C*c1c(*)c(N(*)*)c(*)c(*(*)(*)c2c(*)c(N(*)*)c(*)c(*)c2)c1 Chemical compound C*c1c(*)c(N(*)*)c(*)c(*(*)(*)c2c(*)c(N(*)*)c(*)c(*)c2)c1 0.000 description 5
- 229910052786 argon Inorganic materials 0.000 description 5
- 238000004113 cell culture Methods 0.000 description 5
- 238000010189 synthetic method Methods 0.000 description 5
- OKKJLVBELUTLKV-MZCSYVLQSA-N Deuterated methanol Chemical compound [2H]OC([2H])([2H])[2H] OKKJLVBELUTLKV-MZCSYVLQSA-N 0.000 description 4
- QABCGOSYZHCPGN-UHFFFAOYSA-N chloro(dimethyl)silicon Chemical compound C[Si](C)Cl QABCGOSYZHCPGN-UHFFFAOYSA-N 0.000 description 4
- LIKFHECYJZWXFJ-UHFFFAOYSA-N dimethyldichlorosilane Chemical compound C[Si](C)(Cl)Cl LIKFHECYJZWXFJ-UHFFFAOYSA-N 0.000 description 4
- 238000003384 imaging method Methods 0.000 description 4
- DLEDOFVPSDKWEF-UHFFFAOYSA-N lithium butane Chemical compound [Li+].CCC[CH2-] DLEDOFVPSDKWEF-UHFFFAOYSA-N 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 230000035484 reaction time Effects 0.000 description 4
- 239000012086 standard solution Substances 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical class [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 230000005284 excitation Effects 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 239000010410 layer Substances 0.000 description 3
- 201000007270 liver cancer Diseases 0.000 description 3
- 208000014018 liver neoplasm Diseases 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 101100042630 Caenorhabditis elegans sin-3 gene Proteins 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- CWNKQMWSSLBTSU-UHFFFAOYSA-N N1=CC=CC2=CC=CC=C12.C(C)N1C=CC2=CC=C(C=C12)Br Chemical compound N1=CC=CC2=CC=CC=C12.C(C)N1C=CC2=CC=C(C=C12)Br CWNKQMWSSLBTSU-UHFFFAOYSA-N 0.000 description 2
- BLRPTPMANUNPDV-UHFFFAOYSA-N Silane Chemical compound [SiH4] BLRPTPMANUNPDV-UHFFFAOYSA-N 0.000 description 2
- 238000000862 absorption spectrum Methods 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 229910002091 carbon monoxide Inorganic materials 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- QTMDXZNDVAMKGV-UHFFFAOYSA-L copper(ii) bromide Chemical compound [Cu+2].[Br-].[Br-] QTMDXZNDVAMKGV-UHFFFAOYSA-L 0.000 description 2
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- 229910000077 silane Inorganic materials 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 239000013589 supplement Substances 0.000 description 2
- CMWKITSNTDAEDT-UHFFFAOYSA-N 2-nitrobenzaldehyde Chemical compound [O-][N+](=O)C1=CC=CC=C1C=O CMWKITSNTDAEDT-UHFFFAOYSA-N 0.000 description 1
- RCBPVESMGNZMSG-UHFFFAOYSA-N 4-bromo-2-methylbenzaldehyde Chemical compound CC1=CC(Br)=CC=C1C=O RCBPVESMGNZMSG-UHFFFAOYSA-N 0.000 description 1
- CVMJZUGRIUPKOL-UHFFFAOYSA-N Bc1cc(N(CC)CC2)c2cc1 Chemical compound Bc1cc(N(CC)CC2)c2cc1 CVMJZUGRIUPKOL-UHFFFAOYSA-N 0.000 description 1
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- 241001465754 Metazoa Species 0.000 description 1
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- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium on carbon Substances [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 206010034972 Photosensitivity reaction Diseases 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- 241000405217 Viola <butterfly> Species 0.000 description 1
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- BFMYDTVEBKDAKJ-UHFFFAOYSA-L disodium;(2',7'-dibromo-3',6'-dioxido-3-oxospiro[2-benzofuran-1,9'-xanthene]-4'-yl)mercury;hydrate Chemical compound O.[Na+].[Na+].O1C(=O)C2=CC=CC=C2C21C1=CC(Br)=C([O-])C([Hg])=C1OC1=C2C=C(Br)C([O-])=C1 BFMYDTVEBKDAKJ-UHFFFAOYSA-L 0.000 description 1
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- BTFQKIATRPGRBS-UHFFFAOYSA-N o-tolualdehyde Chemical compound CC1=CC=CC=C1C=O BTFQKIATRPGRBS-UHFFFAOYSA-N 0.000 description 1
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- 229940049954 penicillin Drugs 0.000 description 1
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- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Substances [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 description 1
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- QTENRWWVYAAPBI-YCRXJPFRSA-N streptomycin sulfate Chemical compound OS(O)(=O)=O.OS(O)(=O)=O.OS(O)(=O)=O.CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](N=C(N)N)[C@H](O)[C@@H](N=C(N)N)[C@H](O)[C@H]1O.CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](N=C(N)N)[C@H](O)[C@@H](N=C(N)N)[C@H](O)[C@H]1O QTENRWWVYAAPBI-YCRXJPFRSA-N 0.000 description 1
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- 235000001892 vitamin D2 Nutrition 0.000 description 1
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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
- C07F7/00—Compounds containing elements of Groups 4 or 14 of the Periodic Table
- C07F7/02—Silicon compounds
- C07F7/08—Compounds having one or more C—Si linkages
- C07F7/10—Compounds having one or more C—Si linkages containing nitrogen having a Si-N linkage
-
- 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
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/33—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using ultraviolet light
-
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
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- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Physics & Mathematics (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Pathology (AREA)
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Abstract
The invention relates to a silicon-based rhodamine derivative, which is represented by a biological structural formula shown in the description. The invention also relates to a preparation method of the silicon-based rhodamine derivative, and applications of the silicon-based rhodamine derivative in preparation of cell dye, biological dye, biological molecule/particle fluorescent marker, and the like. The provided synthesis method of silicon-based rhodamine derivative has the advantages of high yield, simple reaction conditions, convenient operation, capability for massive production, and wide substrate selection range.
Description
Technical field
The present invention relates to chemical field, specifically, is a kind of silica-based Rhodamine Derivatives and its preparation method and application.
Background technology
Rhodamine is the one in xanthene compounds, has the advantages such as good light stability, fluorescence quantum yield is high, chemically modified is simple, is a kind of fluorescent small molecule probe dye preferably, in the dyeing being widely used in biological sample and fluorescent mark.But rhodamine in application process, when being particularly applied to biological sample, also also exists many deficiencies as fluorescent probe.The excitation wavelength of such as rhdamine B is positioned at ultraviolet-visible district, and biological sample can produce serious background signal interference in ultraviolet-visible district.On the other hand, the penetrativity of ultraviolet-visible to tissue is poor, larger to the phototoxicity of biological sample.These factors have impact on rhodamine probe greatly in biological sample fluorometric analysis medium sensitivity and accuracy, significantly limit its further application in biological sample fluorescence imaging.
Silica-based rhodamine is the fluorochrome that new development is in recent years got up.It is advantageous that by the Sauerstoffatom in rhodamine molecule is replaced with Siliciumatom, while remaining rhodamine compound advantage, its spectral range red shift, reaches near-infrared region, thus meets the requirement of biological sample analysis imaging.Constantly there is the Patent Publication of silica-based rhodamine aspect in recent years, such as patent WO 2010/126077 A1, patent WO 2012/111818 A1 and patent WO 2013/029650 A1 etc.These patents describe the character of silica-based rhodamine, synthetic method and application etc.In these patents, be all adopt the method for active metal reagent and carbonyl nucleophilic addition(Adn) to prepare silica-based Rhodamine Derivatives, step is more on the one hand for this synthetic method, severe reaction conditions, and productive rate is low, can not be used for a large amount of synthesis; The range of choice some of them that on the other hand use of active metal reagent greatly limit substrate replace silica-based rhodamine and even cannot synthesize, and have impact on its application in biological sample imaging.Therefore, design that a kind of step is few, reaction conditions is simple, productive rate is high, substrate select various, fit the method that realizes preparing in a large number be very necessary for the synthesis of silica-based Rhodamine Derivatives.
Summary of the invention
The object of the invention is for deficiency of the prior art, a kind of silica-based Rhodamine Derivatives is provided.
Of the present invention again one object be that the preparation method of above-mentioned silica-based Rhodamine Derivatives is provided.
Another object of the present invention provides the purposes of above-mentioned silica-based Rhodamine Derivatives.
For achieving the above object, the technical scheme that the present invention takes is:
A kind of silica-based Rhodamine Derivatives, its general structure is as shown in (I):
Especially, R is worked as
11or R
15wherein any one or two are-COOH or-COO
-time, the corresponding lactone volution form of general formula (I) can be formed, there is the compound of general formula (II) structure:
Wherein, R
2, R
3be respectively alkyl or the phenyl of C1-C6 straight or branched, described alkyl is selected from: methyl, ethyl, n-propyl, sec.-propyl, normal-butyl, isobutyl-, sec-butyl, the tertiary butyl, n-pentyl, isopentyl, sec.-amyl sec-pentyl secondary amyl, neo-pentyl, n-hexyl, preferably methyl, ethyl and phenyl, most preferably methyl.
R
4, R
5, R
6, R
7, R
8, R
9be respectively the alkyl of the straight or branched of hydrogen or C1-C6, described alkyl is selected from: methyl, ethyl, n-propyl, sec.-propyl, normal-butyl, isobutyl-, sec-butyl, the tertiary butyl, n-pentyl, isopentyl, sec.-amyl sec-pentyl secondary amyl, neo-pentyl, n-hexyl, preferably methyl, ethyl and phenyl, most preferably hydrogen.
R
20, R
21, R
22, R
23be respectively the saturated of C1-C6 straight or branched or undersaturated alkyl or cycloalkyl, described alkyl is selected from: methyl, ethyl, n-propyl, sec.-propyl, allyl group, normal-butyl, isobutyl-, sec-butyl, the tertiary butyl, n-pentyl, isopentyl, sec.-amyl sec-pentyl secondary amyl, neo-pentyl, n-hexyl, preferably methyl, ethyl, allyl group, most preferably ethyl.
R
20or R
21can with R
5be connected with carbochain, form five yuan or six-membered cyclic structure with parent benzene, preferred five-ring.
R
22or R
23can with R
8be connected with carbochain, form five yuan or six-membered cyclic structure with parent benzene, preferred five-ring.
Preferably, substituting group-NR
20r
21can with-R
5formation has the structure of following general formula (III):
。
Preferably, described substituting group-NR
22r
23can with-R
8formation has the structure of following general formula (IV):
.
Preferably, described substituent R
24, R
25for hydrogen, methyl, ethyl or allyl group, be more preferably ethyl.
R
11, R
12, R
13, R
14, R
15, R
16, R
17, R
18, R
19can be identical or different any substituting groups, described substituting group be selected from: alkyl, halogen, cyano group, nitro, alkoxyl group, haloalkyl.Preferred structure be following in one:
R
11or R
15one is hydrogen, and another is-CN ,-NO
2,-SO
3h ,-SO
3 -,-OH ,-OCH
3,-NH
2,-N
3,-NCS ,-CH
3,-F ,-Cl, in-Br ,-I group one, R
12, R
13, R
14be hydrogen.
R
11or R
15one is hydrogen, and another is-CH
3, R
12, R
13, R
14in any one is-CN ,-NO
2,-SO
3h ,-SO
3 -,-OH ,-OCH
3,-NH
2,-CH
3,-N
3,-NCS,
,-CONH
2,-CH
2nH
2,-F ,-Cl ,-Br ,-I ,-COOH ,-COO
-in any one, all the other two is hydrogen.
R
11or R
15one is hydrogen, and another is-SO
3h or-SO
3 -, R
12, R
13, R
14in any one be-SO
3h ,-SO
3 -one in group, all the other two is hydrogen.
R
11or R
15one is hydrogen, and another is-COOH or-COO
-, R
12, R
13, R
14in any one is-CN ,-NO
2,-SO
3h ,-SO
3 -,-OH ,-OCH
3,-NH
2,-CH
3,-N
3,-NCS,
,-CONH
2,-CH
2nH
2,-F ,-Cl ,-Br ,-I ,-COOH ,-COO
-one in group, all the other two is hydrogen.
R
11or R
15when having at least one to be-COOH or-COO-, in the lactone spirane structure of formation, R
16, R
17, R
18, R
19in any one is-CN ,-NO
2,-SO
3h ,-SO
3 -,-OH ,-OCH
3,-NH
2,-CH
3,-N
3,-NCS,
,-CONH
2,-CH
2nH
2,-F ,-Cl ,-Br ,-I ,-COOH ,-COO
-one in group, its excess-three is hydrogen.
For realizing above-mentioned second object, the technical scheme that the present invention takes is.:
Described silica-based Rhodamine Derivatives preparation method carries out as follows:
The first step: the synthesis with the silica-based intermediate of key of logical formula V structure;
。
As-NR
20r
21with-NR
22r
23, R
4with R
7, R
5with R
8, R
6with R
9all there is same structure, adopt and synthesize with the following method:
N, N-dialkyl m-bromoaniline derivative reacts to n-Butyl Lithium and generates corresponding lithium reagent, then is obtained by reacting crucial silica-based intermediate with dialkyl dichlorosilane further.Its reaction process can be expressed as:
Wherein R
2, R
3, R
4~ R
9, R
20~ R
23implication is the same.
Concrete working method is preferably: 1.0 molar weight N, in the ether of drying, 2 hours are reacted under n-Butyl Lithium 0 ° of C of N-dialkyl m-bromoaniline derivative and 1.05 molar weights, again the dialkyl dichlorosilane of 0.6 molar weight is dissolved in and joins in reaction solution in dry ether, 0 ° of C slowly rises to room temperature, and reaction is spent the night.Add shrend to go out reaction, extraction, steaming desolventizes, and silica gel column chromatography is separated, and eluent is the mixed solvent of sherwood oil and ethyl acetate, and by volume, ethyl acetate: sherwood oil=1:40 ~ 80, obtains the silica-based intermediate of described key.
As-NR
20r
21with-NR
22r
23, R
4with R
7, R
5with R
8, R
6with R
9in have one or more pairs of when there is different structure, adopt and synthesize with the following method:
A kind of N, N-dialkyl m-bromoaniline derivative reacts to n-Butyl Lithium and generates corresponding lithium reagent, then reacts with dialkyl dichlorosilane, replaces one of them chlorine, generates the product of monochlorosilane.Another kind of N, N-dialkyl m-bromoaniline derivative reacts to butyllithium and generates corresponding lithium reagent, then is obtained by reacting crucial silica-based intermediate with monochlorosilane product further.Its reaction process can be expressed as:
Wherein R
2, R
3, R
4~ R
9, R
20~ R
23implication is the same.
Concrete operation method is preferably: a kind of 1.0 molar weight N, in the ether of drying, 2 hours are reacted under n-Butyl Lithium 0 ° of C of N-dialkyl m-bromoaniline derivative and 1.05 molar weights, added in the dry ether of the dialkyl dichlorosilane containing 5.0 molar weights, react 2 hours under 0 ° of C, obtained monochloro is for silane product; The N of another kind of 0.5 molar weight, in the ether of drying, 2 hours are reacted under n-Butyl Lithium 0 ° of C of N-dialkyl m-bromoaniline derivative and 0.525 molar weight, under 0 ° of C, this reaction solution is slowly joined monochloro in silane product, slowly rise to room temperature, continue reaction 12 hours, add shrend to go out reaction, extraction, steaming desolventizes, silica gel column chromatography is separated, and eluent is the mixed solvent of sherwood oil and ethyl acetate, by volume, ethyl acetate: sherwood oil=1: 60, obtains crucial silica-based intermediate.
Second step: crucial silica-based intermediate and different substituted benzaldehyde generate the reaction of silica-based Rhodamine Derivatives;
The benzaldehyde derivative of crucial silica-based intermediate and different replacement is under catalysts conditions, tube sealing is placed in shield cap, react under 140 ° of C, be oxidized through tetrachlorobenzoquinone again, or be not oxidized, finally carry out column chromatography for separation, carry out wash-out with methylene dichloride and the mixed system of methyl alcohol or the mixed system of ethyl acetate, sherwood oil and triethylamine, obtain silica-based Rhodamine Derivatives.Its reaction process can be expressed as:
When there is carboxyl at the ortho position of substituted benzaldehyde, its reaction process can also be expressed as:
Wherein R
2, R
3, R
4~ R
9, R
11-R
19, R
20~ R
23implication is the same.
The described silica-based intermediate of key with logical formula V structure is 1: 5 from the molar ratio of the benzaldehyde derivative of different replacement.
CuBr selected by described catalyzer
2or tosic acid monohydrate; .
Described have the silica-based intermediate of key of logical formula V structure and the mol ratio of catalyzer is 1:0.1 ~ 1
The eluent of described column chromatography for separation, by volume, the mixed solvent of methyl alcohol and methylene dichloride is methyl alcohol: methylene dichloride=1:5 ~ 40, ethyl acetate and sherwood oil mixed solvent are ethyl acetate: sherwood oil=1:40 ~ 80, the mixed solvent of ethyl acetate, sherwood oil and triethylamine is ethyl acetate: sherwood oil=1:10 ~ 80, and the add-on of triethylamine is 1% of ethyl acetate and petroleum ether mixtures volume.
Concrete operation method is preferably: the substituted benzaldehyde of silica-based for the key of 1.0 molar weights intermediate, 5.0 molar weights, the catalyzer of 0.1 ~ 1.0 molar weight are added in the withstand voltage reaction tubes of heavy wall, tube sealing is placed in shield cap, reacts 2-24 hour under 140 ° of C.Naturally cooling, organic solvent dissolution, if the substituted benzaldehyde ortho position added is not carboxyl, then adds oxidizing 10 minutes, if the substituted benzaldehyde ortho position added is carboxyl, is oxidized without oxidizer.Steaming desolventizes, and silica gel column chromatography is separated.If the substituted benzaldehyde ortho position added is not carboxyl, eluent is the mixed solvent of methylene dichloride and methyl alcohol, and by volume, methyl alcohol: methylene dichloride=1: 5 ~ 40, obtains target compound.If the substituted benzaldehyde ortho position added is carboxyl, eluent is the mixed solvent of sherwood oil, ethyl acetate and triethylamine, by volume, and ethyl acetate: sherwood oil=1: 10 ~ 80, the add-on of triethylamine is 1% of ethyl acetate and petroleum ether mixtures volume, obtains target compound.
The silica-based Rhodamine Derivatives of the target compound obtained can also obtain new silica-based Rhodamine Derivatives by the further derivative reaction of substituted radical, namely can also comprise following 3rd step.
The further derivative reaction of the silica-based Rhodamine Derivatives of 3rd step;
Under normal conditions, described silica-based Rhodamine Derivatives carries out as the reduction of the complete hydrolysis of the partial hydrolysis of cyano group, cyano group, cyano group, the reduction of nitro, amino azido reaction etc.
For realizing above-mentioned 3rd object, the technical scheme that the present invention takes is:
The application of described silica-based Rhodamine Derivatives in preparation cell dye, biological stain, biomolecules or biomone fluorescent mark.Described cell dye, biological stain can be used for laboratory animal, viable cell or fix the staining agent of rear cell.
A kind of brand-new synthetic method of the present invention, can synthesis of silica-base Rhodamine Derivatives easily, can improve combined coefficient by this method, can realize a large amount of preparations of silica-based Rhodamine Derivatives.Most importantly this synthetic method has greatly enriched the range of choice of substrate, can synthesize some and adopt front method to be difficult to the silica-based Rhodamine Derivatives synthesized, be conducive to further modification and the application of silica-based Rhodamine Derivatives.
By the some novel silica-based Rhodamine Derivatives of method of the present invention synthesis, to meet its application in different field.
The present invention also carries out the investigation and application of spectroscopic properties to the silica-based Rhodamine Derivatives of synthesis, and for active somatic cell imaging research.
The invention has the advantages that:
The present invention devises a brand-new synthetic route, and before obtaining a series of employing by a kind of brand-new method synthesis, method is difficult to the novel silicon base rhodamine derivative synthesized, and its structure is as shown in table 1 below.The method also has that overall yield is high, reaction conditions is simple, easy to operate, can prepare in a large number, the advantage such as substrate range of choice is wide.
The silica-based rhodamine derivant structure formula that table 1 synthesizes first
Accompanying drawing explanation
Fig. 1 is the abosrption spectrogram of compound in the present invention.
Fig. 2 is the fluorescence emission spectrogram of compound in the present invention.
Fig. 3 is that in the present invention, compound 2-COOH SiR to dye laser co-focusing fluorescence imaging design sketch to mouse brain capillary endothelium (bEND3).
Fig. 4 is that 2-COOH SiR prepared by the embodiment of the present invention 13 to dye laser co-focusing fluorescence imaging design sketch to human liver cancer cell (HepG2).
Fig. 5 is 2,4-DiSO of the embodiment of the present invention 13 preparation
3h SiR to dye laser co-focusing fluorescence imaging design sketch to mouse brain capillary endothelium (bEND3).
Fig. 6 is 2,4-DiSO of the embodiment of the present invention 13 preparation
3h SiR to dye laser co-focusing fluorescence imaging design sketch to human liver cancer cell (HepG2).
In Fig. 3 ~ 6, laser co-focusing fluorescence imaging design sketch when (a) all represents that exciting light is 633 nm; Laser co-focusing fluorescence imaging design sketch when () all represents that exciting light is 488 nm b; C () all represents cell bright field design sketch; D () all represents the Overlay figure of (a), (b), (c).
Embodiment
Below with specific embodiment to the synthetic method of the silica-based Rhodamine Derivatives in the present invention, adopt the silica-based Rhodamine Derivatives with brand new synthesized by the method and be applied to the imaging of viable cell laser confocal microscope and be described in further details, the explanation of the invention is not limited.
embodiment 1:
In the 500 mL long-neck reaction flasks that magnetic stir bar is housed, add N, N-diethyl m-bromoaniline 27.376 g (120.0 mmol).Under argon shield condition, add anhydrous diethyl ether 120 mL, 0 ° of C lower magnetic force stirs 5 min.Will
n-BuLi 52.5 mL (2.4 M in
n-hexane, 126.0 mmol) be slowly added drop-wise in reaction solution, after dropwising, under 0 ° of C, continue reaction 2 h.Dimethyldichlorosilane(DMCS) 8.8 mL (72.0 mmol) is dissolved in 30 mL anhydrous diethyl ethers, is slowly added drop-wise in above-mentioned reaction solution.After dropwising, sluggish rises to room temperature, and stirring is spent the night.Add water 150 mL in reaction solution, separating funnel divides gets organic layer, water layer extracted with diethyl ether (50 mL
3), merge organic layer, 150 mL water washings once, the saturated NaCl solution washing of 150 mL once, anhydrous Na
2sO
4drying, steaming desolventizes, purification by silica gel column chromatography, and eluent is by volume of sherwood oil: ethyl acetate=80: 1, obtains yellow oil SiN1 17.507g, productive rate 82%.
