EP0591489A1 - Substituierte thiazolin-dioxetan-substrate, verfahren zur herstellung und verwendung - Google Patents
Substituierte thiazolin-dioxetan-substrate, verfahren zur herstellung und verwendungInfo
- Publication number
- EP0591489A1 EP0591489A1 EP93907842A EP93907842A EP0591489A1 EP 0591489 A1 EP0591489 A1 EP 0591489A1 EP 93907842 A EP93907842 A EP 93907842A EP 93907842 A EP93907842 A EP 93907842A EP 0591489 A1 EP0591489 A1 EP 0591489A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- compound
- compounds
- alkyl
- group
- straight
- 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.)
- Withdrawn
Links
- 238000000034 method Methods 0.000 title claims description 17
- 239000000758 substrate Substances 0.000 title abstract description 4
- 150000001875 compounds Chemical class 0.000 claims abstract description 39
- BVTJGGGYKAMDBN-UHFFFAOYSA-N Dioxetane Chemical group C1COO1 BVTJGGGYKAMDBN-UHFFFAOYSA-N 0.000 claims abstract description 16
- 125000000217 alkyl group Chemical group 0.000 claims abstract description 11
- 239000001257 hydrogen Substances 0.000 claims abstract description 10
- 229910052739 hydrogen Inorganic materials 0.000 claims abstract description 10
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims abstract description 8
- 230000000087 stabilizing effect Effects 0.000 claims abstract description 4
- 125000004432 carbon atom Chemical group C* 0.000 claims abstract 4
- -1 alkyl carboxy ester Chemical class 0.000 claims description 10
- 150000002576 ketones Chemical class 0.000 claims description 8
- 150000001299 aldehydes Chemical class 0.000 claims description 7
- 238000001816 cooling Methods 0.000 claims description 7
- 108090000790 Enzymes Proteins 0.000 claims description 5
- 102000004190 Enzymes Human genes 0.000 claims description 5
- 230000007717 exclusion Effects 0.000 claims description 5
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 5
- 238000002360 preparation method Methods 0.000 claims description 5
- RFKAGFYMWORAGT-UHFFFAOYSA-N 4,5-dihydro-1,3-thiazole-2-carboxylic acid Chemical compound OC(=O)C1=NCCS1 RFKAGFYMWORAGT-UHFFFAOYSA-N 0.000 claims description 4
- 229910019142 PO4 Inorganic materials 0.000 claims description 4
- 239000002253 acid Substances 0.000 claims description 4
- 150000007857 hydrazones Chemical class 0.000 claims description 4
- 239000010452 phosphate Substances 0.000 claims description 4
- 125000005073 adamantyl group Chemical group C12(CC3CC(CC(C1)C3)C2)* 0.000 claims description 3
- 125000003545 alkoxy group Chemical group 0.000 claims description 3
- 125000003118 aryl group Chemical group 0.000 claims description 3
- 239000003054 catalyst Substances 0.000 claims description 3
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 claims description 3
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 3
- 150000003839 salts Chemical class 0.000 claims description 3
- 125000002769 thiazolinyl group Chemical group 0.000 claims description 3
- 238000003747 Grignard reaction Methods 0.000 claims description 2
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 claims description 2
- UCKMPCXJQFINFW-UHFFFAOYSA-N Sulphide Chemical compound [S-2] UCKMPCXJQFINFW-UHFFFAOYSA-N 0.000 claims description 2
- 239000007795 chemical reaction product Substances 0.000 claims description 2
- 125000000113 cyclohexyl group Chemical group [H]C1([H])C([H])([H])C([H])([H])C([H])(*)C([H])([H])C1([H])[H] 0.000 claims description 2
- 125000004435 hydrogen atom Chemical group [H]* 0.000 claims description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 2
- 239000003960 organic solvent Substances 0.000 claims description 2
- 239000012491 analyte Substances 0.000 claims 1
- 238000010324 immunological assay Methods 0.000 claims 1
- 239000012190 activator Substances 0.000 abstract 1
