CN109053790A - 一种溶酶体靶向的次氯酸近红外荧光探针及其制备方法和应用 - Google Patents
一种溶酶体靶向的次氯酸近红外荧光探针及其制备方法和应用 Download PDFInfo
- Publication number
- CN109053790A CN109053790A CN201811001966.1A CN201811001966A CN109053790A CN 109053790 A CN109053790 A CN 109053790A CN 201811001966 A CN201811001966 A CN 201811001966A CN 109053790 A CN109053790 A CN 109053790A
- Authority
- CN
- China
- Prior art keywords
- milliliters
- hypochlorous acid
- fluorescent probe
- compound
- near infrared
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- QWPPOHNGKGFGJK-UHFFFAOYSA-N hypochlorous acid Chemical compound ClO QWPPOHNGKGFGJK-UHFFFAOYSA-N 0.000 title claims abstract description 65
- 239000007850 fluorescent dye Substances 0.000 title claims abstract description 52
- 210000003712 lysosome Anatomy 0.000 title claims abstract description 35
- 230000001868 lysosomic effect Effects 0.000 title claims abstract description 33
- 230000008685 targeting Effects 0.000 title claims abstract description 15
- 238000002360 preparation method Methods 0.000 title claims abstract description 8
- 239000000523 sample Substances 0.000 claims abstract description 31
- 210000004027 cell Anatomy 0.000 claims abstract description 17
- 238000001514 detection method Methods 0.000 claims abstract description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 12
- 150000001875 compounds Chemical class 0.000 claims description 50
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 34
- 239000000243 solution Substances 0.000 claims description 31
- 239000012043 crude product Substances 0.000 claims description 19
- 238000006243 chemical reaction Methods 0.000 claims description 18
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 16
- 239000000741 silica gel Substances 0.000 claims description 16
- 229910002027 silica gel Inorganic materials 0.000 claims description 16
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-dimethylformamide Substances CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 14
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 13
- 238000003384 imaging method Methods 0.000 claims description 13
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 12
- 238000000746 purification Methods 0.000 claims description 12
- 239000002904 solvent Substances 0.000 claims description 12
- 210000001519 tissue Anatomy 0.000 claims description 11
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 9
- 239000007864 aqueous solution Substances 0.000 claims description 9
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 9
- 238000005406 washing Methods 0.000 claims description 7
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 claims description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- 150000002170 ethers Chemical class 0.000 claims description 6
- 239000011780 sodium chloride Substances 0.000 claims description 6
- 239000007832 Na2SO4 Substances 0.000 claims description 5
- 229910052938 sodium sulfate Inorganic materials 0.000 claims description 5
- DYNFCHNNOHNJFG-UHFFFAOYSA-N 2-formylbenzoic acid Chemical compound OC(=O)C1=CC=CC=C1C=O DYNFCHNNOHNJFG-UHFFFAOYSA-N 0.000 claims description 4
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 claims description 4
- 229940125904 compound 1 Drugs 0.000 claims description 4
- 229940126214 compound 3 Drugs 0.000 claims description 4
- 229920006395 saturated elastomer Polymers 0.000 claims description 4
- 238000003756 stirring Methods 0.000 claims description 4
- NWZSZGALRFJKBT-KNIFDHDWSA-N (2s)-2,6-diaminohexanoic acid;(2s)-2-hydroxybutanedioic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O.NCCCC[C@H](N)C(O)=O NWZSZGALRFJKBT-KNIFDHDWSA-N 0.000 claims description 3
- KKASGUHLXWAKEZ-UHFFFAOYSA-N 1-isothiocyanatopropane Chemical compound CCCN=C=S KKASGUHLXWAKEZ-UHFFFAOYSA-N 0.000 claims description 3
- ONMSBNJJCUCYED-UHFFFAOYSA-N 2-bromo-n,n-dimethylaniline Chemical compound CN(C)C1=CC=CC=C1Br ONMSBNJJCUCYED-UHFFFAOYSA-N 0.000 claims description 3
- -1 3- (4- morpholinyl) propyl Chemical group 0.000 claims description 3
- 238000013019 agitation Methods 0.000 claims description 3
- 229910052786 argon Inorganic materials 0.000 claims description 3
- ODWXUNBKCRECNW-UHFFFAOYSA-M bromocopper(1+) Chemical compound Br[Cu+] ODWXUNBKCRECNW-UHFFFAOYSA-M 0.000 claims description 3
- 229940125782 compound 2 Drugs 0.000 claims description 3
- LIKFHECYJZWXFJ-UHFFFAOYSA-N dimethyldichlorosilane Chemical compound C[Si](C)(Cl)Cl LIKFHECYJZWXFJ-UHFFFAOYSA-N 0.000 claims description 3
- 235000019441 ethanol Nutrition 0.000 claims description 3
- 239000007789 gas Substances 0.000 claims description 3
- 239000011521 glass Substances 0.000 claims description 3
- 238000010438 heat treatment Methods 0.000 claims description 3
- IKDUDTNKRLTJSI-UHFFFAOYSA-N hydrazine monohydrate Substances O.NN IKDUDTNKRLTJSI-UHFFFAOYSA-N 0.000 claims description 3
- 238000007689 inspection Methods 0.000 claims description 3
- DLEDOFVPSDKWEF-UHFFFAOYSA-N lithium butane Chemical compound [Li+].CCC[CH2-] DLEDOFVPSDKWEF-UHFFFAOYSA-N 0.000 claims description 3
- MZRVEZGGRBJDDB-UHFFFAOYSA-N n-Butyllithium Substances [Li]CCCC MZRVEZGGRBJDDB-UHFFFAOYSA-N 0.000 claims description 3
- 238000010791 quenching Methods 0.000 claims description 3
- 230000000171 quenching effect Effects 0.000 claims description 3
- 239000011541 reaction mixture Substances 0.000 claims description 3
- 230000006837 decompression Effects 0.000 claims description 2
- 238000004090 dissolution Methods 0.000 claims description 2
- 238000000605 extraction Methods 0.000 claims description 2
- 238000001917 fluorescence detection Methods 0.000 claims description 2
- 239000012266 salt solution Substances 0.000 claims description 2
- 238000005292 vacuum distillation Methods 0.000 claims description 2
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 claims 2
- NEHMKBQYUWJMIP-UHFFFAOYSA-N chloromethane Chemical class ClC NEHMKBQYUWJMIP-UHFFFAOYSA-N 0.000 claims 1
- 125000003963 dichloro group Chemical group Cl* 0.000 claims 1
- 230000035945 sensitivity Effects 0.000 abstract description 2
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 9
- 238000000799 fluorescence microscopy Methods 0.000 description 9
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- 239000000975 dye Substances 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- JQDZUSDVVHXANW-UHFFFAOYSA-N C1=CC(=C23)C4=NC5=CC=CC=C5N4C(=O)C2=CC=CC3=C1NCCN1CCOCC1 Chemical compound C1=CC(=C23)C4=NC5=CC=CC=C5N4C(=O)C2=CC=CC3=C1NCCN1CCOCC1 JQDZUSDVVHXANW-UHFFFAOYSA-N 0.000 description 5
- 238000002189 fluorescence spectrum Methods 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 4
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 4
- 238000004847 absorption spectroscopy Methods 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 239000003480 eluent Substances 0.000 description 4
- 239000002245 particle Substances 0.000 description 4
- 230000035515 penetration Effects 0.000 description 4