Its nuclear-magnetism
1h NMR (300 MHz, CDCl
3): δ 0.64 (s, 6H), 1.23 (t, 12H,
j=7.2 Hz), 3.43 (q, 8H,
j=7.2 Hz), 6.79-7.35 (m, 8H); Nuclear-magnetism
13c NMR (75 MHz, CDCl
3): δ-2.13,12.67,44.45,112.79,117.75,121.47,128.76,139.11,147.10; Mass spectrum MS (ESI) calcd. for C
22h
34n
2si [M+H]
+: 355.26, found:355.32.
embodiment 2:
In the 250 mL long-neck reaction flasks that magnetic stir bar is housed, add 1-ethyl-6-bromo indole quinoline 4.070 g (18.0 mmol).Under argon shield condition, add anhydrous diethyl ether 50 mL, 0 ° of C lower magnetic force stirs 5 min.Will
n-BuLi 11.8 mL (1.6 M in
n-hexane, 18.9 mmol) be slowly added drop-wise in reaction solution, after dropwising, under 0 ° of C, continue reaction 2 h.Dimethyldichlorosilane(DMCS) 1.3 mL (10.8 mmol) is dissolved in 10 mL anhydrous diethyl ethers, is slowly added drop-wise in above-mentioned reaction solution.After dropwising, sluggish rises to room temperature, and stirring is spent the night.Add water 50 mL in reaction solution, divide and get organic layer, water layer extracted with diethyl ether (30 mL
3), merge organic layer, 50mL water washing once, the saturated NaCl solution washing of 50 mL once, anhydrous Na
2sO
4drying, revolve steaming and desolventize, purification by silica gel column chromatography, eluent is by volume of sherwood oil: ethyl acetate=40: 1, obtains yellow oil SiN2,2.423g, productive rate 77%.
Its nuclear-magnetism
1h NMR (300 MHz, CDCl
3): δ 0.49 (s, 6H), 1.16 (t, 6H,
j=7.2 Hz), 2.94 (t, 4H,
j=8.1 Hz), 3.12 (q, 4H,
j=7.2 Hz), 3.30 (t, 4H,
j=8.4 Hz), 6.65-7.08 (m, 6H); Nuclear-magnetism
13c NMR (75 MHz, CDCl
3): δ-1.80,12.09,28.57,43.17,52.20,112.39,124.00,124.07,131.64,137.17,151.80; Mass spectrum MS (ESI) calcd. for C
22h
30n
2si [M+H]
+: 351.23, found:351.26.
embodiment 3:
In the 100 mL long-neck reaction flasks that magnetic stir bar is housed, add N, N-diethyl m-bromoaniline 2.738 g (12 .0 mmol).Under argon shield condition, add anhydrous diethyl ether 10 mL, 0 ° of C lower magnetic force stirs 5 min.Will
n-BuLi 7.9 mL (1.6 M in
n-hexane, 12.6 mmol) be slowly added drop-wise in reaction solution, after dropwising, under 0 ° of C, continue reaction 2 h.Dimethyldichlorosilane(DMCS) 7.3 mL (60.0 mmol) is dissolved in 30 mL anhydrous diethyl ethers, slowly imports in above-mentioned reaction solution.After dropwising, sluggish rises to room temperature, and reacts 2 hours under room temperature.Except desolventizing and unreacted dimethyldichlorosilane(DMCS) under reduced pressure, obtain SiNCl crude product, add anhydrous diethyl ether 5 mL under argon shield and dissolve SiNCl crude product, gained solution for standby.
Be equipped with in 50 mL long-neck reaction flasks of magnetic stir bar at another, add 1-ethyl-6-bromo indole quinoline 1.357 g (6.0 mmol).Under argon shield condition, add anhydrous diethyl ether 10 mL, 0 ° of C lower magnetic force stirs 5 min.Will
n-BuLi 4.0 mL (1.6 M in
n-hexane, 6.3 mmol) be slowly added drop-wise in reaction solution, after dropwising, under 0 ° of C, continue reaction 2 h.Under ice bath, in the SiNCl solution of preparation before being slowly added drop-wise to by this reaction solution, after dropwising, slowly rise to room temperature, stirring is spent the night.Add water 30 mL in reaction solution, separating funnel divides gets organic layer, water layer extracted with diethyl ether (20 mL
3), merge organic layer, 50 mL water washings once, the saturated NaCl solution washing of 50 mL once, anhydrous Na
2sO
4drying, steaming desolventizes.Purification by silica gel column chromatography, eluent is by volume of sherwood oil: ethyl acetate=60: 1, obtains yellow oil SiN3
,1.116 g, productive rate 53%.
Its nuclear-magnetism
1h NMR (300 MHz, CDCl
3): δ 0.50 (s, 6H), 1.12 (t, 6H,
j=7.2 Hz), 1.16 (t, 3H,
j=7.2 Hz), 2.93 (t, 2H,
j=8.1 Hz), 3.11 (q, 2H,
j=7.2 Hz), 3.30 (t, 2H,
j=8.1 Hz), 3.32 (q, 4H,
j=7.2 Hz), 6.70-7.23 (m, 7H); Nuclear-magnetism
13c NMR (75 MHz, CDCl
3): δ-2.01,12.13,12.60,28.56,43.17,44.38,52.22,112.34,112.73,117.72,121.45,123.99,124.02,128.68,131.64,137.00,139.20,147.05,151.84; Mass spectrum MS (ESI) calcd. for C
22h
32n
2si [M+H]
+: 353.60, found:353.49.
embodiment 4:
In the withstand voltage reaction tubes of 15 mL heavy wall that magnetic stir bar is housed, add raw material SiN1 106 mg (0.3 mmol), phenyl aldehyde 159 mg (1.5 mmol), tosic acid monohydrate 57 mg (0.3 mmol), tube sealing is placed in protective guard, 140 ° of C oil baths react 12 hours, naturally cool to room temperature, add methylene dichloride and each 2 mL dissolvings of methyl alcohol, add the oxidation of enough tetrachlorobenzoquinones, filtering insolubles, purification by silica gel column chromatography, eluent is by volume of methylene dichloride: methyl alcohol=40: 1, obtain target compound 2-H SiR, productive rate 40%.
Its nuclear-magnetism
1h NMR (300 MHz, CD3OD) δ 0.59 (s, 6H), 1.29 (t, 12H, J=6.9 Hz), 3.70 (q, 8H, J=6.6 Hz), 6.74-7.55 (m, 10H); Nuclear-magnetism
13c NMR (75 MHz, CD3OD) δ-2.67,11.72,45.41,113.42,120.65,127.36,127.91,128.37,129.01,139.17,142.03,148.15,152.50,169.07; High resolution mass spectrum HRMS (ESI) calcd. for C
29h
37n
2si
+[M]
+: 441.2721, found:441.2765.
embodiment 5:
Phenyl aldehyde in raw material, with embodiment 4, is just changed to o-tolualdehyde by method.Obtain target compound 2-Me SiR productive rate 34%.
Its nuclear-magnetism
1h NMR (600 MHz, CD
3oD) δ 0.51 (s, 3H), 0.63 (s, 3H), 1.31 (t, 12H,
j=7.2 Hz), 2.06 (s, 3H), 3.73 (q, 8H,
j=7.2 Hz), 6.77-7.48 (m, 10H); Nuclear-magnetism
13c NMR (150 MHz, CD
3oD) δ-2.90 ,-2.65,11.71,18.09,45.41,113.74,120.60,125.39,126.94,128.62,128.76,129.92,135.58,138.66,141.13,148.09,152.64,168.96; High resolution mass spectrum HRMS (ESI) calcd. for C
30h
39n
2si
+[M]
+: 455.2877, found:455.2888.
embodiment 6:
Phenyl aldehyde in raw material, with embodiment 4, is just changed to o-bromobenzaldehye by method.Obtain target compound 2-Br SiR, productive rate 43%.
