Nucleotide with an alpha-phosphate mimetic

• US 8,153,779 B2
• Filed: 10/16/2009
• Issued: 04/10/2012
• Est. Priority Date: 04/18/2007
• Status: Expired due to Fees

• Current Assignee: Roche Diagnostics Operations Incorporated (Roche Holding AG)
• Original Assignee: Roche Diagnostics Operations Incorporated (Roche Holding AG)
• Inventors: Heindl, Dieter
• Primary Examiner(s): Crane, Lawrence E
• Application Number:
  US12/580,445
• Publication Number:
  US 20100093990A1
• Time in Patent Office: 907 Days
• Field of Search: None
• US Class Current: 536/26.23
• 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. The nucleotides have a structure

in which B is a purine or pyrimidine, S is a sugar unit, Y is an OH, a monophosphate, or a diphosphate, and Acc is an electron acceptor selected from the group consisting of (a) —CN and (b) —SO₂—R′ in which R′ contains one amino-substituted alkyl or one optionally substituted aryl.

Claims

1. An optionally labeled nucleotide, oligonucleotide, or polynucleotide having the structure

2. The optionally labeled nucleotide, oligonucleotide, or polynucleotide according to claim 1 wherein the ribosyl moiety is a 1,5-disubstituted ribosyl, 1,5-disubstituted deoxyribosyl, or 1,5-disubstituted dideoxyribosyl.

3. The optionally labeled nucleotide, oligonucleotide, or polynucleotide according to claim 2, wherein the optional 3′

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

4. A process for producing a nucleotide according to claim 1 comprising the step of reacting a trivalent phosphorus atom located at the 5′

position of a nucleotide, the phosphorus atom having at least one protected hydroxyl group, 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 and (b) —

SO₂—

R′

in which R′

contains one amino-substituted alkyl or one optionally substituted aryl.

5. A process for producing a nucleotide according to claim 1 comprising the step of reacting a 5′

phosphoramidite nucleoside with an azide of the structure

N═

N═

N-Acc

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

CN and (b) —

SO₂—

R′

in which R′

contains one amino-substituted alkyl- or one optionally substituted aryl.