Labeling reagents comprising aphenylic analogs of rhodamine dyes

• US 7,256,291 B2
• Filed: 01/22/2004
• Issued: 08/14/2007
• Est. Priority Date: 03/12/2002
• Status: Expired due to Term

• Current Assignee: Enzo Life Sciences Incorporated (Enzo Biochem Incorporated)
• Original Assignee: Enzo Life Sciences Incorporated (Enzo Biochem Incorporated)
• Inventors: Stavrianopoulos, Jannis G., Rabbani, Elazar
• Primary Examiner(s): Riley; Jezia
• Application Number:
  US10/763,076
• Publication Number:
  US 20040254355A1
• Time in Patent Office: 1,300 Days
• Field of Search: 536/26.6, 536/23.1, 546/16, 546/165, 546/270, 546/271.1
• US Class Current: 546/16
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C09B 23/02 the polymethine chain containing an odd number of >CH- or >C[alkyl]- groups
• CPC Class Codes: C09B 47/00 Porphines; Azaporphines non-dyeing compounds C07D487/22
• CPC Class Codes: C09B 57/00 Other synthetic dyes of known constitution
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine

Abstract

This invention provides for labeling reagents, labeled targets and processes for preparing labeling reagents. The labeling reagents can take the form of cyanine dyes, xanthene dyes, porphyrin dyes, coumarin dyes or composite dyes. These labeling reagents are useful for labeling probes or targets, including nucleic acids and proteins. These reagents can be usefully applied to protein and nucleic acid probe based assays. They are also applicable to real-time detection processes.

Claims

1. A labeling reagent comprising an aphenylic analog of a rhodamine dye, said analog comprising at least one reactive group for attaching said labeling reagent to a target, said at least one reactive group being attached directly to said analog or indirectly through a linker arm.

2. The labeling reagent of claim 1, said reagent having the structure

3. The labeling reagent of claim 1, said reagent having the structure

4. The labeling reagent of claim 1, 2 or 3, wherein said reactive group comprises sulfhydryl, hydroxyl, amine, isothiocyanate, isocyanate, monochlorotriazine, dichlorotriazine, mono- or di-halogen substituted pyridine, mono- or di-halogen substituted diazine, maleimide, aziridine, sulfonylhalide, acid halide, hydroxysuccinimide ester, hydroxysulfosuccinimide ester, imidoester, hydrazine, azidonitrophenyl, azide, 3-(2-pyridyl dithio)-proprionamide, glyoxal or aldehyde.

5. The labeling reagent of claim 1, 2 or 3, wherein said reactive group is capable of forming a carbon-carbon linkage with a target.

6. The labeling reagent of claim 5, wherein said reactive group comprises an alkene group, an alkyne group, a halogenated compound or a metallo-organic compound.

7. The labeling reagent of claim 6, wherein said metallo-organic compound comprises mercury, zinc, copper or platinum.

8. The labeling reagent of claim 6, wherein said metallo-organic compound comprises an alkene group or an alkyne group.

9. The labeling reagent of claim 1, wherein said aphenylic linker arm has a backbone comprising one or more carbon atoms.

10. The labeling reagent of claim 1, wherein said aphenylic linker arm has a backbone comprising at least one non-carbon atom.

11. A labeled nucleotide comprising an aphenylic analog of a rhodamine dye, wherein said dye is attached directly to said nucleotide or indirectly through a linker.

12. The labeled nucleotide of claim 11, said labeled nucleotide having the structure:

13. The labeled nucleotide of claim 11, said labeled nucleotide having the structure:

14. The labeled nucleotide of claims 11, 12 or 13, wherein said nucleotide comprises a mono-phosphate, a di-phosphate or a tri-phosphate.

15. The labeled nucleotide of claims 11, 12 or 13, wherein said linker arm has a backbone comprising one or more carbon atoms.

16. The labeled nucleotide of claims 11, 12 or 13, wherein said linker arm has a backbone comprising at least one non-carbon atom.

17. The labeled nucleotide of claims 11, 12 or 13, wherein said linker arm comprises two or more consecutive peptide bonds.