Antisense oligonucleotide inhibition of ras

• US 6,784,290 B1
• Filed: 05/22/2000
• Issued: 08/31/2004
• Est. Priority Date: 10/05/1992
• Status: Expired due to Fees

• Current Assignee: ISIS Pharmaceuticals Incorporated
• Original Assignee: ISIS Pharmaceuticals Incorporated
• Inventors: Monia, Brett P., Cowsert, Lex M., Manoharan, Muthiah
• Primary Examiner(s): FREDMAN, JEFFREY NORMAN
• Application Number:
  US09/575,554
• Time in Patent Office: 1,562 Days
• Field of Search: 536/24.5, 536/231, 536/243, 514/44, 435/6, 435/91.2
• US Class Current: 536/24.5
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: C07K 14/82 Translation products from oncogenes
• CPC Class Codes: C12N 15/1135 against oncogenes or tumor suppressor genes
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 2'-R Modification
• CPC Class Codes: C12N 2310/333 Modified A
• CPC Class Codes: C12N 2310/3521 Methyl
• CPC Class Codes: C12N 2310/3525 MOE, methoxyethoxy
• CPC Class Codes: C12N 2310/3527 Other alkyl chain
• CPC Class Codes: C12N 2310/3533 Halogen
• CPC Class Codes: C12Q 2525/125 incorporating agents resulting in resistance to degradation

Abstract

Compositions and methods are provided for the modulation of ras expression. Oligonucleotides are provided which are targeted to nucleic acids encoding human ras. Oligonucleotides specifically hybridizable with mRNA encoding human H-ras, Ki-ras and N-ras are provided. Such oligonucleotides can be used for therapeutics and diagnostics as well as for research purposes. Methods are also disclosed for modulating ras gene expression in cells and tissues using the oligonucleotides provided, and for specific modulation of expression of activated ras. Methods for diagnosis, detection and treatment of conditions, or particular cancers, associated with ras are also disclosed.

Claims

1. An oligonucleotide 8 to 30 nucleotides in length which is targeted to a nucleic acid encoding human Ki-ras, wherein said oligonucleotide is capable of inhibiting Ki-ras expression, and wherein said oligonucleotide comprises at least an 8-nucleobase portion of SEQ ID NO:

20, 21, 22, 31, 32 or 33.

2. The oligonucleotide of claim 1 which comprises at least one backbone modification.

3. The oligonucleotide of claim 1 wherein at least one of the nucleotide units of said oligonucleotide is modified at the 2′

position of the sugar.

4. The oligonucleotide of claim 1 which is a chimeric oligonucleotide.

5. The oligonucleotide of claim 1 in a pharmaceutically acceptable carrier.

6. A method of modulating the expression of human Ki-ras comprising contacting tissues or cells containing a human Ki-ras gene with an effective amount of an oligonucleotide of 8 to 30 nucleotides in length which is targeted to a nucleic acid encoding human Ki-ras, wherein said oligonucleotide is capable of inhibiting Ki-ras expression, wherein said oligonucleotide comprises at least an 8-nucleobase portion of SEQ ID NO:

20, 21, 22, 26, 28, 31, 32 or 33 and whereby expression of Ki-ras is modulated.

13. The method of claim 6 wherein the cells are cancer cells.

14. The method of claim 6 wherein the cells are blood cells.

15. The method of claim 6 wherein the cells are peripheral blood mononuclear cells.

7. A method of inhibiting the proliferation of cancer cells comprising contacting cancer cells with an effective amount of an oligonucleotide 8 to 30 nucleotides in length which is targeted to a nucleic acid encoding human Ki-ras, wherein said oligonucleotide is capable of inhibiting Ki-ras expression, wherein said oligonucleotide comprises at least an 8-nucleobase portion of SEQ ID NO:

20, 21, 22, 26, 28, 31, 32 or 33 and, whereby proliferation of the cancer cells is inhibited.

8. A method of preventing or treating a condition arising from the activation of a Ki-ras oncogene comprising contacting an animal suspected of having a condition arising from the activation of a Ki-ras oncogene with an effective amount of an oligonucleotide 8 to 30 nucleotides in length which is targeted to a nucleic acid encoding human Ki-ras, wherein said oligonucleotide is capable of inhibiting Ki-ras expression, wherein said oligonucleotide comprises at least an 8-nucleobase portion of SEQ ID NO:

20, 21, 22, 26, 28, 31, 32 or 33 and, whereby said condition is prevented or treated.

9. The method of claim 8 wherein said activation of a Ki-ras oncogene is abnormal expression of a Ki-ras oncogene.

10. The method of claim 8 wherein said condition is a hyperproliferative condition.

11. The method of claim 8 wherein the condition is cancer.

12. The method of claim 8 wherein the condition is colorectal cancer, melanoma, liposarcoma, mesothelioma, sarcoma, colon cancer, or pancreatic cancer.