Labeling reagent

• US 20070196852A1
• Filed: 02/16/2007
• Published: 08/23/2007
• Est. Priority Date: 02/20/2006
• Status: Active Grant

• Current Assignee: Roche Diagnostics Operations Incorporated (Roche Holding AG)
• Original Assignee: Roche Diagnostics Operations Incorporated (Roche Holding AG)
• Inventors: Heindl, Dieter, Josel, Hans-Peter, Bergmann, Frank, Kessler, Dirk, Weilke, Christian, Lampert, Gunter
• Granted Patent: US 7,759,470 B2
• Time in Patent Office: Days
• US Class Current: 435/6.12
• CPC Class Codes: C07H 19/04 Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
• CPC Class Codes: C07H 19/23 Heterocyclic radicals containing two or more heterocyclic rings condensed among themselves or condensed with a common carbocyclic ring system, not provided for in groups C07H19/14 - C07H19/22
• 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: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: Y02P 20/55 Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Abstract

The current invention restates substituted indole nucleosides as both terminal as well as internal building blocks of labeled oligonucleotide probes for the detection, analysis and quantitation of nucleic acids. The substituent comprises a linker and a detectable group or a linker and a reactive group for post synthesis coupling. These modified nucleosides grant access to a wide application area. These new substituted indole nucleosides can be used as labeling reagents for the facile preparation of e.g. optimized hybridization probes, Taqman-probes, or molecular beacon probes.

Claims

1. A compound having the formula

2. The compound of claim 1 wherein X is selected from the group consisting of a reactive group, a protected reactive group, a linker group, a protected linker group, and a signal entity.

3. The compound of claim 1 wherein R═

H.

4. The compound of claim 1 wherein B is selected from the group consisting of a phosphoramidate group, an H-phosphonate, and a CPG and A is a protective group.

5. The compound of claim 1 wherein A is selected from the group consisting of a phosphoramidate group, an H-phosphonate, and a CPG and B is a protective group.

6. An oligonucleotide comprising a compound having the formula:

7. The oligonucleotide of claim 6 wherein said oligonucleotide further comprises a second compound having said formula.

8. The oligonucleotide of claim 6 wherein said oligonucleotide comprises at least two signal entities.

9. The oligonucleotide of claim 6 wherein said signal entity is a fluorescent entity.

10. A method for synthesis of the oligonucleotide of claim 6 comprising the step of incorporating during the synthesis a compound having the formula

11. A method for synthesis of the oligonucleotide of claim 6 comprising the steps of

incorporating during synthesis a compound having the formula

wherein

A is a protecting group and B is selected from the group consisting of a phosphoramidate group, an H-phosphonate, and a CPG,

R is selected from the group consisting of H, OH, O-alkyl, O-alkenyl, O-alkinyl, O-protective group, and F, and

X is a reactive group or a linker with a reactive group, and

coupling a signal entity to said reactive group.

12. A hybridization probe comprising a compound having the formula: