• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    Beansprucht werden Glycoside von Resorufin-Derivater der allgemeinen Formeln la bzw lb in denen R1 Wasserstoff, R2, R3 und R5, die gleich oder verschieden sein können, Wasserstoff, Halogen oder eine Niederalkylgruppe, R4 und R6, die gleich oder verschieden sein können, Wasserstoff, Halogen, eine Cyano-, Niederalkyl-, Niederalkoxy-, Carboxy-, Niederalkoxycarbonyl-, Carboxyniederalkyl-, Niederalkoxycarbonyl-niederalkyl- oder eine gegebenenfalls ein-oder zweifach substituierte Carboxamidogruppe oder die Gruppe in der R7 Wasserstoff oder eine Niederalkyl-Gruppe und n eine ganze Zahl von 1 bis 4 bedeuten, wobei R6 zusätzlich eine Sulfonyl oder Nitrogruppe vorstellen kann, und Y ein Stickstoff oder die Gruppe N O bedeuten. Diese Verbindungen eignen sich besonders als chromogene und fluorogene Substrate zur Bestimmung der Aktivität von entsprechenden Glycosidasen bzw. Oligosaccharid spaltenden Enzymen. Die beanspruchten Glycoside erhält man, wenn Resorufin-Derivate mit einem Mono- oder Oligosaccharid oder einem 1-Halogeno-Derivat hiervon, deren Hydroxygruppen mit üblichen Schutzgruppen substituiert sind, umgesezt werden. Zunächst werden dabei per-0-substituierte Glycoside erhalten, die durch Abspalten der Schutzgruppen in die beanspruchten Verbindungen übergeführt werden können. Ein weiterer Gegenstand der Anmeldung sind diagnostische Mittel zum Nachweis von Glycosidasen bzw. von Oligosaccharid spaltenden Enzymen, die als chromogene und fluorogene Substrate die beanspruchten Glycoside enthalten. Beansprucht werden femer die als Ausgangsprodukte bei der Herstellung der erfindungsgemäßen Verbindungen eingesetzten Resarufin-Derivate.
    公开号:SU1574173A3
    申请号:SU864027681
    申请日:1986-06-24
    公开日:1990-06-23
    发明作者:Кляйн Кристиан；Батц Ханс-Георг；Сернетц Манфред；Хофманн Юрген
    申请人:Берингер Маннхайм Гмбх (Фирма)；
    IPC主号:
    专利说明:

    This invention relates to a process for the preparation of new derivatives of resazurin or resorufin in the form of tautomers, which are the starting compounds for the preparation of new derivatives of glycoside-containing resazurin or
    resorufin, which are used as an agent for the detection of glycosidases.
    The aim of the invention is the creation of new intermediates for the synthesis
     about
    new indicator enzymes for the detection of glycosidases based on derivatives of glycoside containing reaaurin or resorufin with increased efficiency.
    Example 1. Resorufin-1-carboxylic acid methyl ester.
    5b g of nitrosorezorcin, 53.3 g of methyl ester of 3,5-dioxybenzoic acid and 28.0 g of pyrolusite are mixed with 500 ml of methanol. Under ice cooling at C, 34.3 ml of concentrated sulfuric acid are added dropwise. After removal of the ice bath, continue stirring for 2 hours. Then, with cooling, add 200 ml of an aqueous solution of ammonia. The precipitate formed is filtered through a glass filter. To the filtrate 10 g of zinc powder is added in portions at 5-10 ° C. Stir at room temperature until the completion of the reduction process (monitored by thin layer chromatography, ethyl acetate: methanol 4: 1 as solvent, reaction time 1.5 h). On a rotary evaporator at a bath temperature of 25 ° C concentrate to 1/3 of the volume. By acidifying with concentrated hydrochloric acid to color change and drying in vacuum over calcium chloride, resorufin-1-carboxylic acid methyl ester is obtained. The output of 30.9 g (30% of theory). (DMSO), a: 3.98 (singlet, 3N), 6, (doublet, J 2.2 Hz, 1H) {6.85-6.95 (multiplet, 2HJ; 7.09 (doublet, J "2 , 2 Hz, 1H); 7.60 (doublet, J 9.6 Hz
    1H).
