JPH0625146B2 - Novel imidazole derivative and measuring method using the same as a coloring component - Google Patents
Novel imidazole derivative and measuring method using the same as a coloring componentInfo
- Publication number
- JPH0625146B2 JPH0625146B2 JP60068315A JP6831585A JPH0625146B2 JP H0625146 B2 JPH0625146 B2 JP H0625146B2 JP 60068315 A JP60068315 A JP 60068315A JP 6831585 A JP6831585 A JP 6831585A JP H0625146 B2 JPH0625146 B2 JP H0625146B2
- Authority
- JP
- Japan
- Prior art keywords
- group
- substituent
- substituted
- phenyl
- hydrogen peroxide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
- 238000000034 method Methods 0.000 title claims description 26
- 150000002460 imidazoles Chemical class 0.000 title claims description 17
- 239000006103 coloring component Substances 0.000 title claims description 6
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical group OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims description 37
- 125000001424 substituent group Chemical group 0.000 claims description 35
- -1 phenyl-substituted carbamoyl group Chemical group 0.000 claims description 29
- 125000003277 amino group Chemical group 0.000 claims description 19
- 239000000126 substance Substances 0.000 claims description 18
- 238000011002 quantification Methods 0.000 claims description 14
- 102000003992 Peroxidases Human genes 0.000 claims description 12
- 108040007629 peroxidase activity proteins Proteins 0.000 claims description 12
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 12
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 claims description 10
- 102000004190 Enzymes Human genes 0.000 claims description 10
- 108090000790 Enzymes Proteins 0.000 claims description 10
- 229910001413 alkali metal ion Inorganic materials 0.000 claims description 10
- 125000000217 alkyl group Chemical group 0.000 claims description 10
- 239000012472 biological sample Substances 0.000 claims description 10
- 125000003917 carbamoyl group Chemical class [H]N([H])C(*)=O 0.000 claims description 10
- 238000006911 enzymatic reaction Methods 0.000 claims description 8
- 102000004316 Oxidoreductases Human genes 0.000 claims description 7
- 108090000854 Oxidoreductases Proteins 0.000 claims description 7
- 125000003545 alkoxy group Chemical group 0.000 claims description 7
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 7
- 125000005843 halogen group Chemical group 0.000 claims description 7
- 239000000758 substrate Substances 0.000 claims description 6
- 102000001554 Hemoglobins Human genes 0.000 claims description 5
- 108010054147 Hemoglobins Proteins 0.000 claims description 5
- 125000003710 aryl alkyl group Chemical group 0.000 claims description 5
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 claims description 5
- 239000007800 oxidant agent Substances 0.000 claims description 5
- 238000011161 development Methods 0.000 claims description 4
- 230000000694 effects Effects 0.000 claims description 4
- 230000001590 oxidative effect Effects 0.000 claims description 4
- 238000003556 assay Methods 0.000 claims description 2
- 238000004445 quantitative analysis Methods 0.000 claims 4
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 16
- 150000001875 compounds Chemical class 0.000 description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 15
- 239000003153 chemical reaction reagent Substances 0.000 description 14
- LEHOTFFKMJEONL-UHFFFAOYSA-N Uric Acid Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 description 10
- 239000000975 dye Substances 0.000 description 10
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 9
- 238000002835 absorbance Methods 0.000 description 9
- 229940088598 enzyme Drugs 0.000 description 9
- 238000003786 synthesis reaction Methods 0.000 description 9
- 230000015572 biosynthetic process Effects 0.000 description 8
- 238000001816 cooling Methods 0.000 description 8
- 239000013078 crystal Substances 0.000 description 8
- TVWHNULVHGKJHS-UHFFFAOYSA-N Uric acid Natural products N1C(=O)NC(=O)C2NC(=O)NC21 TVWHNULVHGKJHS-UHFFFAOYSA-N 0.000 description 7
- 238000006243 chemical reaction Methods 0.000 description 7
- 229940116269 uric acid Drugs 0.000 description 7
- QGJOPFRUJISHPQ-UHFFFAOYSA-N Carbon disulfide Chemical compound S=C=S QGJOPFRUJISHPQ-UHFFFAOYSA-N 0.000 description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 238000011088 calibration curve Methods 0.000 description 6
- 238000004040 coloring Methods 0.000 description 6
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 6
- 238000005259 measurement Methods 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 6
- 239000011734 sodium Substances 0.000 description 6
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 5
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 5
- 238000007254 oxidation reaction Methods 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 5
- 210000002966 serum Anatomy 0.000 description 5
- RLFWWDJHLFCNIJ-UHFFFAOYSA-N 4-aminoantipyrine Chemical compound CN1C(C)=C(N)C(=O)N1C1=CC=CC=C1 RLFWWDJHLFCNIJ-UHFFFAOYSA-N 0.000 description 4
- BPYKTIZUTYGOLE-IFADSCNNSA-N Bilirubin Chemical compound N1C(=O)C(C)=C(C=C)\C1=C\C1=C(C)C(CCC(O)=O)=C(CC2=C(C(C)=C(\C=C/3C(=C(C=C)C(=O)N\3)C)N2)CCC(O)=O)N1 BPYKTIZUTYGOLE-IFADSCNNSA-N 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 4
- VSCWAEJMTAWNJL-UHFFFAOYSA-K aluminium trichloride Chemical compound Cl[Al](Cl)Cl VSCWAEJMTAWNJL-UHFFFAOYSA-K 0.000 description 4
- 125000000484 butyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 4
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 4
- 239000012153 distilled water Substances 0.000 description 4
- LEQAOMBKQFMDFZ-UHFFFAOYSA-N glyoxal Chemical compound O=CC=O LEQAOMBKQFMDFZ-UHFFFAOYSA-N 0.000 description 4
- CTSLXHKWHWQRSH-UHFFFAOYSA-N oxalyl chloride Chemical compound ClC(=O)C(Cl)=O CTSLXHKWHWQRSH-UHFFFAOYSA-N 0.000 description 4
- 230000003647 oxidation Effects 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- 210000002700 urine Anatomy 0.000 description 4
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 description 3
- 239000005695 Ammonium acetate Substances 0.000 description 3
- WKBOTKDWSSQWDR-UHFFFAOYSA-N Bromine atom Chemical compound [Br] WKBOTKDWSSQWDR-UHFFFAOYSA-N 0.000 description 3
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 3
- 235000019257 ammonium acetate Nutrition 0.000 description 3
- 229940043376 ammonium acetate Drugs 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- GDTBXPJZTBHREO-UHFFFAOYSA-N bromine Substances BrBr GDTBXPJZTBHREO-UHFFFAOYSA-N 0.000 description 3
- 229910052794 bromium Inorganic materials 0.000 description 3
- 229910052801 chlorine Inorganic materials 0.000 description 3
- 239000000460 chlorine Substances 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N glycerol Substances OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- YVALKTNKAGDHTF-UHFFFAOYSA-N 4-[4,5-bis[4-(diethylamino)phenyl]-1-(methylcarbamoyl)imidazol-2-yl]benzoic acid Chemical compound C1=CC(N(CC)CC)=CC=C1C1=C(C=2C=CC(=CC=2)N(CC)CC)N(C(=O)NC)C(C=2C=CC(=CC=2)C(O)=O)=N1 YVALKTNKAGDHTF-UHFFFAOYSA-N 0.000 description 2
- GOUHYARYYWKXHS-UHFFFAOYSA-N 4-formylbenzoic acid Chemical compound OC(=O)C1=CC=C(C=O)C=C1 GOUHYARYYWKXHS-UHFFFAOYSA-N 0.000 description 2
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 2
- PXGOKWXKJXAPGV-UHFFFAOYSA-N Fluorine Chemical compound FF PXGOKWXKJXAPGV-UHFFFAOYSA-N 0.000 description 2
- 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 2
- 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 2
- WHXSMMKQMYFTQS-UHFFFAOYSA-N Lithium Chemical compound [Li] WHXSMMKQMYFTQS-UHFFFAOYSA-N 0.000 description 2
- 102000010909 Monoamine Oxidase Human genes 0.000 description 2
- 108010062431 Monoamine oxidase Proteins 0.000 description 2
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 2