- 238000003018 immunoassay Methods 0.000 abstract 1
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 45
- 239000000203 mixture Substances 0.000 description 23
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 18
- 239000000243 solution Substances 0.000 description 17
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 16
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 15
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 15
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 12
- 238000006243 chemical reaction Methods 0.000 description 10
- CDMKLKAZVMTVHX-UHFFFAOYSA-N 4,5-dihydro-1,3-thiazol-3-ium-4-carboxylate Chemical compound OC(=O)C1CSC=N1 CDMKLKAZVMTVHX-UHFFFAOYSA-N 0.000 description 9
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 9
- CGWAQZQRRWLHGO-UHFFFAOYSA-N 5,5-dimethyl-4h-1,3-thiazole Chemical compound CC1(C)CN=CS1 CGWAQZQRRWLHGO-UHFFFAOYSA-N 0.000 description 8
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 8
- 239000012074 organic phase Substances 0.000 description 8
- 229910052938 sodium sulfate Inorganic materials 0.000 description 8
- 235000011152 sodium sulphate Nutrition 0.000 description 8
- 230000015572 biosynthetic process Effects 0.000 description 7
- 239000002904 solvent Substances 0.000 description 7
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 6
- 238000004440 column chromatography Methods 0.000 description 6
- 239000003208 petroleum Substances 0.000 description 6
- 239000000741 silica gel Substances 0.000 description 6
- 229910002027 silica gel Inorganic materials 0.000 description 6
- 238000003786 synthesis reaction Methods 0.000 description 6
- CBDKQYKMCICBOF-UHFFFAOYSA-N thiazoline Chemical compound C1CN=CS1 CBDKQYKMCICBOF-UHFFFAOYSA-N 0.000 description 6
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 5
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 4
- 239000011541 reaction mixture Substances 0.000 description 4
- LJRDOKAZOAKLDU-UDXJMMFXSA-N (2s,3s,4r,5r,6r)-5-amino-2-(aminomethyl)-6-[(2r,3s,4r,5s)-5-[(1r,2r,3s,5r,6s)-3,5-diamino-2-[(2s,3r,4r,5s,6r)-3-amino-4,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-6-hydroxycyclohexyl]oxy-4-hydroxy-2-(hydroxymethyl)oxolan-3-yl]oxyoxane-3,4-diol;sulfuric ac Chemical compound OS(O)(=O)=O.N[C@@H]1[C@@H](O)[C@H](O)[C@H](CN)O[C@@H]1O[C@H]1[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](N)C[C@@H](N)[C@@H]2O)O[C@@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)N)O[C@@H]1CO LJRDOKAZOAKLDU-UDXJMMFXSA-N 0.000 description 3
- IGXWBGJHJZYPQS-SSDOTTSWSA-N D-Luciferin Chemical compound OC(=O)[C@H]1CSC(C=2SC3=CC=C(O)C=C3N=2)=N1 IGXWBGJHJZYPQS-SSDOTTSWSA-N 0.000 description 3
- CYCGRDQQIOGCKX-UHFFFAOYSA-N Dehydro-luciferin Natural products OC(=O)C1=CSC(C=2SC3=CC(O)=CC=C3N=2)=N1 CYCGRDQQIOGCKX-UHFFFAOYSA-N 0.000 description 3
- BJGNCJDXODQBOB-UHFFFAOYSA-N Fivefly Luciferin Natural products OC(=O)C1CSC(C=2SC3=CC(O)=CC=C3N=2)=N1 BJGNCJDXODQBOB-UHFFFAOYSA-N 0.000 description 3
- DDWFXDSYGUXRAY-UHFFFAOYSA-N Luciferin Natural products CCc1c(C)c(CC2NC(=O)C(=C2C=C)C)[nH]c1Cc3[nH]c4C(=C5/NC(CC(=O)O)C(C)C5CC(=O)O)CC(=O)c4c3C DDWFXDSYGUXRAY-UHFFFAOYSA-N 0.000 description 3
- 230000003213 activating effect Effects 0.000 description 3
- 238000009835 boiling Methods 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 229960001639 penicillamine Drugs 0.000 description 3
- 238000010992 reflux Methods 0.000 description 3
- 150000003549 thiazolines Chemical class 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 2
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 2
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- MZRVEZGGRBJDDB-UHFFFAOYSA-N N-Butyllithium Chemical compound [Li]CCCC MZRVEZGGRBJDDB-UHFFFAOYSA-N 0.000 description 2
