WO2021068828A1 - 一类化学发光强度高、波长长、稳定性好的化学发光底物及其制备方法和应用 - Google Patents
一类化学发光强度高、波长长、稳定性好的化学发光底物及其制备方法和应用 Download PDFInfo
- Publication number
- WO2021068828A1 WO2021068828A1 PCT/CN2020/119273 CN2020119273W WO2021068828A1 WO 2021068828 A1 WO2021068828 A1 WO 2021068828A1 CN 2020119273 W CN2020119273 W CN 2020119273W WO 2021068828 A1 WO2021068828 A1 WO 2021068828A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- chemiluminescence
- reaction
- formula
- long wavelength
- intensity
- Prior art date
Links
- 239000000758 substrate Substances 0.000 title claims abstract description 26
- 238000002360 preparation method Methods 0.000 title claims abstract description 6
- 238000004020 luminiscence type Methods 0.000 title abstract description 9
- 239000000523 sample Substances 0.000 claims abstract description 26
- 238000001514 detection method Methods 0.000 claims abstract description 20
- 238000006243 chemical reaction Methods 0.000 claims description 33
- 230000015572 biosynthetic process Effects 0.000 claims description 14
- 239000000543 intermediate Substances 0.000 claims description 14
- 238000003786 synthesis reaction Methods 0.000 claims description 14
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 12
- 150000003384 small molecules Chemical class 0.000 claims description 6
- -1 adamantane Ketone Chemical class 0.000 claims description 5
- 150000001336 alkenes Chemical class 0.000 claims description 5
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 claims description 5
- JRZJOMJEPLMPRA-UHFFFAOYSA-N olefin Natural products CCCCCCCC=C JRZJOMJEPLMPRA-UHFFFAOYSA-N 0.000 claims description 5
- 229910019142 PO4 Inorganic materials 0.000 claims description 4
- 125000004435 hydrogen atom Chemical group [H]* 0.000 claims description 4
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 4
- 239000010452 phosphate Substances 0.000 claims description 4
- NPAWNPCNZAPTKA-UHFFFAOYSA-M sodium;propane-1-sulfonate Chemical compound [Na+].CCCS([O-])(=O)=O NPAWNPCNZAPTKA-UHFFFAOYSA-M 0.000 claims description 4
- WKBOTKDWSSQWDR-UHFFFAOYSA-N Bromine atom Chemical group [Br] WKBOTKDWSSQWDR-UHFFFAOYSA-N 0.000 claims description 3
- 125000003277 amino group Chemical group 0.000 claims description 3
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 3
- 238000002038 chemiluminescence detection Methods 0.000 claims description 3
- 150000001875 compounds Chemical class 0.000 claims description 3
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 3
- 238000000034 method Methods 0.000 claims description 3
- SCRQAWQJSSKCFN-UHFFFAOYSA-N 2-bromo-5-hydroxybenzaldehyde Chemical compound OC1=CC=C(Br)C(C=O)=C1 SCRQAWQJSSKCFN-UHFFFAOYSA-N 0.000 claims description 2
- 102000004190 Enzymes Human genes 0.000 claims description 2
- 108090000790 Enzymes Proteins 0.000 claims description 2
- 238000006000 Knoevenagel condensation reaction Methods 0.000 claims description 2
- DHKHKXVYLBGOIT-UHFFFAOYSA-N acetaldehyde Diethyl Acetal Natural products CCOC(C)OCC DHKHKXVYLBGOIT-UHFFFAOYSA-N 0.000 claims description 2
- 239000003153 chemical reaction reagent Substances 0.000 claims description 2
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 2
- 125000002524 organometallic group Chemical group 0.000 claims description 2
- 239000007858 starting material Substances 0.000 claims description 2
- 238000006467 substitution reaction Methods 0.000 claims description 2
- ORILYTVJVMAKLC-UHFFFAOYSA-N Adamantane Natural products C1C(C2)CC3CC1CC2C3 ORILYTVJVMAKLC-UHFFFAOYSA-N 0.000 claims 1
- 125000002777 acetyl group Chemical class [H]C([H])([H])C(*)=O 0.000 claims 1
- 238000003018 immunoassay Methods 0.000 claims 1
- 238000003384 imaging method Methods 0.000 abstract description 9
- BVTJGGGYKAMDBN-UHFFFAOYSA-N Dioxetane Chemical group C1COO1 BVTJGGGYKAMDBN-UHFFFAOYSA-N 0.000 abstract description 3
- 238000012360 testing method Methods 0.000 abstract description 2
- 230000000149 penetrating effect Effects 0.000 abstract 1
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 39
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 31
- 239000000243 solution Substances 0.000 description 18
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 15
- 238000003756 stirring Methods 0.000 description 13
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 12
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 12
- 239000012043 crude product Substances 0.000 description 11
- 239000000203 mixture Substances 0.000 description 10
- 239000000126 substance Substances 0.000 description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 9
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 8
- 0 CC(N(*)c1ccc(*)cc11)=CC1=C(C#N)C#N Chemical compound CC(N(*)c1ccc(*)cc11)=CC1=C(C#N)C#N 0.000 description 7
- 239000007983 Tris buffer Substances 0.000 description 6
- 239000011550 stock solution Substances 0.000 description 6
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 6
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 5
- 238000001727 in vivo Methods 0.000 description 5
- 239000002243 precursor Substances 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 238000010992 reflux Methods 0.000 description 5
- 239000012265 solid product Substances 0.000 description 5
- OIXUJRCCNNHWFI-UHFFFAOYSA-N 1,2-dioxane Chemical compound C1CCOOC1 OIXUJRCCNNHWFI-UHFFFAOYSA-N 0.000 description 4
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 4
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 4
- 206010028980 Neoplasm Diseases 0.000 description 4
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 4
- 230000005284 excitation Effects 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- HWYHZTIRURJOHG-UHFFFAOYSA-N luminol Chemical compound O=C1NNC(=O)C2=C1C(N)=CC=C2 HWYHZTIRURJOHG-UHFFFAOYSA-N 0.000 description 4
- 238000002390 rotary evaporation Methods 0.000 description 4
- 238000001228 spectrum Methods 0.000 description 4
- DSDAICPXUXPBCC-MWDJDSKUSA-N trimethyl-β-cyclodextrin Chemical compound COC[C@H]([C@H]([C@@H]([C@H]1OC)OC)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](COC)[C@H]([C@@H]([C@H]3OC)OC)O[C@H]3O[C@H](COC)[C@H]([C@@H]([C@H]3OC)OC)O[C@H]3O[C@H](COC)[C@H]([C@@H]([C@H]3OC)OC)O[C@H]3O[C@H](COC)[C@H]([C@@H]([C@H]3OC)OC)O3)[C@H](OC)[C@H]2OC)COC)O[C@@H]1O[C@H]1[C@H](OC)[C@@H](OC)[C@@H]3O[C@@H]1COC DSDAICPXUXPBCC-MWDJDSKUSA-N 0.000 description 4
- CCGXUHXFCISVNC-UHFFFAOYSA-N 4-bromo-3-(dimethoxymethyl)phenol Chemical compound COC(OC)C1=CC(O)=CC=C1Br CCGXUHXFCISVNC-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 241000699666 Mus <mouse, genus> Species 0.000 description 3
- 241000699660 Mus musculus Species 0.000 description 3
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 3
- XYIPYISRNJUPBA-UHFFFAOYSA-N [3-(3'-methoxyspiro[adamantane-2,4'-dioxetane]-3'-yl)phenyl] dihydrogen phosphate Chemical compound O1OC2(C3CC4CC(C3)CC2C4)C1(OC)C1=CC=CC(OP(O)(O)=O)=C1 XYIPYISRNJUPBA-UHFFFAOYSA-N 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000000284 extract Substances 0.000 description 3
- 238000002189 fluorescence spectrum Methods 0.000 description 3
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 3
- 238000011503 in vivo imaging Methods 0.000 description 3
- 239000012046 mixed solvent Substances 0.000 description 3
- 238000011580 nude mouse model Methods 0.000 description 3
- 239000012044 organic layer Substances 0.000 description 3
- 239000001301 oxygen Substances 0.000 description 3
- 229910052760 oxygen Inorganic materials 0.000 description 3
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 3
- 239000001632 sodium acetate Substances 0.000 description 3
- 235000017281 sodium acetate Nutrition 0.000 description 3
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- SIKJAQJRHWYJAI-UHFFFAOYSA-N Indole Chemical compound C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- SMWDFEZZVXVKRB-UHFFFAOYSA-N Quinoline Chemical compound N1=CC=CC2=CC=CC=C21 SMWDFEZZVXVKRB-UHFFFAOYSA-N 0.000 description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- 150000001299 aldehydes Chemical class 0.000 description 2
- 238000004440 column chromatography Methods 0.000 description 2
- 239000000975 dye Substances 0.000 description 2
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-M hydroxide Chemical compound [OH-] XLYOFNOQVPJJNP-UHFFFAOYSA-M 0.000 description 2
- 238000004949 mass spectrometry Methods 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- AHHWIHXENZJRFG-UHFFFAOYSA-N oxetane Chemical compound C1COC1 AHHWIHXENZJRFG-UHFFFAOYSA-N 0.000 description 2
- WDXARTMCIRVMAE-UHFFFAOYSA-N quinoline-2-carbonitrile Chemical compound C1=CC=CC2=NC(C#N)=CC=C21 WDXARTMCIRVMAE-UHFFFAOYSA-N 0.000 description 2
- 238000007920 subcutaneous administration Methods 0.000 description 2
- JOXIMZWYDAKGHI-UHFFFAOYSA-N toluene-4-sulfonic acid Chemical compound CC1=CC=C(S(O)(=O)=O)C=C1 JOXIMZWYDAKGHI-UHFFFAOYSA-N 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- MXALRLXMXCYNGF-UHFFFAOYSA-N 1h-indole-2-sulfonic acid Chemical class C1=CC=C2NC(S(=O)(=O)O)=CC2=C1 MXALRLXMXCYNGF-UHFFFAOYSA-N 0.000 description 1
- UOQZATKCYBGHSE-UHFFFAOYSA-N 3-[2-adamantylidene(methoxy)methyl]phenol Chemical compound C1C2CC(C3)CC1CC3C2=C(OC)C1=CC=CC(O)=C1 UOQZATKCYBGHSE-UHFFFAOYSA-N 0.000 description 1
- MPCCNXGZCOXPMG-UHFFFAOYSA-N 4-bromobenzene-1,3-diol Chemical compound OC1=CC=C(Br)C(O)=C1 MPCCNXGZCOXPMG-UHFFFAOYSA-N 0.000 description 1
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical class [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 1
- JTTQQRVUAZYMFP-DGWALLRLSA-N CC(N(C(/C=C/c1ccc(-[n]2c3ccccc3c3c2cccc3)[s]1)=C1)c2ccccc2)=C/C1=C(/C(c1ccccc1)=C(C#N)C#N)\C#N Chemical compound CC(N(C(/C=C/c1ccc(-[n]2c3ccccc3c3c2cccc3)[s]1)=C1)c2ccccc2)=C/C1=C(/C(c1ccccc1)=C(C#N)C#N)\C#N JTTQQRVUAZYMFP-DGWALLRLSA-N 0.000 description 1
- DDZDITBWHBYABS-UHFFFAOYSA-N CC(O1)=CC(c2ccccc2)=[O][B]1(F)F Chemical compound CC(O1)=CC(c2ccccc2)=[O][B]1(F)F DDZDITBWHBYABS-UHFFFAOYSA-N 0.000 description 1
- YNDKHIKHIBQQTR-UHFFFAOYSA-N CCC(c(cc(cc1)Br)c1N(C)CC)=C(C#N)C#N Chemical compound CCC(c(cc(cc1)Br)c1N(C)CC)=C(C#N)C#N YNDKHIKHIBQQTR-UHFFFAOYSA-N 0.000 description 1
- ZXEZVPJXSLFKNU-ISLYRVAYSA-N CO/C(/c1c(C=O)ccc(O)c1)=C1\C2CC34C1CC3CC4C2 Chemical compound CO/C(/c1c(C=O)ccc(O)c1)=C1\C2CC34C1CC3CC4C2 ZXEZVPJXSLFKNU-ISLYRVAYSA-N 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- PIWKPBJCKXDKJR-UHFFFAOYSA-N Isoflurane Chemical compound FC(F)OC(Cl)C(F)(F)F PIWKPBJCKXDKJR-UHFFFAOYSA-N 0.000 description 1
- WHXSMMKQMYFTQS-UHFFFAOYSA-N Lithium Chemical compound [Li] WHXSMMKQMYFTQS-UHFFFAOYSA-N 0.000 description 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- KEAYESYHFKHZAL-UHFFFAOYSA-N Sodium Chemical compound [Na] KEAYESYHFKHZAL-UHFFFAOYSA-N 0.000 description 1
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical class [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- YARFTSQNOOFGPY-UHFFFAOYSA-N [3-[2-adamantylidene(methoxy)methyl]-4-bromophenoxy]-tert-butyl-dimethylsilane Chemical compound C12C(C3CC(CC(C1)C3)C2)=C(C=2C=C(O[Si](C)(C)C(C)(C)C)C=CC2Br)OC YARFTSQNOOFGPY-UHFFFAOYSA-N 0.000 description 1
- CMPRPSXFFCAHMN-UHFFFAOYSA-N [4-bromo-3-(dimethoxymethyl)phenoxy]-tert-butyl-dimethylsilane Chemical compound COC(OC)C1=CC(O[Si](C)(C)C(C)(C)C)=CC=C1Br CMPRPSXFFCAHMN-UHFFFAOYSA-N 0.000 description 1
- UGJVZXBXVRMUSG-UHFFFAOYSA-K [B+3].[F-].[F-].[F-] Chemical compound [B+3].[F-].[F-].[F-] UGJVZXBXVRMUSG-UHFFFAOYSA-K 0.000 description 1
- LIQLLTGUOSHGKY-UHFFFAOYSA-N [B].[F] Chemical compound [B].[F] LIQLLTGUOSHGKY-UHFFFAOYSA-N 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 150000001241 acetals Chemical class 0.000 description 1
- DZBUGLKDJFMEHC-UHFFFAOYSA-N acridine Chemical class C1=CC=CC2=CC3=CC=CC=C3N=C21 DZBUGLKDJFMEHC-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- IYKFYARMMIESOX-SPJNRGJMSA-N adamantanone Chemical compound C([C@H](C1)C2)[C@H]3C[C@@H]1C(=O)[C@@H]2C3 IYKFYARMMIESOX-SPJNRGJMSA-N 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 238000003975 animal breeding Methods 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 238000000149 argon plasma sintering Methods 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical group OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- HQABUPZFAYXKJW-UHFFFAOYSA-N butan-1-amine Chemical compound CCCCN HQABUPZFAYXKJW-UHFFFAOYSA-N 0.000 description 1
- FJDQFPXHSGXQBY-UHFFFAOYSA-L caesium carbonate Chemical compound [Cs+].[Cs+].[O-]C([O-])=O FJDQFPXHSGXQBY-UHFFFAOYSA-L 0.000 description 1
- 229910000024 caesium carbonate Inorganic materials 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000002242 deionisation method Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 239000010432 diamond Substances 0.000 description 1
- 229910003460 diamond Inorganic materials 0.000 description 1
- YLFBFPXKTIQSSY-UHFFFAOYSA-N dimethoxy(oxo)phosphanium Chemical compound CO[P+](=O)OC YLFBFPXKTIQSSY-UHFFFAOYSA-N 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000012847 fine chemical Substances 0.000 description 1
- 238000001917 fluorescence detection Methods 0.000 description 1
- 239000003517 fume Substances 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- PZOUSPYUWWUPPK-UHFFFAOYSA-N indole Natural products CC1=CC=CC2=C1C=CN2 PZOUSPYUWWUPPK-UHFFFAOYSA-N 0.000 description 1
- RKJUIXBNRJVNHR-UHFFFAOYSA-N indolenine Natural products C1=CC=C2CC=NC2=C1 RKJUIXBNRJVNHR-UHFFFAOYSA-N 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 229960002725 isoflurane Drugs 0.000 description 1
- 239000012263 liquid product Substances 0.000 description 1
- 229910052744 lithium Inorganic materials 0.000 description 1
- 201000005202 lung cancer Diseases 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 125000004108 n-butyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 239000010453 quartz Substances 0.000 description 1
- 238000010791 quenching Methods 0.000 description 1