- 239000003613 bile acid Substances 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 125000004432 carbon atom Chemical group C* 0.000 description 2
- 235000012000 cholesterol Nutrition 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- GGSUCNLOZRCGPQ-UHFFFAOYSA-N diethylaniline Chemical compound CCN(CC)C1=CC=CC=C1 GGSUCNLOZRCGPQ-UHFFFAOYSA-N 0.000 description 2
- DMBHHRLKUKUOEG-UHFFFAOYSA-N diphenylamine Chemical class C=1C=CC=CC=1NC1=CC=CC=C1 DMBHHRLKUKUOEG-UHFFFAOYSA-N 0.000 description 2
- 239000003480 eluent Substances 0.000 description 2
- 125000001301 ethoxy group Chemical group [H]C([H])([H])C([H])([H])O* 0.000 description 2
- 229910052731 fluorine Inorganic materials 0.000 description 2
- 239000011737 fluorine Substances 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 229910052744 lithium Inorganic materials 0.000 description 2
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 2
- HAMGRBXTJNITHG-UHFFFAOYSA-N methyl isocyanate Chemical compound CN=C=O HAMGRBXTJNITHG-UHFFFAOYSA-N 0.000 description 2
- 150000003904 phospholipids Chemical class 0.000 description 2
- 229910052700 potassium Inorganic materials 0.000 description 2
- 239000011591 potassium Substances 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 238000010992 reflux Methods 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 239000012488 sample solution Substances 0.000 description 2
- 239000000741 silica gel Substances 0.000 description 2
- 229910002027 silica gel Inorganic materials 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 230000006641 stabilisation Effects 0.000 description 2
- 238000011105 stabilization Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 2
- AAAQKTZKLRYKHR-UHFFFAOYSA-N triphenylmethane Chemical compound C1=CC=CC=C1C(C=1C=CC=CC=1)C1=CC=CC=C1 AAAQKTZKLRYKHR-UHFFFAOYSA-N 0.000 description 2
- 230000002485 urinary effect Effects 0.000 description 2
- ZTOJFFHGPLIVKC-YAFCTCPESA-N (2e)-3-ethyl-2-[(z)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound S\1C2=CC(S(O)(=O)=O)=CC=C2N(CC)C/1=N/N=C1/SC2=CC(S(O)(=O)=O)=CC=C2N1CC ZTOJFFHGPLIVKC-YAFCTCPESA-N 0.000 description 1
- HSINOMROUCMIEA-FGVHQWLLSA-N (2s,4r)-4-[(3r,5s,6r,7r,8s,9s,10s,13r,14s,17r)-6-ethyl-3,7-dihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1h-cyclopenta[a]phenanthren-17-yl]-2-methylpentanoic acid Chemical compound C([C@@]12C)C[C@@H](O)C[C@H]1[C@@H](CC)[C@@H](O)[C@@H]1[C@@H]2CC[C@]2(C)[C@@H]([C@H](C)C[C@H](C)C(O)=O)CC[C@H]21 HSINOMROUCMIEA-FGVHQWLLSA-N 0.000 description 1
- GEYOCULIXLDCMW-UHFFFAOYSA-N 1,2-phenylenediamine Chemical compound NC1=CC=CC=C1N GEYOCULIXLDCMW-UHFFFAOYSA-N 0.000 description 1
- SXGZJKUKBWWHRA-UHFFFAOYSA-N 2-(N-morpholiniumyl)ethanesulfonate Chemical compound [O-]S(=O)(=O)CC[NH+]1CCOCC1 SXGZJKUKBWWHRA-UHFFFAOYSA-N 0.000 description 1
- PLLCRLNWJDUVLS-UHFFFAOYSA-N 2-[4-(diethylamino)-3-methoxyphenyl]-4,5-bis[4-(diethylamino)-2-methylphenyl]-n-methylimidazole-1-carboxamide Chemical compound CC1=CC(N(CC)CC)=CC=C1C1=C(C=2C(=CC(=CC=2)N(CC)CC)C)N(C(=O)NC)C(C=2C=C(OC)C(N(CC)CC)=CC=2)=N1 PLLCRLNWJDUVLS-UHFFFAOYSA-N 0.000 description 1
- 125000000094 2-phenylethyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])C([H])([H])* 0.000 description 1
- NUIURNJTPRWVAP-UHFFFAOYSA-N 3,3'-Dimethylbenzidine Chemical class C1=C(N)C(C)=CC(C=2C=C(C)C(N)=CC=2)=C1 NUIURNJTPRWVAP-UHFFFAOYSA-N 0.000 description 1
- ABXUVAZZPRIREH-UHFFFAOYSA-N 4,5-bis[4-(diethylamino)-2-methylphenyl]-N-methylimidazole-1-carboxamide Chemical compound CNC(=O)N1C=NC(=C1C1=C(C=C(C=C1)N(CC)CC)C)C1=C(C=C(C=C1)N(CC)CC)C ABXUVAZZPRIREH-UHFFFAOYSA-N 0.000 description 1
- VNYMBBCHHWJOOP-UHFFFAOYSA-N 4-(diethylamino)-3-methoxybenzaldehyde Chemical compound CCN(CC)C1=CC=C(C=O)C=C1OC VNYMBBCHHWJOOP-UHFFFAOYSA-N 0.000 description 1
- ANFPCIQHFZVDNO-UHFFFAOYSA-N 4-[4,5-bis[4-(diethylamino)-2-methylphenyl]-1H-imidazol-2-yl]-N,N-diethyl-2-methoxyaniline Chemical compound CCN(CC)C1=CC(=C(C=C1)C2=C(N=C(N2)C3=CC(=C(C=C3)N(CC)CC)OC)C4=C(C=C(C=C4)N(CC)CC)C)C ANFPCIQHFZVDNO-UHFFFAOYSA-N 0.000 description 1
- BTBUEUYNUDRHOZ-UHFFFAOYSA-N Borate Chemical compound [O-]B([O-])[O-] BTBUEUYNUDRHOZ-UHFFFAOYSA-N 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- 102000003914 Cholinesterases Human genes 0.000 description 1
- 108090000322 Cholinesterases Proteins 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- BDAGIHXWWSANSR-UHFFFAOYSA-N Formic acid Chemical compound OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 1
- 239000006173 Good's buffer Substances 0.000 description 1
- 108010012029 Guanine Deaminase Proteins 0.000 description 1
- 102000013587 Guanine deaminase Human genes 0.000 description 1
- PAYRUJLWNCNPSJ-UHFFFAOYSA-N N-phenyl amine Natural products NC1=CC=CC=C1 PAYRUJLWNCNPSJ-UHFFFAOYSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 108010092464 Urate Oxidase Proteins 0.000 description 1
- ZTOJFFHGPLIVKC-CLFAGFIQSA-N abts Chemical compound S/1C2=CC(S(O)(=O)=O)=CC=C2N(CC)C\1=N\N=C1/SC2=CC(S(O)(=O)=O)=CC=C2N1CC ZTOJFFHGPLIVKC-CLFAGFIQSA-N 0.000 description 1
- 150000001299 aldehydes Chemical class 0.000 description 1
- HFACYLZERDEVSX-UHFFFAOYSA-N benzidine Chemical class C1=CC(N)=CC=C1C1=CC=C(N)C=C1 HFACYLZERDEVSX-UHFFFAOYSA-N 0.000 description 1
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 229940048961 cholinesterase Drugs 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 125000004093 cyano group Chemical group *C#N 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 235000021588 free fatty acids Nutrition 0.000 description 1
- 235000001727 glucose Nutrition 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- MHAJPDPJQMAIIY-UHFFFAOYSA-M hydrogenperoxide(1-) Chemical compound [O-]O MHAJPDPJQMAIIY-UHFFFAOYSA-M 0.000 description 1
- 125000002883 imidazolyl group Chemical group 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- PNDPGZBMCMUPRI-UHFFFAOYSA-N iodine Chemical compound II PNDPGZBMCMUPRI-UHFFFAOYSA-N 0.000 description 1
- 239000012948 isocyanate Substances 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- CIPVVROJHKLHJI-UHFFFAOYSA-N n,n-diethyl-3-methylaniline Chemical compound CCN(CC)C1=CC=CC(C)=C1 CIPVVROJHKLHJI-UHFFFAOYSA-N 0.000 description 1
- INJANJYXXUDIGU-UHFFFAOYSA-N n,n-diethyl-4-(1h-imidazol-2-yl)-3-methylaniline Chemical compound CC1=CC(N(CC)CC)=CC=C1C1=NC=CN1 INJANJYXXUDIGU-UHFFFAOYSA-N 0.000 description 1
- RTVALBYJOGQHDQ-UHFFFAOYSA-N n,n-diethyl-4-(1h-imidazol-2-yl)aniline Chemical compound C1=CC(N(CC)CC)=CC=C1C1=NC=CN1 RTVALBYJOGQHDQ-UHFFFAOYSA-N 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 125000001147 pentyl group Chemical group C(CCCC)* 0.000 description 1
- DGTNSSLYPYDJGL-UHFFFAOYSA-N phenyl isocyanate Chemical compound O=C=NC1=CC=CC=C1 DGTNSSLYPYDJGL-UHFFFAOYSA-N 0.000 description 1
- OJUGVDODNPJEEC-UHFFFAOYSA-N phenylglyoxal Chemical compound O=CC(=O)C1=CC=CC=C1 OJUGVDODNPJEEC-UHFFFAOYSA-N 0.000 description 1
- 125000004344 phenylpropyl group Chemical group 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 210000002706 plastid Anatomy 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 125000002572 propoxy group Chemical group [*]OC([H])([H])C(C([H])([H])[H])([H])[H] 0.000 description 1
- 239000012264 purified product Substances 0.000 description 1
- 238000001953 recrystallisation Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 229960001922 sodium perborate Drugs 0.000 description 1
- YKLJGMBLPUQQOI-UHFFFAOYSA-M sodium;oxidooxy(oxo)borane Chemical compound [Na+].[O-]OB=O YKLJGMBLPUQQOI-UHFFFAOYSA-M 0.000 description 1
- 125000001174 sulfone group Chemical group 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Description
【発明の詳細な説明】 〔発明の利用分野〕 本発明は、新規なイミダゾール誘導体、及び該化合物を
発色成分として用いる酸化性物質の定量方法並びにペル
オキシダーゼ様物質の定量方法に関する。TECHNICAL FIELD The present invention relates to a novel imidazole derivative, a method for quantifying an oxidizing substance and a method for quantifying a peroxidase-like substance using the compound as a color-forming component.