- OFBQJSOFQDEBGM-UHFFFAOYSA-N Pentane Chemical compound CCCCC OFBQJSOFQDEBGM-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- WQDUMFSSJAZKTM-UHFFFAOYSA-N Sodium methoxide Chemical compound [Na+].[O-]C WQDUMFSSJAZKTM-UHFFFAOYSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 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 2
- 239000008346 aqueous phase Substances 0.000 description 2
- 102000005936 beta-Galactosidase Human genes 0.000 description 2
- 108010005774 beta-Galactosidase Proteins 0.000 description 2
- 238000006114 decarboxylation reaction Methods 0.000 description 2
- FAMRKDQNMBBFBR-BQYQJAHWSA-N diethyl azodicarboxylate Substances CCOC(=O)\N=N\C(=O)OCC FAMRKDQNMBBFBR-BQYQJAHWSA-N 0.000 description 2
- FAMRKDQNMBBFBR-UHFFFAOYSA-N ethyl n-ethoxycarbonyliminocarbamate Chemical compound CCOC(=O)N=NC(=O)OCC FAMRKDQNMBBFBR-UHFFFAOYSA-N 0.000 description 2
- 150000008195 galaktosides Chemical class 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000006386 neutralization reaction Methods 0.000 description 2
- CTSLXHKWHWQRSH-UHFFFAOYSA-N oxalyl chloride Chemical compound ClC(=O)C(Cl)=O CTSLXHKWHWQRSH-UHFFFAOYSA-N 0.000 description 2
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 description 2
- 150000003254 radicals Chemical class 0.000 description 2
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Chemical compound [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 description 2
- 230000006641 stabilisation Effects 0.000 description 2
- 238000011105 stabilization Methods 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- CSRZQMIRAZTJOY-UHFFFAOYSA-N trimethylsilyl iodide Chemical compound C[Si](C)(C)I CSRZQMIRAZTJOY-UHFFFAOYSA-N 0.000 description 2
- RIOQSEWOXXDEQQ-UHFFFAOYSA-N triphenylphosphine Chemical compound C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1 RIOQSEWOXXDEQQ-UHFFFAOYSA-N 0.000 description 2
- UIQGEWJEWJMQSL-UHFFFAOYSA-N 2,2,4,4-tetramethylpentan-3-one Chemical compound CC(C)(C)C(=O)C(C)(C)C UIQGEWJEWJMQSL-UHFFFAOYSA-N 0.000 description 1
- ZUYKJZQOPXDNOK-UHFFFAOYSA-N 2-(ethylamino)-2-thiophen-2-ylcyclohexan-1-one;hydrochloride Chemical class Cl.C=1C=CSC=1C1(NCC)CCCCC1=O ZUYKJZQOPXDNOK-UHFFFAOYSA-N 0.000 description 1
- FPZMXOBOWKUAIY-UHFFFAOYSA-N 2-adamantylidenehydrazine Chemical compound C1C(C2)CC3CC1C(=NN)C2C3 FPZMXOBOWKUAIY-UHFFFAOYSA-N 0.000 description 1
- ICGLPKIVTVWCFT-UHFFFAOYSA-N 4-methylbenzenesulfonohydrazide Chemical compound CC1=CC=C(S(=O)(=O)NN)C=C1 ICGLPKIVTVWCFT-UHFFFAOYSA-N 0.000 description 1
- DEWDWBYQOFXKIH-UHFFFAOYSA-N 6-methoxy-1,3-benzothiazole-2-carbonitrile Chemical compound COC1=CC=C2N=C(C#N)SC2=C1 DEWDWBYQOFXKIH-UHFFFAOYSA-N 0.000 description 1
- JHAWWJQGHKGXHA-UHFFFAOYSA-N 6-tert-butylquinoline Chemical compound N1=CC=CC2=CC(C(C)(C)C)=CC=C21 JHAWWJQGHKGXHA-UHFFFAOYSA-N 0.000 description 1
- WKBOTKDWSSQWDR-UHFFFAOYSA-N Bromine atom Chemical compound [Br] WKBOTKDWSSQWDR-UHFFFAOYSA-N 0.000 description 1
- 239000004215 Carbon black (E152) Substances 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 239000003298 DNA probe Substances 0.000 description 1
- KRHYYFGTRYWZRS-UHFFFAOYSA-M Fluoride anion Chemical compound [F-] KRHYYFGTRYWZRS-UHFFFAOYSA-M 0.000 description 1
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 1
- 108060001084 Luciferase Proteins 0.000 description 1
- 239000005089 Luciferase Substances 0.000 description 1
- CIUQDSCDWFSTQR-UHFFFAOYSA-N [C]1=CC=CC=C1 Chemical group [C]1=CC=CC=C1 CIUQDSCDWFSTQR-UHFFFAOYSA-N 0.000 description 1
- 229960000583 acetic acid Drugs 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- IYKFYARMMIESOX-UHFFFAOYSA-N adamantanone Chemical compound C1C(C2)CC3CC1C(=O)C2C3 IYKFYARMMIESOX-UHFFFAOYSA-N 0.000 description 1
- 125000001931 aliphatic group Chemical group 0.000 description 1
- 150000001344 alkene derivatives Chemical class 0.000 description 1
- 150000001336 alkenes Chemical class 0.000 description 1
- SMZOGRDCAXLAAR-UHFFFAOYSA-N aluminium isopropoxide Chemical compound [Al+3].CC(C)[O-].CC(C)[O-].CC(C)[O-] SMZOGRDCAXLAAR-UHFFFAOYSA-N 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 125000004429 atom Chemical group 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 150000003935 benzaldehydes Chemical class 0.000 description 1
- IOJUPLGTWVMSFF-UHFFFAOYSA-N benzothiazole Chemical group C1=CC=C2SC=NC2=C1 IOJUPLGTWVMSFF-UHFFFAOYSA-N 0.000 description 1
- GDTBXPJZTBHREO-UHFFFAOYSA-N bromine Substances BrBr GDTBXPJZTBHREO-UHFFFAOYSA-N 0.000 description 1
- 229910052794 bromium Inorganic materials 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- CGZZMOTZOONQIA-UHFFFAOYSA-N cycloheptanone Chemical compound O=C1CCCCCC1 CGZZMOTZOONQIA-UHFFFAOYSA-N 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- 239000012380 dealkylating agent Substances 0.000 description 1
- 230000020335 dealkylation Effects 0.000 description 1
- 238000006900 dealkylation reaction Methods 0.000 description 1
- SBZXBUIDTXKZTM-UHFFFAOYSA-N diglyme Chemical compound COCCOCCOC SBZXBUIDTXKZTM-UHFFFAOYSA-N 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 229930195733 hydrocarbon Natural products 0.000 description 1
- 150000002430 hydrocarbons Chemical class 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- SCEZYJKGDJPHQO-UHFFFAOYSA-M magnesium;methanidylbenzene;chloride Chemical compound [Mg+2].[Cl-].[CH2-]C1=CC=CC=C1 SCEZYJKGDJPHQO-UHFFFAOYSA-M 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- CXKWCBBOMKCUKX-UHFFFAOYSA-M methylene blue Chemical compound [Cl-].C1=CC(N(C)C)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 CXKWCBBOMKCUKX-UHFFFAOYSA-M 0.000 description 1
- 229960000907 methylthioninium chloride Drugs 0.000 description 1
- FZFLTDNAHASQQC-RVDMUPIBSA-N n-[(e)-benzylideneamino]-4-methylbenzenesulfonamide Chemical compound C1=CC(C)=CC=C1S(=O)(=O)N\N=C\C1=CC=CC=C1 FZFLTDNAHASQQC-RVDMUPIBSA-N 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 238000007124 photooxygenation reaction Methods 0.000 description 1
- 239000003495 polar organic solvent Substances 0.000 description 1
- 239000002798 polar solvent Substances 0.000 description 1
- 229910000027 potassium carbonate Inorganic materials 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000012266 salt solution Substances 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 235000010288 sodium nitrite Nutrition 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 238000010792 warming Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D513/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00
- C07D513/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00 in which the condensed system contains two hetero rings
- C07D513/10—Spiro-condensed systems
-
- 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
- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
- C07F9/02—Phosphorus compounds
- C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
- C07F9/6561—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing systems of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring or ring system, with or without other non-condensed hetero rings
-
- 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
- C09K11/07—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials having chemically interreactive components, e.g. reactive chemiluminescent compositions
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
-
- 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
- G01N33/582—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with fluorescent label
Definitions
- the invention relates to thiazoline-dioxetane substrates, processes for the production and use in enzymatic analysis processes.