Its nuclear-magnetism
1h NMR (300 MHz, CDCl
3) δ 0.56 (s, 3H), 0.61 (s, 3H), 1.16 (t, 12H,
j=7.2 Hz), 3.37 (q, 8H,
j=7.2 Hz), 6.59-7.74 (m, 10H); Nuclear-magnetism
13c NMR (75 MHz, CDCl
3) δ-1.68 ,-0.92,12.84,46.08,113.91,120.70,122.69,126.99,127.48,130.48,130.74,133.04,139.55,141.45,148.39,152.51,166.83; High resolution mass spectrum HRMS (ESI) calcd. for C
29h
36brN
2si
+[M]
+: 519.1826, found:519.1827,521.1810.
embodiment 7:
Phenyl aldehyde in raw material, with embodiment 4, is just changed to Ortho Nitro Benzaldehyde by method.Obtain target compound 2-NO
2siR, productive rate 47%.
Its nuclear-magnetism
1h NMR (300 MHz, CD
3oD) δ 0.58 (s, 3H), 0.63 (s, 3H), 1.28 (t, 12H,
j=7.2 Hz), 3.70 (q, 8H,
j=6.9 Hz), 6.73-8.41 (m, 10H); Nuclear-magnetism
13c NMR (75 MHz, CD
3oD) δ-3.31 ,-2.45,11.70,45.46,113.92,120.64,124.89,126.68,130.39,131.79,133.65,134.42,140.04,147.87,148.37,152.62,165.10; High resolution mass spectrum HRMS (ESI) calcd. for C
29h
36n
3o
2si
+[M]
+: 486.2571, found:486.2588.
embodiment 8:
Method is with embodiment 4, and just phenyl aldehyde in raw material is changed to phenyl aldehyde-2-sodium sulfonate, eluent is by volume of methylene dichloride: methyl alcohol=10:1.Obtain target compound 2-SO
3h SiR, productive rate 16%.
Its nuclear-magnetism
1h NMR (600 MHz, CD
3oD) δ 0.60 (s, 6H), 1.30 (t, 12H,
j=7.2 Hz), 3.70 (q, 8H,
j=7.2 Hz), 6.70-8.17 (m, 10H); Nuclear-magnetism
13c NMR (150 MHz, CD
3oD) δ-2.93 ,-2.62,11.72,45.23,113.01,119.78,127.84,128.15,128.52,129.27,130.11,136.40,142.58,144.09,148.02,152.39,170.25; High resolution mass spectrum HRMS (ESI) calcd. for C
29h
37n
2o
3sSi
+[M+H]
+: 521.2289, found:521.2301; C
39h
36n
2o
3sSiNa
+[M+Na]
+: 543.2108, found:543.2116.
embodiment 9:
Phenyl aldehyde in raw material, with embodiment 4, is just changed to 2-methyl-4 bromobenzaldehyde by method.Obtain target compound 2-Me-4-Br SiR, productive rate 43%.
Its nuclear-magnetism
1h NMR (300 MHz, CD
3oD) δ 0.59 (s, 3H), 0.60 (s, 3H), 1.29 (t, 12H,
j=7.2 Hz), 2.04 (s, 3H), 3.71 (q, 8H,
j=7.2 Hz), 6.77-7.61 (m, 9H); Nuclear-magnetism
13c NMR (75 MHz, CD
3oD) δ-2.91 ,-2.70,11.70,17.92,45.44,113.91,120.78,122.25,126.63,128.59,130.53,132.74,137.76,138.30,140.79,148.01,152.65,166.93; High resolution mass spectrum HRMS (ESI) calcd. for C
30h
38brN
2o
2si
+[M]
+: 533.1982, found:533.1978,535.1966.
embodiment 10:
Phenyl aldehyde in raw material, with embodiment 4, is just changed to 2-methyl-5-nitro phenyl aldehyde by method.Obtain target compound 2-Me-5-NO
2siR, productive rate 59%.
Its nuclear-magnetism
1h NMR (300 MHz, CD
3oD) δ 0.61 (s, 3H), 0.64 (s, 3H), 1.30 (t, 12H,
j=7.2 Hz), 2.18 (s, 3H), 3.73 (q, 8H,
j=7.2 Hz), 6.77-8.36 (m, 9H); Nuclear-magnetism
13c NMR (75 MHz, CD
3oD) δ-2.91 ,-2.70,11.70,18.25,45.51,114.17,121.13,123.29,123.63,126.39,131.35,139.97,140.40,143.89,146.24,148.10,152.71,164.49; High resolution mass spectrum HRMS (ESI) calcd. for C
30h
38n
3o
2si
+[M]
+: 500.2728, found:500.2729.
embodiment 11:
Phenyl aldehyde in raw material, with embodiment 4, is just changed to 3-methyl-4 aldehyde benzoic acid by method.Obtain target compound 2-Me-4-COOH SiR, productive rate 33%.
Its nuclear-magnetism
1h NMR (300 MHz, CD
3oD) δ 0.60 (s, 3H), 0.62 (s, 3H), 1.29 (t, 12H,
j=7.2 Hz), 2.12 (s, 3H), 3.72 (q, 8H,
j=7.2 Hz), 6.76-8.07 (m, 9H); Nuclear-magnetism
13c NMR (75 MHz, CD
3oD) δ-2.92 ,-2.69,11.69,18.04,45.44,113.94,120.82,126.34,126.72,129.16,131.08,131.15,136.29,140.70,143.43,148.00,152.68,167.18,167.84; High resolution mass spectrum HRMS (ESI) calcd. for C
31h
39n
2o
2si
+[M]
+: 499.2775, found:499.2759.
embodiment 12:
Method is with embodiment 4, and just phenyl aldehyde in raw material is changed to phenyl aldehyde-2,4-sodium disulfonate, eluent is by volume of methylene dichloride: methyl alcohol=5:1, obtains target compound 2,4-DiSO
3h SiR, productive rate 10%.
Its nuclear-magnetism
1h NMR (300 MHz, CD
3oD) δ 0.57 (s, 3H), 0.58 (s, 3H), 1.25 (t, 12H,
j=7.2 Hz), 2.02 (s, 1H), 3.67 (q, 8H,
j=6.9 Hz), 6.71-8.65 (m, 9H); Nuclear-magnetism
13c NMR (75 MHz, CD
3oD) δ-2.98 ,-2.60,11.70,45.75,113.65,120.49,125.70,126.87,127.88,130.63,135.62,137.01,138.25,141.47,142.42,147.89,152.00 high resolution mass spectrum HRMS (ESI) calcd. for C
29h
36n
2o
6s
2si [M+H]
+: 601.1857, found:601.1867; [M+Na]
+: 623.1676, found:623.1681.
embodiment 13:
In the withstand voltage reaction tubes of 15 mL heavy wall that magnetic stir bar is housed, add raw material SiN1 106 mg (0.3 mmol), adjacent aldehyde benzoic acid 225 mg (1.5 mmol), cupric bromide 7 mg (0.03 mmol), tube sealing is placed in shield cap, 140 ° of C oil baths react 5 hours, naturally cool to room temperature, add methylene dichloride and each 2 mL dissolvings of methyl alcohol, purification by silica gel column chromatography, eluent is by volume of sherwood oil: ethyl acetate: triethylamine=80: 1: 1, obtains target compound 2-COOH SiR, productive rate 52%.