- LISFMEBWQUVKPJ-UHFFFAOYSA-N quinolin-2-ol Chemical compound C1=CC=C2NC(=O)C=CC2=C1 LISFMEBWQUVKPJ-UHFFFAOYSA-N 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 239000012312 sodium hydride Substances 0.000 description 1
- 229910000104 sodium hydride Inorganic materials 0.000 description 1
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical class O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- BCNZYOJHNLTNEZ-UHFFFAOYSA-N tert-butyldimethylsilyl chloride Chemical compound CC(C)(C)[Si](C)(C)Cl BCNZYOJHNLTNEZ-UHFFFAOYSA-N 0.000 description 1
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 1
- CYTQBVOFDCPGCX-UHFFFAOYSA-N trimethyl phosphite Chemical compound COP(OC)OC CYTQBVOFDCPGCX-UHFFFAOYSA-N 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
- C07D215/02—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom
- C07D215/12—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom with substituted hydrocarbon radicals attached to ring carbon atoms
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
- C07D209/04—Indoles; Hydrogenated indoles
- C07D209/10—Indoles; Hydrogenated indoles with substituted hydrocarbon radicals attached to carbon atoms of the hetero ring
- C07D209/12—Radicals substituted by oxygen atoms
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
- C07D409/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing three or more hetero rings
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F5/00—Compounds containing elements of Groups 3 or 13 of the Periodic Table
- C07F5/02—Boron compounds
- C07F5/022—Boron compounds without C-boron linkages
-
- 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
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1003—Carbocyclic compounds
- C09K2211/1007—Non-condensed systems
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1003—Carbocyclic compounds
- C09K2211/1011—Condensed systems
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1029—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1092—Heterocyclic compounds characterised by ligands containing sulfur as the only heteroatom
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1096—Heterocyclic compounds characterised by ligands containing other heteroatoms
Definitions
- the invention belongs to the field of fine chemicals, and specifically relates to a synthesis method and biological application of a new type of electron-rich methoxy-alkenyl-based chemiluminescence probe.
- Chemiluminescence is a phenomenon of light radiation accompanying the chemical reaction of substances. Because it does not require external excitation light, chemiluminescence detection can effectively overcome the difficult problems of photobleaching, light scattering and autofluorescence faced in traditional fluorescence detection methods (Talanta 2000, 51, 415-439). In addition, chemiluminescence detection also has significant advantages such as low detection limit, high sensitivity, and a wide range of analyte detection concentrations. Therefore, chemiluminescence probes have attracted the attention of chemists and biologists. In 1987, Paul Schaap reported for the first time that 1,2-dioxane was used as a high-energy structure of chemiluminescent substrates.
- chemiluminescent probes suitable for a variety of active substances can be constructed.
- Schaap-type chemiluminescence probes have the advantages of long luminescence half-life (hour level) and relatively stable signal intensity, weak luminescence intensity is the main bottleneck faced by this type of chemiluminescence probes. When applied to biological sample imaging, Schaap-type chemiluminescence probes often cannot meet the signal intensity requirements of the detector.
- alkaline phosphatase chemiluminescence probe AMPPD
- the detection mechanism is as follows: (1) Alkaline phosphatase induces AMPPD to undergo a hydrolysis reaction, and the phosphate group leaves to form an unstable intermediate Body AMPPD - ; (2) The high-energy state 1,2-dioxane cleavage, and then show the chemiluminescence signal.
- the 1,2-dioxane structure is not stable, so the probe has low luminescence intensity and is susceptible to environmental interference.
- Schaap-type chemiluminescence probes have short luminescence wavelengths and are difficult to be used in in vivo imaging. Therefore, how to develop a new type of chemiluminescence substrate with long emission wavelength, high intensity, good stability, and promotion of the detection substance has become an urgent problem to be solved.
- the present invention aims to provide a class of chemiluminescence probes with long wavelength, high intensity and excellent stability.
- Chemiluminescence probe Make full use of the chemical modifiability of this type of substance: extend the ⁇ system by introducing electron-attracting fluorescent units to extend its chemical emission wavelength to the near-infrared region; optimize the fluorescent unit through molecular engineering, improve energy conversion efficiency, and increase the intensity of chemiluminescence signals; Use electron-rich double bonds to replace the 1,2-dioxane structure to improve the stability of chemiluminescence probes.
- the present invention provides a general preparation method for the above-mentioned chemiluminescence probe with long wavelength, high intensity and excellent stability, as well as partial absorption, fluorescence, and autoluminescence fluorescence spectra, and its application in biological detection.
- the chemical composition and functions of this type of chemiluminescence probe are as follows: (1) Fluorophore unit, which expands the wavelength of chemiluminescence and enhances the intensity of chemiluminescence; (2) The electron-rich double bond unit can controllably generate high-energy states 1,2-two The structure of oxetane; (3) the response unit, which specifically recognizes the detection substance.
- this type of probe responds specifically, the detection unit is removed, and a chemiluminescent precursor with phenolic hydroxyl anion is generated. Subsequently, under the excitation of white light, the electron-rich double bond part of the chemiluminescent precursor reacts with oxygen to form a 1,2-dioxetane structure, which is cleaved under the stimulation of phenolic hydroxyl anions, which is shown as A significant chemiluminescence signal.
- the present invention is realized through the following schemes:
- the chemiluminescence substrate with high chemiluminescence intensity, long wavelength and oxidation-free according to the present invention has the structure shown in formula I
- R 1 is independently selected from any one of the small molecule fluorophores shown in formula II-V (where the curve mark is the substitution position, the same below); in formula II, R 3 is a hydrogen atom or a bromine atom , Amine group, carboxyl group, in formula II and III, R 4 is one of ethyl or sodium propyl sulfonate.
