Fluorogenic hydrazine-substituted compounds

Patent metadata

Field	Value
Publication number	US-9389183-B2
Application number	US-201313793517-A
Country	US
Kind code	B2
Filing date	Mar 11, 2013
Priority date	Jan 24, 2008
Publication date	Jul 12, 2016
Grant date	Jul 12, 2016

Abstract

The present disclosure is directed to fluorogenic schiff base-forming dyes capable of detecting analytes containing aldehyde and ketone groups. The dyes contain nucleophilic hydrazinyl appendages and are capable of binding and detecting analytes in situ.

First claim

The invention claimed is:

1. A hydrazinyl-substituted compound of Formula I:

or a stereoisomer, tautomer, hydrate, solvate, or salt thereof;
wherein,
X is H or an analyte selected from the group consisting of an amino acid, a peptide, a protein, a carbohydrate, a polysaccharide, a nucleoside, a nucleotide, an oligonucleotide, a nucleic acid polymer, a drug, a lipid, a synthetic polymer, malondialdehyde, and 4-hydroxynonenal;
R 1 is selected from the group consisting of H, alkyl and substituted alkyl;
R 1a is H;
R 2 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 and R 10 are each independently selected from the group consisting of H, alkyl, substituted alkyl, alkoxy, substituted alkoxy, acyl, acylamino, acyloxy, amino, substituted amino, aminocarbonyl, aminothiocarbonyl, aminocarbonylamino, aminothiocarbonylamino, aminocarbonyloxy, aminosulfonyl, aminosulfonyloxy, aminosulfonylamino, amidino, carboxyl, carboxyl ester, (carboxyl ester)amino, (carboxyl ester)oxy, cyano, halo, hydroxy, nitro, SO 3 − , sulfonyl, substituted sulfonyl, sulfonyloxy, thioacyl, thiol, alkylthio, substituted alkylthio, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl; and
R 3 is selected from the group consisting of alkoxy, substituted alkoxy, amino, substituted amino, hydrazinyl, substituted hydrazinyl, analyte substituted hydrazinyl, alkyl, substituted alkyl, acyl, acylamino, acyloxy, aminocarbonyl, aminothiocarbonyl, aminocarbonylamino, aminothiocarbonylamino, aminocarbonyloxy, aminosulfonyl, aminosulfonyloxy, aminosulfonylamino, amidino, carboxyl, carboxyl ester, (carboxyl ester)amino, (carboxyl ester)oxy, cyano, halo, hydroxy, nitro, SO 3 − , sulfonyl, substituted sulfonyl, sulfonyloxy, thioacyl, thiol, alkylthio, substituted alkylthio, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl; or
R 2 and R 3 , R 3 and R 4 , or R 2 , R 3 and R 4 are taken together to form a fused heterocyclyl group, a fused substituted heterocyclyl group, a fused aryl group, a fused substituted aryl group, a fused heteroaryl group or a fused substituted heteroaryl group.

2. The compound of claim 1 , wherein R 2 , R 3 and R 4 are taken together to form a fused heterocyclyl group or substituted fused heterocyclyl group of the formula:

and
R 12 and R 13 are each independently selected from the group consisting of H, alkyl, substituted alkyl, alkoxy, substituted alkoxy, acyl, acylamino, acyloxy, amino, substituted amino, aminocarbonyl, aminothiocarbonyl, aminocarbonylamino, aminothiocarbonylamino, aminocarbonyloxy, aminosulfonyl, aminosulfonyloxy, aminosulfonylamino, amidino, carboxyl, carboxyl ester, (carboxyl ester)amino, (carboxyl ester)oxy, cyano, halo, hydroxy, nitro, SO 3 − , sulfonyl, substituted sulfonyl, sulfonyloxy, thioacyl, thiol, alkylthio, substituted alkylthio, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl.