- the object of the present invention was to provide a 1,2-dioxetane based on the activated intermediate of thiazolines and preferably luciferin, which is stable and only decomposes by reaction with an activating agent with the formation of chemiluminescence.
- This object is achieved by a compound of the general formula III,
- R- [and R2 are in each case identical or different and represent hydrogen or lower alkyl C 1 -C 4, straight-chain or branched
- X is a cleavable group which can be cleaved off by an activating agent and at least one of the groups R4 or R5 represents a group stabilizing the dioxetane structure and at most one of the groups R4 or R5 represents hydrogen.
- Acids, bases, salts, enzymes, inorganic or organic catalysts and Electron donors used are Acids, bases, salts, enzymes, inorganic or organic catalysts and Electron donors used.
- the group -O-X can be located at any position on the phenyl ring.
- R is a benzothiazole group
- the group -O-X is preferably arranged in the 5-position.
- R is a phenyl group
- the group -O-X is preferably arranged in the 3-position. The same applies to the group -OR3, from which the group -O-X arises.
- -O-X is preferably the hydroxy salt of an oxyacid, phosphate, aryl or alkyl carboxyl ester, alkyloxy or arylsilyloxy, sulfate, oxypyranoside.
- composition of the aryl and alkyl radicals is not critical. Any person skilled in the art can select suitable radicals X without further ado. Solubility and cleavage of X by the activating agents must only be possible.
- R1 and R2 are preferably hydrogen, methyl or ethyl groups. If, for example, phosphate is used as -O-X, the chemiluminescence reaction can be induced by adding alkaline phosphatase. When using the galactoside, the chemiluminescence reaction can be induced by ß-galactosidase. If a silyloxy radical is used as -O-X, the addition of fluoride can induce chemiluminescence.
- Groups which protect the dioxetane group from reactions are suitable as groups R4 and / or R5 which stabilize the dioxetane structure. This stabilization is preferably carried out by sterically shielding this group.
- R4 and R5 can be the same or different.
- the adamantyl residue it is preferably bound in such a way that R4 and R5 represent parts of the ring structure and are therefore bridged (formula purple):
- the compounds of general formula III are new. A synthetic method for these compounds is not yet known.
- the invention accordingly also relates to a process for the preparation of the compounds of the formula III, which is characterized in that a compound of the formula IV or IVa.
- R3 alkyl straight-chain or branched with 1 - 6 C- Represents atoms, preferably methyl or ethyl, with a compound of formula V.
- R1 and R2 have the meanings given above, with the exclusion of water and air in the presence of an alkyllithium compound with aldehydes or ketones, decarboxylated the reaction product, dealkylated the alkoxy group, there the chemically labile group according to methods familiar to the skilled worker -OX introduces and photo-oxygenates to the dioxetane.
- reaction of the compound of the formula IV (alkoxybenzothiazole-2-carboxylic acid nitrile) or IVa with the compound of the formula V (preferably penicillamine or cysteine) is preferably carried out in a polar solvent, such as water or water / alcohol mixtures, at room temperature with the exclusion of light.
- a polar solvent such as water or water / alcohol mixtures
- Suitable compounds as aldehydes and ketones are those which, as radical R4 and / or R5, stabilize the dioxetane structure of the compounds of the formula III. Such stabilization can be achieved by steric shielding of the dioxetane structure.
- Suitable ketones and aldehydes are accordingly, for example, adamantanone, benzaldehydes, cyclohexanones, secondary and tertiary aliphatic aldehydes and ketones, such as. B. Di-t-butyl ketone.
- R3 is preferably water-free in an aprotic organic solvent such as.
- the alkyl lithium compound is preferably added as a solution in a hydrocarbon (pentane, hexane). Butyllithium is particularly preferably used. After the reaction has taken place, the compound obtained is precipitated from the reaction mixture, preferably by adding an aqueous salt solution at room temperature.