- 230000004044 response Effects 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- 206010061218 Inflammation Diseases 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 206010028980 Neoplasm Diseases 0.000 description 3
- 208000027418 Wounds and injury Diseases 0.000 description 3
- 230000001093 anti-cancer Effects 0.000 description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 3
- 239000012472 biological sample Substances 0.000 description 3
- 201000011510 cancer Diseases 0.000 description 3
- XTEGARKTQYYJKE-UHFFFAOYSA-N chloric acid Chemical compound OCl(=O)=O XTEGARKTQYYJKE-UHFFFAOYSA-N 0.000 description 3
- 229940005991 chloric acid Drugs 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 230000005284 excitation Effects 0.000 description 3
- 230000004054 inflammatory process Effects 0.000 description 3
- 208000014674 injury Diseases 0.000 description 3
- 150000002500 ions Chemical class 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 239000001301 oxygen Substances 0.000 description 3
- 229910052760 oxygen Inorganic materials 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 239000011550 stock solution Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000005160 1H NMR spectroscopy Methods 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 102000004317 Lyases Human genes 0.000 description 2
- 108090000856 Lyases Proteins 0.000 description 2
- 102000003896 Myeloperoxidases Human genes 0.000 description 2
- 108090000235 Myeloperoxidases Proteins 0.000 description 2
- 206010057249 Phagocytosis Diseases 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 239000001569 carbon dioxide Substances 0.000 description 2
- 229910002092 carbon dioxide Inorganic materials 0.000 description 2
- 239000000460 chlorine Substances 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- WQYVRQLZKVEZGA-UHFFFAOYSA-N hypochlorite Chemical compound Cl[O-] WQYVRQLZKVEZGA-UHFFFAOYSA-N 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 230000008782 phagocytosis Effects 0.000 description 2
- 239000012071 phase Substances 0.000 description 2
- 230000035479 physiological effects, processes and functions Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 239000012086 standard solution Substances 0.000 description 2
- BWGRDBSNKQABCB-UHFFFAOYSA-N 4,4-difluoro-N-[3-[3-(3-methyl-5-propan-2-yl-1,2,4-triazol-4-yl)-8-azabicyclo[3.2.1]octan-8-yl]-1-thiophen-2-ylpropyl]cyclohexane-1-carboxamide Chemical compound CC(C)C1=NN=C(C)N1C1CC2CCC(C1)N2CCC(NC(=O)C1CCC(F)(F)CC1)C1=CC=CS1 BWGRDBSNKQABCB-UHFFFAOYSA-N 0.000 description 1
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 1
- 208000017667 Chronic Disease Diseases 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 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 1
- LFZAGIJXANFPFN-UHFFFAOYSA-N N-[3-[4-(3-methyl-5-propan-2-yl-1,2,4-triazol-4-yl)piperidin-1-yl]-1-thiophen-2-ylpropyl]acetamide Chemical compound C(C)(C)C1=NN=C(N1C1CCN(CC1)CCC(C=1SC=CC=1)NC(C)=O)C LFZAGIJXANFPFN-UHFFFAOYSA-N 0.000 description 1
- 208000037273 Pathologic Processes Diseases 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 206010003246 arthritis Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 125000001309 chloro group Chemical group Cl* 0.000 description 1
- 238000007334 copolymerization reaction Methods 0.000 description 1
- 108010037721 cytase Proteins 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 239000012216 imaging agent Substances 0.000 description 1
