Label target and labeling reagents comprising backbones with at least two consecutive peptide bonds

• US 8,357,793 B2
• Filed: 01/22/2004
• Issued: 01/22/2013
• Est. Priority Date: 03/12/2002
• Status: Active Grant

• Current Assignee: Enzo Life Sciences Incorporated (Enzo Biochem Incorporated)
• Original Assignee: Enzo Life Sciences Incorporated (Enzo Biochem Incorporated)
• Inventors: Stavrianopoulos, Jannis G., Rabbani, Flazar
• Primary Examiner(s): Riley, Jezia
• Application Number:
  US10/763,104
• Publication Number:
  US 20110098441A1
• Time in Patent Office: 3,288 Days
• Field of Search: 436/544, 536/23.1, 536/25.3, 536/26.6, 435/6
• US Class Current: 536/26.6
• 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 labeled target comprising

T-L-M

wherein T is a target, M is a marker moiety and L is a chemical group covalently linking said M to T, said chemical group L comprising a backbone that comprises at least two consecutive peptide bonds, wherein L further comprises at least one additional moiety comprsing a peptide bond, an amino acid, a C₁-C₂₀ aliphatic chain an alkene group, an alkene group, a saturated or unsaturated or partially saturated ring, a heterocyclic ring or a sugar.

2. The labeled target of claim 1, wherein said target T comprises a protein, a peptide, a nucleic acid, a nucleotide or a nucleotide analog, a receptor, a natural or synthetic drug, a synthetic oligomer, a synthetic polymer, a hormone, a lymphokine, a cytokine, a toxin, a ligand, an antigen, a hapten, an antibody, a carbohydrate, a sugar or an oligo- or polysaccharide.

3. The labeled target of claim 1, wherein said target T comprises a nucleic acid, nucleotide, or nucleotide analog.

4. The labeled target of claim 1, wherein said ligand comprises biotin, iminobiotin, digoxygenin or fluorescein.

5. The labeled target of claim 1, wherein said marker moiety M comprises a ligand that comprises biotin, iminobiotin, digoxigenin or fluorescein.

6. The labeled target of claim 1, wherein said marker moiety M comprises a fluorescent dye.

7. The labeled target of claim 6, wherein said fluorescent dye comprises a xanthene dye, a cyanine dye, a coumarin dye, a porphyrin dye or a composite dye.

8. The labeled target of claim 7, wherein said fluorescent dye comprises a xanthene dye which comprises fluorescein, rhodamine or rhodol, or derivatives thereof.

9. The labeled target of claim 1, wherein at least one of said two consecutive peptide bonds are separated by a single atom.

10. The labeled target of claim 9, wherein said single atom comprises C, N, S, O or P.

11. The labeled target of claim 1, wherein said backbone further comprises one or more carbon atoms.

12. The labeled target of claim 1, wherein said backbone further comprises at least one non-carbon atom.

13. The labeled target of claim 12, wherein said non-carbon atom is a sulfur, oxygen or nitrogen.

14. The labeled target of claim 1, wherein said chemical group L comprises a di-peptide or an oligo-peptide.

15. The labeled target of claim 14, wherein said di-peptide or oligo-peptide comprises (glycine)₂ or (glycine)₄.

16. A labeling reagent comprising

R-L-M

wherein R is a reactive group, M is a marker moiety and L is a chemical group covalently linking said M to R, said chemical group L comprising a backbone that comprises at least two consecutive peptide bonds, wherein at least one of said two consecutive peptide bonds are separated by a single atom.

17. The labeling reagent of claim 16, wherein said reactive group R 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.

18. The labeling reagent of claim 16, wherein said reactive group R is capable of forming a carbon-carbon linkage with a target.

19. The labeling reagent of claim 16, wherein said reactive group R comprises an alkene group, an alkyne group, a halogenated compound or a metallo-organic compound.

20. The labeling reagent of claim 16, wherein said reactive group R comprises a metallo-organic compound comprising mercury, zinc, copper or platinum.

21. The labeling reagent of claim 16, wherein said reactive group R comprises a metallo-organic compound comprising an aromatic group, an aromatic heterocyclic group, an alkene group or an alkyne group.

22. The labeling reagent of claim 16, wherein said marker moiety M comprises a ligand that comprises biotin, iminobiotin, digoxigenin or fluorescein.

23. The labeling reagent of claim 16, wherein said marker moiety M comprises a fluorescent dye.

24. The labeling reagent of claim 23, wherein said fluorescent dye comprises a xanthene dye, a cyanine dye, a coumarin dye, a porphyrin dye or a composite dye.

25. The labeling reagent of claim 23, wherein said fluorescent dye comprises a xanthene dye comprising fluorescein, rhodamine or rhodol, or derivatives thereof.

26. The labeling reagent of claim 16, wherein said single atom comprises C, N, S, O or P.

27. The labeling reagent of claim 16, wherein said backbone further comprises one or more carbon atoms.

28. The labeling reagent of claim 16, wherein said backbone further comprises at least one non-carbon atom.

29. The labeling reagent of claim 28, wherein said non-carbon atom comprises sulfur, oxygen or nitrogen.

30. The labeling reagent of claim 16, wherein said chemical group L further comprises at least one additional moiety comprising a peptide bond, an amino acid, a C₁-C₂₀ aliphatic chain, an alkene group, an alkyne group, a saturated or unsaturated or partially saturated ring, a heterocyclic ring, or a sugar.

31. The labeling reagent of claim 16, wherein said chemical group L comprises a di-peptide or an oligo-peptide.

32. The labeling reagent of claim 31, wherein said di-peptide or oligo-peptide comprises (glycine)₂ or (glycine)₄.gs and sugars.