生体成分、例えば血液や尿などの体液成分を測定するこ
とは、その変動が疾病と大きく関連しているため、疾患
の診断、病態の解明、治療経過の判定を行なう上で、必
須なものとなっている。例えば、血液中のコレステロー
ル、トリグリセライド、グルコース、尿酸、リン脂質、
胆汁酸、モノアミンオキシダーゼなどを始め、非常に多
種類の微量成分の測定法が開発されており、疾病の診断
上役立っていることは周知の通りである。The measurement of biological components, such as body fluid components such as blood and urine, is essential for diagnosing diseases, elucidating the pathological condition, and determining the course of treatment, because the fluctuations are greatly related to diseases. Has become. For example, cholesterol in blood, triglyceride, glucose, uric acid, phospholipids,
It is well known that assay methods for very various kinds of trace components such as bile acid and monoamine oxidase have been developed and are useful for diagnosis of diseases.
現在、血清成分の測定法としては、それが酵素以外のも
のである場合には、目的成分に特異的に作用する酵素を
用い、また、目的成分が酵素の場合には、その基質とな
るべき化合物を用いて、夫々酵素反応を行ない、これに
よる生成物を測定して目的成分量を求める、所謂“酵素
法”が一般に広く普及している。なかでも、H2O2生成酵
素、例えば、オキシダーゼを働かせて目的成分に相当す
るH2O2を生成させ、これをペルオキシダーゼ、及び発色
成分である被酸化性呈色試薬を用いて発色系に導き、こ
れを比色定量することにより目的成分量を求める方法
が、被酸化性呈色試薬の開発と相まって増加しつつあ
る。例えば、コレステロール−コレステロールオキシダ
ーゼ、トリグリセライド−リポプロテインリパーゼ−グ
リセーロルオキシダーゼ、尿酸−ウリカーゼなどの組合
せで発生するH2O2を、ペルオキシダーゼ（ＰＯＤ）、被
酸化性呈色試薬を用いて発色系に導き、その呈色の吸光
度を測定することにより目的成分量を求める方法であ
る。この方法に於て用いられる発色成分である被酸化性
呈色試薬の代表的なものとしては、４−アミノアンチピ
リンと、フェノール系化合物又はN,N−ジ置換アニリン
系化合物とを組合せた被酸化性呈色試薬、３−メチルベ
ンゾチアゾリノンヒドラゾン（ＭＢＴＨ）とアニリン系
化合物との組合せ試薬、2,2′−アジノビス（３−エチ
ルベンゾチアゾリン−６−スルホン酸）（ＡＢＴＳ）、
トリフェニルメタン系ロイコ色素、ベンジジン誘導体、
ｏ−トリジン誘導体、ジフェニルアミン誘導体、ｏ−フ
ェニレンジアミン等が挙げられる。しかしながら、これ
ら従来から用いられている被酸化性呈色試薬は、ジフェ
ニルアミン誘導体を除いていずれもその呈色波長が700n
m以下であり、ビリルビン、ヘモグロビン等の血清成分
の影響を受け易く（尿中成分測定時には尿中の色素体の
影響を受け易い）、又、４−アミノアンチピリンとの組
合せ試薬やトリフェニルメタン系ロイコ色素の一部を除
いて、いずれも色原体の安定性が低い等の問題点を有す
る。一方、比較的色原体の安定性が良く、又呈色波長が
比較的長波長側にある色原体として染料前駆体（ロイコ
色素）のトリアリルイミダゾール誘導体が開示されてい
る（特公昭５７−５５１９号公報、特公昭５７−２６１
１８号公報、特開昭５８−４５５５７号公報、米国特許
第３２９７７１０号明細書等）。Currently, as a method for measuring serum components, when it is something other than an enzyme, an enzyme that acts specifically on the target component is used, and when the target component is an enzyme, it should be the substrate The so-called "enzyme method", in which a compound is used to carry out an enzyme reaction with each other, and the resulting product is measured to determine the amount of the target component, is generally widespread. Among them, H 2 O 2 producing enzyme, for example, oxidase is activated to produce H 2 O 2 corresponding to the target component, and this is converted into a coloring system by using peroxidase and an oxidizable coloring reagent which is a coloring component. A method for obtaining the amount of a target component by conducting a colorimetric quantification of this is being increased along with the development of an oxidizable color reagent. For example, H 2 O 2 generated by a combination of cholesterol-cholesterol oxidase, triglyceride-lipoprotein lipase-glycerol oxidase, uric acid-uricase, etc. is converted into a coloring system using peroxidase (POD) and an oxidizable color reagent. It is a method of obtaining the amount of a target component by deriving and measuring the absorbance of the color. A typical example of an oxidizable color reagent that is a color-forming component used in this method is a combination of 4-aminoantipyrine and a phenol compound or an N, N-disubstituted aniline compound. Sex color reagent, combination reagent of 3-methylbenzothiazolinone hydrazone (MBTH) and aniline compound, 2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS),
Triphenylmethane leuco dye, benzidine derivative,
Examples thereof include o-tolidine derivative, diphenylamine derivative, o-phenylenediamine and the like. However, all of these conventionally used oxidizable color reagents have a coloring wavelength of 700 n except for diphenylamine derivatives.
It is less than m and is easily affected by serum components such as bilirubin and hemoglobin (it is easily affected by urinary plastids when measuring urinary components), and is a combination reagent with 4-aminoantipyrine or triphenylmethane Except for some leuco dyes, all have problems such as low stability of chromogen. On the other hand, a triallylimidazole derivative of a dye precursor (leuco dye) is disclosed as a chromogen having relatively good stability of the chromogen and having a coloration wavelength on the relatively long wavelength side (JP-B-57). -5519, Japanese Patent Publication No. 57-261
18, JP-A-58-45557, US Pat. No. 3,297,710, etc.).
しかしながら、これら既存のトリアリルイミダゾール誘
導体は、いずれもそのフェニル基の一つに-OH基を有
し、 となることにより呈色するものであって、その呈色波長
はいずれも依然として700nm以下である。従って、これ
らのトリアリルイミダゾール誘導体にしても、血液や尿
など生体試料中の微量成分の測定に於ける発色成分とし
て用いて未だ充分満足のいくものであるとは云えない。However, these existing triallylimidazole derivatives each have an -OH group in one of the phenyl groups, The coloration wavelength is 700 nm or less. Therefore, it cannot be said that even these triallylimidazole derivatives are still sufficiently satisfactory to be used as color-forming components in the measurement of trace components in biological samples such as blood and urine.
本発明の目的は、上記した如き従来法の問題点を解決し
た、極大吸収波長が700nm以上で、且つ色原体が安定で
ある新規な被酸化性呈色試薬の開発と該化合物を発色成
分として用いることにより、ヘモグロビン、ビリルビン
等有色の共存物質の影響を回避した、精度の高い生体試
料中の微量成分の測定法を実現することにある。The object of the present invention is to solve the problems of the conventional methods as described above, to develop a novel oxidizable color reagent having a maximum absorption wavelength of 700 nm or more, and a stable chromogen, and to develop the compound as a coloring component. It is intended to realize a highly accurate method for measuring a trace component in a biological sample by avoiding the influence of colored coexisting substances such as hemoglobin and bilirubin.
