Boehringer Mannheim Gmbh

Elli Rauscher

Wolfgang Gruber

August W Wahlefeld

Ulrich Neumann

Axel Schmidt

Eugen Schaich

Herbert Von Der Eltz

Gunter Weimann

Joachim Ziegenhorn

Ulfert Deneke

Alexander Hagen

Leonard F Dixon

Bernhard Empl

Hans-Georg Batz

Erich Bernt

Martina Junius-Comer

Gerhard Michal

Helmut Determann

Michal Gerhard

The invention concerns new indophenyl-substituted maltose derivatives, their use for the determination of the enzymatic activity of .alpha.-amylase and reagents for the determination of .alpha.-amylase which contain the new indophenyl-substituted maltose derivatives as the enzyme substrate.

Indophenol substituted maltooligosides as .alpha.-amylase substrates

The present invention provides a process for the specific determination of pancreatic .alpha.-amylase in the presence of salivary .alpha.-amylase in body fluids by reaction with a system for the detection of .alpha.-amylase with the use of an inhibitor for salivary .alpha.-amylase, wherein, as substrate, there is used a compound of the general formula:-- ##STR1## in which R.sub.1 is a straight-chained or branched alkyl or alkoyl radical containing up to 6 carbon atoms, a cycloalkyl or cycloalkoxyl radical containing 3 to 6 carbon atoms or a benzoyl, benzyl or phenyl radical which is optionally hydrophilically substituted, R.sub.2 is a hydrogen atom or in which R.sub.1 and R.sub.2 together form a methylene bridge, the hydrogen atoms of which, independently of one another, can each be substituted by an alkyl radical containing up to 5 carbon atoms or a phenyl radical, n is 1, 2 or 3 and X is an optically determinable residue. The present invention also provides a reagent for the specific determination of pancreatic .alpha.-amylase, as well as new compounds for use in the above process and reagent.

Process and reagent for the specific determination of pancreatic

The present invention provides a process for the specific determination of pancreatic .alpha.-amylase in the presence of salivary .alpha.-amylase in body fluids by reaction with a system for the detection of .alpha.-amylase with the use of an inhibitor for salivary .alpha.-amylase, wherein, as substrate, there is used a compound of the general formula: ##STR1## in which R.sub.1 is a straight-chained or branched alkyl or alkoyl radical containing up to 6 carbon atoms, a cycloalkyl or cycloalkoxyl radical containing 3 to 6 carbon atoms or a benzoyl, benzyl or phenyl radical which is optionally hydrophilically substituted, R.sub.2 is a hydrogen atom or in which R.sub.1 and R.sub.2 together form a methylene bridge, the hydrogen atoms of which, independently of one another, can each be substituted by an alkyl radical containing up to 5 carbon atoms or a phenyl radical, Gluc is a glucose molecule, n is 1, 2 or 3 and X is an optically determinable residue. The present invention also provides a reagent for the specific determination of pancreatic .alpha.-amylase, as well as new compounds for use in the above process and reagent.

.alpha.-Amylase is determined by the enzymatic splitting of an .alpha.-amylase substrate and measurement of a fission product, wherein there is used as a substrate a maltoheptaose compound of the formula ##STR1## wherein R is a glucoside, phenylglucoside, mononitrophenylglucoside, dinitrophenylglucoside, sorbitol or gluconic acid group. Reagents comprising such a substrate and a system for the determination of a fission product formed from the amylase substrate by .alpha.-amylase, are also provided. A process for the preparation of a maltoheptaose will Bacillus macerans amylase, free of p-nitrophenyl-alpha-glucoside splitting activity is disclosed.

