Detection of biologically active compounds

• US 20070042412A1
• Filed: 08/18/2006
• Published: 02/22/2007
• Est. Priority Date: 02/19/2004
• Status: Abandoned Application

• Current Assignee: University College Cork- National University of Ireland
• Original Assignee: University College Cork- National University of Ireland
• Inventors: O'Riordan, Tomas, Papkovsky, Dmitri, O'Sullivan, Paul, Burke, Martina
• Application Number:
  US11/506,209
• Publication Number:
  US 20070042412A1
• Time in Patent Office: Days
• US Class Current: 435/6
• 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

Abstract

A probe comprises a supramolecular structure having a chemical or biological recognition moiety; a phosphorescent reporter label; and an effector which interacts with the label so that the probe alters its phosphorescent characteristics on recognition of a target. The phosphorescent reporter label may have an emission lifetime in the order of 1 μs to 10 ms and may be selected from phosphorescent tetrapyrrolic compounds and their metallocomplexes.

Claims

1-52. -52. (canceled)

53. A probe comprising a supramolecular structure having:

a chemical or biological recognition moiety;

a phosphorescent reporter label; and

an effector moiety,

in which probe the label interacts with the effector so that the probe alters its phosphorescent characteristics upon recognition of a target.

54. The probe as claimed in claim 53 wherein the phosphorescent reporter label has an emission lifetime in the order of 1 μ

s to 10 ms.

55. The probe as claimed in claim 53 wherein the phosphorescent reporter label has an emission lifetime in the order of 10 μ

s to 1000 μ

s.

56. The probe as claimed in claim 53 wherein the phosphorescent reporter label is selected from a group of phosphorescent tetrapyrrolic compounds and their metallocomplexes.

57. The probe as claimed in claim 56 wherein the phosphorescent reporter label is selected from any one or more of phosphorescent metallocomplexes of porphyrins, chlorins, porphyrin-ketones and related structures.

58. The probe as claimed in claim 57 wherein the phosphorescent label is platinum(II)-porphyrin.

59. The probe as claimed in claim 57 wherein the phosphorescent label is platinum(II)-coproporphyrin.

60. The probe as claimed in claim 57 wherein the phosphorescent label is palladium(II)-porphyrin.

61. The probe as claimed in claim 57 wherein the phosphorescent label is palladium(II)-coproporphyrin.

62. The probe as claimed in claim 53 wherein the phosphorescent label is in the form of a monofunctional labelling reagent.

63. The probe as claimed in claim 53 wherein the effector moiety is selected from any one or more of dabcyl, QSY-7™

, ‘

black hole quenches’

™

, rhodamine green, FITC, Cy5™

, and analogs thereof.

64. The probe as claimed in claim 53 wherein the effector moiety comprises a small-size chemical structure.

65. The probe as claimed in claim 64 wherein the effector moiety comprises a chemical structure less than 300 Daltons in size.

66. The probe as claimed in claim 64 wherein the effector moiety is selected from any one or more of dinitrophenol, a nitrophenol moiety and derivatives thereof.

67. The probe as claimed in claim 53 wherein the effector moiety is a modified nucleotide base.

68. The probe as claimed in claim 53 wherein the phosphorescent reporter label and the effector are both provided by the same chemical structure.

69. The probe as claimed in claim 68 wherein the reporter label and the effector both comprise a phosphorescent metalloporphyrin label.

70. The probe as claimed in claim 53 wherein the recognition moiety is a common biomolecular structure or a biopolymer.

71. The probe as claimed in claim 53 further comprising a spacer(s) linking the recognition moiety, the reporter label and the effector.

72. The probe as claimed in claim 71 wherein the spacer(s) is 2 to 18 atoms in length.

73. The probe as claimed in claim 53 wherein the reporter label is attached to one of the termini of a biopolymer acting as recognition moiety.

74. The probe as claimed in claim 53 wherein the recognition moiety comprises a biopolymer with the reporter label attached to one of its termini and the effector attached to the other termini.

75. The probe as claimed in claim 53 wherein the recognition moiety comprises a biopolymer with the reporter label attached to one of its termini and the effector attached internally.

76. The probe as claimed in claim 53 wherein the recognition moiety comprises a biopolymer with the effector attached to one of its termini and the reporter label attached internally.

77. The probe as claimed in claim 53 wherein the probe is quenched in its free form in solution.

78. The probe as claimed in claim 53 wherein the chemical or biological recognition moiety comprises a single-stranded oligonucleotide sequence.

79. The probe as claimed in claim 78 wherein the probe produces a phosphorescent signal response upon recognition of a complementary target, hybridisation and formation of a double-stranded structure with the target.

80. The probe as claimed in claim 78 wherein the reporter label and the effector are attached to the 5′

- and 3′

-ends respectively of the specific nucleic acid sequence.

81. The probe as claimed in claim 78 wherein the reporter label is attached to the 5′

-end of the probe and the effector is incorporated internally or attached to one of the bases inside the probe sequence.

82. The probe as claimed in claim 78 wherein the probe is 15 to 100 bases long.

83. The probe as claimed in claim 82 wherein the probe is 20 to 50 bases long.

84. The probe as claimed in claim 78 wherein the probe has the ability to hybridise to a target and act as a primer in the process of elongation of the polynucleotide chain by a polymerase enzyme with the target acting as a template.

85. The probe as claimed in claim 78 wherein the reporter label is Pt-porphyrin and the internal effector is a modified nucleotide base.

86. The probe as claimed in claim 53 wherein the chemical or biological recognition moiety comprises an oligopeptide sequence.

87. The probe as claimed in claim 84 wherein quenching of the reporter label is affected by probe cleavage associated with the recognition process.

88. The probe as claimed in claim 87 wherein the probe is cleaved or modified by a specific enzyme.

89. The probe as claimed in claim 53 wherein the chemical or biological recognition moiety comprises a structure acting as an intrinsic quencher for the reporter label.

90. The phosphorescent probe as claimed in claim 89 wherein the intrinsic quencher for the phosphorescent metalloporphyrin label is a histidine residue within an oligopeptide sequence.

91. The phosphorescent probe as claimed in claim 90 wherein the intrinsic quencher for the phosphorescent porphyrin label is a tyrosine residue within an oligopeptide sequence.

92. The probe as claimed in claim 53 wherein the chemical or biological recognition moiety comprises a polysaccharide or a peptide nucleic acid.

102. The probe as claimed in claim 53 wherein the reporter label has two distinct excitation bands.

105. The assay utilising a probe as claimed in claim 53 wherein the assay is selected from hybridisation, binding and enzymatic assays.

106. The assay as claimed in claim 105 wherein the assay is based on the use of close proximity quenching of a long-decay phosphorescent label.