Dyes for the detection or quantification of desirable target molecules

• US 7,569,695 B2
• Filed: 05/24/2005
• Issued: 08/04/2009
• Est. Priority Date: 05/24/2005
• Status: Active Grant

• Current Assignee: Enzo Life Sciences Incorporated (Enzo Biochem Incorporated)
• Original Assignee: Enzo Life Sciences Incorporated (Enzo Biochem Incorporated)
• Inventors: Rabbani, Elazar, Cheng, Wei, Pande, Praveen, Liu, Dakai, Xiang, Yuejun, Khazanchi, Rajesh
• Primary Examiner(s): SEAMAN, D MARGARET M
• Application Number:
  US11/137,771
• Publication Number:
  US 20060269926A1
• Time in Patent Office: 1,533 Days
• Field of Search: 546/270.1
• US Class Current: 546/270.1
• CPC Class Codes: C07D 209/14 Radicals substituted by nitrogen atoms, not forming part of a nitro radical
• CPC Class Codes: C07D 235/20 Two benzimidazolyl-2 radicals linked together directly or via a hydrocarbon or substituted hydrocarbon radical
• CPC Class Codes: C07D 401/14 containing three or more hetero rings
• CPC Class Codes: C07D 405/14 containing three or more hetero rings
• CPC Class Codes: C07D 417/06 linked by a carbon chain containing only aliphatic carbon atoms
• CPC Class Codes: C07D 417/14 containing three or more hetero rings
• CPC Class Codes: C07H 19/10 with the saccharide radical esterified by phosphoric or polyphosphoric acids
• 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: C07K 14/00 Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C09B 23/0091 having only one heterocyclic ring at one end of the methine chain, e.g. hemicyamines, hemioxonol styryl dyes see C09B23/14
• CPC Class Codes: C09B 23/04 one >CH- group, e.g. cyanines, isocyanines, pseudocyanines
• CPC Class Codes: C09B 23/06 three >CH- groups, e.g. carbocyanines
• CPC Class Codes: C09B 23/083 five >CH- groups
• CPC Class Codes: C09B 23/105 two >CH- groups
• CPC Class Codes: C09B 23/145 the ethylene chain carrying an heterocyclic residue, e.g. heterocycle-CH=CH-C6H5
• CPC Class Codes: C09B 23/146 (Benzo)thiazolstyrylamino dyes

Abstract

The present invention provides dyes, reactive dyes and labeled reagents that may be used in the detection or quantification of desirable target molecules, such as proteins and nucleic acids. Dyes are provided that may be used free in solution where the binding of the dye to the target molecule provides signal generation. Dyes are also provided that comprise reactive groups that may be used to attach the dyes to probes that will bind to desirable target molecules. The novel dyes of the present invention have been modified by the addition of charged and polar groups to provide beneficial properties.

Claims

1. A dye having the formula:

2. The dye of claim 1 wherein at least one of R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹³, R¹⁴, or R¹⁵ comprises a reactive group.

3. The dye of claim 2 wherein said reactive group comprises a nucleophilic reactive group, an electrophilic reactive group, a terminal alkene, a terminal alkyne, a coordinate group or an alkylating agent.

4. The dye of claim 3 wherein said nucleophilic reactive group comprises a thiol, amine or hydroxyl group.

5. The dye of claim 3 wherein said electrophilic reactive group comprises an isocyanate, isothiocyanate, monochlorotriazine, dichlorotriazine, 4,6,-dichloro-1 ,3,5-triazines, mono- or di-halogen substituted pyridine, mono- or di-halogen substituted diazine, maleimide, haloacetamide, aziridine, sulfonyl halide, acid halide, hydroxysuccin im ide ester, hydroxysulfosuccinimide ester, imido ester, hydrazine, azidonitrophenol, azide, 3-(2-pyridyl dithio)-proprionamide glyoxal or aldehyde group.

6. The dye of claim 1, having the following structure:

7. The dye of claim 6, having the following structure:

8. The dye of claim 6, having the following structure:

9. The dye of claim 8, having the following structure:

10. The dye of claim 8, having the following structure:

11. The dye of claim 10, having the following structure:

12. The dye of claim 1, having the following structure:

13. The dye of claim 1, wherein said dye further comprises one or more charged groups or polar groups.

14. The dye of claim 13, wherein at least one of said one or more charged groups or polar groups is monomeric.

15. The dye of claim 13, wherein at least one of said one or more charged groups or polar groups is polymeric.

16. The dye of claim 13, wherein at least one of said one or more charged groups or polar groups is anionic.

17. The dye of claim 13, wherein at least one of said one or more charged groups or polar groups is cationic.

18. The dye of claim 1, wherein said dye further comprises one or more anionic charged groups or polar groups and one or more cationic charged groups or polar groups.

19. The dye of claim 9, having the following structure:

20. A dye having the following structure:

21. The dye of claim 9, having the following structure:

22. A dye having the following structure:

23. The dye of claim 1, wherein said alkyl or alkoxy groups independently comprise from 1-18 carbons.

24. The dye of claim 23, wherein said alkyl or alkoxy groups independently comprise from 1-6 carbons.

25. A dye having the formula:

26. The dye of claim 25, wherein said nucleophilic reactive group comprises a thiol, amine or hydroxyl group.

27. The dye of claim 25, wherein said electrophilic reactive group comprises an isocyanate, isothiocyanate, monochlorotriazine, dichiorotriazine, 4,6,-dichloro-1 ,3,5-triazines, mono- or di-halogen substituted pyridine, mono- or di-halogen substituted diazine, maleimide, haloacetamide, aziridine, sulfonyl halide, acid halide, hydroxysuccinimide ester, hyd roxysulfosucoinimide ester, imido ester, hydrazine, azidonitrophenol, azide, 3-(2-pyridyl dithio)-proprionamide, glyoxal or aldehyde group.