- R 2 is independently selected from any one of the detection groups shown in formula VI-IX.
- the phosphate intermediate is obtained through the acetal reaction, hydroxyl protection and phospholipidation reaction in turn; the phosphate intermediate is further combined with diamond Alkanone compound reaction (Hornall-Wadsworth-Emmons reaction) to prepare olefin intermediates; olefin intermediates are activated by organometallic reagents and react with N,N-dimethylformamide to prepare olefins Aldehyde intermediate; Enal intermediate is further reacted with a small molecule fluorophore with active methyl group (Knoevenagel condensation reaction) to prepare a chemiluminescent substrate; the substrate is further connected to various detection units to obtain the final chemiluminescent probe.
- This type of chemiluminescence probe first interacts with the detected substance (such as enzyme or active small molecule), the detection group leaves, and the chemiluminescence precursor with phenolic hydroxyl anion is released; then, under the excitation of white light, the chemiluminescence precursor It rapidly reacts with oxygen to form a 1,2-dioxetane structure; this structure is cleaved under the action of phenolic hydroxyl anions to release a chemiluminescent signal.
- the detected substance such as enzyme or active small molecule
- the abscissa is the wavelength (nm)
- the left ordinate is the absorbance
- the right ordinate is the relative intensity of fluorescence and chemiluminescence.
- the upper part is the image collected by the Imaging Quant 4000 system
- the lower part is the quantification of the light intensity in the image.
- the water content in the image is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% and 99%, the three groups are parallel;
- the abscissa in the quantitative histogram It is the percentage of DMSO (%), and the ordinate is the number of photons per unit area and unit time (p/s/cm2/sr).
- R 1 is independently selected from any one of the small molecule fluorophores shown in II-VII;
- R 2 is independently selected from any one of the detection groups shown in formulas VIII-XI;
- R 3 is one of a hydrogen atom, a bromine atom, an amino group, and a carboxyl group
- R 4 is one of ethyl or sodium propyl sulfonate
- R 1 is independently selected from any one of the groups shown in II, and R 2 is a borate group;
- R 3 is independently selected from hydrogen atoms, and R 4 is one of ethyl and sodium propyl sulfonate;
- R 4 is ethyl
- the F-QM-OH prepared in Example 1 was dissolved in analytically pure dimethyl sulfoxide to prepare a 1.0 ⁇ 10 -2 M stock solution. Then, 2 mL of a mixed solvent of DMSO/H 2 O with a water (H 2 O) content of 99% was prepared. Take 20 ⁇ L of the above stock solution and add it to the prepared DMSO/H 2 O mixed solvent, mix it evenly and transfer it to an optical quartz cuvette (10 ⁇ 10 mm) to test its fluorescence spectrum.
- the maximum emission peak of the F-QM-OH substrate is located at approximately 600nm in the near-infrared region, and the Stokes shift is 120nm; F-QM-OH chemiluminescence spectrum and fluorescence The spectra are basically the same.
- Chemiluminescence probe F-QM-B applied to hydrogen peroxide detection
- All in-vivo experiments in the present invention comply with the rules and regulations of the breeding and use of laboratory animals, and have been approved by the Animal Breeding and Use Committee of East China University of Science and Technology.
- the tumor-bearing nude mice for the experiment were purchased from Shanghai Slack Animal Experiment Co., Ltd., and were raised in a sterile mouse cage in a laminar flow fume hood in a sterile room, and fed with food and water treated with high-pressure steam.
- chemiluminescence probe F-QM-B In order to evaluate the performance of the chemiluminescence probe F-QM-B in vivo, tumor-bearing nude mice were used as the imaging object, and the traditional chemiluminescence dye luminol was used as a reference. As shown in Figure 4, three A549 (human lung cancer cells) subcutaneous tumor mice were injected in situ with F-QM-B trimethyl- ⁇ -cyclodextrin solution (F-QM-B concentration 1.0 ⁇ 10 -4 M, trimethyl- ⁇ -cyclodextrin concentration 1.0 ⁇ 10 -3 M), F-QM-B trimethyl- ⁇ -cyclodextrin solution and luminol solution.
- mice were anesthetized with 2.5% isoflurane gas.
- the mice in the illuminated group had obvious chemiluminescence signals, while the non-illuminated group (left 2) did not show obvious chemiluminescence signals, which indicates that F-QM-B can only be used with Hydrogen oxide responds, and the chemiluminescence signal is displayed only after light is applied.