Its nuclear-magnetism
1h NMR (300 MHz, CDCl
3) δ 0.64 (s, 3H), 0.66 (s, 3H), 1.18 (t, 12H,
j=7.2 Hz), 3.39 (q, 8H,
j=7.2 Hz), 6.50-8.01 (m, 10H); Nuclear-magnetism
13c NMR (75 MHz, CDCl
3) δ-1.78,0.44,12.55,44.28,92.30,112.45,115.94,124.86,125.58,127.48,128.46,128.65,130.79,133.50,137.30,146.58,154,15,170.63; High resolution mass spectrum HRMS (ESI) calcd. for C
30h
36n
2o
2si [M+H]
+: 485.2619, found:485.2623.
embodiment 14:
Method is with embodiment 13, and just SiN1 in raw material is changed to SiN2, eluent is by volume of sherwood oil: ethyl acetate: triethylamine=70: 1: 1, obtains target compound 2-COOH SiR
2, productive rate 16%.
Its nuclear-magnetism
1h NMR (300 MHz, CDCl
3): δ 0.54 (s, 3H), 0.58 (s, 3H), 1.18 (t, 12H,
j=7.2 Hz), 2.70-2.88 (m, 4H), 3.18 (q, 4H,
j=7.2 Hz), 3.29 (t, 4H,
j=8.4 Hz), 6.63-7.96 (m, 8H);
13c NMR (75 MHz, CDCl
3): δ-0.39,0.06,12.04,28.37,42.67,51.70,92.06,109.97,123.47,123.83,125.49,125.77,128.37,132.54,133.03,133.95,134.07,151.37,156.39,171.46; High resolution mass spectrum HRMS (ESI) calcd. for C
30h
32n
2o
2si [M+H]
+: 481.2306, found:481.2307.
embodiment 15:
Method is with embodiment 13, and just SiN1 in raw material is changed to SiN3, eluent is by volume of sherwood oil: ethyl acetate: triethylamine=70: 1: 1, obtains target compound 2-COOH SiR
3, productive rate 31%.
Its nuclear-magnetism
1h NMR (300 MHz, CDCl
3): δ 0.54 (s, 3H), 0.58 (s, 3H), 1.18 (t, 12H,
j=7.2 Hz), 2.70-2.88 (m, 4H), 3.18 (q, 4H,
j=7.2 Hz), 3.29 (t, 4H,
j=8.4 Hz), 6.63-7.96 (m, 8H); Nuclear-magnetism
13c NMR (75 MHz, CDCl
3): δ-0.39,0.06,12.04,28.37,42.67,51.70,92.06,109.97,123.47,123.83,125.49,125.77,128.37,132.54,133.03,133.95,134.07,151.37,156.39,171.46; High resolution mass spectrum HRMS (ESI) calcd. for C
30h
32n
2o
2si [M+H]
+: 481.2306, found:481.2307.
embodiment 16:
Aldehyde benzoic acid adjacent in raw material, with embodiment 13, is just changed to 2-aldehyde radical-4-bromo-benzoic acid, 8 hours reaction times by method.Obtain target compound 2-COOH-4-Br SiR, productive rate 40%.
Its nuclear-magnetism
1h NMR (300 MHz, CDCl
3) δ 0.59 (s, 3H), 0.63 (s, 3H), 1.17 (t, 12H,
j=7.2 Hz), 3.37 (q, 8H,
j=7.2 Hz), 6.51-7.83 (m, 9H); Nuclear-magnetism
13c NMR (75 MHz, CDCl
3) δ-1.36,0.26,12.58,44.30,91.75,112.78,115.82,125.93,126.80,127.79,128.49,128.94,129.87,132.20,136.84,146.67,156.71,169.95; High resolution mass spectrum HRMS (ESI) calcd. for C
30h
35brN
2o
4si [M+H]
+: 563.1724, found:563.1740,565.1725.
embodiment 17:
Method is with embodiment 13, and just aldehyde benzoic acid adjacent in raw material is changed to 2-aldehyde radical-4-cyanobenzoic acid, in 3 hours reaction times, eluent is by volume of sherwood oil: ethyl acetate: triethylamine=60: 1: 1.Obtain target compound 2-COOH-4-CN SiR, productive rate 31%.
Its nuclear-magnetism
1h NMR (300 MHz, CDCl
3): δ 0.63 (s, 3H), 0.68 (s, 3H), 1.21 (t, 12H,
j=6.9 Hz), 3.41 (q, 8H,
j=6.9 Hz), 6.50-8.01 (m, 9H); Nuclear-magnetism
13c NMR (75 MHz, CDCl
3): δ-1.46,0.27,12.54,44.31,92.59,112.68,115.95,117.16,117.77,126.43,128.32,128.65,129.01,130.54,132.22,137.03,146.80,155.29,168.96; High resolution mass spectrum HRMS (ESI) calcd. for C
31h
35n
3o
2si [M+H]
+: 510.2571, found:510.2581.
embodiment 18:
Method is with embodiment 13, and just aldehyde benzoic acid adjacent in raw material is changed to 2-aldehyde radical-5-nitrobenzoic acid, in 3 hours reaction times, eluent is by volume of sherwood oil: ethyl acetate: triethylamine=70: 1: 1.Obtain target compound 2-COOH-4-NO
2siR, productive rate 31%.
Its nuclear-magnetism
1h NMR (300 MHz, CDCl
3): δ 0.59 (s, 3H), 0.63 (s, 3H), 1.16 (t, 12H,
j=7.2 Hz), 3.37 (q, 8H,
j=7.2 Hz), 6.48-8.80 (m, 9H); Nuclear-magnetism
13c NMR (75 MHz, CDCl
3): δ-1.36,0.18,12.52,44.27,92.56,112.73,115.88,121.31,125.70,128.38,128.52,128.73,136.82,146.84,148.57,160.51,168.49; High resolution mass spectrum HRMS (ESI) calcd. for C
30h
35n
3o
4si [M+H]
+: 530.2470, found:530.2481.
embodiment 19:
Aldehyde benzoic acid adjacent in raw material, with embodiment 13, is just changed to 2-aldehyde radical-4-alkynyl phenylformic acid, 2 hours reaction times by method.Obtain target compound 2-COOH-5-CCH SiR, productive rate 16%.
Its nuclear-magnetism
1h NMR (300 MHz, CDCl
3) δ 0.59 (s, 3H), 0.62 (s, 3H), 1.15 (t, 12H,
j=7.2 Hz), 3.22 (s, 1H), 3.36 (q, 8H,
j=7.2 Hz), 6.48-7.92 (m, 9H); Nuclear-magnetism
13c NMR (75 MHz, CDCl
3) δ-1.55,0.30,12.55,44.25,80.57,82.69,92.10,112.60,115.82,125.42,127.14,127.50,128.28,128.48,130.12,132.47,136.98,146.62,154.60,169.94; High resolution mass spectrum HRMS (ESI) calcd. for C
32h
36n
2o
2si [M+H]
+: 509.2619, found:509.2614.
embodiment 20:
2-COOH-5-CN SiR 51 mg (0.10 mmol) is added, salt of wormwood 17 mg (0.12 mmol), DMSO 5 mL in the 10 mL eggplant-shape bottles that magnetic stir bar is housed.Magnetic agitation, then add hydrogen peroxide (30% aqueous solution) 200mL.60 ° of C react 1 hour, reaction solution is poured in 25 mL frozen water, dichloromethane extraction three times, merge organic layer, organic layer washes three times, anhydrous sodium sulfate drying, steaming desolventizes, silica gel column chromatography, eluent is by volume of sherwood oil: ethyl acetate: triethylamine=40: 1: 1, obtains target compound 2-COOH-5-CONH
2siR, productive rate 67%.