本発明は、下記一般式〔Ｉ〕 ［式中、Ｒ１は低級アルキル基，低級アルコキシ基，ハ
ロゲン原子，ニトロ基，置換基を有していてもよいアミ
ノ基，-SO3H基又は-SO3M1基（但し、Ｍ１はアルカリ金
属イオン又はアンモニウムイオンを表わす。），-COOH
基又は-COOM2基（但し、Ｍ２はアルカリ金属イオン又は
アンモニウムイオンを表わす。）を置換基として有して
いてもよいフェニル基又はアラルキル基を表わし、
Ｒ２，Ｒ３は夫々独立して、少くともそのオルト位又は
パラ位のどちらかが置換基を有していてもよいアミノ基
で置換された置換フェニル基を表わし、Ｒ４はアルキル
置換カルバモイル基、又は置換基を有していてもよいフ
ェニル置換カルバモイル基を表わす。］ で示されるイミダゾール誘導体及び該化合物を発色成分
として用いる酸化性物質並びにペルオキシダーゼ様物質
の定量法である。The present invention has the following general formula [I] [Wherein R 1 is a lower alkyl group, a lower alkoxy group, a halogen atom, a nitro group, an amino group which may have a substituent, a —SO 3 H group or a —SO 3 M 1 group (provided that M 1 Represents an alkali metal ion or ammonium ion), -COOH
Group or a —COOM 2 group (wherein M 2 represents an alkali metal ion or an ammonium ion) represents a phenyl group or an aralkyl group which may have as a substituent,
R 2 and R 3 each independently represent a substituted phenyl group substituted with an amino group which may have a substituent in at least the ortho position or the para position, and R 4 represents an alkyl-substituted carbamoyl group. Or a phenyl-substituted carbamoyl group which may have a substituent. ] The imidazole derivative shown by these, and the quantification method of an oxidizing substance and a peroxidase-like substance which use this compound as a coloring component.
即ち、本発明は上記一般式〔Ｉ〕で示されるイミダゾー
ル誘導体が、いずれもその呈色波長が700nm以上の長波
長側にあり、しかも、色原体として極めて安定であるこ
とを本発明者らが初めて見出し、これを血清や尿など生
体試料中の微量成分の測定に於ける発色成分として用い
ることにより、上記した本発明の目的を達成し得ること
を見出して本発明を完成するに到ったものである。That is, the present inventors have found that the imidazole derivative represented by the above general formula [I] has a coloring wavelength on the long wavelength side of 700 nm or more and is extremely stable as a chromogen. Was found for the first time, and by using this as a color-forming component in the measurement of a trace component in a biological sample such as serum or urine, the inventors have found that the above-mentioned object of the present invention can be achieved and completed the present invention. It is a thing.
一般式〔Ｉ〕で示される本発明のイミダゾール誘導体に
於て、Ｒ１で表わされる置換基を有していてもよいフェ
ニル基の置換基としては、例えば、メチル基，エチル
基，プロピル基，ブチル基，ペンチル基等炭素数１〜５
の低級アルキル基（直鎖状、分枝状のいずれにしても
可。）、例えばメトキシ基，エトキシ基，プロポキシ
基，ブトキシ基等炭素数１〜４の低級アルコキシ基（直
鎖状、分枝状のいずれにても可。）塩素，臭素，弗素，
沃素等のハロゲン原子、ニトロ基、アミノ基、例えばメ
チル基，エチル基，プロピル基，ブチル基等の低級アル
キル基又は例えば-C2H4OH基，-C3H6OH基，-C2H4NHSO2CH
3基，-C2H4NHCOCH3基，-C2H4SO3H基（又は-C2H4SO3Na基
等），-C3H6SO3H基（又は-C3H6SO3Na基等）， （又は 等）等の置換低級アルキル基等で置換されたアミノ基、
-SO3H基又は-SO3M1基（但し、Ｍ１はナトリウム、カリ
ウム、リチウム等のアルカリ金属イオン又はアンモニウ
ムイオンを表わす。）、-COOH基又は-COOM2基（但し、
Ｍ２はナトリウム，カリウム，リチウム等のアルカリ金
属イオン又はアンモニウムイオンを表わす。）等が挙げ
られるが、水酸基はこれに含まれない。又、Ｒ１で表わ
されるアラルキル基としては、例えばベンジル基、フェ
ネチル基、フェニルプロピル基等が挙げられる。Ｒ２，
Ｒ３で表わされる、少なくともそのオルト位又はパラ位
のどちらかが置換基を有していてもよいアミノ基で置換
された置換フェニル基に於ける置換アミノ基の置換基と
しては、例えば、メチル基，エチル基，プロピル基，ブ
チル基等の低級アルキル基、-C2H4OH基，-C3H6OH基，-C
2H4NHSO2CH3基，-C2H4NHCOCH3基，-C2H4SO3H基（又は-C
2H4SO3Na基等），-C3H6SO3H基（又は-C3H6SO3Na基
等）， （又は 基等）等の置換低級アルキル基等が挙げられるが、これ
らに限定されるものではない。又、Ｒ２，Ｒ３に於ける
アミノ基又は置換アミノ基以外の置換基としては、例え
ば、メチル基，エチル基，プロピル基，ブチル基等の低
級アルキル基、メトキシ基，エトキシ基，プロポキシ基
等の低級アルコキシ基、塩素，臭素，弗素，沃素等のハ
ロゲン原子、ニトロ基、シアノ基、カルボキシル基、ス
ルホン基等が挙げられるが、水酸基はこれに含まれな
い。又、Ｒ４で表わされるアルキル置換カルバモイル基
としては、例えばＮ−メチルカルバモイル基，Ｎ−エチ
ルカルバモイル基，Ｎ−イソプロピルカルバモイル基，
N,N−ジメチルカルバモイル基，N,N−ジエチルカルバモ
イル基，N,N−ジイソプロピルカルバモイル基等の低級
アルキル置換カルバモイル基が挙げられ、置換基を有し
ていてもよいフェニル置換カルバモイル基の置換基とし
ては、例えば、メチル基，エチル基，プロピル基等の低
級アルキル基、例えばメトキシ基，エトキシ基等の低級
アルコキシ基、塩素，臭素等のハロゲン原子等が挙げら
れ、これらの置換基を有していてもよいフェニル基で置
換したカルバモイル基が用いられる。又、Ｎ−アルキル
−Ｎ−フェニル−カルバモイル基も同様に用い得ること
は云うまでもない。In the imidazole derivative of the present invention represented by the general formula [I], the substituent of the phenyl group which may have a substituent represented by R 1 is, for example, a methyl group, an ethyl group, a propyl group, Butyl group, pentyl group, etc. 1 to 5 carbon atoms
Lower alkyl groups (both linear and branched) such as methoxy, ethoxy, propoxy, butoxy, etc. lower alkoxy groups having 1 to 4 carbon atoms (linear or branched) Any of the above.) Chlorine, bromine, fluorine,
Halogen atom such as iodine, nitro group, amino group, lower alkyl group such as methyl group, ethyl group, propyl group, butyl group or the like, or -C 2 H 4 OH group, -C 3 H 6 OH group, -C 2 H 4 NHSO 2 CH
3 groups, -C 2 H 4 NHCOCH 3 group, -C 2 H 4 SO 3 H group (or -C 2 H 4 SO 3 Na group, etc.), -C 3 H 6 SO 3 H group (or -C 3 H 6 SO 3 Na group, etc.), (Or Etc.) etc., an amino group substituted with a substituted lower alkyl group etc.,
-SO 3 H group or -SO 3 M 1 group (provided that M 1 represents an alkali metal ion such as sodium, potassium or lithium or ammonium ion), -COOH group or -COOM 2 group (provided that
M 2 represents an alkali metal ion such as sodium, potassium or lithium, or an ammonium ion. ) And the like, but does not include hydroxyl groups. Examples of the aralkyl group represented by R 1 include benzyl group, phenethyl group, phenylpropyl group and the like. R 2 ,
The substituent of the substituted amino group in the substituted phenyl group represented by R 3 which is substituted with an amino group which may have a substituent in at least the ortho position or the para position is, for example, methyl. Group, lower alkyl group such as ethyl group, propyl group, butyl group, -C 2 H 4 OH group, -C 3 H 6 OH group, -C
2 H 4 NHSO 2 CH 3 group, -C 2 H 4 NHCOCH 3 group, -C 2 H 4 SO 3 H group (or -C
2 H 4 SO 3 Na group, etc.), -C 3 H 6 SO 3 H group (or -C 3 H 6 SO 3 Na group, etc.), (Or Groups, etc.), but not limited thereto. The substituents other than the amino group or the substituted amino group in R 2 and R 3 include, for example, lower alkyl groups such as methyl group, ethyl group, propyl group and butyl group, methoxy group, ethoxy group, propoxy group. Examples thereof include lower alkoxy groups such as, halogen atoms such as chlorine, bromine, fluorine, iodine, etc., nitro groups, cyano groups, carboxyl groups, sulfone groups and the like, but hydroxyl groups are not included therein. Examples of the alkyl-substituted carbamoyl group represented by R 4 include N-methylcarbamoyl group, N-ethylcarbamoyl group, N-isopropylcarbamoyl group,
Examples include lower alkyl-substituted carbamoyl groups such as N, N-dimethylcarbamoyl group, N, N-diethylcarbamoyl group, N, N-diisopropylcarbamoyl group, etc. Substituents of phenyl-substituted carbamoyl group which may have a substituent Examples thereof include lower alkyl groups such as methyl group, ethyl group and propyl group, lower alkoxy groups such as methoxy group and ethoxy group, halogen atoms such as chlorine and bromine, and the like. A carbamoyl group substituted with an optionally substituted phenyl group is used. Needless to say, an N-alkyl-N-phenyl-carbamoyl group can be used as well.