- decarboxylation of the resulting ⁇ -hydroxycarboxylic acid to introduce a double bond is preferably carried out under dehydrating conditions, e.g. B. with diethyl azodicarboxylate and triphenylphosphine.
- the dealkylation at the 6 'position is expediently carried out by adding a dealkylating agent such as trimethyl iodosilane. It is preferred to work anhydrous.
- the thiazoline carboxylic acid can preferably be converted to a thiazolin-4-one via the intermediate stage of the amine and alcohol and then to a thiazolin-4-thione by reaction with sulfide.
- This thiazolin-4-thione is reacted with a hydrazone of the aldehyde or ketone, preferably with adamantanone-2-hydrazone or benzaldehyde hydrazone, which can contain a removable group (adamantanontosyl hydrazone or benzaldehyde tosyl hydrazone) to give the compounds of the formula III according to the invention.
- R4 and / or R5 represents a phenyl radical, an aliphatic radical or cycloalkyl radical, a substituted 2- Phenyl-5,5-dialkyl-thiazolin-4-one in a Grignard reaction to the 4-substituted derivative with subsequent introduction of a double bond at the 4-position of the thiazoline. Then, as described below, the dioxetane group is introduced.
- the dioxetane group is expediently introduced by reacting the alkene derivative (double bond at the 2-position of the thiazoline), which is preferably dissolved in a polar organic solvent, such as methylene chloride or chloroform, at a temperature of -100-50 ° C and irradiation with visible light using a sensitizer (for example Rose Bengal, methylene blue, cf. Tetra ⁇ hedron Letters (1988) 3137 - 3140).
- a sensitizer for example Rose Bengal, methylene blue, cf. Tetra ⁇ hedron Letters (1988) 3137 - 3140.
- the supernatant contains the desired compound of formula III.
- Another object of the invention is a method for determining an acid, base of a salt, enzyme, inorganic or organic catalyst and electron donor by reacting this compound with a compound of general formula III and measuring the light emitted as a measure of the content of the connection to be determined.
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Hematology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Microbiology (AREA)
- Urology & Nephrology (AREA)
- Wood Science & Technology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Food Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Materials Engineering (AREA)
- Biophysics (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Cell Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Plural Heterocyclic Compounds (AREA)
- Heterocyclic Compounds That Contain Two Or More Ring Oxygen Atoms (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Nitrogen And Oxygen Or Sulfur-Condensed Heterocyclic Ring Systems (AREA)
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE4210759A DE4210759A1 (de) | 1992-04-01 | 1992-04-01 | Substituierte Thiazolin-Dioxetan-Substrate, Verfahren zur Herstellung und Verwendung |
| DE4210759 | 1992-04-01 | ||
| PCT/EP1993/000752 WO1993020083A1 (de) | 1992-04-01 | 1993-03-29 | Substituierte thiazolin-dioxetan-substrate, verfahren zur herstellung und verwendung |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| EP0591489A1 true EP0591489A1 (de) | 1994-04-13 |
Family
ID=6455673
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| EP93907842A Withdrawn EP0591489A1 (de) | 1992-04-01 | 1993-03-29 | Substituierte thiazolin-dioxetan-substrate, verfahren zur herstellung und verwendung |
Country Status (5)
| Country | Link |
|---|---|
| US (3) | US5455357A (ja) |
| EP (1) | EP0591489A1 (ja) |
| JP (1) | JPH06102669B2 (ja) |
| DE (1) | DE4210759A1 (ja) |
| WO (1) | WO1993020083A1 (ja) |
