Oligonucleotide N3'-P5' phosphoramidates: hybridization and nuclease resistance properties

• US 5,837,835 A
• Filed: 06/06/1995
• Issued: 11/17/1998
• Est. Priority Date: 03/18/1994
• Status: Expired due to Term

• Current Assignee: Geron Corporation
• Original Assignee: Lynx Therapeutics, Inc. (Illumina Incorporated)
• Inventors: Schultz, Ronald G., Gryaznov, Sergei M., Chen, Jer-kang
• Primary Examiner(s): Chambers, Jasemine C.
• Assistant Examiner(s): PRIEBE, SCOTT DAVID
• Application Number:
  US08/477,306
• Time in Patent Office: 1,260 Days
• Field of Search: 435/6, 435/91.1, 536/23.1, 536/24.5, 536/24.3, 536/24.1, 514/44
• US Class Current: 536/23.1
• CPC Class Codes: C07H 19/06 Pyrimidine radicals
• CPC Class Codes: C07H 19/16 Purine radicals
• 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

Modified oligonucleotides 3'"'"'-NHP(O)(O^-)O-5'"'"' phosphoramidates were synthesized on a solid phase support. The phosphoramidate analogs were found to have significantly increased resistance toward phosphodiesterase digestion. Thermal dissociation experiments demonstrated that these compounds form more stable duplexes than phosphodiesters with complementary DNA and particularly RNA strands. Further, the phosphoramidate analogs can also form stable triplexes with double-stranded DNA target, where under similar conditions parent phosphodiester compounds failed to do so.

Claims

1. An oligodeoxyribonucleotide comprising a defined sequence of nucleoside subunits joined by inter-subunit linkages and defined by the formula:

##STR1## wherein B is a purine or pyrimidine or an analog thereof and n is between 2 and 48.

2. The oligodeoxyribonucleotide of claim 1, wherein B of said oligonucleotide is selected from the group consisting of uracil, thymine, adenine, guanine, cytosine, 5-methylcytosine, 5-bromouracil, and inosine.

3. The oligodeoxyribonucleotide of claim 2, wherein B of said oligodeoxyribonucleotide is selected from the group consisting of uracil, thymine, adenine, guanine, and cytosine.

4. The oligodeoxyribonucleotide of claim 1, said oligodeoxyribonucleotide that hybridizes with a target nucleic acid sequence.

5. The oligodeoxyribonucleotide of claim 3, said oligodeoxyribonucleotide that hybridizes to an RNA target.

6. The oligodeoxyribonucleotide of claim 1, wherein said oligodeoxyribonucleotide carries a reporter moiety.

7. The oligodeoxyribonucleotide of claim 6, wherein said reporter moiety is selected from the group consisting of radioactive labels, biotin labels, and fluorescent labels.