- the luminol group (left 3) also did not show obvious chemiluminescence signals, which shows that short-wavelength chemiluminescence dyes do not meet the requirements of in vivo imaging.
- the chemiluminescence probe prepared by the present invention has significant advantages such as long wavelength, controllable luminescence, and good specificity, and has been successfully applied to the detection of hydrogen peroxide in vivo.
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Physics & Mathematics (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Plasma & Fusion (AREA)
- Health & Medical Sciences (AREA)
- Materials Engineering (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
发明提供了一类化学发光强度高、波长长、稳定性好的化学发光底物及其制备方法和应用。所述化学发光底物具有式Ⅰ所示结构。另外,本发明提供了所述化学发光探针在生理环境下的自发荧光波长及强度,及其在活体成像中的应用。测试结果表明,本发明中提供的的化学发光探针具有长波长的自发荧光(600nm),能有效的穿透皮肤组织,高自发荧光强度(>10 7p/s/cm 2/sr),优异的热稳定性(不含1,2-二氧杂环丁烷结构)及多样化的检测基团(适用于不同检测模型)。
Description
本发明属于精细化工领域,具体涉及一类基于富电子性甲氧基-烯基的新型化学发光探针的合成方法与生物应用。
化学发光是物质在进行化学反应过程中伴随的一种光辐射现象。由于不需要外源激发光,化学发光检测能够有效克服传统荧光检测法中面临的光漂白、光散射和自发荧光等难以解决的难题(Talanta 2000,51,415-439)。除此之外,化学发光检测还具有检测限低、灵敏度高、分析物检测浓度范围广等显著优势,因此化学发光探针备受化学家和生物学家的关注。1987年,Paul Schaap首次报道了以1,2-二氧杂环作为高能态结构的化学发光底物。相比于传统化学发光底物(如鲁米诺、吖啶酯等)检测对象(活性氧)单一的缺陷,基于Schaap型化学发光底物能构建适用于多种活性物质的化学发光探针。
尽管Schaap型化学发光探针具有发光半衰期长(小时级)、信号强度较稳定的优势,但是发光强度弱是该类化学发光探针面临的主要瓶颈。在应用于生物样品成像时,Schaap型化学发光探针往往不能满足检测器的信号强度要求。以碱性磷酸酯酶化学发光探针(AMPPD)为例(如下图所示),其检测机理如下:(1)碱性磷酸酯酶诱导AMPPD发生水解反应,磷酸基团离去生成不稳定中间体AMPPD
-;(2)高能态1,2-二氧杂环发生裂解,进而表现化学发光信号。但是,在生理条件下,1,2-二氧杂环结构并不稳定,因此该探针发光强度低、易受到环境干扰。此外,Schaap型化学发光探针发光波长短,难以应用于活体成像中。因此,如何发展兼具发光波长长、强度高、稳定性好、检测物可推广的新型化学发光底物,成为亟待解决的难题。
碱性磷酸酯酶(ALP)-AMPPD化学发光体系
发明内容
针对现有1,2,-二氧杂环丁烷类型化学发光探针波长短、强度弱、及稳定性差的瓶颈,本发明旨在提供了一类化学发光波长长、强度高、稳定性优异的化学发光探针。充分利用该类物质 的化学可修饰性:通过引入吸电子荧光单元扩展π体系,将其化学发射波长拓展至近红外区域;通过分子工程优化荧光单元,提高能量转换效率,提升化学发光信号强度;通过使用富电子双键替代1,2-二氧杂环结构,提升化学发光探针的稳定性。
本发明提供了上述化学发光长波长、强度高、稳定性优异的化学发光探针的通用制备方法以及部分吸收、荧光、自发光荧光图谱,及其在生物检测中的应用。这类化学发光探针的化学组成与功能如下:(1)荧光团单元,拓展化学发光波长、增强化学发光强度;(2)富电子的双键单元,可控生成高能态1,2-二氧杂环丁烷结构;(3)响应单元,特异性识别检测物质。
在检测物条件下,该类探针发生特异性响应,检测单元离去,生成具有酚羟基负离子的化学发光前体。随后,在白光激发下,化学发光前体中富电性的双键部分与氧气发生加成反应生成1,2-二氧杂环丁烷结构,该结构在酚羟基负离子的刺激下裂解,表现为出显著的化学发光信号。
本发明通过下述方案实现:
一方面,本发明所述化学发光强度高、长波长、免氧化的化学发光底物,其结构为式Ⅰ所示
式Ⅰ中,R
1独立选自式Ⅱ-Ⅴ所示的小分子荧光团(其中曲线标记处为取代位,下同)中的任意一种;在Ⅱ式中R
3为氢原子、溴原子、胺基、羧基的一种,在Ⅱ式和Ⅲ式中R
4为乙基或丙基磺酸钠的一种。
R
2独立选自式Ⅵ-Ⅸ所示的检测基团的任意一种。
通用制备方法:
化合物的合成采用模块化制备方式,以2-溴-5-羟基苯甲醛为起始原料,依次通过缩醛反应、羟基保护及磷脂化反应,得到磷酸酯中间体;磷酸酯中间体进一步与金刚烷酮化合物反应(霍纳尔-沃兹沃思-埃蒙斯反应),制备得到烯烃中间体;烯烃中间体通过金属有机试剂活化,与N,N-二甲基甲酰胺反应,制备得到烯醛中间体;烯醛中间体进一步带有活泼甲基的小分子荧光团反应(Knoevenagel缩合反应),制备得到化学发光底物;底物进一步连接各类检测单元即得到最终的化学发光探针。
通用检测方法
该类化学发光探针首先与被检测物(如酶或活性小分子)作用,检测基团离去,释放出具有酚羟基负离子的化学发光前体;随后,在白光激发下,化学发光前体迅速与氧气发生加成反应,生成1,2-二氧杂环丁烷结构;该结构在酚羟基负离子的作用下裂解释放出化学发光信号。
本文发明的化学发光底物激活步骤
图1.F-QM-OH(详见实施例1)在PBS溶液(含1%DMSO)中的紫外吸收(10
-5mol·L
-1)、荧光 及化学发光图谱(10
-4mol·L
-1);
其中,横坐标为波长(nm),左侧纵坐标为吸光度、右侧纵坐标为荧光及化学发光相对强度。
图2.F-QM-OH(详见实施例1)在水和DMSO混合溶剂中不断增加水含量的化学发光图像及强度(5*10
-5mol·L
-1);
其中,上方为Imaging Quant 4000系统采集图像,下方是图像中光强度的定量。图像中从左到右水含量依次为10%、20%、30%、40%、50%、60%、70%、80%、90%及99%,三组平行;定量柱状图中横坐标是DMSO的百分含量(%),纵坐标是单位面积及单位时间内的光子数(p/s/cm2/sr)。
图3.自发光底物F-QM-B(浓度为5*10
-5mol·L
-1)在Tris缓冲液(含10%DMSO)中随过氧化氢及是否光照化学发光图像。
图4.自发光底物F-QM-B在A549皮下瘤模型小鼠上的活体成像应用。
在本发明一个优选的技术方案中:
R
1独立选自Ⅱ-Ⅶ所示的小分子荧光团的任意一种;
R
2独立选自式Ⅷ-Ⅺ所示的检测基团中的任意一种;
R
3为氢原子、溴原子、胺基、羧基的一种;
R
4为乙基或丙基磺酸钠的一种;
在进一步优选的技术方案中,R
1独立选自Ⅱ所示基团的任意一种,R
2为硼酸酯基团;
更进一步优选的R
3独立选自氢原子,R
4为乙基、丙基磺酸钠中的一种;
更进一步优选的R
4为乙基;
下面通过实施例对本发明作进一步的阐述,其目的仅在于更好地理解本发明的内容。因此,所举之例并不限制本发明的保护范围:
实施例1
以喹啉腈化学发光底物为例,具体合成路线如下:
1. 4-溴-3-(二甲氧基甲基)苯酚的合成
在50mL干燥的单口瓶中加入4-溴-3-羟基-苯酚(1g,4.55mmol)与20mL甲醇,搅拌溶解;后加入对甲基苯磺酸(171.3mg,0.91mmol),回流6h;反应完毕,将反应液倒入200ml稀碳酸钠溶液(2g)中,并用乙酸乙酯(200mL×2)萃取,合并萃取液,无水硫酸钠干燥,旋干,柱层析纯化,淡粉色液体0.7g,产率57%。
1H NMR(400MHz,CDCl
3-d
1,ppm):δ=7.40-7.38(d,J=8.4Hz,1H,Ph-H),δ=7.12-7.11(d,J=3.2Hz,1H,Ph-H),δ=6.73-6.70(d-d,J
1=8.8Hz,J
2=3.2Hz,1H,Ph-H),δ=5.84(s,1H,Ph-OH),δ=5.50(s,1H,-CH-O-),δ=3.41(s,6H,-O-CH
3).
13C NMR(100MHz,CDCl
3-d
1,ppm):155.51,133.74,117.82,115.34,112.71,103.37,54.35
2.(4-溴-3-(二甲氧基甲基)苯氧基)(叔丁基)二甲基硅烷的合成
在50mL圆底烧瓶中加入4-溴-3-(二甲氧基甲基)苯酚(500mg,2.02mmol)与咪唑(368mg,4.04mmol)及15mL二氯甲烷,搅拌溶解;冰浴下滴加TBSCl(411mg,6.06mmol)与二氯甲烷(5ml)的混合液,滴加完毕常温搅拌;点板监控反应进程,反应完毕,加入50ml二氯甲烷,并用水洗(100ml×5),无水硫酸钠干燥有机层,旋干,得淡粉色液体0.6g,产率82%。
1H NMR(400MHz,CDCl
3-d
1,ppm):δ=7.40-7.37(d,J=8.6Hz,1H,Ph-H),δ=7.10-7.09(d,J=3.0Hz,1H,Ph-H),δ=6.71-6.68(d-d,J
1=8.6Hz,J
2=3.0Hz,1H,Ph-H),δ=5.48(s,1H,-CH-O-),δ=3.38(s,6H,-O-CH
3),δ=0.97(s,9H,-Si-C-(CH
3)
3),δ=0.20(s,6H,-Si-CH
3).
13C NMR(100MHz,CDCl
3-d
1,ppm):155.02,137.71,133.47,121.91,120.17,114.02,102.79,53.86,25.64,18.20,-4.48.