Families Citing this family (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE4210759A1 (de) * | 1992-04-01 | 1993-10-07 | Boehringer Mannheim Gmbh | Substituierte Thiazolin-Dioxetan-Substrate, Verfahren zur Herstellung und Verwendung |
| DE19538708A1 (de) * | 1995-10-18 | 1997-04-24 | Boehringer Mannheim Gmbh | Heterocyclische Dioxetan-Substrate, Verfahren zur Herstellung und Verwendung |
| DE69720332T2 (de) * | 1996-01-16 | 2003-12-04 | Lumigen Inc | Verbindungen, zusammensetzungen und methoden zur erzeugung von chemilumineszenz mit phosphatase-enzymen |
| US5876995A (en) | 1996-02-06 | 1999-03-02 | Bryan; Bruce | Bioluminescent novelty items |
| US6247995B1 (en) | 1996-02-06 | 2001-06-19 | Bruce Bryan | Bioluminescent novelty items |
| US6416960B1 (en) | 1996-08-08 | 2002-07-09 | Prolume, Ltd. | Detection and visualization of neoplastic tissues and other tissues |
| CA2271717A1 (en) | 1996-12-12 | 1998-06-18 | Prolume, Ltd. | Apparatus and method for detecting and identifying infectious agents |
| EP1925320A3 (en) | 1998-03-27 | 2008-09-03 | Prolume, Ltd. | Luciferases, fluorescent proteins, nucleic acids encoding the luciferases and fluorescent proteins and the use thereof in diagnostics |
| DE69938293T2 (de) | 1998-03-27 | 2009-03-12 | Bruce J. Beverly Hills Bryan | Luciferase, gfp fluoreszenzproteine, kodierende nukleinsaüre und ihre verwendung in der diagnose |
| US7390670B2 (en) | 2003-02-20 | 2008-06-24 | Lumigen, Inc. | Signalling compounds and methods for detecting hydrogen peroxide |
| SG11201903523YA (en) | 2016-10-20 | 2019-05-30 | Harvard College | In vitro and cell based assays for measuring the activity of botulinum neurotoxins |
Family Cites Families (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5068339A (en) * | 1986-07-17 | 1991-11-26 | Board Of Governors Of Wayne State University | Enhanced chemiluminescence from 1,2-dioxetanes through energy transfer to tethered fluorescers |
| US5004565A (en) * | 1986-07-17 | 1991-04-02 | The Board Of Governors Of Wayne State University | Method and compositions providing enhanced chemiluminescence from 1,2-dioxetanes |
| US5707559A (en) * | 1986-07-17 | 1998-01-13 | Tropix, Inc. | Chemiluminescent 1,2-dioxetane compounds |
| US4983779A (en) * | 1986-07-17 | 1991-01-08 | The Board Of Governors Of Wayne State University | Process for the preparation of vinyl ethers |
| US4959182A (en) * | 1986-07-17 | 1990-09-25 | Board Of Governors Of Wayne State University | Method and compositions providing enhanced chemiluminescence from 1,2-dioxetanes |
| US5013827A (en) * | 1986-07-17 | 1991-05-07 | Board Of Governors Of Wayne State University | Enhanced chemiluminescence from 1,2-dioxetanes through energy transfer to tethered fluorescers |
| US4857652A (en) * | 1986-07-17 | 1989-08-15 | Board Of Governors Of Wayne State University | Chemiluminescent 1,2-dioxetane compounds |
| US4962192A (en) * | 1986-07-17 | 1990-10-09 | Board Of Governors Of Wayne State University | Chemiluminescent 1,2-dioxetane compounds |
| US5089630A (en) * | 1987-12-31 | 1992-02-18 | Bronstein Irena Y | Dioxetanes for use in assays |
| DE68917387T2 (de) * | 1987-12-31 | 1994-12-22 | Tropix Inc | Dioxetane zur verwendung bei immuntests. |
| DE4210759A1 (de) * | 1992-04-01 | 1993-10-07 | Boehringer Mannheim Gmbh | Substituierte Thiazolin-Dioxetan-Substrate, Verfahren zur Herstellung und Verwendung |
-
1992
- 1992-04-01 DE DE4210759A patent/DE4210759A1/de not_active Withdrawn
-
1993
- 1993-03-29 WO PCT/EP1993/000752 patent/WO1993020083A1/de not_active Application Discontinuation
- 1993-03-29 US US08/150,188 patent/US5455357A/en not_active Expired - Fee Related
- 1993-03-29 EP EP93907842A patent/EP0591489A1/de not_active Withdrawn
- 1993-03-29 JP JP5517071A patent/JPH06102669B2/ja not_active Expired - Fee Related
-
1995
- 1995-05-25 US US08/450,211 patent/US5712106A/en not_active Expired - Fee Related
-
1997
- 1997-10-08 US US08/947,065 patent/US5994552A/en not_active Expired - Fee Related
Non-Patent Citations (1)
| Title |
|---|
| See references of WO9320083A1 * |
Also Published As
| Publication number | Publication date |
|---|---|
| US5994552A (en) | 1999-11-30 |
| WO1993020083A1 (de) | 1993-10-14 |
| US5455357A (en) | 1995-10-03 |
| JPH06503844A (ja) | 1994-04-28 |
| DE4210759A1 (de) | 1993-10-07 |
| US5712106A (en) | 1998-01-27 |
| JPH06102669B2 (ja) | 1994-12-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP0040177A1 (de) | Verfahren zur Herstellung von Alkenylbenzol- oder -naphthalincarbonsäurederivaten | |
| EP0591489A1 (de) | Substituierte thiazolin-dioxetan-substrate, verfahren zur herstellung und verwendung | |
| DE69909611T2 (de) | Reagenz zum nachweis von singulett-sauerstoff | |
| EP0167973B1 (de) | Redox-Indikatoren | |
| EP0007019A1 (de) | Neue 5-Phenylpyrazol-Derivate und ihre Salze; Verfahren zu deren Herstellung und diese Verbindungen enthaltende Arzneimittel | |
| EP0218541A2 (de) | 3-Vinyl- und 3-Ethinyl-beta-carbolinderivate, ihre Herstellung und ihre Verwendung als Arzneimittel | |
| DE60125958T2 (de) | Reagenzien zur bestimmung von atomaren sauerstoff | |
| DE69910642T2 (de) | 1,2-Dioxetanderivate | |
| EP0273270A1 (de) | Verfahren zur Herstellung von (+)-Biotin | |
| DE2003461A1 (de) | Verfahren zur Herstellung von 2-Alkylpyridaziniumverbindungen | |
| EP0863906B1 (de) | Heterocyclische dioxetan-substrate, verfahren zur herstellung und verwendung | |
| DE1795147A1 (de) | Verfahren zur Herstellung von Sauerstoffderivaten des Indols | |
| DE2814747C2 (de) | Verfahren zur Herstellung von Androsta-4,9(11)-dien-3,17-dionen durch saure Dehydratisierung von 9α-Hydroxyandrostendionen | |
| DE60003598T2 (de) | Mit einem Pyranring kondensierte 1,2-Dioxetan-Derivate und ihre Verwendung | |
| EP0513700B1 (de) | Verfahren zur Herstellung von Imidazothiazolon-Derivaten | |
| DE2652892A1 (de) | Substituierte aminobenzaldehydverbindungen | |
| Shalaby et al. | Reactivity of the Exocyclic Double Bond in 5‐arylidene derivatives of barbituric and thiobarbituric acids | |
| DE2644789A1 (de) | Alkylen-di-phenylalk(en)ylcarbonsaeuren und ihre herstellung und verwendung | |
| DE1620023A1 (de) | Verfahren zum Herstellen von Indolyl-essigsaeure-Verbindungen | |
| AT404724B (de) | Neue zwischenverbindung und verfahren zur herstellung von antihypercholesterinämisch wirkenden tetrazolverbindungen | |
| DE1643226A1 (de) | Photochrome Verbindungen und ihre Herstellung | |
| DE2820862A1 (de) | Chromanderivate | |
| EP0036159A1 (de) | 4-(2,5-Dihydroxyphen-1-yl)-crotonsäure und deren Derivate sowie Verfahren zur Herstellung dieser Verbindungen | |
| EP0019132A1 (de) | Verfahren zur Herstellung von alpha-Halogencyclobutanonen | |
| JPH05117241A (ja) | ４−ヒドロペルオキシ−２−（ｎ，ｎ−ジ置換アミノフエニル）−４，５−ジフエニル−４ｈ−イソイミダゾール |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
| 17P | Request for examination filed |
Effective date: 19931027 |
|
| AK | Designated contracting states |
Kind code of ref document: A1 Designated state(s): AT BE CH DE DK ES FR GB GR IE IT LI LU NL PT SE |
|
| 17Q | First examination report despatched |
Effective date: 19970807 |
|
| RAP3 | Party data changed (applicant data changed or rights of an application transferred) |
Owner name: ROCHE DIAGNOSTICS GMBH |
|
| STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN |
|
| 18D | Application deemed to be withdrawn |
Effective date: 20001130 |