    Fluorescence at absorbance max L70 nm, with emission dxs 588 nm.
    Similarly receive the following derivatives of resorufin:
    ResoruFin-1-carboxylic acid with PP (visible region, 0.1 m potassium phosphate buffer, pH 7.5) L ma) s - 569 nm;
    methyl eFir-4-methoxyresorufin-1-carboxylic acid with UV (visible region, 0.1 F potassium phosphate buffer, pH 7.5) 7k mako 592 nm, methyl ester 8-bromo-4-methoxy-resor-Fin 1-carboxylic acid with UV (visible region, 0.1 M potassium phosphate buffer, pH 7.5) / XMOIKC 569 HM
    methyl 8-bromo-4-methoxy resorufin-t-carboxylic acid with UV
    ,
    JQ 15 - 20. 25 30 rd c,
    40
    - MJ
    5741734
    (visible region, 0.1 N potassium-Phosphate buffer, pH 7, 5) l, ax 588 nm
    1-nitroresorufin with UV (visible region, 0.1 V potassium-phosphate buffer, pH 7.5) L max 578 nm,
    Resorufin-1-sulfonic acid with UV (visible region, 0.1 M potassium phosphate buffer, pH 7, 5) 7i M0se 570 nm;
    1-methylresorufin-4-carboxylic acid with UV (visible region, 0.2 M potassium phosphate buffer, pH 7.5) = 571 nm1,
    morFolidresorufin-4-carboxylic acid with UV (visible region, 0.1 M potassium phosphate buffer, pH 7.7) is 575 nm.
    NMR-n (- DMSO), o: 3.3-3.8 (multiplet, 8H); 6.50 (doublet, J 2 Hz, 1H), 6.64 (doublet, J 10 Hz, 1H); 6.76 (double doublet, J 10 and 2 Hz, 1H); 7.44 and 7.51 (doublet each time, J 10 Hz, 2H).
    Example 2. Resorufin-4-carboxylic acid.
    1.60 g (10.5 mmol) of nitrosorezhorescin, 1.55 g (10.0 mmol) of 2,6-dioxibenzoic acid and 0.86 g (10 mmol)
    35
    45
    50
    55
    pyrolusite is taken up in 20 ml of methanol and cooled to 0 ° C. To the mixture obtained in this way is 1.06 ml of concentrated sulfuric acid is added dropwise. Continue to stir for another 2 hours without refrigeration. The precipitated red color is filtered off, washed with methanol and dried. The output of 2.3 g (85% of theory). UV (visible region, 0.1 M potassium phosphate buffer, pH 7.5) max 614 nm (5 48 cm), after 2 acidification max 522 nm (Ј 32 cm x x).
    Example 3. Resorufin-4-carboxylic acid.
    2.3 g of resorufin-4-carboxylic acid obtained in Example 2 is dissolved in 20 ml of water and 5 ml of 25% ammonia. While cooling with ice, 5 g of zinc dust are added to the blue solution, and then the ice-cooling is removed, so that the solution is gradually heated to room temperature. Recovery can be easily monitored by changing color from blue to dark purple and using thin layer chromatography (solvent: methanol / ethyl acetate, 1: 1). Excess zinc powder is filtered off. The reaction solution is acidified.
    5 ml of glacial acetic acid and concentrated hydrochloric acid. The precipitated product is filtered, washed with dilute hydrochloric acid and dried in vacuo over calcium chloride. Output 1.8 g (82% of theory), UV (visible region, 0.1 In potassium phosphate buffer, pH 7.5): A max 579.4 nm (Ј 48.6 cm mol 1);
    after acidification, max 485.9 nm (Ј 34, 7 cm2).
    Example 4. 8-Chlororesazurin-4-carboxylic acid.
    1.83 g of 4-chloro-6-nitrosorezhorescin, 1.56 g of 2,6-dioxybenzoic acid, 0.86 g of pyrolusite and 1.06 ml of sulfuric acid are fed into 20 ml of methanol and stirred for 2 hours at 0 ° C, and also
    14 h at room temperature. A red precipitate is obtained which is filtered and dried.
