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US 6,388,061 B1Granted
Monomeric building blocks for labeling peptide nucleic acids
AssigneeDako A/S
InventorsFrank Bergmann, Rupert Herrmann, Christoph Seidel, Troels Koch
Life of this patent
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Abstract
The invention relates to novel monomeric building blocks for labeling peptide nucleic acids and similarly constructed nucleic acid-binding oligomers possessing groups which are coupled to a nitrogen base and/or to the peptide backbone of the peptide nucleic acid. The invention furthermore relates to peptide nucleic acids which contain at least one labelled monomeric building block.
Claims35
Compound of the formula (I)
- in which:
- B is a natural or unnatural nitrogen base which optionally carries a protecting group,
- L is a labeling group with the labeling group being ruthenium (bipyridyl)3 bonded to B,
- A, C and D are in each case, independently of each other, chemical bonds or organic radicals,
- E is a group which is selected from N, R1N+ or CH, where R1 is an organic radical or hydrogen,
- Q is a group NR2Y, where R2 is an organic radical or hydrogen, and Y is a protecting group or a carrier,
- I is a group which is selected from COX, CSX, SOX or SO2X, where X is OH, SH, OM, SM or a protecting group,
- M is a cation, and
- n is an integer from 1 to 3.
6 more elements
A nucleic acid-binding oligomer which has a structure of the general formula (III):
- in which:
- n is an integer of at least 3,
- x is an integer from 2 to n−1,
- each of the groups B1 to Bn is a nitrogen base as previously defined,
- each of the groups C1-Cn has the meaning (CR6R7)y, where R6 is hydrogen and R7 is selected from the group consisting of the side chains of naturally occurring alpha-amino acids, or R6 and R7 are independently selected from the group consisting of hydrogen, C1-C6-alkyl, aryl, aralkyl, heteroaryl, hydroxyl, C1-C6-alkoxy, C1-C6-alkylthio, NR3R4 and SR5, where R3 and R4 are as defined below and R5 is hydrogen, C1-C6-alkyl or C1-C6-alkyl which is substituted by hydroxyl, C1-C6-alkoxy or C1-C6-alkylthio, or R6 and R7 together form an alicyclic system or heterocyclic system, or C1-Cn is CO, CS or CNR3;
- each of the radicals D1-Dn has the meaning (CR6R7)2, where R6 and R7 are as defined before, and y and z are integers of from 0 to 10, where the sum of y+z is at least 2;
- each of the radicals G1-Gn−1 has the meaning —NR3CO, NR3CS—, —NR3—SO— or —NR3SO2 in any orientation, where R3 is as defined below;
- each of the radicals A1-An and E1-En is selected such that:
- (a) A1-An is a group of the formula (IIIa), (IIIb), (IIIc) or (IIId), and E1-En is N or R3N+, or
- (b) A1-An is a group of the formula (IIId), and E1-En is CH:
- in which:
- X is 0, S, Se, NR3, CH2 or C(CH3)2,
- Y is a single bond, O, S or NR4,
- p and q are in each case an integer of from 0 to 5,
- r and s are in each case integers of from 0 to 5,
- each of the radicals R8 and R9 is selected independently from the group consisting of hydrogen, hydroxyl, amine, halogen, C1-C4-alkoxy, C1-C4-alkylthio and unsubstituted or substituted C1-C4-alkyl, where the substituents are selected from hydroxyl, C1-C4-alkoxy or C1-C4-alkylthio groups;
- each of the radicals R3 and R4 is selected independently from the group consisting of hydrogen, C1-C4-alkyl which is unsubstituted or substituted by hydroxyl or C1-C4-alkoxy or C1-C4-alkylthio, hyroxyl, C1-C6-alkoxy, C1-C6-alkylthio and amine;
- Q′ and I′ are selected independently from the group consisting of NH2, CONH2, COOH, hydrogen, C1-C6-alkyl, O(C1-C6)-alkyl, an amine which is blocked by a protecting group, labeling groups, intercalators, chelators, peptides, proteins, carbohydrates, lipids, steroids, nucleosides, nucleotides, nucleoside-diphosphates, nucleosidetriphosphates, oligo-nucleotides, including oligoribonucleotides and oligodeoxyribonucleotides, oligonucleosides and soluble and insoluble polymers and also nucleic acid-binding groups, and
- X1 and Y1 is in each case an integer of from 0 to 10, with the compound being such that ruthenium (bipyridyl)3 is present at at least one nitrogen base and/or at a position in the peptide backbone.
16 more elements
Dependencies are inferred from the claim text.