- 210000005007 innate immune system Anatomy 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 210000005228 liver tissue Anatomy 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 230000004770 neurodegeneration Effects 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 239000012074 organic phase Substances 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000009054 pathological process Effects 0.000 description 1
- 210000001539 phagocyte Anatomy 0.000 description 1
- 238000011897 real-time detection Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 1
- CIHOLLKRGTVIJN-UHFFFAOYSA-N tert‐butyl hydroperoxide Chemical compound CC(C)(C)OO CIHOLLKRGTVIJN-UHFFFAOYSA-N 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
- 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
- C07F7/0803—Compounds with Si-C or Si-Si linkages
- C07F7/081—Compounds with Si-C or Si-Si linkages comprising at least one atom selected from the elements N, O, halogen, S, Se or Te
- C07F7/0812—Compounds with Si-C or Si-Si linkages comprising at least one atom selected from the elements N, O, halogen, S, Se or Te comprising a heterocyclic ring
- C07F7/0816—Compounds with Si-C or Si-Si linkages comprising at least one atom selected from the elements N, O, halogen, S, Se or Te comprising a heterocyclic ring said ring comprising Si as a ring atom
-
- 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
- C07F7/0803—Compounds with Si-C or Si-Si linkages
- C07F7/0825—Preparations of compounds not comprising Si-Si or Si-cyano linkages
- C07F7/083—Syntheses without formation of a Si-C bond
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N21/643—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" non-biological material
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1029—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1029—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
- C09K2211/1033—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom with oxygen
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1029—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
- C09K2211/104—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom with other heteroatoms
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Optics & Photonics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Materials Engineering (AREA)
- Molecular Biology (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
本发明公开了一种溶酶体靶向的次氯酸近红外荧光探针及其制备方法和应用,属于分析化学技术领域。本发明的技术方案要点为:一种溶酶体靶向的次氯酸近红外荧光探针,其结构式为:本发明还具体公开了该溶酶体靶向的次氯酸近红外荧光探针的制备方法及其在水环境或生物细胞体系检测中的应用。本发明的荧光探针具有近红外发射、灵敏度高、选择性好及响应迅速等特点。
Description
技术领域
本发明属于分析化学技术领域,具体涉及一种溶酶体靶向的次氯酸近红外荧光探针及其制备方法和应用。
背景技术
活性氧(ROS)与各种生理和病理进程有关,如病原体反应进程、机体衰老进程以及抵抗炎症进程。次氯酸(HClO)是活性氧中一种高效的氧化剂,通常在细胞吞噬和炎症反应过程中产生,在先天免疫系统中起着重要作用。次氯酸主要分布在吞噬细胞的酸性溶酶体中,通常由在吞噬酶体中的髓过氧化物酶(MPO)催化的氯离子过氧化产生。有证据表明,生命体内错位或过量的HClO浓度表达与多种疾病有关,如神经变性、关节炎和动脉粥样硬化。例如,HClO在溶酶体中的异常积累可诱发慢性疾病,这是因为过量的次氯酸会诱导溶酶体的破裂而导致细胞死亡。此外,近来溶酶体被认为是选择性杀伤癌细胞的有力靶点,可以作为一种新型的抗癌途径(溶酶体靶向抗癌,LCD)。在诱导LCD的各种方法中,活性氧(ROS)是最常见的。然而,关于包括HClO在内的ROS诱导的癌细胞LCD的细节仍知之甚少。因此,实时监测及影像癌细胞溶酶体中HClO对于阐明LCD相关的抗癌机制和评价新的抗癌药物具有重要意义。总之,开发合适的化学工具来直接实时监测溶酶体中的次氯酸显得尤为必要。
近年来,次氯酸荧光探针的研制取得了显著进展,其中一些已成功地应用于溶酶体中次氯酸的实时检测和成像。然而,由于这些探针的短波长激发和发射(一般为λex<600nm和λem<650nm)而限制了它们在体内的实际应用,这也引发了许多问题,包括体内自发荧光的干扰、成像试剂的光漂白、对生物样品的光损伤现象。近红外荧光探针(λex>600nm和λem>650nm)能较好地解决上述问题,使生物样品的光损伤最小、组织穿透深度增加及降低背景自体荧光的干扰。然而,据我们所知,目前还没有检测溶酶体中HClO的近红外荧光探针的相关报道,导致在组织或活体中溶酶体次氯酸检测成像方面存在一定的局限性。
发明内容
针对目前还没有溶酶体靶向近红外次氯酸荧光探针报道的现状以及溶酶体内次氯酸检测所面临的问题,本发明提供了一种溶酶体靶向的次氯酸近红外荧光探针,该荧光探针具有近红外发射、灵敏度高、选择性好及响应迅速等特点。
本发明还提供了上述溶酶体靶向的近红外次氯酸荧光探针的制备方法及其在水环境或生物细胞体系检测中的应用。
本发明为解决上述技术问题采用如下技术方案,一种溶酶体靶向的次氯酸近红外荧光探针,其特征在于该荧光探针的结构式如下:
本发明所述的溶酶体靶向的次氯酸近红外荧光探针的制备方法,其特征在于具体步骤为:
步骤S1:于-78℃,在氩气保护下,将6克3-溴-N,N-二甲基苯胺与60毫升无水乙醚加入至干燥的250毫升圆底烧瓶中,磁力搅拌5分钟使其溶解,随后将13.1毫升摩尔浓度为2.4mol/L的正丁基锂的正己烷溶液滴加至反应液中,滴加完毕后于0℃反应2小时,再将2.2毫升二氯二甲基硅烷溶于10毫升无水乙醚中,然后滴加至上述反应液中,滴加完毕后反应至室温并搅拌过夜,加50毫升水猝灭反应,并将反应液用乙醚萃取、用水洗涤后,用饱和NaCl水溶液洗涤,无水硫酸钠干燥,减压旋干溶剂后得到粗产品,将粗产品用硅胶柱纯化得到化合物1,其结构式如下:
步骤S2:将500mg化合物1、1260mg 2-羧基苯甲醛和37.5mg溴化铜加入到100毫升玻璃厚壁耐压管中,于140℃加热搅拌反应5小时后自然冷却至室温,然后将反应混合物溶于50毫升二氯甲烷中,用质量浓度为10%的NaOH溶液洗涤三遍,回收并旋干二氯甲烷相得到粗产品,将粗产品用硅胶柱纯化得到化合物2,其结构式如下:
步骤S3:将428.6mg化合物2、50毫升乙醇和2毫升质量浓度为85%的水合肼加入至100毫升的圆底烧瓶,于78℃反应4小时后将反应液减压蒸馏除去溶剂,将残余物用50毫升二氯甲烷溶解后,用饱和食盐水水洗二氯甲烷相,并用无水硫酸钠干燥,减压蒸馏除去二氯甲烷相得到粗产物,将粗产物用硅胶柱纯化得到化合物3,其结构式如下:
步骤S4:将221.3mg化合物3、15毫升DMF和2毫升3-(4-吗啉基)异硫氰酸丙酯加入至50毫升的圆底烧瓶,于80℃搅拌反应72小时,将反应液减压蒸馏除去溶剂后得到残余物,饱和NaCl水溶液洗涤,无水Na2SO4干燥,再将残余物用硅胶柱纯化得到目标荧光探针化合物Lyso-NIR-HClO。
本发明所述的溶酶体靶向的次氯酸近红外荧光探针在水环境或生物细胞体系选择性检测次氯酸中的应用,其中检测包括水溶液中荧光检测、细胞成像检测、组织成像检测。
本发明与现有技术相比具有以下有益效果:(1)该荧光探针的合成相对比较容易,且后处理过程相对简单;(2)该荧光探针实现了次氯酸分子高选择性高灵敏度快速检测,具有抵抗生命体内其它分子干扰的能力;(3)该荧光探针可以应用于细胞溶酶体中次氯酸的检测,通过与商业化溶酶体染料进行比较,发现此荧光探针对溶酶体具有较高的溶酶体靶向能力,两种染料的共定位系数为0.93,说明此荧光探针可以作为细胞溶酶体内次氯酸的检测探针;(4)该荧光探针具有近红外发射,能够应用于组织或者活体中次氯酸的成像检测,通过降低生命体内自发荧光背景干扰、降低对生物样品的光损伤、提高组织穿透深度等特点,以获得更加准确及稳定的光学信号及成像效果。故而,本发明中的荧光探针在次氯酸检测领域具有广阔的应用前景,对次氯酸在生物体生理和病理过程的作用机制以及溶酶体在炎症反应中的作用等研究具有重要意义。
附图说明
图1是实施例1制得的荧光探针化合物Lyso-NIR-HClO在加入不同浓度次氯酸后的荧光光谱图;
图2是实施例1制得的荧光探针化合物Lyso-NIR-HClO在加入不同浓度次氯酸后的紫外可见吸收光谱图;
图3是实施例1制得的荧光探针化合物Lyso-NIR-HClO在发射波长为680nm处的的荧光强度随次氯酸浓度(0-40μM)变化的关系曲线图,插图为荧光探针化合物Lyso-NIR-HClO在发射波长为680nm处的的荧光强度随次氯酸浓度(0-10μM)变化的关系曲线图;
图4是实施例1制得的荧光探针化合物Lyso-NIR-HClO对不同离子和分子的选择性柱状图,其中1、PBS;2、K+;3、Ca2+;4、Mg2+;5、Zn2+;6、Fe3+;7、Cu2+;8、CH3COO-;9、NO3 -;10、Cl-;11、F-;12、I-;13、Cys;14、GSH;15、Glucose;16、ATP;17、ADP;18、t-BuOO.;19、NO2 -;20、H2O2;21、ONOO-;22、O2 .-;23、.OH;24、1O2;25、NO;26、t-BuOOH;27、NaClO;
图5是pH对实施例1制得的荧光探针化合物Lyso-NIR-HClO检测次氯酸的影响图;
图6是实施例1制得的荧光探针化合物Lyso-NIR-HClO在体系中含有不同浓度的次氯酸(1μM,5μM,10μM)时,溶液在680nm处的荧光强度随时间变化关系曲线图;
图7是在HeLa细胞中实施例1制得的荧光探针化合物Lyso-NIR-HClO检测外源性次氯酸荧光成像图与商业化溶酶体定位染料LysoSensorTM Green DND-189的共定位成像图,其中a)探针浓度为5μM与LysoSensorTM Green DND-189(1μM)加入到HeLa细胞中培养30分钟后明场图,b)商业染料LysoSensorTM Green DND-189绿色通道荧光成像图,c)加入次氯酸后探针分子红色通道荧光成像图,d)绿色通道与红色通道叠加图,e)图d中粉红色椭圆内区域的绿色通道与红色通道叠加强度散点图;f)图d中粉红色直线绿色通道与红色通道叠加荧光强度比较;
图8是实施例1制得的荧光探针化合物Lyso-NIR-HClO检测HeLa中不同浓度外源性次氯酸(0μM,10μM,20μM)荧光成像图;
图9是实施例1制得的荧光探针化合物Lyso-NIR-HClO检测小鼠肝脏组织中外源性次氯酸(200μM)荧光成像图,其中a)与c)探针浓度为20μM,未加次氯酸;b)与d)探针浓度为20μM,加200μM次氯酸,成像深度均为38μm。
具体实施方式
以下通过实施例对本发明的上述内容做进一步详细说明,但不应该将此理解为本发明上述主题的范围仅限于以下的实施例,凡基于本发明上述内容实现的技术均属于本发明的范围。
实施例1
荧光探针化合物Lyso-NIR-HClO的合成
(1)化合物1的合成
于-78℃,在氩气保护下,将6克3-溴-N,N-二甲基苯胺与60毫升无水乙醚加入至干燥的250毫升圆底烧瓶中,磁力搅拌5分钟使其溶解,随后将13.1毫升摩尔浓度为2.4mol/L的正丁基锂的正己烷溶液缓慢滴加至反应液中,滴加完毕后于0℃反应2小时,再将2.2毫升二氯二甲基硅烷溶于10毫升无水乙醚中,然后缓慢滴加至上述反应液中,滴加完毕后反应至室温搅拌过夜,加50毫升水猝灭反应,并将反应液用乙醚萃取,分液漏斗分取有机相,用水洗涤后(50毫升×2),饱和NaCl水溶液洗涤(50毫升×1),无水Na2SO4干燥,旋转蒸发仪蒸除溶剂得粗产品,将粗产品用硅胶柱纯化,硅胶颗粒大小为200-300目,洗脱剂体积配比为石油醚/乙酸乙酯=80:1,得到化合物1,黄色油状物,3.35g,产率75%,其合成路线如下:
(2)化合物2的合成
将500mg化合物1、1260mg 2-羧基苯甲醛和37.5mg溴化铜加入到100毫升玻璃厚壁耐压管中,封管后放置到油浴锅中于140℃加热搅拌5小时后自然冷却至室温,然后将反应混合物溶解在50毫升二氯甲烷中,用质量浓度为10%的NaOH溶液洗涤(50毫升×3),除去未反应的2-羧基苯甲醛等酸性副产物,将得到的二氯甲烷相用无水Na2SO4干燥,旋转蒸发仪蒸除溶剂得粗产品,将粗产品用硅胶柱纯化,硅胶颗粒大小为200-300目,洗脱剂体积配比为石油醚/乙酸乙酯=2:1,得到化合物2,绿色固体,0.33g,产率45%,其合成路线如下:
(3)化合物3的合成
将428.6mg化合物2、50毫升乙醇和2毫升质量浓度为85%的水合肼加入至100毫升的圆底烧瓶,于78℃反应4小时后减压除去溶剂得残余物,用50毫升CH2Cl2溶解残余物,饱和NaCl水溶液洗涤(50毫升×3),无水Na2SO4干燥,旋转蒸发仪蒸除溶剂得粗产品,将粗产品用硅胶柱纯化,硅胶颗粒大小为200-300目,洗脱剂体积配比为石油醚/乙酸乙酯=1:3,得到化合物3,象牙白固体化合物,250.2mg,产率56.5%。1H NMR(400MHz,CDCl3)δ7.93-7.89(m,1H),7.34-7.29(m,2H),6.91-6.90(m,2H),6.89-6.87(m,1H),6.76(s,1H),6.73(s,1H),6.66-6.65(d,J=3.2Hz,1H),6.64-6.63(d,J=2.8Hz,1H),3.69(s,2H),2.96(s,12H),0.61(s,3H),0.59(s,3H).HRMS(ESI):calcd for[M+H]+443.2262,found 443.2264,其合成路线如下:
