N- and O-substituted aminophenols, method and use for diagnosis

A process for colorimetric determination of a hydrolase enzyme by reaction of a chromogenic enzyme substrate with the enzyme, oxidation of a leuko colored material liberated from the chromogenic enzyme substrate by the enzyme and determination of the resulting colored material as a measure for the amount of enzyme, comprising

• reacting the hydrolase enzyme with an N- and O-substituted aminophenyl derivative as the chromogenic enzyme substrate of the formula:
• ##STR99## wherein G is the residue of an organic or inorganic acid or a glycoside residue,
• R.sup.1 and R.sup.2, which are the same or different, are hydrogen, halogen, SO.sub.3 H, PO.sub.3 H.sub.2 or a salt of the acid residues of SO.sub.3 H or PO.sub.3 H.sub.2, hydroxyl, nitro, carboxyl, carboxamido, cyano, alkyl, alkenyl, alkoxy, alkylsulphinyl, alkylsulphonyl, alkoxycarbonyl, alkylcarbonyl, aryl or aralkyl unsubstituted or substituted at least once by hydroxyl, carboxyl, halogen, cyano, SO.sub.3 H, PO.sub.3 H.sub.2 or a salt of one of the acid residues of SO.sub.3 H or PO.sub.3 H.sub.2 or,
• when two substituents of R.sup.1 and R.sup.2 are present on neighboring carbon atoms, R.sup.1 and R.sup.2 together represent a 1,4-butadiendiyl radical which is unsubstituted or substituted at least once by SO.sub.3 H, PO.sub.3 H.sub.2 or a salt of the acid groups of SO.sub.3 H or PO.sub.3 H.sub.2, an alkyl or a carboxyl group,
• R.sup.3 is hydrogen, --CO--COOH, SO.sub.3 H, PO.sub.3 H.sub.2 or a salt of the acid groups of SO.sub.3 H or PO.sub.3 H.sub.2, an alkylcarbonyl radical unsubstituted or substituted at least once by halogen, COOH, SO.sub.3 H or PO.sub.3 H.sub.2 or a salt of the acid groups of SO.sub.3 H or PO.sub.3 H.sub.2 or an arylcarbonyl radical unsubstituted or substituted at least once by SO.sub.3 H, PO.sub.3 H.sub.2 or a salt of the acid groups of SO.sub.3 H or PO.sub.3 H.sub.2, and
• L is a radical of the formula:
• ##STR100## wherein R.sup.4 and R.sup.5, which are the same or different, are alkyl or together represent a saturated 3-6 member hydrocarbon chain which is uninterrupted or interrupted by oxygen, sulphur or nitrogen and wherein alkyl or the hydrocarbon chain is unsubstituted or substituted at least once by hydroxyl, carboxyl, alkoxycarbonyl, alkoxy, SO.sub.3 H, PO.sub.3 H.sub.2 or a salt of one of the acid groups of SO.sub.3 H or PO.sub.3 H.sub.2 or halogen, and
• R.sup.6 and R.sup.7, which can be the same or different, are hydrogen, halogen, hydroxyl, carboxamido, an alkyl, alkoxy, alkylcarbonyl, alkoxycarbonyl, aryl or aralkyl radical unsubstituted or substituted at least once by hydroxyl, carboxyl, halogen, SO.sub.3 H, PO.sub.3 H.sub.2 or a salt of one of the acid groups of SO.sub.3 H or PO.sub.3 H.sub.2, or
• L is a pyrazolo-heterocylic radical of the formula:
• ##STR101## wherein X-Y signifies NR.sup.8 --CO or N.dbd.CR.sup.9, wherein R.sup.8 is hydrogen or an alkyl and R.sup.9 is alkyl, alkenyl, alkoxy, alkylthio, aryl, aralkyl, in each case unsubstituted or substituted by hydroxyl, dialkylphosphinyl, carboxyl, SO.sub.3 H, PO.sub.3 H.sub.2 or a salt of one of the acid groups of SO.sub.3 H or PO.sub.3 H.sub.2, alkoxycarbonyl, amino, wherein the amino is unsubstituted or substituted by one or two alkyl radicals which one or two alkyl radicals are unsubstituted or substituted by at least one hydroxyl, carboxyl or alkoxycarbonyl radical and wherein when amino is substituted by two alkyl radicals, the alkyl radicals can be joined to form a ring which, apart from the first nitrogen atom of the amino group, is uninterrupted or is interrupted by oxygen, sulphur or a second nitrogen atom, or the amino is unsubstituted or substituted by one or two acyl radicals, alkoxy- or aralkoxycarbonyl radicals, H.sub.2 N--CO, alkyl-, aralkyl- or arylcarbamoyl radicals;
• or R.sup.9 is hydrogen, carboxyl, alkoxycarbonyl, carboxamido or halogen, and
• Z signifies NR.sup.10 --N.dbd.N, wherein R.sup.10 is hydrogen or an alkyl or aralkyl radical, or
• Z is an unsaturated 3-5 member chain of nitrogen atoms or of carbon atoms, or said chain having at least one nitrogen or sulphur atom, whereby carbon atoms in the chain are unsubstituted or substituted by alkyl, alkoxy, hydroxyalkyl, alkylthio, hydroxyl, aralkyl, aryl, carboxyl, carboxamido, alkoxycarbonyl, cyano, amino, wherein the amino is unsubstituted or substituted by one or two alkyl radicals which one or two alkyl radicals are unsubstituted or substituted by at least one hydroxyl, carboxyl, alkoxycarbonyl, halogen, or nitrogen which is not attached via a double bond and nitrogen is unsubstituted or substituted by alkyl or aralkyl or two neighboring chain substituents form an alkylene which alkylene is unsubstituted or substituted or anellated with aryl and tautomers thereof.