Chemiluminescent enhancers
AssigneeBritish Technology Group Limited
InventorLarry J. Kricka
Life of this patent
Estimating the term…
Abstract
Method of increasing the light output and/or signal:background ratio of light output from a chemiluminescent reaction of a dihydrophthalazinedione, a peroxidase enzyme catalyst and an oxidant, by carrying out the reaction in the presence of an enhancer which is an aromatic organo boron compound. Kits suitable for use in diagnositc assays comprising such enhancers are also described.
Claims20
A method of increasing the light output and/or signal:background ratio of light output from a chemiluminescent reaction of a dihydrophthalazinedione (DPD), a peroxidase enzyme catalyst and an oxidant, said method comprising the step of carrying out said chemiluminescent reaction in the presence of an enhancer comprising a compound selected from the group consisting of compounds of formula (I) ##STR4## in which the R groups are the same and each is selected from the group consisting of hydrogen, n-butyl, 4'-chlorophenyl and 3',5'-dichlorophenyl;
- or the Rs together are O,O-propylene (thereby forming with the boron atom, a cyclic ether);
- W is selected from the group consisting of hydrogen, methyl, methoxy, hydroxy and chloro;
- X is selected from the group consisting of hydrogen, chloro, amino and nitro;
- Y is selected from the group consisting of hydrogen, methyl, carboxy, chloro, bromo, iodo, phenyl, phenoxy, 4'-chloroanilino, 4'-boronylphenyl, 4'-bromophenyl, 2'-carboxyethenyl and trimethylsilyl;
- Z is selected from the group consisting of hydrogen, 5-chloro, 5-bromo, 5-(3'-trifluorormethyl)phenylazo and 6-chloro;
- or
- W and X together may represent a fused benzene ring and X and Y together may represent a fused benzene ring substituted by hydroxy in the 6-position of the naphthalene ring numbering, provided that
- (1) when each R is hydrogen:
- (a) W, X, Y, Z are each hydrogen;
- or
- (b) W, X and Z are each hydrogen and Y is selected from the group consisting of iodo, bromo, chloro, trimethylsilyl, phenoxy, phenyl, 4'-chloroanilino, methyl, 4'-boronylphenyl and 2'-carboxyethenyl;
- or
- (c) W and Z are each hydrogen and:
- (i) X and Y together represent a fused benzene ring substituted by hydroxy in the 6-position of the naphthalene ring numbering;
- or
- (ii) X is either nitro or chloro and Y is chloro;
- or
- (iii) X is nitro and Y is carboxy;
- or
- (d) W, Y and Z are each hydrogen and X is amino, chloro or nitro;
- or
- (e) W and X together represent a fused benzene ring and Y and Z are each hydrogen;
- or
- (f) X and Y are each hydrogen and:
- (i) W is methoxy and Z is 5-bromo;
- or
- (ii) W is hydroxy and Z is 5-(3'-trifluoromethyl)phenylazo;
- or
- (iii) W is methyl and Z is hydrogen;
- or
- (g) W is chloro, X is chloro and Y and Z are each hydrogen;
- or
- (h) W and Y are each chloro, X is amino and Z is 6-chloro;
- (2) when each R is n-butyl, W, X and Z are each hydrogen and Y is bromo or 4'-bromophenyl;
- (3) when each R is 4'-chlorophenyl, W, X and Z are each hydrogen and Y is chloro;
- (4) when each R is 3',5'-dichlorophenyl, W and Y are each hydrogen, X is chloro and Z is 5-chloro;
- and
- (5) when the Rs together represent O,O-propylene, X, Y and Z are each hydrogen;
- and the compounds bis(catechol) borate, boroglycine, pentaerythritol borate, 4-(3'-borono-4'-hydroxyphenylazo) benzoic acid, diphenylisobutoxyborane, diphenylboronic anhydride and dimethylphenylboronic acid.
A method of diagnostic assay of a substance, wherein the presence or amount of said substance is related to the presence or amount of a reaction partner selected from the group consisting of a dihydrophthalazinedione (DPD), a peroxidase enzyme, an oxidant and an enhancer which together react in a chemiluminescent reaction, wherein the reaction is carried out, the light output is detected or measured and the presence or amount of the substance to be assayed is related to the light output, and wherein the enhancer comprises a compound selected from the group consisting of compounds of formula (I) ##STR5## in which the R groups are the same and each is selected from the group consisting of hydrogen, n-butyl, 4'-chlorophenyl and 3',5'-dichlorophenyl;
- or the Rs together are O,O-propylene (thereby forming with the boron atom, a cyclic ether);
- W is selected from the group consisting of hydrogen, methyl, methoxy, hydroxy and chloro;
- X is selected from the group consisting of hydrogen, chloro, amino and nitro;
- Y is selected from the group consisting of hydrogen, methyl, carboxy, chloro, bromo, iodo, phenyl, phenoxy, 4'-chloroanilino, 4'-boronylphenyl, 4'-bromophenyl, 2'-carboxyethenyl and trimethylsilyl;
- Z is selected from the group consisting of hydrogen, 5-chloro, 5-bromo, 5-(3'-trifluoromethyl)phenylazo and 6-chloro;
- or
- W and X together may represent a fused benzene ring and X and Y together may represent a fused benzene ring substituted by hydroxy in the 6-position of the naphthalene ring numbering, provided that
- (1) when each R is hydrogen:
- (a) W, X, Y, Z are each hydrogen;
- or
- (b) W, X and Z are each hydrogen and Y is selected from the group consisting of iodo, bromo, chloro, trimethylsilyl, phenoxy, phenyl, 4'-chloroanilino, methyl, 4'-boronylphenyl and 2'-carboxyethenyl;
- or
- (c) W and Z are each hydrogen and:
- (i) X and Y together represent a fused benzene ring substituted by hydroxy in the 6-position of the naphthalene ring numbering;
- or
- (ii) X is either nitro or chloro and Y is chloro;
- or
- (iii) X is nitro and Y is carboxy;
- or
- (d) W, Y and Z are each hydrogen and X is amino, chloro or nitro;
- or
- (e) W and X together represent a fused benzene ring and Y and Z are each hydrogen;
- or
- (p) X and Y are each hydrogen and:
- (i) W is methoxy and Z is 5-bromo;
- or
- (ii) W is hydroxy and Z is 5-(3'-trifluoromethyl)phenylazo;
- or
- (iii) W is methyl and Z is hydrogen;
- or
- (g) W is chloro, X is chloro and Y and Z are each hydrogen;
- or
- (h) W and Y are each chloro, X is amino and Z is 6-chloro;
- (2) when each R is n-butyl, W, X and Z are each hydrogen and Y is bromo or 4'-bromophenyl;
- (3) when each R is 4'-chlorophenyl, W, X and Z are each hydrogen and Y is chloro;
- (4) when each R is 3',5'-dichlorophenyl, W and Y are each hydrogen, X is chloro and Z is 5-chloro;
- and
- (5) when the Rs together represent O,O-propylene, X, Y and Z are each hydrogen;
- and the compounds bis(catechol) borate, boroglycine, pentaerythritol borate, 4-(3'-borono-4'-hydroxyphenylazo) benzoic acid, diphenylisobutoxyborane, diphenylboronic anhydride and dimethylphenylboronic acid.
Dependencies are inferred from the claim text.