3.((2-溴-5-((叔丁基二甲基甲硅烷基)氧基)苯基)(甲氧基)甲基)膦酸二甲酯的合成
在25mL反应管中加入上一步产物(2.5g,6.29mmol)与3mL N,N-二甲基甲酰胺,搅拌混合均匀,随后加入亚磷酸三甲酯(1.03g,7.55mmol)与三氟化硼乙醚的二氯甲烷溶液(8.3mL,7.55mmol),滴加完毕,室温反应16h;反应完毕,在反应液中加入适量饱和的碳酸氢钠水溶液洗涤,后用乙酸乙酯萃取(150mL×3),合并萃取液,无水硫酸钠干燥,旋蒸,得淡黄色液体粗产物,这一步产品是活泼中间体因此未经进一步纯化即投入下一步反应。
4.(3-(金刚烷-2-亚基(甲氧基)甲基)-4-溴苯氧基)(叔丁基)二甲基硅烷的合成
在100mL反应管中加入上一步产物(500mg,1.14mmol)、氢化钠(82mg,3.42mmol)与30mL THF;逐滴滴加金刚烷酮(170mg,1.14mmol),室温搅拌1小时;加入去离子水淬灭反应,乙酸乙酯萃取(30mL×5),无水硫酸钠干燥,旋蒸,得淡黄色液体产物300mg,产率57%。
1H NMR(400MHz,CDCl
3-d
1,ppm):δ=7.44-7.42(d,J
3=8.6Hz,1H,Ph-H),δ=6.74-6.73(d,J=2.9Hz,1H,Ph-H),δ=6.70-6.68(d-d,J
1=8.6Hz,J
2=3.0Hz,1H,Ph-H),δ=3.32(s,3H,-O-CH
3),δ=3.26(s,1H,-Adamantane-H),δ=2.09(s,1H,-Adamantane-H),δ=2.00-1.79(m,12H,-Adamantane-H),δ=0.97(s,9H,-Si-C-(CH
3)
3),δ=0.19(s,6H,-Si-CH
3).
5. 2-(金刚烷-2-亚基(甲氧基)甲基)-4-羟基苯的合成
在25mL反应管中加入上一步产物(500mg,1.08mmol)与5mL四氢呋喃,搅拌混合均匀,在-78℃的乙醇中冻抽3次;在-78℃搅拌的条件下,逐滴注入正丁基锂溶液(1mL,2.4mmol),滴加完毕,继续在-78℃下反应2h;然后滴加N,N-二甲基甲酰胺(0.4ml),持续搅拌1h;后转入室温反应0.5h;反应完毕,向反应液注入1mL去离子水淬灭反应;后加入乙酸乙酯(50mL), 用饱和食盐水洗涤(50mL×3),无水硫酸钠干燥有机层,旋蒸,得黄色液体粗产物;将粗产物用flash柱纯化,的白色固体产物280mg,产率87%。
1H NMR(400MHz,CDCl
3-d
1,ppm):δ=10.13(s,1H,-CHO),δ=7.95-7.93(d,J=8.4Hz,1H,Ph-H),δ=6.92-6.90(d-d,J
1=8.8Hz,J
2=2.4Hz,1H,Ph-H),δ=6.80-6.79(d,J=2.4Hz,1H,Ph-H),δ=3.32(s,6H,-O-CH
3),δ=3.32(s,1H,-Adamantane-H),δ=1.97(s,1H,-Adamantane-H),δ=1.94-1.66(m,12H,-Adamantane-H).
Mass spectrometry(ESI-MS,m/z):[M-H
+]calcd.for[C
19H
22O
3-H
+]297.1791;found 297.1490.
6. F-QM-OH的合成
在100mL圆底烧瓶中加入A(222mg,0.74mmol)、喹啉睛(208mg,0.89mmol)与50ml乙腈,搅拌混合均匀;后加入醋酸钠(73mg,0.89mmol);加热回流10h,点板监控反应进程。反应完毕,旋蒸得棕黄色粗产物,将粗产物用flash柱纯化,得淡红色固体产物80mg,产率18%。
1H NMR(400MHz,CDCl
3-d
1,ppm):δ=9.14(d,J=0.8Hz,1H,=CH-),δ=7.77-7.73(m,1H,Ph-H),δ=7.61-7.59(d,J=8Hz,1H,Ph-H),δ=7.54-7.52(d,J=8Hz,1H,Ph-H),δ=7.48-7.44(t,J=8Hz,1H,Ph-H),δ=7.45-7.41(d,J=16Hz,1H,Alkene-H),δ=7.09(s,1H,Ph-H),δ=7.02-6.98(d,J=16Hz,1H,Alkene-H),δ=6.90-6.87(d-d,J
1=8.4Hz,J
2=2.8Hz,1H,Ph-H),δ=6.78-6.77(d,J=2.8Hz,1H,Ph-H),δ=4.40-4.34(q,J=6.8Hz,2H,-N-CH
2-CH
3),δ=3.30(s,3H,-O-CH
3),δ=3.30(s,1H,-Adamantane-H),δ=2.22(s,1H,-Adamantane-H),δ=1.96-1.75(m,12H,-Adamantane-H),δ=1.55-1.51(t,J=7.2Hz,2H,-N-CH
2-CH
3).
13C NMR(100MHz,CDCl
3-d
1,ppm):164.03,157.37,154.93,145.77,143.13,142.90,142.38,135.84,130.51,125.85,122.79,122.49,121.19,61.52,48.96,43.75,41.70,37.44,34.26,18.88
Mass spectrometry(ESI-MS,m/z):[M-H
+]calcd.for[C
34H
33N
3O
2-H
+]514.2495;found 514.2491.
7. F-QM-B的合成
在100mg圆底烧瓶中加入自发光喹啉底物(150mg,0.29mmol)、碳酸铯(472mg,1.45mmol)与30ml乙腈,搅拌混合均匀,随后加入对苄溴硼酸酯(255mg,0.87mmol),常温反应3小时;反应完毕,饱和氯化铵溶液水洗(100mL×3),无水硫酸钠干燥有机,柱层析分离,得红色产物130mg,产率61%。
1H NMR(400MHz,CDCl
3-d
1,ppm):δ=9.14-9.17(d-d,J
1=8.8Hz,J
2=1.2Hz,1H,=CH-),δ=7.85-7.83(d,J=8Hz,2H,Ph-H),δ=7.77-7.72(m,1H,Ph-H),δ=7.60-7.56(t,J=8Hz,2H,Ph-H),δ=7.47-7.38(m,4H,Ph-H),δ=7.09(s,1H,Ph-H),δ=7.01-6.97(m,2H,Alkene-H),δ=6.87-6.86(d,J=2.8Hz,1H,Alkene-H),δ=5.15(s,2H,-O-CH
2-Ph),δ=4.39-4.33(q,J=7.2Hz,2H,-N-CH
2-CH
3),δ=3.28(s,3H,-O-CH
3),δ=3.28(s,1H,-Adamantane-H),δ=2.16(s,1H,-Adamantane-H),δ=1.95-1.73(m,12H,-Adamantane-H),δ=1.55-1.51(t,J=7.2Hz,2H,-N-CH
2-CH
3),δ=1.35(s,12H,-C-(CH
3)
2).
