Method for making chromogenic and/or fluorogenic substrates for use in monitoring catalytic or enzymatic activity

Abstract From the Canadian patent

A method for preparing a substrate capable of undergoing catalytic-induced hydrolysis of the phosphate ester at the 3' position, to yield a species capable of being monitored spectrophotometrically or fluorometrically and thus useful in carrying out various non-isotopic immunoassays comprises, (a) forming a mononucleotide 2' 3' cyclic phosphate, of the formula:, wherein 8 is a nucleotide base and wherein the CH20H group at, the 4'-position is either cis or trans to the cyclic phosphate; (b) blocking said mononucleotide 2', 3'-cyclic phosphate at the 5'-hydroxyl with a suitable 5'-blocking member to form a mononucleotide 5'-O-blocked-2', 3'-cyclic phosphate; (c) opening said cyclic phosphate …