Antisense inhibition of ras gene with chimeric and alternating oligonucleotides

• US 6,359,124 B1
• Filed: 02/12/1999
• Issued: 03/19/2002
• Est. Priority Date: 04/03/1995
• Status: Expired due to Term

• Current Assignee: ISIS Pharmaceuticals Incorporated (Ionis Pharmaceuticals, Inc.)
• Original Assignee: ISIS Pharmaceuticals Incorporated (Ionis Pharmaceuticals, Inc.)
• Inventors: Monia, Brett P., Freier, Susan M., Cook, Phillip Dan, Ecker, David J., Sanghvi, Yogesh S.
• Primary Examiner(s): Fredman, Jeffrey
• Assistant Examiner(s): CHAKRABARTI, ARUN K
• Application Number:
  US09/248,386
• Time in Patent Office: 1,131 Days
• Field of Search: 536/23.1, 536/24.5, 536/27.21, 536/28.4, 536/28.5, 435/6, 514/44
• US Class Current: 536/23.1
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• 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: C12N 15/1135 against oncogenes or tumor suppressor genes
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/318 where the PO2 is completely replaced, e.g. MMI or formacetal
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 2'-R Modification
• CPC Class Codes: C12N 2310/341 Gapmers, i.e. of the type ===---===
• CPC Class Codes: C12N 2310/345 having at least two different backbone modifications
• CPC Class Codes: C12N 2310/346 having a combination of backbone and sugar modifications
• CPC Class Codes: C12N 2310/3525 MOE, methoxyethoxy
• CPC Class Codes: C12Q 1/6876 Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
• CPC Class Codes: C12Q 2600/158 Expression markers

Abstract

Compositions and methods are provided for the modulation of expression of the human ras gene in both the normal and activated forms. Oligonucleotides are provided that have methylene(methylimino) linkages alternating with phosphorothioate or phosphodiester linkages. Further oligonucleotides are provide that have a first region having a methylene(methylimino) linkage alternating with a phosphorothioate or phosphodiester linkage and a second region having phosphorothioate linkages. Such oligonucleotides can be used for diagnostics as well as for research purposes including methods for diagnosis, detection and treatment of conditions arising from the activation of the H-ras gene.

Claims

1. An oligonucleotide comprising from 8 to 30 nucleotide units specifically hybridizable with selected DNA or mRNA deriving from the human H-ras gene and containing a first region having at least one nucleosidic unit modified at the 2′

position with a 2′

-O-methoxyethyl substituent and a second region having at least one phosphorothioate linkage.

2. The oligonucleotide of claim 1 wherein said first region has at least three nucleosidic units modified at the 2′

position with 2′

-O-methoxyethyl substituents.

3. The oligonucleotide of claim 1 wherein said second region has at least four nucleosidic units linked together with phosphorothioate linkages.

4. The oligonucleotide of claim 2 wherein each of the nucleosidic units of said second region are 2′

-deoxynucleotides.

5. The oligonucleotide of claim 4 wherein said second region is flanked by two of said first regions each of which includes at least one nucleosidic unit modified at the 2′

position with a 2′

-methoxyethyl substituent.

6. The oligonucleotide of claim 1 comprising SEQ ID No:

2.

7. An oligonucleotide comprising from 8 to 30 nucleotide units hybridizable with selected DNA or mRNA deriving from the human H-ras gene and containing at least one nucleosidic unit modified at the 2′

position with a 2′

-O-methoxyethyl substituent.

8. The oligonucleotide of claim 7 comprising SEQ ID No:

2.