3. The compound of claim 1 , wherein X is H.

4. The compound of claim 1 , wherein X is an analyte.

5. The compound of claim 4 , wherein the analyte is bound through an imine.

6. The compound of claim 4 , wherein the analyte comprises an aldehyde group.

7. The compound of claim 1 , wherein R 3 is analyte substituted hydrazinyl.

8. The compound of claim 7 , wherein the analyte of R 3 is bound through an imine.

9. The compound of claim 7 , wherein the analyte of R 3 is selected from the group consisting of an amino acid, a peptide, a protein, a carbohydrate, a polysaccharide, a nucleoside, a nucleotide, an oligonucleotide, a nucleic acid polymer, a drug, a lipid, and a synthetic polymer.

10. The compound of claim 7 , wherein the analyte of R 3 comprises a carbonyl group or an aldehyde group.

11. A compound selected from the group consisting of:

12. A method for determining the presence of an analyte of interest in a sample, wherein the analyte of interest is selected from the group consisting of an amino acid, a peptide, a protein, a carbohydrate, a polysaccharide, a nucleoside, a nucleotide, an oligonucleotide, a nucleic acid polymer, a drug, a lipid, a synthetic polymer, malondialdehyde, and 4-hydroxynonenal, wherein the method comprises:
contacting the sample with a compound having Formula I:

or a stereoisomer, tautomer, hydrate, solvate, or salt thereof;
wherein,
X is H;
R 1 is selected from the group consisting of H, alkyl and substituted alkyl; R 1a is H;
R 2 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 and R 10 are each independently selected from the group consisting of H, alkyl, substituted alkyl, alkoxy, substituted alkoxy, acyl, acylamino, acyloxy, amino, substituted amino, aminocarbonyl, aminothiocarbonyl, aminocarbonylamino, aminothiocarbonylamino, aminocarbonyloxy, aminosulfonyl, aminosulfonyloxy, aminosulfonylamino, amidino, carboxyl, carboxyl ester, (carboxyl ester)amino, (carboxyl ester)oxy, cyano, halo, hydroxy, nitro, SO 3 − ,sulfonyl, substituted sulfonyl, sulfonyloxy, thioacyl, thiol, alkylthio, substituted alkylthio, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl;
R 3 is selected from the group consisting of alkoxy, substituted alkoxy, amino, substituted amino, hydrazinyl, alkyl, substituted alkyl, acyl, acylamino, acyloxy, aminocarbonyl, aminothiocarbonyl, aminocarbonylamino, aminothiocarbonylamino, aminocarbonyloxy, aminosulfonyl, aminosulfonyloxy, aminosulfonylamino, amidino, carboxyl, carboxyl ester, (carboxyl ester)amino, (carboxyl ester)oxy, cyano, halo, hydroxy, nitro, SO 3 − , sulfonyl, substituted sulfonyl, sulfonyloxy, thioacyl, thiol, alkylthio, substituted alkylthio, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl; or
R 2 and R 3 , R 3 and R 4 , or R 2 , R 3 and R 4 are taken together to form a fused heterocyclyl group, a fused substituted heterocyclyl group, a fused aryl group, a fused substituted aryl group, a fused heteroaryl group or a fused substituted heteroaryl group;
incubating the sample and the compound for a sufficient amount of time to form a complex between the compound and analyte;
illuminating the complex with an appropriate wavelength to form an illuminated complex; and
detecting emissions from the illuminated complex.

13. The method of claim 12 , wherein the analyte and the compound are connected by a covalent bond.

14. The method of claim 13 , wherein the analyte is bound to the compound through an imine.

15. The method of claim 12 , wherein the compound becomes fluorescent after formation of the complex.

16. The method of claim 12 , wherein the analyte comprises an aldehyde group.

17. The method of claim 12 , wherein the sample comprises live ce

Assignees

• Life Technologies Corp

Inventors

• Chen Aimei
• Gee Kyle
• Kang Hee Chol

Classifications

• C09K11/06

  containing organic luminescent materials · CPC title

• C07F9/5442

  Aromatic phosphonium compounds (P-C aromatic linkage) · CPC title

• C07D311/82 Primary

  Xanthenes · CPC title

• G01N33/52

  Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper {and including single- and multilayer analytical elements (immunological elements G01N33/54386; involving labelled immunochemicals G01N33/58; for haemoglobin or occult blood G01N33/72)} · CPC title

• C09K2211/10

  Non-macromolecular compounds · CPC title