一般式〔Ｉ〕で示される本発明のイミダゾール誘導体
は、例えば一般式〔Ｉ〕に於て、 （但し、 は前記、置換基を有していてもよいアミノ基を表わし、
Ｘ１はその他の置換基を表わす。） （但し、 は前記、置換基を有していてもよいアミノ基を表わし、
Ｘ２はその他の置換基を表わす。） とした場合、酸化により次の如き構造の染料を生成す
る。The imidazole derivative of the present invention represented by the general formula [I] is, for example, in the general formula [I]: (However, Represents an amino group which may have a substituent,
X 1 represents another substituent. ) (However, Represents an amino group which may have a substituent,
X 2 represents another substituent. ), A dye having the following structure is formed by oxidation.
又、一般式〔Ｉ〕に於て、 （但し、 は置換基を有していてもよいアミノ基を表わし、Ｘ３は
その他の置換基を表わす。） （但し、 は置換基を有していてもよいアミノ基を表わし、Ｘ１は
その他の置換基を表わす。） （但し、 は置換基を有していてもよいアミノ基を表わし、Ｘ２は
その他の置換基を表わす。） とした場合は酸化により次の如き構造の染料を生成す
る。 Further, in the general formula [I], (However, Represents an amino group which may have a substituent, and X 3 represents another substituent. ) (However, Represents an amino group which may have a substituent, and X 1 represents another substituent. ) (However, Represents an amino group which may have a substituent, and X 2 represents another substituent. ), A dye having the following structure is formed by oxidation.
即ち、いずれにしても、Ｒ４で表わされるアルキル置換
カルバモイル基、又は置換基を有していてもよいフェニ
ル置換カルバモイル基は、酸化反応の際にイミダゾール
基より外れて、例えば過酸化水素で酸化した場合にはRN
HCOOHとなり、酸化により生成した色素とは全く別の存
在となる。即ち、Ｒ４で表わされるアルキル置換カルバ
モイル基、又は置換基を有していてもよいフェニル置換
カルバモイル基は、色原体の安定化（ブランクの安定
化）にのみ関与し、発色には何ら影響を与えない。 That is, in any case, the alkyl-substituted carbamoyl group represented by R 4 or the phenyl-substituted carbamoyl group which may have a substituent is removed from the imidazole group during the oxidation reaction and is oxidized by, for example, hydrogen peroxide. RN if done
It becomes HCOOH, which is completely different from the dye produced by oxidation. That is, the alkyl-substituted carbamoyl group represented by R 4 or the phenyl-substituted carbamoyl group which may have a substituent is involved only in the stabilization of the chromogen (blank stabilization) and has no influence on the color development. Don't give.
一般式〔Ｉ〕で示される本発明のイミダゾール誘導体が
酸化されて生成する上記〔II〕又は〔III〕で示される
色素はいずれも、既存のトリアリルイミダゾール誘導体
に於て が になることにより生ずる色素よりもその呈色波長が更に
長波長側にシフトし、極大吸収波長はいずれも700nm以
上となる。The dyes represented by the above [II] or [III], which are produced by the oxidation of the imidazole derivative of the present invention represented by the general formula [I], are the same as existing triallylimidazole derivatives. But The coloration wavelength of the resulting dye shifts to a longer wavelength side than that of the dye, and the maximum absorption wavelength is 700 nm or more.
表１に、一般式〔Ｉ〕で示される本発明化合物の具体例
数例と、その呈色時の極大吸収波長を示すが、本発明化
合物はこれらに限定されるものではない。Table 1 shows some specific examples of the compound of the present invention represented by the general formula [I] and the maximum absorption wavelength at the time of coloring, but the compound of the present invention is not limited thereto.
一般式〔Ｉ〕で示される本発明化合物は、公知の方法に
より容易に合成することができる。即ち、例えば、Orga
nic Syntheses Vol.5,111頁1973年に記載の方法に準じ
て、式〔IV〕で示されるエタンジオンを合成し、 （式中、Ｒ２，Ｒ３は前記と同じ。） 次いで、これを例えば米国特許第３２９７７１０号明細
書に記載の方法に準じてR1-CHO（Ｒ１は前記と同じ）な
るアルデヒド類及び酢酸アンモンと、酢酸溶媒中数時間
加熱（要すれば還流）反応させれば式〔Ｖ〕で示される
イミダゾール誘導体が得られるから、 （式中、Ｒ１，Ｒ２，Ｒ３は前記と同じ。） 得られたイミダゾール誘導体を適当な溶媒の存在下、室
温乃至溶媒の沸点でイソシアン酸アルキル（又はイソシ
アン酸フェニル）と数時間乃至十数時間反応させれば、
目的物が収率よく得られる。要すればこれを、適当な精
製方法、例えば再結晶、カラムクロマトグラフィー等に
より精製すれば精製品が容易に得られる。 The compound of the present invention represented by the general formula [I] can be easily synthesized by a known method. That is, for example, Orga
nic Syntheses Vol. 5, page 111, according to the method described in 1973, to synthesize the ethanedione represented by the formula [IV], (In the formula, R 2 and R 3 are the same as the above.) Then, according to the method described in, for example, US Pat. No. 3,297,710, aldehydes such as R 1 -CHO (R 1 is the same as above) and When the reaction is carried out with ammonium acetate in an acetic acid solvent for several hours while heating (reflux if necessary), an imidazole derivative represented by the formula [V] can be obtained. (In the formula, R 1 , R 2 and R 3 are the same as above.) The obtained imidazole derivative is treated with an alkyl isocyanate (or phenyl isocyanate) at room temperature to the boiling point of the solvent for several hours to several hours. If you react for more than 10 hours,
The desired product is obtained in good yield. If necessary, a purified product can be easily obtained by purifying it by an appropriate purification method such as recrystallization or column chromatography.
本発明のイミダゾール誘導体は、酸化性物質の定量やペ
ルオキシダーゼ様物質の定量に於ける発色成分として有
効に用い得るが、とりわけ酵素反応により生成した過酸
化水素をペルオキシダーゼの存在下発色系に導き、その
呈色を比色定量することにより行う生体試料中の微量成
分の定量に於ける発色成分として特に有効に使用し得
る。The imidazole derivative of the present invention can be effectively used as a color-forming component in the quantification of oxidative substances and peroxidase-like substances, but especially hydrogen peroxide produced by an enzymatic reaction is led to a chromogenic system in the presence of peroxidase, It can be used particularly effectively as a color-forming component in the quantification of trace components in a biological sample by colorimetrically quantifying coloration.
即ち、本発明の酸化性物質の定量法は、基質、又は酵素
反応により生成した物質に酸化酵素を作用させ、生成す
る過酸化水素を定量することにより行う生体試料中の基
質又は酵素活性の定量法として特に効果的に使用し得
る。That is, the method for quantifying an oxidative substance of the present invention is a quantification of a substrate or an enzyme activity in a biological sample, which is performed by causing an oxidase to act on a substrate or a substance produced by an enzymatic reaction, and quantifying produced hydrogen peroxide. It can be used particularly effectively as a method.
本発明の方法により測定可能な生体試料中の微量成分と
しては、例えば、コレステロール、グルコース、グリセ
リン、トリグリセライド、遊離脂肪酸、尿酸、リン脂
質、胆汁酸、モノアミンオキシダーゼ、グアナーゼ、コ
リンエステラーゼ等が挙げられるが、これらに限定され
るものではなく、酵素反応により生成する過酸化水素を
定量することによって測定が可能な生体成分は全て定量
可能である。Examples of trace components in a biological sample that can be measured by the method of the present invention include cholesterol, glucose, glycerin, triglyceride, free fatty acids, uric acid, phospholipids, bile acids, monoamine oxidase, guanase, and cholinesterase. The present invention is not limited to these, and all biological components that can be measured by quantifying hydrogen peroxide generated by an enzymatic reaction can be quantified.
