Patent ID: 6346615
Filing Date: 2002-02-12
Classification: C07D

Abstract:
A method of detecting the presence of a target substance in a sample, wherein said target substance is a ligand which is bound by a second member of a ligand binding pair, comprising:combining said sample with a complex of said second member of said ligand binding pair and an enzyme, under conditions which permit binding of said ligand binding pair if said target is present in said sample, removing from said sample all said complex which is not bound, adding a dioxetane of the formula I: â€ƒwherein Y1 and Y2 are independently H, a hydroxyl group, a halogen, an unsubstituted lower alkyl group, a hydroxy lower alkyl group, a halo lower alkyl group, a phenyl group, a halo phenyl group, an alkoxy phenyl group, an alkoxy phenoxy group, a hydroxy alkoxy group, a cyano group, an amide group, an alkoxy group or a carboxyl group, wherein R is C1-12 alkyl, aryl or aralkyl, wherein X is an enzyme-labile group selected from the group consisting of a phosphate, galactoside, acetate, 1-phospho-2,3-diacylglyceride, 1-thio-D-glucoside, adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, adenosine, &agr;-D-glucoside,&bgr;-D-glucoside, &bgr;-D-glucuronide, &agr;-D-mannoside, &bgr;-D-mannoside, &bgr;-D-fructofuranoside, &bgr;-glucosiduronate, P-toluenesulfonyl-L-arginine ester, and P-toluenesulfonyl-L-arginine amide, and wherein Z is an electron-active group selected from the group consisting of electron withdrawing groups and electrondonating groups and occupies the four or five position on the phenyl ring, to said sample, and monitoring said sample for chemiluminescence emission, wherein said enzyme cleaves said enzyme-labile group and said emission is indicative of the presence of said target compound in said sample.