    The yield of the desired product is 2.45 g (80% of theory). UV (visible region 0.1 N potassium phosphate buffer, pH 7.5) mox 621, 5 nm.
    The following compounds are prepared in a similar manner:
    6-Methylresazurin-4-carboxylic acid with UV (visible region, 0.1 M potassium phosphate buffer, pH 7.5) mothers 612 nm,
    8-methylresazurin-4-carboxylic acid with UV (visible region, 0.1 M potassium phosphate buffer, pH 7.5) max 618 Mi
    I
    8-bromresazurin-4-carboxylic acid with UV (visible region, 0.1 W. potassium phosphate buffer, pH 7.5) max 609 nm.
    Example 5. 8-chlororesorufin-4-carboxylic acid.
    2 g of the 8-chloro-azazurin-4-carboxylic acid obtained in Example 4 is dissolved in 20 ml of water and 5 ml of 25% ammonia. Under ice cooling, zinc powder is added, up to
    until complete color change to red-violet. Excess zinc is filtered off. After acidification, the product is filtered off and dried in vacuo at 40 ° C over calcium chloride. The yield of the desired product 1.5 g (79% of theory). UV (sparing area, 0.1 M calium phosphate buffer, pH 7.5) b max 585.5 nm.
    Similarly to Examples 1-5, more compounds are obtained, summarized in the table.

    The structure and spectra of the derivatives of resazurin or resorufin in the form of tautomers of formula (I) are listed in the table.
    权利要求:
    Claims (1)
    [1]
    The invention The method of obtaining derivatives of re-1 zazurin or resorFin in the form of tautomers of the general formulas
    ten
    R
    15
    v U)
    20
    five
    Where
    4iV K9iK4
    hydrogen, halogen,
    0
    five
    C, -C5-elkyl, C-Cd-alkoxycarbonyl, SCh-C-alkoxy or carboxyl; hydrogen, halogen or
    alkyl
    hydrogen, halogen, kil, C, -C5-alkoxycarbonyl, C ,, -C5-alkoxy, carb xygroup or CO-morphinogroupЈ
     hydrogen, halogen, carboxyl, C1-C5-alkyl, C1-C5-alkoxycarbonyl, C, -C5-alkoxygroup, nitro-group, sulfogroup or a group of the formula
    -So
    -0-С, 4Р4-ОС Н5 or
    0
    Y o tl and h
    -so-in- (c4nu),
    nitrogen or the group N0, and the radicals are not hydrogen at the same time, if Y is nitrogen, but u and u so that
    dinene general formula 1
    to
    (Ii)
    50
    where R ,, - R3 have the indicated meanings, are reacted with a compound of the general formula
    55
    (Iii)
    where have the indicated meanings
    in the presence of pyrolusite and sulfuric acid in an alcoholic medium at a temperature from 0 ° C to room temperature to obtain the desired product of the general formulas (I), where Y is the group N0, or it is treated
     powdered zinc in an aqueous medium in the presence of ammonia at a temperature of from 0 ° C to room temperature to obtain the target product of general formulas (I), where Y is nitrogen.
    UV (vvidima region; potassium phosphate buffer with a pH of 7.5).
