N-alkylacridan carboxyl derivatives useful for chemiluminescent detection
AssigneeLumigen, Inc.
InventorsHashem Akhavan-Tafti, Renuka Desilva, Katsuaki Sugioka
Life of this patent
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Abstract
N-alkylacridan carboxylic acid derivative compounds (I) are used to generate chemiluminescence by the action of a peroxidase enzyme and an oxidant. The compounds I are useful in assays of all types.
Claims68
An acridan of the formula ##STR11## wherein R.sub.l is selected from the group consisting of alkyl, heteroalkyl and aralkyl groups, wherein R.sub.3 and R.sub.4 are selected from the group consisting of substituted and unsubstituted aryl groups and alkyl, heteroalkyl and aralkyl groups and wherein R.sub.5 and R.sub.6 are selected from the group consisting of hydrogen and non-interfering substituents and wherein ##STR12## is a leaving group which allows the production of light from the acridan by reaction with a peroxide and a peroxidase.
The acridan of the formula ##STR14##
A method for producing chemiluminescence which comprises reacting a peroxide compound and a peroxidase with an acridan of the formula ##STR15## wherein R.sub.1 is selected from the group consisting of alkyl, heteroalkyl and aralkyl groups, wherein R.sub.5 and R.sub.6 are selected from the group consisting of hydrogen and non-interfering substituents, wherein Y is a leaving group which allows the production of light from the acridan by reaction with a peroxide and a peroxidase.
A reagent composition which generates light in the presence of a peroxidase which comprises:
- (a) an acridan of the formula:
- ##STR18## wherein R.sub.1 is selected from the group consisting of alkyl, heteroalkyl and aralkyl groups, wherein R.sub.5 and R.sub.6 are selected from the group consisting of hydrogen and non-interfering substituents and wherein Y is a leaving group which allows the production of light from the acridan by reaction with a peroxide and a peroxidase;
- (b) optionally a phenolic compound which enhances light production from the acridan;
- (c) a peroxide compound which participates in the reaction of the acridan with the peroxidase;
- (d) a chelating agent which prevents the peroxide compound from reacting prior to addition of the peroxidase to the composition;
- and
- (e) a non-ionic surfactant.
A reagent composition which generates light in the presence of a peroxidase which comprises:
- (a) an acridan having a formula selected from the group consisting of:
- ##STR20## (b) optionally, a phenolic compound which enhances light production from the acridan;
- (c) a peroxide compound which participates in the reaction of the acridan with the peroxidase;
- (d) a chelating agent which prevents the peroxide compound from reacting prior to addition of the peroxidase to the composition;
- and
- (e) a non-ionic surfactant.
A method for detecting an analyte selected from the group consisting of hydrogen peroxide, hydrogen peroxide generated by an enzyme, peroxidase alone, organic molecules labeled with the peroxidase and biological molecules labeled with the peroxidase in an assay procedure by a chemiluminescent reaction comprising reacting an acridan with a peroxide and a peroxidase to produce light for detecting the analyte wherein the acridan is of the following formula ##STR21## wherein R.sub.1 is selected from the group consisting of alkyl, heteroalkyl and aralkyl groups, wherein R.sub.5 and R.sub.6 are selected from the group consisting of hydrogen and non-interfering substituents and wherein Y is a leaving group which allows the production of light from the acridan by reaction with the peroxide and the peroxidase to detect the analyte.
A method for detecting an analyte selected from the group consisting of peroxidase alone, organic molecules labeled with the peroxidase and biological molecules labeled with the peroxidase in an assay procedure by a chemiluminescent reaction comprising:
- (a) providing a reagent composition which generates light in the presence of the peroxidase which comprises:
- an acridan of the formula:
- ##STR27## wherein R.sub.1 is selected from the group consisting of alkyl, heteroalkyl and aralkyl groups, wherein R.sub.5 and R.sub.6 are selected from the group consisting of hydrogen and non-interfering substituents and wherein Y is a leaving group which allows the production of light from the acridan by reaction with a peroxide and a peroxidase;
- optionally a phenolic compound which enhances light production from the acridan;
- a peroxide compound which participates in the reaction of the acridan with the peroxidase;
- a chelating agent which prevents the peroxide compound from reacting prior to the addition of the peroxidase to the composition;
- and a non-ionic surfactant;
- and
- (b) reacting the peroxidase with the reagent composition so that light is produced for detecting the analyte.