实施例2
其他化学发光羟基底物,具体合成路线如下:
1.自发光吲哚底物的合成
在100mL圆底烧瓶中加入磺酸基吲哚盐(322mg,1.08mmol)、喹啉睛(300mg,0.90mmol)与50ml乙腈,搅拌混合均匀;后加入醋酸钠(74mg,0.90mmol);加热回流10h,点板监控反应进程。反应完毕,旋蒸得红色粗产物,将粗产物用flash柱纯化,得淡红色固体产物270mg,产率53%。
1H NMR(400MHz,CDCl
3-d
1,ppm):δ=7.82-7.79(m,1H,Ph-H),δ=7.59-7.57(d,J=8Hz,1H,Ph-H),δ=7.53-7.49(m,3H,Ph-H),δ=7.48-7.44(d,J=16Hz,1H,Alkene-H),δ=7.12(s,1H,Ph-H),δ=7.02-6.98(d,J=16Hz,1H,Alkene-H),δ=6.92-6.89(d-d,J
1=8.4Hz,J
2=2.8Hz,1H,Ph-H),δ=6.81-6.80(d,J=2.8Hz,1H,Ph-H),δ=4.42-4.36(q,J=6.8Hz,2H,-N-CH
2-CH
2-),δ=3.30(s,3H,-O-CH
3),δ=3.30(s,1H,-Adamantane-H),δ=3.10-3.04(m,2H,-CH
2-SO
3
-)δ=2.22(s,1H,-Adamantane-H),δ=1.96-1.75(m,12H,-Adamantane-H),δ=1.62-1.58(m,J=7.2Hz,2H,-N-CH
2-CH
2-CH
3).
2.自发光TCM底物的合成
在100ml干燥的双口瓶中加入单边TCM底物(150mg,0.24mmol)与自发光醛(100mg,1.5eq),并加入35ml乙腈搅拌溶解,后入醋酸钠(36mg,0.26mmol),回流约6h;点板监控反应进程,反应完毕,旋蒸得红色粗产物,将粗产物用flash柱纯化,得红色固体产物70mg,产率32%。
1H NMR(400MHz,CDCl
3-d
1,ppm):δ=8.09-8.08(d,J=8.0Hz,1H,Ph-H),δ=7.67-7.41(m,14H,Ph-H),δ=7.35-7.39(m,4H,Ph-H),δ=7.23-7.22(d,J=4Hz,1H,Ph-H),δ=7.13-7.12(d,J=4Hz,1H,Ph-H),δ=6.97-6.95(d,J=8.4Hz,1H,Ph-H),δ=6.16-6.12(d,J=16Hz,1H,Alkene-H),δ=5.99-5.95(d,J=16Hz,1H,Alkene-H),δ=3.23(s,1H,-Adamantane-H),δ=3.16(s,3H,-O-CH
3),δ=2.08(s,1H,-Adamantane-H),δ=1.93-1.68(m,12H,-Adamantane-H).
3.自发光BF
2底物的合成
在100mL圆底烧瓶中加入氟硼化合物(200mg,0.95mmol)、自发光醛(341mg,1.14mmol)与50ml乙腈,搅拌混合均匀;后加入正丁胺(0.5mL);加热回流10h,点板监控反应进程。反应完毕,旋蒸得红色粗产物,将粗产物用flash柱纯化,得淡红色固体产物120mg,产率26%。
1H NMR(400MHz,CDCl
3-d
1,ppm):δ=7.77-7.73(m,1H,Ph-H),δ=7.65-7.63(d,J=8Hz,2H,Ph-H),δ=7.59-7.57(d,J=8Hz,2H,Ph-H),δ=7.50-7.46(t,J=8Hz,1H,Ph-H),δ=7.38-7.34(d,J=16Hz,1H,Alkene-H),δ=7.18-7.12(m,2H,Ph-H),δ=6.58-6.54(d,J=16Hz,1H,Alkene-H),δ=6.12(s,1H,=CH-CO-),δ=3.30(s,3H,-O-CH
3),δ=3.30(s,1H,-Adamantane-H),δ=2.22(s,1H,-Adamantane-H),δ=1.96-1.75(m,12H,-Adamantane-H).
实施例3
F-QM-OH在聚集态的吸收、荧光光谱及化学发光光谱
取实施例1制备的F-QM-OH溶于分析纯二甲基亚砜中,制成1.0×10
-2M的储备液。然后制备水(H
2O)含量为99%的DMSO/H
2O混合溶剂2mL。取20μL上述储备液加入到已制备的DMSO/H
2O混合溶剂中,混合均匀后转移至光学石英比色皿(10×10mm)中测试其荧光光谱。如图1所示,以480nm作为激发波长,F-QM-OH底物的最大发射峰大约位于600nm处位于近红外区域,斯托克斯位移为120nm;F-QM-OH化学发光光谱与荧光光谱基本一致。
实施例4
白光激活化学发光前体F-QM-OH
取实施例1制备的F-QM-OH溶于分析纯二甲基亚砜中,制成1.0×10
-2M的储备液,同时制备1.0×10
-2M三甲基-β-环糊精的Tris缓冲液。然后取1μL上述储备液加入到10μL三甲基环糊精溶液中,再加入不同比例的Tris及DMSO溶液189μL,最终得到不同比例的DMSO:Tris溶液同时含有5.0×10
-5M的F-QM-OH与5.0×10
-4M的三甲基-β-环糊精。混合均匀后统一使用白光(200mW/cm
2)光照2分钟,并采用Imaging Quant 4000系统统一采集化学发光强度。结果如图2所示,在不同比例的Tris-DMSO缓冲液中,F-QM-OH均表现显著的化学发光信号。
实施例5
化学发光探针F-QM-B应用于过氧化氢检测
取F-QM-B(通过F-QM-OH进一步制备得到)溶于分析纯二甲基亚砜中,制成1.0×10
-3M的储备液。然后取10μL上述储备液加入到180μL Tris溶液中,混合均匀;取两组上述溶液,一组加入10μL过氧化氢溶液(1.0×10
-2M),另一组加入10μL Tris缓冲液作为对照;混合均匀后统一使用白光(100mW/cm
2)光照2分钟,并采用Imaging Quant 4000系统统一采集化学发光强度。结果如图3所示,过氧化氢组表现为明显的自发光信号,而对照组几乎没有自发光信号。
实施例6
化学发光探针F-QM-B应用活体检测
本发明中所有活体实验均遵守实验室动物饲养和使用的规章制度,并得到华东理工大学大学动物饲养和使用委员会批准。实验用荷瘤裸鼠购自上海斯莱克动物实验有限公司,饲养在无 菌室中层流通风橱内的无菌鼠笼中,使用高压蒸汽处理过的食物和水进行喂食。
为评估化学发光探针F-QM-B活体应用性能,采用荷瘤裸鼠作为成像对象,传统化学发光染料鲁米诺作为参比。如图4所示,从左至右依次对3只A549(人肺癌细胞)皮下瘤小鼠原位注射F-QM-B的三甲基-β-环糊精溶液(F-QM-B浓度1.0×10
-4M,三甲基-β-环糊精浓度1.0×10
-3M)、F-QM-B的三甲基-β-环糊精溶液以及鲁米诺溶液。注射后,对左1小鼠进行2分钟光照(白光,400mW/cm
2),并使用Perkin Elmer In-Vivo Professional Imaging System对小鼠进行整体的化学发光成像。成像实验前,对裸鼠进行含2.5%的异氟烷气体麻醉。
如图4所示,光照组(左1)小鼠具具有明显的化学发光信号,而未光照组(左2)并不表现明显的化学发光信号,这说明F-QM-B仅当与过氧化氢发生响应,并施以光照后才表现化学发光信号。此外,鲁米诺组(左3)同样并不表现明显的化学发光信号,这说明短波长化学发光染料并不满足活体成像要求。综上,本发明所制备化学发光探针具有波长长、发光可控、特异性好等显著优势,成功应用于过氧化氢活体检测。
Claims (4)
- 一种如权利要求1所述的自发荧光强度高、长波长、稳定性好的化学发光底物的制备方法,步骤如下:化合物的合成采用模块化制备方式,以2-溴-5-羟基苯甲醛为起始原料,依次通过缩醛反应、羟基保护及磷脂化反应,得到磷酸酯中间体;磷酸酯中间体进一步金刚烷酮反应(霍纳尔-沃兹沃思-埃蒙斯反应),制备得到烯烃中间体;烯烃中间体通过金属有机试剂活化,与N,N-二甲基甲酰胺反应,制备得到烯醛中间体;烯醛中间体进一步带有活泼甲基的小分子荧光团反应(Knoevenagel缩 合反应),制备得到化学发光底物。
- 一种权利要求1所述的自发荧光强度高、长波长、稳定性好的化学发光底物在化学发光探测及化学发光酶免疫分析中的应用。
- 根据权利要求3所述的应用,其特征在于,将权利要求2制备得到的底物连接各类检测单元得到化学发光探针。
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201910956954.2 | 2019-10-10 | ||
CN201910956954.2A CN110804009A (zh) | 2019-10-10 | 2019-10-10 | 一类化学发光强度高、波长长、稳定性好的化学发光底物及其制备方法和应用 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2021068828A1 true WO2021068828A1 (zh) | 2021-04-15 |
Family
ID=69488109
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/CN2020/119273 WO2021068828A1 (zh) | 2019-10-10 | 2020-09-30 | 一类化学发光强度高、波长长、稳定性好的化学发光底物及其制备方法和应用 |
Country Status (2)
Country | Link |
---|---|
CN (1) | CN110804009A (zh) |
WO (1) | WO2021068828A1 (zh) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115947946A (zh) * | 2022-12-13 | 2023-04-11 | 中山大学 | 一种肾清除型双通道光学纳米探针及其制备方法和应用 |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113061128B (zh) * | 2021-03-19 | 2022-12-09 | 华东理工大学 | 具有聚集诱导发光和宽光谱吸收特性的光敏剂及其制备方法和应用 |
CN115932284B (zh) * | 2023-03-10 | 2023-07-21 | 江苏奥雅生物科技有限公司 | 一种吖啶酯偶联免疫蛋白的方法及其应用 |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1990002742A1 (en) * | 1988-09-14 | 1990-03-22 | Tropix, Inc. | Purification of stable water-soluble dioxetanes |
US5981207A (en) * | 1998-12-18 | 1999-11-09 | Pharmacopeia, Inc. | Caged enzyme substrates as probes for reporter enzyme activity |
CN103209970A (zh) * | 2010-07-08 | 2013-07-17 | 生命科技公司 | 原位化学发光底物和测定 |
CN108884386A (zh) * | 2016-01-26 | 2018-11-23 | 特拉维夫大学拉玛特有限公司 | 用于诊断和体内成像的化学发光探针 |
WO2018216013A1 (en) * | 2017-05-24 | 2018-11-29 | Ramot At Tel-Aviv University Ltd. | Near-infrared chemiluminescent probes for in-vivo imaging |
-
2019
- 2019-10-10 CN CN201910956954.2A patent/CN110804009A/zh active Pending
-
2020
- 2020-09-30 WO PCT/CN2020/119273 patent/WO2021068828A1/zh active Application Filing
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1990002742A1 (en) * | 1988-09-14 | 1990-03-22 | Tropix, Inc. | Purification of stable water-soluble dioxetanes |
US5981207A (en) * | 1998-12-18 | 1999-11-09 | Pharmacopeia, Inc. | Caged enzyme substrates as probes for reporter enzyme activity |
CN103209970A (zh) * | 2010-07-08 | 2013-07-17 | 生命科技公司 | 原位化学发光底物和测定 |
CN108884386A (zh) * | 2016-01-26 | 2018-11-23 | 特拉维夫大学拉玛特有限公司 | 用于诊断和体内成像的化学发光探针 |
WO2018216013A1 (en) * | 2017-05-24 | 2018-11-29 | Ramot At Tel-Aviv University Ltd. | Near-infrared chemiluminescent probes for in-vivo imaging |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115947946A (zh) * | 2022-12-13 | 2023-04-11 | 中山大学 | 一种肾清除型双通道光学纳米探针及其制备方法和应用 |
CN115947946B (zh) * | 2022-12-13 | 2023-11-07 | 中山大学 | 一种肾清除型双通道光学纳米探针及其制备方法和应用 |
Also Published As
Publication number | Publication date |
---|---|
CN110804009A (zh) | 2020-02-18 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
WO2021068828A1 (zh) | 一类化学发光强度高、波长长、稳定性好的化学发光底物及其制备方法和应用 | |
JP5517624B2 (ja) | 耐酸化性指示薬分子 | |
JP5319714B2 (ja) | 過酸化水素検出方法に用いるシグナリング化合物 | |
JPS6393777A (ja) | 化学発光性1,2−ジオキセタン化合物およびその製造法と発光法 | |
CN102146284B (zh) | 一种比率型荧光探针及其应用 | |
KR20180108697A (ko) | 진단 및 인비보 영상용 화학발광 프로브 | |
KR20010041855A (ko) | 형광성 란타니드 킬레이트에 의한 검체의 분석 | |
CN102924524B (zh) | 一种具有活体细胞显影功能的锰配合物双光子吸收材料及其合成方法 | |
JPH0269590A (ja) | 化学発光法と発光用組成物 | |
WO2009107769A1 (ja) | 活性酸素測定用試薬 | |
CN108727256A (zh) | 一种基于三苯胺多吡啶盐的光敏剂及其制备方法与应用 | |
CN114031614B (zh) | 一种双细胞器成像、细胞活力评估和光动力癌细胞消融的荧光探针及制备和应用 | |
WO1999051586A1 (fr) | Reactif de determination d'oxygene singulet | |
CN113861076B (zh) | 一种aie型三苯胺衍生物荧光探针及其制备方法和其在水合肼检测中的应用 | |
CN106008435B (zh) | 一种用于Au3+检测的荧光增强型荧光探针及其制备方法 | |
CN111423439B (zh) | 一种检测Fe(Ⅲ)的香豆素类共轭杂环荧光探针 | |
CN111303111B (zh) | 一类黄菁近红外二区染料、制备方法及荧光成像应用 | |
CN111423467A (zh) | 一种检测氟离子用荧光探针及其制备方法和应用 | |
CN116606220A (zh) | 一种单组分有机余辉化合物及其生物检测的应用 | |
CN110615755A (zh) | 一种控释单线态氧的近红外荧光分子及其制备方法 | |
CN104804463B (zh) | 用于靶向肿瘤组织的近红外荧光染色剂及制备方法和应用 | |
CN115011141A (zh) | 一类基于氯取代硅罗丹明类近红外染料母体、制备方法及其应用 | |
CN115215833B (zh) | 化学发光探针及其应用 | |
CN1986550A (zh) | 一种基于铕配合物的单线态氧荧光探针及其应用 | |
CN117024332A (zh) | 一种具有高信噪比、高稳定性的近红外单线态氧发光探针及其制备方法与应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 20874008 Country of ref document: EP Kind code of ref document: A1 |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
122 | Ep: pct application non-entry in european phase |
Ref document number: 20874008 Country of ref document: EP Kind code of ref document: A1 |