Its nuclear-magnetism
1h NMR (300 MHz, CDCl
3) δ 0.58 (s, 3H), 0.63 (s, 3H), 1.15 (t, 12H,
j=6.9 Hz), 3.35 (q, 8H,
j=6.9 Hz), 6.46-8.01 (m, 9H); Nuclear-magnetism
13c NMR (75 MHz, CDCl
3) δ-1.52,0.30,12.54,44.25,92.55,112.67,115.83,123.55,125.85,127.93,128.55,129.83,130.01,137.06,138.40,146.68,155.15,168.09,169.81; High resolution mass spectrum HRMS (ESI) calcd. for C
31h
37n
3o
3si [M+H]
+: 528.2677, found:528.2676.
embodiment 21:
In the 50 mL reaction flasks that stirrer is housed, add 2-COOH-4-NO
2siR106 mg (0.20 mmol), methyl alcohol 15 mL, methylene dichloride 10 mL, 10% Pd/C 50 mg.At H
2room temperature reaction 4 hours in atmosphere, filtration under diminished pressure removing insolubles, react 10 minutes under adding enough tetrachlorobenzoquinone room temperatures, filter, steaming desolventizes, purification by silica gel column chromatography, eluent is by volume of sherwood oil: ethyl acetate: triethylamine=100: 10: 1, obtains target compound 2-COOH-4-NH
2siR, productive rate 79%.
Its nuclear-magnetism
1h NMR (300 MHz, CDCl
3) δ 0.60 (s, 6H), 1.14 (t, 12H,
j=6.9 Hz), 3.35 (q, 8H,
j=6.9 Hz), 3.97 (s, 2H), 6.44-7.25 (m, 9H); Nuclear-magnetism
13c NMR (75 MHz, CDCl
3) δ-2.18,0.58,12.56,44.26,92.46,109.53,112.22,116.06,121.11,125.72,128.46,129.24,131.62,137.75,143.44,146.52,147.21,170.70; High-resolution mass spectrometer HRMS (ESI) calcd. for C
30h
35n
3o
2si [M+H]
+: 500.2728, found:500.2712.
embodiment 22: compound light property testing
1. solution preparation
Accurately take 2-H SiR respectively, 2-Me SiR, 2-Br SiR, 2-NO
2siR, 2-SO
3h SiR, 2-Me-4-Br SiR, 2-Me-5-NO
2siR, 2-Me-4-COOH SiR, 2,4-DiSO
3h SiR 1 ~ 3 mg, is settled to dissolve with ethanol the high standard solution that 10 mL obtain sample, measures the high standard solution of certain volume, be settled to 10 mL with ethanol, control its concentration between 2.4 mM ~ 3.2 mM with micropipet, for subsequent use.
Accurately take 2-COOH SiR respectively, 2-COOH SiR2,2-COOH SiR3,2-COOH-4-Br SiR, 2-COOH-4-CN SiR, 2-COOH-4-NO
2siR, 2-COOH-5-CCH SiR, 2-COOH-5-CONH
2siR, 2-COOH-4-NH
2siR 1 ~ 3 mg, is settled to dissolve with ethanol the high standard solution that 10 mL obtain sample, measures the high standard solution of certain volume with micropipet, add the ethanolic soln 80mL containing 1.0 ‰ hydrochloric acid, be settled to 10 mL with ethanol, control its concentration between 2.4 mM ~ 3.2 mM, for subsequent use.
2. the mensuration of sample absorption spectrum
By sample solution ultraviolet spectrophotometer (Agilent the Cary 100) (quartz container prepared, optical path length 1 cm) measure its ultra-violet absorption spectrum, measurement range 450 nm ~ 750 nm, measure its maximum absorption wavelength, calculate the molar absorptivity of its sample, the results are shown in Table 2, abosrption spectrogram is shown in accompanying drawing 1.
3. the mensuration of electromagnetic radiation spectrum
The sample solution fluorescence spectrophotometer prepared is measured its fluorescence emission spectrum, excitation wavelength 610 nm, measurement range 620 nm ~ 750 nm, measure its maximum emission wavelength, take Viola crystallina as reference substance, calculate its fluorescence quantum yield, the results are shown in following table 2, the fluorescence emission spectrum of sample is shown in accompanying drawing 2.
Table 2 compound spectra data
embodiment 23: the cell dyeing of compound and laser co-focusing fluorescence imaging experiments
1. cell culture processes:
Test cell: select mouse brain capillary endothelium (bEND3) and human liver cancer cell (HepG2);
Cell culture condition: the DMEM substratum using the penicillin containing 10% FBS, 0.1 mg/mL Streptomycin sulphate and 100 U/mL, is containing 5% CO
2, 95% air, 37 DEG C of constant temperature, culturing cell in the cell culture incubator of saturated humidity.When cell aggregation degree reaches 90%, sucking-off substratum also cleans cell 2 times by PBS solution, uses 37 DEG C, 0.25% trypsinase to digest 1 minute, add 2 times of volume medium neutralizations, centrifugal 5 minutes of 1000rpm, to go down to posterity cell culture dish with 1: 3 ratio, within every 2 days, changes a subculture.
2. the configuration of test fluorescence dye
2-COOH SiR DMSO is configured to 1.0 mM solution for standby; 2,4-DiSO
3h SiR PBS damping fluid is configured to 1.0 mM solution for standby.
3. cell dyeing method
By bEND3 cell with 5 × 10
4density is inoculated in laser co-focusing special culture dish, is containing 5% CO
2, 95% air, 37 DEG C of constant temperature, in the cell culture incubator of saturated humidity after culturing cell 24 h, discard substratum clean 2 times with precooling PBS.2-COOH SiR solution 100 mL is added in 1.0 mL fresh cultures, add after mixing in culture dish, 40 minutes are hatched in incubator, DAPI staining agent is added afterwards in this substratum, continue in incubator to hatch 20 minutes, with PBS buffer solution three times, observe under laser confocal microscope, excitation light source 488 nm and 633 nm.Laser co-focusing fluorescence imaging figure is shown in accompanying drawing 3.2-COOH SiR is same to consistent to bEND3 cell dyeing method to HepG2 cell dyeing method, and laser co-focusing fluorescence imaging figure is shown in accompanying drawing 4.2,4-DiSO
3h SiR is consistent with 2-COOH SiR with HepG2 cell dyeing method to bEND3 cell, and laser co-focusing fluorescence imaging figure is shown in accompanying drawing 5, accompanying drawing 6.
4. cell dyeing experimental result
By laser co-focusing fluorescence imaging to cell dyeing effect observation, find 2-COOH SiR and 2,4-DiSO
3the cytolemma that H SiR all can pass through viable cell enters in cell, and is uniformly distributed in kytoplasm, but can not dye to nucleus.As Suo Shi accompanying drawing 3 ~ 6 laser co-focusing fluorescence imaging design sketch (photo is converted into grayscale mode).
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the prerequisite not departing from the inventive method; can also make some improvement and supplement, these improve and supplement and also should be considered as protection scope of the present invention.
Claims (10)
1. a silica-based Rhodamine Derivatives, is characterized in that, described silica-based Rhodamine Derivatives is the compound with general formula (I) structure:
R
11, R
15wherein any one for or two be-COOH or-COO
-time, the corresponding lactone volution form of general formula (I) can be formed, there is the compound of general formula (II) structure:
Wherein:
R
2, R
3be respectively alkyl or the phenyl of C1-C6 straight or branched, described alkyl is selected from: methyl, ethyl, n-propyl, sec.-propyl, normal-butyl, isobutyl-, sec-butyl, the tertiary butyl, n-pentyl, isopentyl, sec.-amyl sec-pentyl secondary amyl, neo-pentyl, n-hexyl;
R
4, R
5, R
6, R
7, R
8, R
9be respectively the alkyl of the straight or branched of hydrogen or C1-C6, described alkyl is selected from: methyl, ethyl, n-propyl, sec.-propyl, normal-butyl, isobutyl-, sec-butyl, the tertiary butyl, n-pentyl, isopentyl, sec.-amyl sec-pentyl secondary amyl, neo-pentyl, n-hexyl;
R
20, R
21, R
22, R
23be respectively the saturated of C1-C6 straight or branched or undersaturated alkyl or cycloalkyl, described alkyl is selected from: methyl, ethyl, n-propyl, sec.-propyl, allyl group, normal-butyl, isobutyl-, sec-butyl, the tertiary butyl, n-pentyl, isopentyl, sec.-amyl sec-pentyl secondary amyl, neo-pentyl, n-hexyl;
R
11, R
12, R
13, R
14, R
15, R
16, R
17, R
18, R
19for identical or different substituting groups, described substituting group is selected from: alkyl, halogen, cyano group, nitro, alkoxyl group, haloalkyl.
