Carboxy X rhodamine analogs

• US 10,139,400 B2
• Filed: 05/28/2015
• Issued: 11/27/2018
• 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.
• Primary Examiner(s): Jarrell, Noble E
• Application Number:
  US14/724,111
• Publication Number:
  US 20150260725A1
• Time in Patent Office: 1,279 Days
• Field of Search: None
• 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 a selected molecule in a sample, comprising:

a) contacting a sample suspected of containing the selected molecule with a composition comprising a dye conjugate according to formula (IIIa), (IIIb) or (IIIc) so as to yield a mixture;

2. The method of claim 1, wherein at least one of R^2-16 is -L-C_s.

3. The method of claim 1, wherein C_s is a polynucleotide.

16. The method of claim 1, wherein the protein is an antibody.

17. The method of claim 1, wherein the polypeptide is a polypeptide-based toxin.

4. 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 dye conjugate according to formula (IIIa), (IIIb) or (IIIc) so as to yield a mixture;

5. The method of claim 4, wherein more than one nucleic acid polymer in a sample is detected in a single reaction.

6. The method of claim 5, wherein the more than one nucleic acid polymer is detected by capillary electrophoresis.

12. The method of claim 4, wherein L is —

CO—

, —

SCH₂CO—

, or —

SO₂—

.

13. The method of claim 12, wherein L is —

SCH₂CO— and

L is bonded to a nucleotide base in the oligonucleotide to form a group

14. The method of claim 4, wherein L is a group

15. The method of claim 4, wherein L is a self-immolative linker selected from the group consisting of

7. A method of labeling a biomolecule comprising:

a) contacting a sample suspected of containing the biomolecule with a composition comprising a dye conjugate according to formula (IIIa), (IIIb) or (IIIc) so as to yield a mixture;

8. The method of claim 7, wherein the dye conjugate contains a cyanobenzothiazole.

9. The method of claim 7, wherein the biomolecule is a protein.

10. The method of claim 9, wherein the protein comprises an N-terminal cysteine.

11. The method of claim 10, wherein the protein comprising an N-terminal cysteine is labeled with the dye conjugate comprising a cyanobenzothiazole.

18. The method of claim 7, wherein the protein is an antibody.

19. The method of claim 7, wherein the polypeptide is a polypeptide-based toxin.