(4)荧光探针化合物Lyso-NIR-HClO的合成
将化合物3(221.3mg)、15毫升DMF和2毫升3-(4-吗啉基)异硫氰酸丙酯加入至50毫升的圆底烧瓶,于80℃搅拌72小时,在减压条件下除去溶剂后用50毫升CH2Cl2溶解残余物,饱和NaCl水溶液洗涤(50毫升×3),无水Na2SO4干燥,旋转蒸发仪蒸除溶剂得粗产品,将粗产品用硅胶柱纯化,硅胶颗粒大小为200-300目,洗脱剂体积配比为二氯甲/甲醇=15:1,得到荧光探针化合物Lyso-NIR-HClO,淡黄色固体,201.0mg,产率31.9%。1H NMR(600MHz,CDCl3)δ8.00-7.99(d,J=7.2Hz,1H),7.62-7.60(m,1H),7.58-7.55(m,1H),7.15-7.14(d,J=7.2Hz,1H),6.85(d,J=1.8Hz,2H),6.60(s,1H),6.57(d,J=1.8Hz,1H),6.56(d,J=1.8Hz,1H),6.50-6.48(m,2H),5.77-5.75(m,1H),3.64(m,4H),3.22-3.18(m,2H),2.97(s,12H),2.32(s,4H),2.16-2.14(m,2H),1.31-1.26(m,2H),0.57(s,3H),0.54(s,3H).HRMS(ESI):[M+H]+calcd 629.3088,found 629.3083,其合成路线如下:
实施例2
荧光探针化合物Lyso-NIR-HClO与不同浓度次氯酸作用的荧光光谱图的测定
取实施例1制备的荧光探针化合物Lyso-NIR-HClO溶于N,N-二甲基甲酰胺(DMF)中制成10μM的储备液,从储备液中取出2毫升加入到5毫升的离心管当中,加入不同当量(0-8)的次氯酸标准溶液,用PBS缓冲溶液(10mM,pH=7.4)的溶液稀释至4毫升(DMF/PBS体积比为1:1),以620nm为激发光,狭缝宽度设置为5nm/5nm,测量其荧光光谱。荧光光谱如图1所示,随着次氯酸加入,在680nm处的荧光逐渐增强,其荧光强度与次氯酸浓度的关系如图3所示,在5.0×10-8-1.0×10-5mol/L的范围内呈线性关系,所用的荧光测定仪器为Perkin ElmerLS55荧光分光光度计。
实施例3
荧光探针化合物Lyso-NIR-HClO与不同浓度次氯酸作用的紫外可见吸收光谱图的测定
图2为实施例1制得的荧光探针化合物Lyso-NIR-HClO与不同浓度的次氯酸作用后的紫外可见吸收光谱图,次氯酸的加入量为0-40μM。从图中2可以看出,在660nm处可以观察到一个吸收峰,随次氯酸浓度的增加,其660nm处的吸光度逐渐增加。紫外可见吸收光谱测定用的仪器为TU-1900型紫外可见分光光度计(Beijing Purkinje General InstrumentCo.,Ltd.)。
实施例4
荧光探针化合物Lyso-NIR-HClO对不同分子或离子的选择性
从实施例2中的荧光探针储备液中取出2毫升加入到5毫升的离心管当中,加入待考察竞争物质的标准溶液,用PBS缓冲溶液(10mM,pH=7.4)的溶液稀释至4毫升(DMF/PBS体积比为1:1),共考察了26种竞争物质,共配置27个样品,最后一个为次氯酸。以620nm为激发光,狭缝宽度设置为5nm/5nm,测量其荧光光谱,结果如图4所示。由图4可以发现,细胞内金属离子、阴离子、中性分子、活性氮、其他活性氧对荧光探针化合物Lyso-NIR-HClO在680nm处的荧光几乎没有影响,而次氯酸溶液的加入使探针Lyso-NIR-HClO在680nm处的荧光显著增强。
实施例5
溶液pH值对荧光探针化合物Lyso-NIR-HClO测定次氯酸的荧光响应的影响
我们分别考察了pH值对空白荧光探针化合物Lyso-NIR-HClO以及Lyso-NIR-HClO+次氯酸(30μM)的两种情况下荧光强度的影响,结果如图5。在pH=3.5-8.0范围内,pH对空白荧光探针化合物Lyso-NIR-HClO的荧光强度基本无影响。而在次氯酸(30μM)存在的情况下,当pH=5.0时,荧光探针化合物Lyso-NIR-HClO的荧光强度最大,表明此时荧光探针对次氯酸的响应最好。而细胞溶酶体的pH处于4.5-5.5,从图5中可知,在pH处于4.5-5.5范围内,荧光探针化合物Lyso-NIR-HClO对次氯酸都显示出较好的响应,表明荧光探针化合物Lyso-NIR-HClO能够满足溶酶体内次氯酸检测的需求。
实施例6
荧光探针化合物Lyso-NIR-HClO与次氯酸作用的响应时间的测定
在荧光探针化合物Lyso-NIR-HClO溶液中分别加入1μM、5μM、10μM的次氯酸,配制成3个样品,在Perkin Elmer LS 55荧光分光光度计的时间模式下,分别测定含不同浓度的次氯酸样品在680nm处的荧光强度随时间的变化情况,结果如图6。
实施例7
荧光探针化合物Lyso-NIR-HClO与商业化溶酶体染料LysoSensorTM Green DND-189的溶酶体共定位实验
在HeLa细胞中,将荧光探针化合物Lyso-NIR-HClO与商业化的溶酶体染料LysoSensorTM Green DND-189进行共定位实验,说明荧光探针化合物Lyso-NIR-HClO可以定位到溶酶体中,并对溶酶体内的外源性的次氯酸进行荧光成像应用(如图7所示)。具体操作步骤如下:将荧光探针化合物Lyso-NIR-HClO的DMSO溶液和商业化溶酶体染料加入到育有HeLa细胞的培养液中,两者终浓度分别为5μM与1μM。在二氧化碳培养箱中培养30分钟后,向体系中加入一定量次氯酸水溶液,使体系次氯酸浓度为20μM,等待20分钟后用共聚焦显微镜进行成像,同时用405nm与635nm激发,在绿色通道可以观察到有绿色荧光(480-525nm)发出,此为LysoSensorTM Green DND-189发射的光,而在红色通道可以观察到有红色荧光(650-720nm)发出,此为荧光探针化合物Lyso-NIR-HClO与次氯酸响应后发出的红光,将两者用软件进行处理可以得出两种染料的共定位系数为0.93。
实施例8
荧光探针化合物Lyso-NIR-HClO对细胞外源性次氯酸的荧光成像
在HeLa细胞中,考察荧光探针化合物Lyso-NIR-HClO对外源性的次氯酸的荧光成像情况。具体操作步骤如下:将荧光探针化合物Lyso-NIR-HClO的DMSO溶液加入到育有HeLa细胞的培养液中,终浓度为5μM。在二氧化碳培养箱中培养30分钟后,向体系中加入不同量次氯酸水溶液,使体系次氯酸浓度分别为0μM、10μM、20μM,等待20分钟后用共聚焦显微镜进行成像,成像结果如图8所示。从图中可知,随着次氯酸浓度的增加,细胞红色通道的荧光强度逐渐增加,激发波长为635nm,红色通道波长收集范围为650-720nm。
实施例9
荧光探针化合物Lyso-NIR-HClO在组织中的应用
近红外的探针具有体内荧光背景低,组织穿透性强的优点。分别在两个组织样品中加入20μM的荧光探针化合物Lyso-NIR-HClO,其中第一个样品只加荧光探针化合物(图a与图c),另一个继续加入终浓度为200μM次氯酸(图b与图d),如图9所示。图a与图b是两个小鼠肝冷冻切片样品的荧光成像明场图(成像深度为38μm),图c与图d是两个小鼠肝冷冻切片样品的荧光成像荧光通道图(成像深度为38μm)。从图可以看出,荧光探针化合物Lyso-NIR-HClO具有较好的组织穿透能力,能够检测影像组织中的次氯酸。
以上实施例描述了本发明的基本原理、主要特征及优点,本行业的技术人员应该了解,本发明不受上述实施例的限制,上述实施例和说明书中描述的只是说明本发明的原理,在不脱离本发明原理的范围下,本发明还会有各种变化和改进,这些变化和改进均落入本发明保护的范围内。
Claims (3)
1.一种溶酶体靶向的次氯酸近红外荧光探针,其特征在于该荧光探针的结构式如下:
2.一种权利要求1所述的溶酶体靶向的次氯酸近红外荧光探针的制备方法,其特征在于具体步骤为:
步骤S1:于-78℃,在氩气保护下,将6克3-溴-N,N-二甲基苯胺与60毫升无水乙醚加入至干燥的250毫升圆底烧瓶中,磁力搅拌5分钟使其溶解,随后将13.1毫升摩尔浓度为2.4mol/L的正丁基锂的正己烷溶液滴加至反应液中,滴加完毕后于0℃反应2小时,再将2.2毫升二氯二甲基硅烷溶于10毫升无水乙醚中,然后滴加至上述反应液中,滴加完毕后反应至室温并搅拌过夜,加50毫升水猝灭反应,并将反应液用乙醚萃取、用水洗涤后,用饱和NaCl水溶液洗涤,无水硫酸钠干燥,减压旋干溶剂后得到粗产品,将粗产品用硅胶柱纯化得到化合物1,其结构式如下:
步骤S2:将500mg化合物1、1260mg 2-羧基苯甲醛和37.5mg溴化铜加入到100毫升玻璃厚壁耐压管中,于140℃加热搅拌反应5小时后自然冷却至室温,然后将反应混合物溶于二氯甲烷中,用质量浓度为10%的NaOH溶液洗涤三遍,回收并旋干二氯甲烷相得到粗产品,将粗产品用硅胶柱纯化得到化合物2,其结构式如下:
步骤S3:将428.6mg化合物2、50毫升乙醇和2毫升质量浓度为85%的水合肼加入至100毫升的圆底烧瓶,于78℃反应4小时后将反应液减压蒸馏除去溶剂,将残余物用50毫升二氯甲烷溶解后,用饱和食盐水水洗二氯甲烷相,并用无水硫酸钠干燥,减压蒸馏除去二氯甲烷相得到粗产物,将粗产物用硅胶柱纯化得到化合物3,其结构式如下:
步骤S4:将221.3mg化合物3、15毫升DMF和2毫升3-(4-吗啉基)异硫氰酸丙酯加入至50毫升的圆底烧瓶,于80℃搅拌反应72小时,将反应液减压蒸馏除去溶剂后得到残余物,饱和NaCl水溶液洗涤,无水Na2SO4干燥,再将残余物用硅胶柱纯化得到目标荧光探针化合物Lyso-NIR-HClO。
3.权利要求1所述的溶酶体靶向的次氯酸近红外荧光探针在水环境或生物细胞体系选择性检测次氯酸中的应用,其中检测包括水溶液中荧光检测、细胞成像检测、组织成像检测。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201811001966.1A CN109053790A (zh) | 2018-08-30 | 2018-08-30 | 一种溶酶体靶向的次氯酸近红外荧光探针及其制备方法和应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201811001966.1A CN109053790A (zh) | 2018-08-30 | 2018-08-30 | 一种溶酶体靶向的次氯酸近红外荧光探针及其制备方法和应用 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN109053790A true CN109053790A (zh) | 2018-12-21 |
Family
ID=64757860
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201811001966.1A Pending CN109053790A (zh) | 2018-08-30 | 2018-08-30 | 一种溶酶体靶向的次氯酸近红外荧光探针及其制备方法和应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN109053790A (zh) |
Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109851622A (zh) * | 2019-01-22 | 2019-06-07 | 济南大学 | 一种靶向溶酶体的次氯酸根荧光探针 |
CN110156839A (zh) * | 2019-05-24 | 2019-08-23 | 河南师范大学 | 一种线粒体靶向的次氯酸根双光子荧光探针及其制备方法和应用 |
CN110183482A (zh) * | 2019-06-27 | 2019-08-30 | 河南师范大学 | 一种监测溶酶体pH的近红外荧光探针及其制备方法和应用 |
CN114478612A (zh) * | 2022-01-17 | 2022-05-13 | 北京化工大学 | 一种基于硅罗丹明的脑部次氯酸检测荧光探针及其制备方法和应用 |
CN114634464A (zh) * | 2022-03-18 | 2022-06-17 | 南华大学 | 一种用于检测次氯酸的溶酶体靶向近红外荧光探针及其制备方法和应用 |
CN115043893A (zh) * | 2022-05-17 | 2022-09-13 | 河南师范大学 | 一种肝细胞靶向的次氯酸近红外荧光探针及其制备方法和应用 |
CN115466292A (zh) * | 2022-09-19 | 2022-12-13 | 兰州大学 | 一种钌配合物探针及其制备方法和应用 |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN106810561A (zh) * | 2017-01-04 | 2017-06-09 | 济南大学 | 一种溶酶体靶向次氯酸比率荧光探针及其制备方法与应用 |
CN106967078A (zh) * | 2017-03-17 | 2017-07-21 | 济南大学 | 一种溶酶体靶向次氯酸荧光探针及其制备和应用 |
-
2018
- 2018-08-30 CN CN201811001966.1A patent/CN109053790A/zh active Pending
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN106810561A (zh) * | 2017-01-04 | 2017-06-09 | 济南大学 | 一种溶酶体靶向次氯酸比率荧光探针及其制备方法与应用 |
CN106967078A (zh) * | 2017-03-17 | 2017-07-21 | 济南大学 | 一种溶酶体靶向次氯酸荧光探针及其制备和应用 |
Non-Patent Citations (5)
Title |
---|
JONATHAN B. GRIMM ET AL.,: "General Synthetic Method for Si-Fluoresceins and Si-Rhodamines", 《ACS CENTRAL SCIENCE》 * |
LIN YUAN ET AL.,: "Fluorescent Detection of Hypochlorous Acid from Turn-On to FRET-Based Ratiometry by a HOCl-Mediated Cyclization Reaction", 《CHEM. EUR. J.》 * |
TING WANG ET AL.,: "Spirolactonized Si-rhodamine: a novel NIR fluorophore utilized as a platform to construct Si-rhodamine-based probes", 《CHEM. COMMUN.》 * |
YINGYING HUO ET AL.,: "Selective and sensitive visualization of endogenous nitric oxide in living cells and animals by a Sirhodamine deoxylactam-based near-infrared fluorescent probe", 《CHEM. SCI.》 * |
王保刚: "近红外硅基罗丹明一合成方法学及荧光探针的构建与成像研究", 《中国博士学位论文全文数据库》 * |
Cited By (12)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109851622A (zh) * | 2019-01-22 | 2019-06-07 | 济南大学 | 一种靶向溶酶体的次氯酸根荧光探针 |
CN109851622B (zh) * | 2019-01-22 | 2021-08-31 | 济南大学 | 一种靶向溶酶体的次氯酸根荧光探针 |
CN110156839A (zh) * | 2019-05-24 | 2019-08-23 | 河南师范大学 | 一种线粒体靶向的次氯酸根双光子荧光探针及其制备方法和应用 |