A kit for detecting in a sample an analyte selected from the group consisting of hydrogen peroxide, hydrogen peroxide generated by an enzyme, peroxidase alone, organic molecules labeled with the peroxidase and biological molecules labeled with the peroxidase in an assay procedure by a chemiluminescent reaction comprising reacting an acridan with a peroxide and a peroxidase to produce light which comprises in separate containers:
- (a) an acridan of the formula:
- ##STR33## wherein R.sub.1 is selected from the group consisting of alkyl, heteroalkyl and aralkyl groups, wherein R.sub.5 and R.sub.6 are selected from the group consisting of hydrogen and non-interfering substituents and wherein Y is a leaving group which allows the production of light from the acridan by reaction with a peroxide and a peroxidase;
- and
- (b) a peroxidase, wherein the light is detected in the assay procedure by reacting the acridan with a peroxide, the peroxidase and the sample to thereby detect the analyte.
A kit for detecting in a sample an analyte selected from the group consisting of the hydrogen peroxide, hydrogen peroxide generated by an enzyme, peroxidase alone, organic molecules labeled with the peroxidase and biological molecules labeled with the peroxidase in an assay procedure by a chemiluminescent reaction comprising reacting an acridan with a peroxide and a peroxidase to produce light which comprises in separate containers:
- (a) an acridan having a formula selected from the group consisting of:
- ##STR35## in a reagent composition;
- and (b) a peroxidase, wherein the light is detected in the assay procedure by reacting the acridan with the peroxide, the peroxidase and the sample to detect the analyte.
A kit for detecting in a sample an analyte selected from the group consisting of peroxidase alone, organic molecules labeled with the peroxidase and biological molecules labeled with the peroxidase in an assay procedure by a chemiluminescent reaction to produce light which comprises in separate containers:
- (a) a reagent composition the components of which may be in a single or multiple containers which generates light in the presence of a peroxidase which comprises:
- an acridan of the formula:
- ##STR36## wherein R.sub.1 is selected from the group consisting of alkyl, heteroalkyl and aralkyl groups, wherein R.sub.5 and R.sub.6 are selected from the group consisting of hydrogen and non-interfering substituents and wherein Y is a leaving group which allows the production of light from the acridan by reaction with a peroxide and a peroxidase;
- optionally a phenolic compound which enhances light production from the acridan;
- a peroxide compound which participates in the reaction of the acridan with the peroxidase;
- a chelating agent which prevents the peroxide compound from reacting prior to the addition of a peroxidase to the composition;
- and a non-ionic surfactant;
- and
- (b) a peroxidase, wherein the light is detected in the assay procedure by reacting the reagent composition with the peroxidase and the sample to thereby detect the analyte.
A kit for detecting in a sample an analyte selected from the group consisting of peroxidase alone, organic molecules labeled with the peroxidase and biological molecules labeled with the peroxidase in an assay procedure by a chemiluminescent reaction to produce light which comprised in separate containers:
- (a) a reagent composition which may be in a single or multiple containers and which generates light in the presence of a peroxidase which comprises:
- an acridan selected from the group consisting of:
- ##STR38## wherein R.sub.1 is selected from the group consisting of alkyl, heteroalkyl and aralkyl groups, wherein R.sub.5 and R.sub.6 are selected from the group consisting or hydrogen and non-interfering substituents and wherein Y is a leaving group which allows the production of light from the acridan by reaction with a peroxide and a peroxidase;
- optionally a phenolic compound which enhances light production from the acridan;
- a peroxide compound which participates in the reaction of the acridan with the peroxidase;
- a chelating agent which prevents the peroxide compound from reacting prior to the addition of a peroxidase with the composition;
- and a nonionic surfactant;
- and
- (b) a peroxidase, wherein the light is detected in the assay procedure by reacting the reagent composition with the peroxidase and the sample to detect the analyte.
A method for detecting hydrogen peroxide in an assay procedure by a chemiluminescent reaction comprising reacting hydrogen peroxide and a peroxidase with an acridan of the formula:
- ##STR39## wherein R.sub.1 is selected from the group consisting of alkyl, heteroalkyl and aralkyl groups, wherein R.sub.5 and R.sub.6 are selected from the group consisting of hydrogen and non-interfering substituents and wherein Y is a leaving group which allows the production of light from the acridan by reaction with the peroxide and the peroxidase to detect the hydrogen peroxide.
A method for detecting peroxidase selected from the group consisting of the peroxidase alone, organic molecules labeled with the peroxidase and biological molecules labeled with the peroxidase in an assay procedure by a chemiluminescent reaction comprising reacting in the presence of a peroxide an acridan selected from the group consisting of:
- ##STR41## with the peroxidase to produce light.
Dependencies are inferred from the claim text.