本発明の方法による生体成分の定量に於て、過酸化水素
を生成させる酵素として用いられる酸化酵素（オキシダ
ーゼ）及びその他の目的で用いられる酵素類並びに酵素
反応に関与する基質及びその他の物質の種類及び使用量
は被酸化性呈色試薬を用いる自体公知の生体成分の定量
法に準じて夫々測定対象となる物質に応じて適宜選択す
ればよい。又、本発明による過酸化水素の定量に於て用
いられるペルオキシダーゼとしては、その起源、由来に
特に限定はなく、植物、動物、微生物起源のペルオキシ
ダーゼ又はペルオキシダーゼ様物質が、一種若しくは要
すれば二種以上組合せて用いられる。又、その使用量は
目的に応じて適宜定められ、特に限定されない。In the determination of biological components by the method of the present invention, oxidase (oxidase) used as an enzyme for producing hydrogen peroxide, enzymes used for other purposes, and types of substrates and other substances involved in the enzymatic reaction The amount and the amount to be used may be appropriately selected according to the substance to be measured in accordance with a known method for quantifying biological components using an oxidizable color reagent. Further, the peroxidase used in the determination of hydrogen peroxide according to the present invention is not particularly limited in its origin and origin, and peroxidase or peroxidase-like substance of plant, animal or microbial origin may be used in one kind or in two kinds if necessary. The above is used in combination. The amount used is appropriately determined according to the purpose and is not particularly limited.
本発明の方法による生体成分の定量は、通常、pH4.0〜1
0.0、より好ましくはpH6.0〜8.0で実施される。用いら
れる緩衝剤としては、リン酸塩、クエン酸塩、ホウ酸
塩、炭酸塩、トリス緩衝液、グッド（Good's）緩衝液な
どが挙げられるが、特にこれらに限定されない。The quantification of biological components by the method of the present invention is usually pH 4.0-1.
It is carried out at 0.0, more preferably pH 6.0 to 8.0. Examples of the buffer used include phosphate, citrate, borate, carbonate, Tris buffer, Good's buffer and the like, but are not particularly limited thereto.
本発明のイミダゾール誘導体は、過酸化水素等酸化性物
質の定量に有効に用い得るが、又、これと過酸化水素と
を組み合せることによりペルオキシダーゼ様物質の定量
を行うことも可能である。ペルオキシダーゼ様物質とし
ては、ペルオキシダーゼそのものの他、ヘモグロビンそ
の他のヘム化合物が挙げられる。The imidazole derivative of the present invention can be effectively used for quantifying oxidative substances such as hydrogen peroxide, but it is also possible to quantify a peroxidase-like substance by combining this with hydrogen peroxide. Examples of the peroxidase-like substance include hemoglobin and other heme compounds, in addition to peroxidase itself.
即ち、本発明のイミダゾール誘導体は、例えば、ペルオ
キシダーゼを標識化合物に用いた酵素免疫測定法にも応
用可能であり、又、血清中のヘモグロビンを過酸化水素
若しくは過硼素酸ナトリウムのような酸化性物質を用い
て測定する場合などにも有効に使用し得る。That is, the imidazole derivative of the present invention can be applied to, for example, an enzyme immunoassay using peroxidase as a labeling compound, and hemoglobin in serum can be oxidized with hydrogen peroxide or an oxidizing substance such as sodium perborate. It can also be used effectively when measuring using.
以下に実施例を挙げるが、本発明はこれら実施例により
何ら制約を受けるものではない。Examples will be given below, but the present invention is not limited by these examples.
実施例１． ２−（４−カルボキシフェニル）−３−Ｎ−メチルカル
バモイル−４，５−ビス（４−ジエチルアミノフェニ
ル）イミダゾール〔本発明化合物(2)〕の合成 （ｉ）１，２−ビス（４−ジエチルアミノフェニル）エ
タン−１，２−ジオンの合成 無水塩化アルミニウム6.7ｇに二硫化炭素20mlを加え氷
冷下、N,N−ジエチルアニリン20ｇを滴下した。さらに
撹拌下、氷冷しながらオキザリルクロリド1.5ｇを滴下
し、60分間撹拌反応させた。反応後、水50ml及びクロロ
ホルム100mlを加え、分液して得たクロロホルム層を減
圧濃縮して結晶を析出せしめた。析出した結晶を取し
酢酸エチルから再結晶して黄色の目的物5.0ｇを得た。Example 1. Synthesis of 2- (4-carboxyphenyl) -3-N-methylcarbamoyl-4,5-bis (4-diethylaminophenyl) imidazole [the present compound (2)] (i) 1,2-bis (4-diethylamino) Synthesis of (phenyl) ethane-1,2-dione 20 ml of carbon disulfide was added to 6.7 g of anhydrous aluminum chloride, and 20 g of N, N-diethylaniline was added dropwise under ice cooling. Further, 1.5 g of oxalyl chloride was added dropwise with stirring under ice-cooling, and the reaction was stirred for 60 minutes. After the reaction, 50 ml of water and 100 ml of chloroform were added, and the chloroform layer obtained by liquid separation was concentrated under reduced pressure to precipitate crystals. The precipitated crystals were collected and recrystallized from ethyl acetate to obtain 5.0 g of a yellow target product.
（ii）２−（４−カルボキシフェニル）−４，５−ビス
（４−ジエチルアミノフェニル）イミダゾールの合成 （ｉ）で得た１，２−ビス（４−ジエチルアミノフェニ
ル）エタン−１，２−ジオン1.5ｇとｐ−カルボキシベ
ンズアルデヒド0.7ｇ、酢酸アンモニウム５ｇを酢酸30m
l中で２時間加熱還流して反応させた。冷却後、水60ml
を加え氷冷下アンモニア水で中和したところ結晶が析出
した。析出した結晶を取、水洗、乾燥後、ヘキサンで
処理し白色の目的化合物1.1ｇを得た。(Ii) Synthesis of 2- (4-carboxyphenyl) -4,5-bis (4-diethylaminophenyl) imidazole 1,2-bis (4-diethylaminophenyl) ethane-1,2-dione obtained in (i) 1.5 g of p-carboxybenzaldehyde 0.7 g of ammonium acetate 5 g of acetic acid 30 m
The mixture was heated under reflux for 2 hours in 1 to react. After cooling, 60 ml of water
When the mixture was added and neutralized with aqueous ammonia under ice cooling, crystals were precipitated. The precipitated crystals were collected, washed with water, dried and treated with hexane to obtain 1.1 g of the white target compound.
（iii）２−（４−カルボキシフェニル）−３−Ｎ−メ
チルカルバモイル−４，５−ビス（４−ジエチルアミノ
フェニル）イミダゾールの合成 （ii）で得た２−（４−カルボキシフェニル）−４，５
−ビス（４−ジエチルアミノフェニル）イミダゾール1.
1ｇをクロロホルム50mlに溶解し、撹拌下、イソシアン
酸メチル５mlを加え室温で12時間放置した。次いで、メ
タノール50mlを加え反応を停止後濃縮した。濃縮物を、
クロロホルムを溶離液に用いたシリカゲルカラムで精製
し、目的物（殆白色粉末）980mgを得た。(Iii) Synthesis of 2- (4-carboxyphenyl) -3-N-methylcarbamoyl-4,5-bis (4-diethylaminophenyl) imidazole 2- (4-carboxyphenyl) -4, obtained in (ii) 5
-Bis (4-diethylaminophenyl) imidazole 1.
1 g was dissolved in 50 ml of chloroform, 5 ml of methyl isocyanate was added with stirring, and the mixture was left at room temperature for 12 hours. Then, 50 ml of methanol was added to stop the reaction and then concentrated. Concentrate
Purification with a silica gel column using chloroform as an eluent gave 980 mg of the desired product (almost white powder).
ＮＭＲ（CDCl3，ＴＭＳ）ppm：1.2(12H,t,CH3-C-）、1.
9(1H,s, 2.7(3H,s,-N-CH3)、3.6(8H,q.C-CH2-)、7.1〜7.7(12H,b
road,フェニル水素)、11.8(1H,s,-COOH) 実施例２． ２−（３−メトキシ−４−ジエチルアミノフェニル）−
３−Ｎ−メチルカルバモイル−４，５−ビス（２−メチ
ル−４−ジエチルアミノフェニル）イミダゾール〔本発
明化合物(10)〕の合成 （ｉ）１，２−ビス（２−メチル−４−ジエチルアミノ
フェニル）エタン−１，２−ジオンの合成 無水塩化アルミニウム6.7ｇに二硫化炭素20mlを加え氷
冷下、３−メチル−N,N−ジエチルアニリン22ｇを滴下
した。さらに撹拌下、氷冷しながらオキザリルクロリド
1.5ｇを滴下し、60分間撹拌反応させた。反応後、水50m
l及びクロロホルム100mlを加え、分液して得たクロロホ
ルム層を減圧濃縮して結晶を析出せしめた。析出した結
晶を取し酢酸エチルから再結晶して黄色の目的物5.4
ｇを得た。NMR (CDCl 3 , TMS) ppm: 1.2 (12H, t, CH 3 -C-), 1.
