Nucleotide with an Alpha-Phosphate Mimetic

• US 20100093990A1
• Filed: 10/16/2009
• Published: 04/15/2010
• Est. Priority Date: 04/18/2007
• Status: Active Grant

• Current Assignee: Roche Diagnostics Operations Incorporated (Roche Holding AG)
• Original Assignee: Roche Diagnostics Operations Incorporated (Roche Holding AG)
• Inventors: Heindl, Dieter
• Granted Patent: US 8,153,779 B2
• Time in Patent Office: Days
• US Class Current: 536/26.6
• CPC Class Codes: C07H 19/10 with the saccharide radical esterified by phosphoric or polyphosphoric acids
• CPC Class Codes: C07H 19/20 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

Abstract

Described are modified mononucleotides and processes for their production wherein these nucleotides contain at least once the structure P═N-Acc in which Acc is an electron acceptor or an electron acceptor substituted with a residue R, and R is any organic substituent.

Claims

1. A nucleotide having a structure

in which B is a naturally occurring nucleobase, a modified nucleobase or a nucleobase analogue,

S is a sugar unit,

Y is an OH, a monophosphate, or a diphosphate,

wherein an oxygen atom of the alpha-phosphate is replaced by

—

N-Acc

in which Acc is an electron acceptor selected from the group consisting of (a) —

CN, (b) —

SO2-R′

in which R′

contains at least one amino-substituted alkyl, an optionally substituted aryl or an optionally substituted heterocycle, and (c) electron-deficient, six-membered N+-heterocycles selected from a group comprising pyridinium, pyrimidinium, and quinolinium.

2. A nucleotide according to claim 1 wherein R or R′

contains a detectable unit or a functional group.

3. A nucleotide according to claim 1 wherein the sugar unit is a ribose, deoxyribose, or dideoxyribose.

4. A nucleotide according to claim 3, wherein the 3′

position is selected from the group consisting of H, OH, a protective group, a detectable label, a mononucleotide, a polynucleotide, and a solid phase.

5. A process for producing a nucleotide modified at the alpha position wherein a trivalent phosphorus atom located at the 5′

position of a nucleotide and provided with at least one protected hydroxyl group is reacted with an azide of the structure

N═

N═

N-Acc

in which Acc is an electron acceptor selected from the group consisting of (a) —

CN, (b) —

SO2-R′

in which R′

contains at least one amino-substituted alkyl, an optionally substituted aryl or an optionally substituted heterocycle, and (c) electron-deficient, six-membered N+-heterocycles selected from a group comprising pyridinium, pyrimidinium, and quinolinium.

6. The process according to claim 5, wherein a 5′

phosphoramidite nucleoside is reacted with an azide of the structure

N═

N═

N-Acc