    类似技术:
    公开号 | 公开日 | 专利标题
    SU1574173A3|1990-06-23|Method of obtaining derivatives of resazurin or resorufin as tautomers
    SU578887A3|1977-10-30|Method of preparing phthalide ester 6-|-|-penicillanoic acid or its additive salts
    SU625610A3|1978-09-25|Method of producing 3',3",5',5"-tetrachlorophenol-3,4,5,6-tetrabromosulphurphthalein or 3'3"-dibromo-5',5"-dichlorophenol-3,4,5,6-tetrachlorosulphurphthalein
    SU629880A3|1978-10-25|Method of obtaining 7-d-|-2-formyloxy-2-|-cephalosporanic acids or salts thereof
    CN109232626B|2020-08-21|SO based on difluoro boro coumarin2Ratiometric fluorescent probes
    SU458131A3|1975-01-25|The method of obtaining cipher bases of 7-amino-3-desatzfetoxycephalosporanic acid
    SU679135A3|1979-08-05|Method of producing aminopropanol derivatives or salts thereof
    JP2687347B2|1997-12-08|Aromatic phosphine and analytical method using the same
    SU554816A3|1977-04-15|The method of obtaining nitrogen-containing polycyclic compounds or their salts, or racemates, or optical antipodes
    US3670007A|1972-06-13|Aminomalonitrile and method of preparation thereof
    SU1138027A3|1985-01-30|Method of obtaining substituted aurons
    SU803859A3|1981-02-07|Method of preparing omega-thiopropionamides or their acid-additive salts
    SU1498390A3|1989-07-30|Method of producing 6-amino-1,2-dihydro-1-oxy-2-imino-4-piperidinpyrimedine
    JPH07165684A|1995-06-27|Phenethylamine derivative and its production
    RU2132841C1|1999-07-10|Method of purifying oxytetracycline, and intermediate product
    WO1988008415A1|1988-11-03|Reduction of carboxilic esters
    DE2416355A1|1974-10-24|DIRECT SYNTHESIS OF DOPAMINE AMINO ACID AMIDES
    SU1156362A1|1991-07-30|Arylamides p-|-dimethoxybenzylanthranylic acids as intermediate compounds for synthesis of biological active 1-[|-dimethoxybenzyl)-2-methyl-3-aryl-4-|-quinazolinon perchlorates
    Campaigne et al.1974|The preparation and aqueous basic oxidation of diethyl 1‐methyl‐3‐hydroxy‐5‐phenylpyrrole‐2, 4‐dicarboxylate
    US5585512A|1996-12-17|Esters of p-hydroxyaniline mustard
    US3864332A|1975-02-04|Method of preparing {60 -aminobenzlpenicillin
    JPH078853B2|1995-02-01|Method for producing dopamine derivative
    US3076800A|1963-02-05|Jj-lactams unsubstituted at the nitrogen
    US4546203A|1985-10-08|Facile synthesis of β-hydroxy-β-methylglutaric acid
    SU1215622A3|1986-02-28|Method of producing derivatives of n-carbamoylbenzoic sulfimide
    同族专利:
    公开号 | 公开日
    KR870000699B1|1987-04-07|
    ES8603192A1|1985-12-16|
    SU1487824A3|1989-06-15|
    AT66929T|1991-09-15|
    DK140385A|1985-09-30|
    FI851248A0|1985-03-28|
    HU198734B|1989-11-28|
    CS206485A2|1990-07-12|
    JPH02160771A|1990-06-20|
    ZA852333B|1985-12-24|
    JPS60218397A|1985-11-01|
    DK140385D0|1985-03-28|
    EP0156347A3|1988-01-13|
    AU4037585A|1985-10-03|
    EP0156347B1|1991-09-04|
    ES541589A0|1985-12-16|
    AU550958B2|1986-04-10|
    EP0156347A2|1985-10-02|
    DE3583954D1|1991-10-10|
    DD246120A5|1987-05-27|
    DE3411574A1|1985-10-10|
    KR850006427A|1985-10-05|
    DD235648A5|1986-05-14|
    FI81359C|1990-10-10|
    JPH027589B2|1990-02-19|
    FI851248L|1985-09-30|
    HUT38104A|1986-04-28|
    CS273162B2|1991-03-12|
    FI81359B|1990-06-29|
    CA1257253A|1989-07-11|
    JPH0579064B2|1993-11-01|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    US3378463A|1965-06-22|1968-04-16|Army Usa|Method of measuring enzyme activity|