9 (1H, s, 2.7 (3H, s, -N-CH 3 ), 3.6 (8H, qC-CH 2- ), 7.1 ~ 7.7 (12H, b
road, phenyl hydrogen), 11.8 (1H, s, -COOH) Example 2. 2- (3-methoxy-4-diethylaminophenyl)-
Synthesis of 3-N-methylcarbamoyl-4,5-bis (2-methyl-4-diethylaminophenyl) imidazole [the present compound (10)] (i) 1,2-bis (2-methyl-4-diethylaminophenyl) ) Synthesis of ethane-1,2-dione 20 ml of carbon disulfide was added to 6.7 g of anhydrous aluminum chloride, and 22 g of 3-methyl-N, N-diethylaniline was added dropwise under ice cooling. While further stirring and cooling with ice, oxalyl chloride
1.5 g was added dropwise, and the reaction was stirred for 60 minutes. After the reaction, water 50m
l and chloroform (100 ml) were added, and the layers were separated and the resulting chloroform layer was concentrated under reduced pressure to precipitate crystals. The precipitated crystals are collected and recrystallized from ethyl acetate to give the yellow target compound 5.4.
g was obtained.
（ii）２−（３−メトキシ−４−ジエチルアミノフェニ
ル）−４，５−ビス（２−メチル−４−ジエチルアミノ
フェニル）イミダゾールの合成 （ｉ）で得た１，２−ビス（２−メチル−４−ジエチル
アミノフェニル）エタン−１，２−ジオン1.5ｇと３−
メトキシ−４−ジエチルアミノベンズアルデヒド1.0
ｇ、酢酸アンモニウム５ｇを酢酸30ml中で２時間加熱還
流して反応させた。冷却後、水60mlを加え氷冷下アンモ
ニア水で中和したところ結晶が析出した。析出した結晶
を取、水洗、乾燥後、ヘキサンで処理し白色の目的化
合物0.6ｇを得た。(Ii) Synthesis of 2- (3-methoxy-4-diethylaminophenyl) -4,5-bis (2-methyl-4-diethylaminophenyl) imidazole 1,2-bis (2-methyl-) obtained in (i) 4-diethylaminophenyl) ethane-1,2-dione 1.5 g and 3-
Methoxy-4-diethylaminobenzaldehyde 1.0
g and 5 g of ammonium acetate were heated and refluxed in 30 ml of acetic acid for 2 hours to react. After cooling, 60 ml of water was added and neutralized with aqueous ammonia under ice cooling to precipitate crystals. The precipitated crystals were collected, washed with water, dried and treated with hexane to obtain 0.6 g of the white target compound.
（iii）２−（３−メトキシ−４−ジエチルアミノフェ
ニル）−３−Ｎ−メチルカルバモイル−４，５−ビス
（２−メチル−４−ジエチルアミノフェニル）イミダゾ
ールの合成 （ii）で得た２−（３−メトキシ−４−ジエチルアミノ
フェニル）−４，５−ビス（２−メチル−４−ジエチル
アミノフェニル）イミダゾール0.6ｇをクロロホルム50m
lに溶解し、撹拌下、イソシアン酸メチル５mlを加え室
温で12時間放置した。次いで、メタノール50ｍを加え
反応を停止後濃縮した。濃縮物を、クロロホルムを溶離
液に用いたシリカゲルカラムで精製し、目的物（殆白色
粉末）570mgを得た。(Iii) Synthesis of 2- (3-methoxy-4-diethylaminophenyl) -3-N-methylcarbamoyl-4,5-bis (2-methyl-4-diethylaminophenyl) imidazole 2- (2-) obtained in (ii) Chloroform 50 m of 0.6 g of 3-methoxy-4-diethylaminophenyl) -4,5-bis (2-methyl-4-diethylaminophenyl) imidazole
It was dissolved in 1 l, 5 ml of methyl isocyanate was added with stirring, and the mixture was left at room temperature for 12 hours. Then, 50 m of methanol was added to stop the reaction and then concentrated. The concentrate was purified by a silica gel column using chloroform as an eluent to obtain 570 mg of the desired product (almost white powder).
実施例３〜１０ 実施例１又は実施例２と同様の方法により表２(1)〜表
２(2)に記載の本発明化合物を合成した。その構造式と
物性データを表２(1)〜表２(2)に併せて示す。 Examples 3 to 10 The compounds of the present invention shown in Table 2 (1) to Table 2 (2) were synthesized by the same method as in Example 1 or Example 2. The structural formula and physical property data are also shown in Tables 2 (1) and 2 (2).
実施例１１ 尿酸の定量 (1)試薬 50mmol/lＭＥＳ〔２−（Ｎ−モリホリノ）エ
タンスルホン酸〕緩衝液（pH6.5）にウリカーゼ100U/
l、ペルオキシダーゼ2,000U/l、本発明化合物(2)100μm
ol/lの濃度になるように溶解し調製した。 Example 11 Determination of uric acid (1) Reagent 50 mmol / l MES [2- (N-morpholino) ethanesulfonic acid] buffer (pH 6.5) with uricase 100 U /
l, peroxidase 2,000 U / l, compound of the present invention (2) 100 μm
It was prepared by dissolving so as to have a concentration of ol / l.
(2)試料 尿酸を蒸留水で10,7.5,5,2.5mg/dlになるよう
に溶解し調製した。(2) Sample Uric acid was prepared by dissolving it in distilled water to give concentrations of 10,7.5,5,2.5 mg / dl.
(3)測定操作 各試料液及び蒸留水各50μｌに試薬3.0ml
を加え、37℃で５分間加温し740nmの吸光度を盲検を対
照に測定した。(3) Measurement operation 3.0 ml of reagent for each 50 μl of each sample solution and distilled water
Was added and the mixture was heated at 37 ° C. for 5 minutes, and the absorbance at 740 nm was measured using a blind test as a control.
第１図に尿酸濃度と吸光度との関係を示す。第１図より
明らかな如く、各尿酸濃度に対してプロットした吸光度
を結ぶ検量線は原点を通る直線となり、検量線は良好な
定量性を示している。FIG. 1 shows the relationship between uric acid concentration and absorbance. As is clear from FIG. 1, the calibration curve connecting the absorbances plotted for each uric acid concentration is a straight line passing through the origin, and the calibration curve shows good quantification.
実施例１２ 過酸化水素の定量 (1)試薬 50mmol/lリン酸緩衝液（pH7.0）にペルオキシ
ダーゼ2,000U/l、本発明化合物(2)100μmol/lになるよ
うに溶解し調製した。Example 12 Quantification of hydrogen peroxide (1) Reagent was prepared by dissolving it in 50 mmol / l phosphate buffer (pH 7.0) so that peroxidase 2,000 U / l and the compound (2) of the present invention 100 μmol / l.
(2)試料 市販過酸化水素水を蒸留水で希釈し2.0,1.5,
1.0,0.5mmol/lになるように溶解し調製した。(2) Sample Dilute commercially available hydrogen peroxide solution with distilled water to 2.0, 1.5,
It was prepared by dissolving so as to be 1.0, 0.5 mmol / l.
(3)測定操作 各試料液及び蒸留水各20μｌに試薬3.0ml
を加え、37℃で３分間加温し740nmの吸光度を盲検を対
照に測定した。(3) Measurement operation 3.0 ml of reagent for each 20 μl of each sample solution and distilled water
Was added, and the mixture was heated at 37 ° C. for 3 minutes, and the absorbance at 740 nm was measured using a blind test as a control.
第２図に過酸化水素濃度と吸光度との関係を示す。第２
図より明らかな如く、各過酸化水素濃度に対してプロッ
トした吸光度を結ぶ検量線は原点を通る直線となり、検
量線は良好な定量性を示している。FIG. 2 shows the relationship between hydrogen peroxide concentration and absorbance. Second
As is clear from the figure, the calibration curve connecting the absorbances plotted for each hydrogen peroxide concentration is a straight line passing through the origin, and the calibration curve shows good quantification.
以上述べた如く、本発明の新規イミダゾール誘導体は、
いずれもその呈色時の極大吸収波長が、700nm以上と長
波長側にある為、血清，尿等生体試料中の微量成分の定
量に於ける発色成分としてこれを用いた場合には、試料
中に共存する有色の妨害物質の影響を全く受けずに測定
を行うことができるという点、及びイミダゾール環の３
位のＮに置換カルバモイル基をつけたことにより色原体
として安定化されたイミダゾール誘導体となり得た点に
顕著な効果を奏するものであり、斯業に貢献するところ
大なるものである。As described above, the novel imidazole derivative of the present invention is
In both cases, the maximum absorption wavelength at the time of color development is 700 nm or longer, which is on the long wavelength side. Therefore, when it is used as a color-forming component in the determination of trace components in biological samples such as serum and urine, That the measurement can be performed without being affected by the colored interfering substances coexisting in the
It has a remarkable effect in that it can be an imidazole derivative which is stabilized as a chromogen by adding a substituted carbamoyl group to N at position N, which is a great contribution to the art.