2. the silica-based Rhodamine Derivatives according to claims 1, is characterized in that, described silica-based Rhodamine Derivatives replace situation be selected from following in one, two or more:
A) described substituent R
2, R
3for methyl, ethyl or phenyl;
B) described substituent R
4, R
5, R
6, R
7, R
8, R
9for hydrogen;
C) described substituent R
20, R
21, R
22, R
23for methyl, ethyl or allyl group.
3. the silica-based Rhodamine Derivatives according to claims 1, is characterized in that, described silica-based Rhodamine Derivatives possess under conditions one or both:
A) described substituent R
20or R
21with R
5be connected with carbochain, form five yuan or six-membered cyclic structure with parent benzene;
B) described substituent R
22or R
23with R
8be connected with carbochain, form five yuan or six-membered cyclic structure with parent benzene.
4. the silica-based Rhodamine Derivatives according to claims 3, is characterized in that, described silica-based Rhodamine Derivatives possess under conditions one or both:
Described substituting group-NR
20r
21with-R
5formation has the structure of following general formula (III):
Described substituting group-NR
22r
23with-R
8formation has the structure of following general formula (IV):
。
5. the silica-based Rhodamine Derivatives according to claims 4, is characterized in that, described silica-based Rhodamine Derivatives possess under conditions one or both:
A) substituent R described in
24for hydrogen, methyl, ethyl or allyl group;
B) substituent R described in
25for hydrogen, methyl, ethyl or allyl group.
6., according to the arbitrary described silica-based Rhodamine Derivatives of claims 1-4, it is characterized in that, described silica-based Rhodamine Derivatives replace situation be selected from following in one:
A) R
12, R
13, R
14be hydrogen, R
11or R
15wherein any one is-CN ,-NO
2,-SO
3h ,-SO
3 -,-OH ,-OCH
3,-NH
2,-N
3,-NCS ,-CH
3,-F ,-Cl, any one group in-Br ,-I, another is hydrogen;
B) R
11, R
15wherein any one is methyl, and another is hydrogen, R
12, R
13, R
14in any one is-CN ,-NO
2,-SO
3h ,-SO
3 -,-OH ,-OCH
3,-NH
2,-CH
3,-N
3,-NCS,
,-CONH
2,-CH
2nH
2,-F ,-Cl ,-Br ,-I ,-COOH ,-COO
-in any one group, all the other two are hydrogen;
C) R
11, R
15wherein any one is-SO
3h or-SO
3 -, another is hydrogen, R
12, R
13, R
14in any one be-SO
3h or-SO
3 -, two other is hydrogen;
D) R
11or R
15one is-H, and another is-COOH or-COO
-, R
12, R
13, R
14in any one is-CN ,-NO
2,-SO
3h ,-SO
3 -,-OH ,-OCH
3,-NH
2,-CH
3,-N
3,-NCS,
,-CONH
2,-CH
2nH
2,-F ,-Cl ,-Br ,-I ,-COOH ,-COO
-one in group, all the other two is hydrogen;
E) R
16, R
17, R
18, R
19in any one is-CN ,-NO
2,-SO
3h ,-SO
3 -,-OH ,-OCH
3,-NH
2,-CH
3,-N
3,-NCS,
,-CONH
2,-CH
2nH
2,-F ,-Cl ,-Br ,-I ,-COOH ,-COO
-one in group, its excess-three is hydrogen.
7. the preparation method of the arbitrary described silica-based Rhodamine Derivatives of claims 1-6, it is characterized in that, described preparation method comprises the steps:
The first step: the synthesis with the silica-based intermediate of key of logical formula V structure
As-NR
20r
21with-NR
22r
23, R
4with R
7, R
5with R
8, R
6with R
9when all there is same structure between two, adopt and synthesize with the following method;
N, N-dialkyl m-bromoaniline derivative reacts to n-Butyl Lithium and generates corresponding lithium reagent, then reacts with dialkyl dichlorosilane further, silica gel column chromatography, carry out wash-out with ethyl acetate and sherwood oil mixed system, generate the silica-based intermediate of key with logical formula V structure;
As-NR
20r
21with-NR
22r
23, R
4with R
7, R
5with R
8, R
6with R
9have when more than a pair or a pair there is different structure between any two, adopt and synthesize with the following method;
A kind of N, N-dialkyl m-bromoaniline derivative generates corresponding lithium reagent to n-Butyl Lithium, then reacts with dialkyl dichlorosilane, replaces one of them chlorine, generates the product of monochlorosilane; Another kind of N, N-dialkyl m-bromoaniline derivative reacts to n-Butyl Lithium and generates corresponding lithium reagent, react with monochlorosilane product further again, silica gel column chromatography, carry out wash-out with ethyl acetate and sherwood oil mixed system, generate the silica-based intermediate of key with logical formula V structure;
Second step: the synthesis of silica-based Luo Mingdan derivative
The benzaldehyde derivative of the silica-based intermediate of key and different replacement with logical formula V structure is under catalytic condition, tube sealing is placed on 140 ° of C reactions in shield cap, be oxidized through tetrachlorobenzoquinone again or be not oxidized, finally carry out column chromatography for separation, carry out wash-out with methylene dichloride and the mixed system of methyl alcohol or the mixed system of ethyl acetate, sherwood oil and triethylamine, obtain silica-based Rhodamine Derivatives.
8. the preparation method according to claims 7, is characterized in that,
Preparing solvent used when having the silica-based intermediate of key of logical formula V structure is ether; When described preparation has the key of a logical formula V structure silica-based intermediate, temperature of reaction is 0 ° of C;
When described preparation has the key of logical formula V structure silica-based intermediate, if-NR
20r
21with-NR
22r
23, R
4with R
7, R
5with R
8, R
6with R
9when all having same structure between two, the feed ratio of each reactant is N, N-dialkyl m-bromoaniline: butyllithium: dialkyl dichlorosilane=1: 1.05:0.6; If-NR
20r
21with-NR
22r
23, R
4with R
7, R
5with R
8, R
6with R
9have when having different structure more than a pair or a pair between any two, the feed ratio of each reactant is N, N-dialkyl m-bromoaniline: butyllithium: dialkyl dichlorosilane=1: 1.05:5;
The described silica-based intermediate of key with logical formula V structure is 1: 5 from the molar ratio of the benzaldehyde derivative of different replacement;
CuBr selected by described catalyzer
2or tosic acid monohydrate;
Described have the silica-based intermediate of key of logical formula V structure and the mol ratio of catalyzer is 1:0.1 ~ 1
The eluent of described column chromatography for separation, by volume, the mixed solvent of methyl alcohol and methylene dichloride is methyl alcohol: methylene dichloride=1:5 ~ 40, ethyl acetate and sherwood oil mixed solvent are ethyl acetate: sherwood oil=1:40 ~ 80, the mixed solvent of ethyl acetate, sherwood oil and triethylamine is ethyl acetate: sherwood oil=1:10 ~ 80, and the add-on of triethylamine is 1% of ethyl acetate and petroleum ether mixtures volume.
9. preparation method according to claim 7, is characterized in that, described preparation method also comprises the further derivative reaction of the silica-based Rhodamine Derivatives of the 3rd step; Described derivative reaction is: the azido reaction of the partial hydrolysis reaction of cyano group, the complete hydrolysis reaction of cyano group, the reduction reaction of cyano group, the reduction reaction of nitro or amino.
10. the application of the arbitrary described silica-based Rhodamine Derivatives of claims 1-6 in preparation cell dye, biological stain, biomolecules or biomone fluorescent mark.
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| CN104262378B (en) | 2017-05-03 |
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