CARBOXY X RHODAMINE ANALOGS

• US 20150260725A1
• Filed: 05/28/2015
• Published: 09/17/2015
• Est. Priority Date: 11/21/2011
• Status: Active Grant

• Current Assignee: Promega Corporation
• Original Assignee: Promega Corporation
• Inventors: Kirkland, Thomas A., McDougall, Mark G., Dwight, Stephen J.
• Granted Patent: US 10,139,400 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: C07D 223/16 Benzazepines; Hydrogenated benzazepines
• CPC Class Codes: C07D 471/06 Peri-condensed systems
• CPC Class Codes: C07D 491/147 the condensed system containing one ring with oxygen as ring hetero atom and two rings with nitrogen as ring hetero atom
• CPC Class Codes: C07D 491/16 Peri-condensed systems
• CPC Class Codes: C07D 491/22 in which the condensed system contains four or more hetero rings
• CPC Class Codes: C07F 9/65586 at least one of the hetero rings does not contain nitrogen as ring hetero atom
• CPC Class Codes: C07H 19/073 with 2-deoxyribosyl as the saccharide radical
• CPC Class Codes: C07H 19/10 with the saccharide radical esterified by phosphoric or polyphosphoric acids
• CPC Class Codes: C09B 11/24 Phthaleins containing amino groups ; Phthalanes; Fluoranes; Phthalides; Rhodamine dyes; Phthaleins having heterocyclic aryl rings; Lactone or lactame forms of triarylmethane dyes
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: G01N 2333/90241 acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13)
• CPC Class Codes: G01N 33/542 with steric inhibition or signal modification, e.g. fluorescent quenching
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: G01N 33/6845 Methods of identifying protein-protein interactions in protein mixtures

Abstract

The present invention provides novel fluorescent dyes and kits containing the same, which are useful for labeling a wide variety of biomolecules, cells and microorganisms. The present invention also provides various methods of using the fluorescent dyes for research and development, forensic identification, environmental studies, diagnosis, prognosis and/or treatment of disease conditions.

Claims

1. A method to detect an interaction between a biomolecule and a protein of interest comprising:

a) contacting a sample suspected of containing the protein of interest with a composition comprising a conjugate comprising a compound and the biomolecule; and

b) detecting the presence or amount of the compound, thereby detecting the interaction between the protein of interest and the biomolecule;

wherein the compound is a compound according to formula (IIIa), (IIIb) or (IIIc);

2. The method of claim 1, wherein the protein of interest is fused to a reporter protein.

3. The method of claim 2, wherein the reporter protein is a luciferase protein.

4. The method of claim 1, wherein the labeled biomolecule is a tracer.

5. The method of claim 1, wherein the interaction is detected by bioluminescence reasonance energy transfer.

6. The method of claim 1, wherein the protein of interest is expressed in a cell.

7. The method of claim 1, wherein at least one of R2-16 is -L-CS.

8. A method to detect a selected molecule in a sample, comprising:

a) contacting a sample suspected of containing the selected molecule with a composition comprising a conjugate comprising a compound and a ligand for the selected molecule so as to yield a mixture; and

b) detecting the presence or amount of the compound in the mixture, thereby detecting the selected molecule in the sample;

wherein the compound is a compound according to formula (IIIa), (IIIb) or (IIIc);

9. The method of claim 8, wherein at least one of R^2-16 is -L-C_s.

10. The method of claim 8, wherein the ligand is an oligonucleotide.

11. A method of detecting the presence of a nucleic acid polymer in a sample comprising:

a) contacting a sample suspected of containing a nucleic acid polymer with a composition comprising a conjugate comprising a compound and an oligonucleotide; and

b) detecting the presence or amount of the compound in the sample, thereby detecting the presence of the nucleic acid polymer in the sample;

wherein the compound is a compound according to formula (IIIa), (IIIb) or (IIIc);

12. The method of claim 11, wherein more than one nucleic acid polymer in a sample is detected in a single reaction.

13. The method of claim 12, wherein the more than one nucleic acid polymer is detected by capillary electrophoresis.

14. A method of labeling a biomolecule comprising:

a) contacting a sample suspected of containing the biomolecule with a composition comprising a compound so as to yield a mixture; and

b) detecting the presence or amount of the compound, thereby detecting the presence or amount of the labeled biomolecule in the mixture;

wherein the compound is a compound according to formula (IIIa), (IIIb) or (IIIc);

15. The method of claim 14, wherein the compound contains a cyanobenzothiazole.

16. The method of claim 14, wherein the biomolecule is a protein.

17. The method of claim 16, wherein the protein comprises an N-terminal cysteine.

18. The method of claim 17, wherein the protein comprising an N-terminal cysteine is labeled with the compound comprising a cyanobenzothiazole.