Process for preparing a maltoheptaose derivative

The present invention provides compounds of the general formula ##STR1## wherein R.sub.1 is a hydrogen atom, a methyl, ethyl, propyl, isopropyl, butyl, tert. butyl or 1-alkoxyalkyl radical or an optionally hydrophilically substituted cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, phenyl, tetrahydropyranyl, piperidinyl, N-methyl substituted piperidinyl or N-ethyl substituted piperidinyl, pyridinyl, thiophenyl or 1,1-dioxo-tetra- hydrothiopyranyl radical or an amino group which is optionally substituted by the same or different substituents selected from methyl, ethyl, propyl and isopropyl radicals. R.sub.2 is an oligoglucoside residue with 2, 3 or 4 glucose units and X is a hydrogen atom or an optically determinable residue. The present invention also provides processes for the preparation of compounds (I) and reagents for the determination of .alpha.-amylase containing them. Furthermore, the present invention provides a process for the determination of .alpha.-amylase using a compound of general formula (I).

Substrates and reagents useful in determining .alpha.-amylase and a

The present invention provides gamma-glutamyl-4-azoanilides, processes for their preparation and their use as substrates for gamma-glutamyl transferase determination, of the formula ##STR1## wherein X is alkyl, --(CH.sub.2).sub.m --COOH or --O--(CH.sub.2).sub.n --COOH wherein m is 0-4 and n is 1-4; R is optionally substituted p-nitrophenyl; an optionally substituted thiophene residue; an optionally substituted thiazole residue; an optionally substituted benzothiazole residue; an optionally substituted benzoisothiazole residue; an N-alkylthiazole residue; or an optionally substituted 1,3,5-thiodiazole residue; as well as the alkali metal, alkaline earth metal and ammonium salts thereof.

Gamma-glutamyl-4-azoanilides, process for the preparation thereof and

The present invention provides compounds of the general formula: ##STR1## in which R and R.sub.1, independently of one another, each represent a straight-chained or branched alkyl or alkoyl radical containing up to 6 carbon atoms or a phenyl radical or R and R.sub.1 together also form a methylene bridge, the hydrogen atoms of which, independently of one another, can each be substituted by an alkyl radical containing up to 5 carbon atoms or a phenyl radical, R.sub.2 represents an oligoglucoside residue containing 2 to 7 glucose units and X is a hydrogen atom or an optically-determinable residue. The present invention also provides a process for the preparation of these compounds.

Method and reagent compositions for determining alpha amylase using a

Gamma-Glutamyl-4-azoanilides

Heptaose compounds and preparation thereof

.alpha.-Amylase is determined by the enzymatic splitting of an .alpha.-amylase substrate and measurement of a fission product, wherein there is used as a substrate a maltoheptaose compound of the formula ##STR1## wherein R is a glucoside, phenylglucoside, mononitrophenylglucoside, dinitrophenylglucoside, sorbitol or gluconic acid group. Reagents comprising such a substrate and a system for the determination of a fission product formed from the amylase substrate by .alpha.-amylase, are also provided.

Process and reagent for the determination of .alpha.-amylase

Method for determining .alpha.-amylase by incubating with a starch substrate, wherein the substrate is based upon particulate starch derivatized with detectable groups for the determination of .alpha.-amylase, the substrate consisting of superficially cross-linked starch or amylose grains having a regulated, reduced swellability, said grains having a diameter of from 0.01 to 0.20 mm; these grains are prepared by suspending non-swollen grains in a solution with only enough cross-linking agent to achieve superficial cross-linking and then derivatizing the grains with the detectable groups for the determination of .alpha.-amylase.

Method for the determination of .alpha.-amylase

Triglycerides are determined by enzymatic saponification with lipase in a buffer in the presence of a carboxylesterase and an alkali metal or alkaline earth metal (10-15 carbon atom) alkyl sulfate, in the additional presence of a compound of the general formula RO(CH.sub.2 CH.sub.2).sub.n H, in which R is alkyl or alkenyl containing 14 to 20 carbon atoms and n is a number of from 7 to 20; this determination prevents turbidity and does not disadvantageously influence enzyme activity.

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Munchen, Germany

Elli Rauscher