28. The dye of claim 25, wherein said dye further comprises one or more charged groups or polar groups.

29. The dye of claim 28, wherein at least one of said one or more charged groups or polar groups is monomeric.

30. The dye of claim 28, wherein at least one of said one or more charged groups or polar groups is polymeric.

31. The dye of claim 28, wherein at least one of said one or more charged groups or polar groups is anionic.

32. The dye of claim 28, wherein at least one of said one or more charged groups or polar groups is cationic.

33. The dye of claim 25, wherein said dye further comprises one or more anionic charged groups or polar groups and one or more cationic charged groups or polar groups.

34. The dye of claim 25, wherein said alkyl or alkoxy groups independently comprise from 1-18 carbons.

35. The dye of claim 34, wherein said alkyl or alkoxy groups independently comprise from 1-6 carbons.

36. A dye having the formula:

37. The dye of claim 36 wherein at least one of R²,R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹³, R¹⁴, or R¹⁵ comprises a reactive group.

38. The dye of claim 37 wherein said reactive group comprises a nucleophilic reactive group, an electrophilic reactive group, a terminal alkene, a terminal alkyne, a coordinate group or an alkylating agent.

39. The dye of claim 38 wherein said nucleophilic reactive group comprises a thiol, amine or hydroxyl group.

40. The dye of claim 38 wherein said electrophilic reactive group comprises an isocyanate, isothiocyanate, monochlorotriazine, dichlorotriazine, 4,6,-dichloro-1 ,3,5 - triazines, mono- or di-halogen substituted pyridine, mono- or di-halogen substituted diazine, maleimide, haloacetamide, aziridine, sulfonyl halide, acid halide, hydroxysuccinim ide ester, hydroxysu Ifosucoinim ide ester, imido ester, hydrazine, azidonitrophenol, azide, 3-(2-pyridyl dithio)-proprionamide, glyoxal or aldehyde group.

41. The dye of claim 36, having the followina structure:

42. The dye of claim 41, having the following structure:

43. The dye of claim 41, having the following structure:

44. The dye of claim 43, having the following struoture:

45. The dye of claim 43, having the following structure:

46. The dye of claim 45, having the following stucture:

47. The dye of claim 36, having the following structure:

48. The dye of claim 36, wherein said dye further comprises one or more charged groups or polar groups.

49. The dye of claim 48, wherein at least one of said one or more charged groups or polar groups is monomeric.

50. The dye of claim 48, wherein at least one of said one or more charged groups or polar groups is polymeric.

51. The dye of claim 48, wherein at least one of said one or more charged groups or polar groups is anionic.

52. The dye of claim 48, wherein at least one of said one or more charged groups or polar groups is cationic.

53. The dye of claim 36, wherein said dye further comprises one or more anionic charged groups or polar groups and one or more cationic charged groups or polar groups.

54. The dye of claim 41, having the following structure

55. The dye of claim 43, having the following structure:

56. The dye of claim 45, having the following structure:

57. The dye of claim 47, having the following structure:

58. A dye having the following structure:

59. A dye having the following structure:

60. The dye of claim 36, wherein said alkyl or alkoxy groups independently comprise from 1-18 carbons.

61. The dye of claim 56, wherein said alkyl or alkoxy groups independently comprise from 1-6 carbons.

62. A dye having the formula:

63. The dye of claim 62, wherein said nucleophilic reactive group comprises a thiol, amine or hydroxyl group.

64. The dye of claim 62, wherein said electrophilic reactive group comprises an isocyanate, isothiocyanate, monochiorotriazine, diohiorotriazine, 4,6,-dlchloro-1,3,5-triazines, mono- or di-halogen substituted pyridine, mono- or di-halogen substituted diazine, maleimide, haloacetamide, aziridine, sulfonyl halide, acid halide, hydroxysuccinimide ester, hydroxysulfosuccinimid ester, imido ester, hydrazine, azidonitrophenol;

, azide, 3-(2-pyridyl dithio)-proprionamide, glyoxal or aldehyde group.

65. The dye of claim 62, wherein said dye further comprises one or more charged groups or polar groups.

66. The dye of claim 65, wherein at least one of said one or more charged groups or polar groups is monomeric.

67. The dye of claim 65, wherein at least one of said one or more charged groups or polar groups is polymeric.

68. The dye of claim 65, wherein at least one of said one or more charged groups or polar groups is anionic.

69. The dye of claim 65, wherein at least one of said one or more charged groups or polar groups is cationic.

70. The dye of claim 62, wherein said dye further comprises one or more anionic charged groups or polar groups and one or more cationic charged groups or polar groups.

71. The dye of claim 62, having the following structure:

72. The dye of claim 62, having the following structure:

73. The dye of claim 62 having the following structure:

74. The dye of claim 62 having the following structure:

75. The dye of claim 62, wherein said alkyl or alkoxy groups independently comprise from 1-18 carbons.

76. The dye of claim 75 wherein said alkyl or alkoxy groups independently comprise from 1-6 carbons.