    US3391104A|1966-11-14|1968-07-02|Eastman Kodak Co|Light stabilized, poly-alpha-olefin plastic composition|
    US3950322A|1973-08-27|1976-04-13|Research Corporation|Fluorogenic substrate glycosides|
    DE2752501A1|1977-11-24|1979-05-31|Kurt Prof Dr Wallenfels|Alpha-d-malto:dextrin derivs. as indicators - for colorimetric analysis of amylase|
    US4406832A|1981-09-03|1983-09-27|Mallinckrodt, Inc.|Peptide-type substrates useful in the quantitative determination of endotoxin|
    NZ206795A|1983-01-21|1987-08-31|Merck Frosst Canada Inc|Phenothiazines, analogues and pharmaceutical compositions|DE3412939A1|1984-04-06|1985-10-17|Behringwerke Ag, 3550 Marburg|SUBSTRATES FOR HYDROLASES, METHOD FOR THEIR PRODUCTION AND THEIR USE|
    DE3534927A1|1985-10-01|1987-04-02|Boehringer Mannheim Gmbh|PHOSPHATES OF RESORUFIN DERIVATIVES, METHOD FOR THE PRODUCTION THEREOF AND THEIR USE FOR DETERMINING THE ACTIVITY OF PHOSPHATASES|
    DE3629175A1|1986-08-28|1988-03-17|Behringwerke Ag|CHROMOGENIC COMPOUNDS, METHOD FOR THEIR PRODUCTION AND THEIR USE|
    US4810636A|1986-12-09|1989-03-07|Miles Inc.|Chromogenic acridinone enzyme substrates|
    DE3644401A1|1986-12-24|1988-07-07|Boehringer Mannheim Gmbh|NEW HYDROLASE SUBSTRATES|
    ES2097782T3|1987-04-10|1997-04-16|Univ Southern Australia|METHOD AND COMPOSITION FOR THE DETERMINATION OF SODIUM IONS IN FLUIDS.|
    DE3732871A1|1987-09-30|1989-04-20|Behringwerke Ag|CHROMOGENEOUS SUBSTRATES|
    US4932871A|1987-11-20|1990-06-12|Miles Inc.|Rapid method for preparing chromogenic alpha-amylase substrate compounds at high preparative yields|
    DE3743908A1|1987-12-23|1989-07-06|Boehringer Mannheim Gmbh|NEW OLIGOGLUCOSIDE|
    JPH01211595A|1988-02-18|1989-08-24|Kikkoman Corp|Novel n-acetyl-beta-d-glucosamine derivative, production thereof and utilization thereof to reagent for measuring n-acetyl-beta-d-glucosamidase activity|
    US5151348A|1988-12-23|1992-09-29|E. I. Du Pont De Nemours And Company|Enzyme-linked immunoassay for measurement of cyclosporin a levels in whole blood samples|
    US5104980A|1989-06-12|1992-04-14|Miles Inc.|Chromogenic dibenzoxazepinone and dibenzothiazepinone enzyme substrates|
    KR100710561B1|2000-07-12|2007-04-24|에스케이 주식회사|Silent mark for oil product and detection method thereto|
    WO2002100843A1|2001-06-13|2002-12-19|Jianqing Lu|Phenoxazine compounds, the compositions containing them and their medicinal uses|
    FR2854893B1|2003-05-13|2006-07-07|Biomerieux Sa|NOVEL ENZYMATIC SUBSTRATES DERIVED FROM PHENOXAZINONE AND THEIR USE AS DEVELOPER IN THE DETECTION OF MICROORGANISMS WITH PEPTIDASE ACTIVITY|
    FR2875242B1|2004-09-10|2006-11-17|Biomerieux Sa|NOVEL ENZYMATIC SUBSTRATES DERIVED FROM PHENOXAZINONE AND THEIR USE AS DEVELOPER IN THE DETECTION OF MICROORGANISMS WITH PEPTIDASE ACTIVITY|
    EP3242927B1|2015-01-09|2018-10-10|Unilever PLC, a company registered in England and Wales under company no. 41424|Laundry treatment composition comprising a dye|
    EP3932995A1|2020-07-04|2022-01-05|Emulseo SAS|Novel fluorescent substrates and uses thereof in microfluidics|
    法律状态:
    优先权:
    申请号 | 申请日 | 专利标题
    DE19843411574|DE3411574A1|1984-03-29|1984-03-29|GLYCOSIDES OF RESORUFIN DERIVATIVES, METHOD FOR THE PRODUCTION THEREOF AND THE USE THEREOF FOR DETERMINING THE ACTIVITY OF GLYCOSIDASES|
    [返回顶部]