Chromogenic dibenzoxazepinone and dibenzothiazepinone enzyme substrates
AssigneeMiles Inc.
InventorPaul F. Corey
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Abstract
Chromogenic enzyme substrate compounds comprising a dibenz[b,e] [1,4]oxazepinone or dibenzo[b,e] [1,4]thiazepinone nucleus having an enzyme-cleavable group such as a radical of a sugar, carboxylic acid, amino acid, peptide, phosphoric acid, or sulfuric acid. The substrate compounds are, in general, highly soluble in aqueous media and only slightly colored, and produce, upon enzyme cleavage, a chromogen exhibiting a large change in absorbance and a pKa below 7. Such substrates find use as indicators for the determination of enzyme analytes and enzymes used as markers in a variety of assays, including immunoassays.
Claims19
A method for determining a particular enzyme in a liquid test sample, comprising the steps of:
- (a) contacting the test sample with a chromogenic enzyme substrate compound of the formula:
- ##STR8## wherein W is O or S;
- R and R', which can be the same or different, are H, alkyl, or aryl;
- and Y is an enzyme-cleavable group that is
- (i) capable of being cleaved from the nucleus of the enzyme substrate compound by said enzyme, or
- (ii) capable of being modified by said enzyme to produce a secondary substrate compound in which Y is capable of being cleaved from the nucleus of the modified enzyme substrate compound by a secondary enzyme,
- and, in the latter case (ii), the secondary substrate compound is contacted with said secondary enzyme;
- and
- (b) measuring and correlating the resulting color generated by the cleaved nucleus of the enzyme substrate compound to the presence of said enzyme in said liquid test sample.
Dependencies are inferred from the claim text.