CN110156839B (zh) * | 2019-05-24 | 2022-01-04 | 河南师范大学 | 一种线粒体靶向的次氯酸根双光子荧光探针及其制备方法和应用 |
CN110183482A (zh) * | 2019-06-27 | 2019-08-30 | 河南师范大学 | 一种监测溶酶体pH的近红外荧光探针及其制备方法和应用 |
CN110183482B (zh) * | 2019-06-27 | 2022-03-01 | 河南师范大学 | 一种监测溶酶体pH的近红外荧光探针及其制备方法和应用 |
CN114478612A (zh) * | 2022-01-17 | 2022-05-13 | 北京化工大学 | 一种基于硅罗丹明的脑部次氯酸检测荧光探针及其制备方法和应用 |
CN114634464A (zh) * | 2022-03-18 | 2022-06-17 | 南华大学 | 一种用于检测次氯酸的溶酶体靶向近红外荧光探针及其制备方法和应用 |
CN115043893A (zh) * | 2022-05-17 | 2022-09-13 | 河南师范大学 | 一种肝细胞靶向的次氯酸近红外荧光探针及其制备方法和应用 |
CN115043893B (zh) * | 2022-05-17 | 2024-01-09 | 河南师范大学 | 一种肝细胞靶向的次氯酸近红外荧光探针及其制备方法和应用 |
CN115466292A (zh) * | 2022-09-19 | 2022-12-13 | 兰州大学 | 一种钌配合物探针及其制备方法和应用 |
CN115466292B (zh) * | 2022-09-19 | 2023-12-26 | 兰州大学 | 一种钌配合物探针及其制备方法和应用 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN109053790A (zh) | 一种溶酶体靶向的次氯酸近红外荧光探针及其制备方法和应用 | |
Zhao et al. | Transforming the recognition site of 4-hydroxyaniline into 4-methoxyaniline grafted onto a BODIPY core switches the selective detection of peroxynitrite to hypochlorous acid | |
CN109053791A (zh) | 一种基于硅罗丹明的次氯酸近红外荧光探针及其制备方法和应用 | |
Yin et al. | A sensitive and selective fluorescence probe based fluorescein for detection of hypochlorous acid and its application for biological imaging | |
Shu et al. | A novel visual and far-red fluorescent dual-channel probe for the rapid and sensitive detection of hypochlorite in aqueous solution and living cells | |
CN105017271B (zh) | 一种次氯酸荧光比率探针及其应用 | |
Shen et al. | A rhodamine B-based probe for the detection of HOCl in lysosomes | |
CN109081836B (zh) | 一种基于半花菁结构的汞离子近红外荧光探针及其制备方法和应用 | |
CN106220640B (zh) | 一类汞离子荧光探针及其制备方法和应用 | |
Lin et al. | A two-photon near-infrared fluorescent probe for imaging endogenous hypochlorite in cells, tissue and living mouse | |
CN106967053A (zh) | 二价铜离子荧光探针及其制备方法和用途 | |
CN110156839A (zh) | 一种线粒体靶向的次氯酸根双光子荧光探针及其制备方法和应用 | |
CN106810561A (zh) | 一种溶酶体靶向次氯酸比率荧光探针及其制备方法与应用 | |
Zhong et al. | Aggregation-induced fluorescence probe for hypochlorite imaging in mitochondria of living cells and zebrafish | |
CN109266331A (zh) | 一种基于半花菁结构测次氯酸根离子的近红外荧光探针、其制备方法及应用 | |
CN108398409A (zh) | 一种荧光比率检测次氯酸根的方法 | |
Wu et al. | Acylhydrazone as a novel “off–on–off” fluorescence probe for the sequential detection of Al 3+ and F− | |
Hu et al. | A fluorescent probe for hypochlorite with colorimetric and fluorometric characteristics and imaging in living cells | |
Wu et al. | A red-to-near-infrared fluorescent probe for the detection of thiophenol based on a novel hydroxylflavone-quinoline-amino molecular system with large Stokes shift | |
Yu et al. | Quinoline based colorimetric and “turn-off” fluorescent chemosensor for phosgene sensing in solution and vapor phase | |
Ding et al. | The design and synthesis of two imidazole fluorescent probes for the special recognition of HClO/NaHSO 3 and their applications | |
Chen et al. | A pyrene-based ratiometric fluorescent probe with a large Stokes shift for selective detection of hydrogen peroxide in living cells | |
Zeng et al. | A simple highly selective ratiometric fluorescent probe for detection of peroxynitrite and its bioimaging applications | |
CN114672302B (zh) | 一种基于硅罗丹明的近红外mof荧光探针的制备和应用 | |
Huang et al. | Elevated hypochlorous acid levels in asthmatic mice were disclosed by a near-infrared fluorescence probe |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
WD01 | Invention patent application deemed withdrawn after publication | ||
WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20181221 |