第１図は、実施例１１に於て得られた検量線を表わし、
横軸の各尿酸濃度（mg/dl）について得られた吸光度を
縦軸に沿ってプロットした点を結んだものである。 第２図は、実施例１２に於て得られた検量線を表わし、
横軸の各過酸化水素濃度（mmol/l）について得られた吸
光度を縦軸に沿ってプロットした点を結んだものであ
る。FIG. 1 shows the calibration curve obtained in Example 11,
The absorbance obtained for each uric acid concentration (mg / dl) on the horizontal axis is plotted along the vertical axis. FIG. 2 shows the calibration curve obtained in Example 12,
The abscissa represents the absorbance obtained for each hydrogen peroxide concentration (mmol / l), and the points plotted along the ordinate are connected.
───────────────────────────────────────────────────── フロントページの続き (56)参考文献 特開 昭61−176572（ＪＰ，Ａ) Ｃｈｅｍ，Ａｂ，74（22）要約番号 119391Ｗ（1971) Ｃｈｅｍ，Ａｂ，72（17）要約番号 89636Ｕ（1970) ─────────────────────────────────────────────────── ─── Continuation of the front page (56) Reference JP-A-61-176572 (JP, A) Chem, Ab, 74 (22) Abstract number 119391W (1971) Chem, Ab, 72 (17) Abstract number 89636U (1970) )
Claims (10)
ロゲン原子，ニトロ基，置換基を有していてもよいアミ
ノ基，-SO3H基又は-SO3M1基（但し、Ｍ１はアルカリ金
属イオン又はアンモニウムイオンを表わす。），-COOH
基又は-COOM2基（但し、Ｍ２はアルカリ金属イオン又は
アンモニウムイオンを表わす。）を置換基として有して
いてもよいフェニル基又はアラルキル基を表わし、
Ｒ２，Ｒ３は夫々独立して、少くともそのオルト位又は
パラ位のどちらかが置換基を有していてもよいアミノ基
で置換された置換フェニル基を表わし、Ｒ４はアルキル
置換カルバモイル基、又は置換基を有していてもよいフ
ェニル置換カルバモイル基を表わす。］ で示されるイミダゾール誘導体。1. The following general formula [I] [Wherein R 1 is a lower alkyl group, a lower alkoxy group, a halogen atom, a nitro group, an amino group which may have a substituent, a —SO 3 H group or a —SO 3 M 1 group (provided that M 1 Represents an alkali metal ion or ammonium ion), -COOH
Group or a —COOM 2 group (wherein M 2 represents an alkali metal ion or an ammonium ion) represents a phenyl group or an aralkyl group which may have as a substituent,
R 2 and R 3 each independently represent a substituted phenyl group substituted with an amino group which may have a substituent in at least the ortho position or the para position, and R 4 represents an alkyl-substituted carbamoyl group. Or a phenyl-substituted carbamoyl group which may have a substituent. ] The imidazole derivative shown by these.
ロゲン原子，ニトロ基，置換基を有していてもよいアミ
ノ基，-SO3H基又は-SO3M1基（但し、Ｍ１はアルカリ金
属イオン又はアンモニウムイオンを表わす。），-COOH
基又は-COOM2基（但し、Ｍ２はアルカリ金属イオン又は
アンモニウムイオンを表わす。）を置換基として有して
いてもよいフェニル基又はアラルキル基を表わし、
Ｒ２，Ｒ３は夫々独立して、少くともそのオルト位又は
パラ位のどちらかが置換基を有していてもよいアミノ基
で置換された置換フェニル基を表わし、Ｒ４はアルキル
置換カルバモイル基、又は置換基を有していてもよいフ
ェニル置換カルバモイル基を表わす。］ で示されるイミダゾール誘導体を発色成分として用いる
ことを特徴とする酸化性物質の定量法。2. The following general formula [I] [Wherein R 1 is a lower alkyl group, a lower alkoxy group, a halogen atom, a nitro group, an amino group which may have a substituent, a —SO 3 H group or a —SO 3 M 1 group (provided that M 1 Represents an alkali metal ion or ammonium ion), -COOH
Group or a —COOM 2 group (wherein M 2 represents an alkali metal ion or an ammonium ion) represents a phenyl group or an aralkyl group which may have as a substituent,
R 2 and R 3 each independently represent a substituted phenyl group substituted with an amino group which may have a substituent in at least the ortho position or the para position, and R 4 represents an alkyl-substituted carbamoyl group. Or a phenyl-substituted carbamoyl group which may have a substituent. ] The quantification method of the oxidizing substance characterized by using the imidazole derivative shown by these as a coloring component.
の範囲第２項記載の定量法。3. The quantification method according to claim 2, wherein the oxidizing substance is hydrogen peroxide.
化発色させてその呈色を比色定量する特許請求の範囲第
３項記載の定量法。4. The quantitative method according to claim 3, wherein the color-forming component is subjected to oxidative coloration in the presence of peroxidase to colorimetrically quantify the color development.
酸化水素である特許請求の範囲第３項又は第４項記載の
定量法。5. The quantitative method according to claim 3 or 4, wherein the hydrogen peroxide is hydrogen peroxide produced by an enzymatic reaction.
量に於て酵素反応により生成する過酸化水素である特許
請求の範囲第５項記載の定量法。6. The quantification method according to claim 5, wherein the hydrogen peroxide is hydrogen peroxide produced by an enzymatic reaction in the quantification of a trace component in a biological sample.
は酵素反応により生成した物質に酸化酵素を作用させ生
成する過酸化水素を定量することにより行う生体試料中
の基質又は酵素活性の定量である特許請求の範囲第６項
記載の定量法。7. A method for quantifying a trace component in a biological sample by quantifying a substrate or a hydrogen peroxide produced by reacting an oxidase with a substance produced by an enzymatic reaction to determine a substrate or enzyme activity in the biological sample. The quantitative method according to claim 6, which is quantitative.
ロゲン原子，ニトロ基，置換基を有していてもよいアミ
ノ基，-SO3H基又は-SO3M1基（但し、Ｍ１はアルカリ金
属イオン又はアンモニウムイオンを表わす。），-COOH
基又は-COOM2基（但し、Ｍ２はアルカリ金属イオン又は
アンモニウムイオンを表わす。）を置換基として有して
いてもよいフェニル基又はアラルキル基を表わし、
Ｒ２，Ｒ３は夫々独立して、少くともそのオルト位又は
パラ位のどちらかが置換基を有していてもよいアミノ基
で置換された置換フェニル基を表わし、Ｒ４はアルキル
置換カルバモイル基、又は置換基を有していてもよいフ
ェニル置換カルバモイル基を表わす。］ で示されるイミダゾール誘導体を発色成分として用いる
ことを特徴とするペルオキシダーゼ様物質の定量法。8. The following general formula [I] [Wherein R 1 is a lower alkyl group, a lower alkoxy group, a halogen atom, a nitro group, an amino group which may have a substituent, a —SO 3 H group or a —SO 3 M 1 group (provided that M 1 Represents an alkali metal ion or ammonium ion), -COOH
Group or a —COOM 2 group (wherein M 2 represents an alkali metal ion or an ammonium ion) represents a phenyl group or an aralkyl group which may have as a substituent,
R 2 and R 3 each independently represent a substituted phenyl group substituted with an amino group which may have a substituent in at least the ortho position or the para position, and R 4 represents an alkyl-substituted carbamoyl group. Or a phenyl-substituted carbamoyl group which may have a substituent. ] The imidazole derivative shown by these is used as a coloring component, The quantitative method of the peroxidase-like substance characterized by the above-mentioned.
ゼである特許請求の範囲第８項記載の定量法。9. The assay method according to claim 8, wherein the peroxidase-like substance is peroxidase.
又はその他のヘム化合物である特許請求の範囲第８項記
載の定量法。10. The quantification method according to claim 8, wherein the peroxidase-like substance is hemoglobin or another heme compound.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP60068315A JPH0625146B2 (en) | 1985-03-30 | 1985-03-30 | Novel imidazole derivative and measuring method using the same as a coloring component |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP60068315A JPH0625146B2 (en) | 1985-03-30 | 1985-03-30 | Novel imidazole derivative and measuring method using the same as a coloring component |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS61227570A JPS61227570A (en) | 1986-10-09 |
| JPH0625146B2 true JPH0625146B2 (en) | 1994-04-06 |
Family
ID=13370261
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP60068315A Expired - Lifetime JPH0625146B2 (en) | 1985-03-30 | 1985-03-30 | Novel imidazole derivative and measuring method using the same as a coloring component |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0625146B2 (en) |
-
1985
- 1985-03-30 JP JP60068315A patent/JPH0625146B2/en not_active Expired - Lifetime
Non-Patent Citations (2)
| Title |
|---|
| Ｃｈｅｍ，Ａｂ，７２（１７）要約番号８９６３６Ｕ（１９７０） |
| Ｃｈｅｍ，Ａｂ，７４（２２）要約番号１１９３９１Ｗ（１９７１） |
Also Published As
| Publication number | Publication date |
|---|---|
| JPS61227570A (en) | 1986-10-09 |
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