Antisense oligonucleotides against human protein kinase C

• US 5,959,096 A
• Filed: 06/07/1995
• Issued: 09/28/1999
• Est. Priority Date: 03/16/1992
• Status: Expired due to Term

• Current Assignee: ISIS Pharmaceuticals Incorporated (Ionis Pharmaceuticals, Inc.)
• Original Assignee: ISIS Pharmaceuticals Incorporated (Ionis Pharmaceuticals, Inc.)
• Inventors: Bennett, C. Frank, Dean, Nicholas
• Primary Examiner(s): LeGuyader, John L.
• Assistant Examiner(s): SHIBUYA, MARK LANCE
• Application Number:
  US08/481,066
• Time in Patent Office: 1,574 Days
• Field of Search: 514/44, 536/23.1, 536/23.5, 536/24.1, 536/24.31, 536/24.5, 435/91.1, 435/91.2, 435/91.41, 435/91.4, 435/91.5, 435/375, 935/2, 935/5, 935/6, 935/8, 935/9, 935/10, 436/6, 436/69.2, 436/172.3, 436/354, 436/368, 436/371, 436/320.1, 436/375
• 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: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• 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: C07K 14/70564 Selectins, e.g. CD62
• CPC Class Codes: C12N 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• 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 2310/341 Gapmers, i.e. of the type ===---===
• CPC Class Codes: C12N 2310/346 having a combination of backbone and sugar modifications
• CPC Class Codes: C12N 2310/3521 Methyl
• CPC Class Codes: C12N 2310/3527 Other alkyl chain
• CPC Class Codes: C12N 2310/3533 Halogen
• CPC Class Codes: C12N 9/12 transferring phosphorus containing groups, e.g. kinases (2.7)
• 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 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: C12Y 207/11013 Protein kinase C (2.7.11.13)

Abstract

Compositions and methods are provided for the treatment and diagnosis of diseases associated with protein kinase C. Oligonucleotides are provided which are targeted to nucleic acids encoding PKC. Oligonucleotides specifically hybridizable with a translation initiation site, 5'"'"'-untranslated region or 3'"'"'-untranslated region are provided. Oligonucleotides specifically hybridizable with a particular PKC isozyme or set of isozymes are also provided. In preferred embodiments, the oligonucleotides contain one or more chemical modifications. Methods of modulating PKC expression and of treating animals suffering from disease amenable to therapeutic intervention by modulating protein kinase C expression using oligonucleotides targeted to PKC are disclosed.

Claims

1. An antisense oligonucleotide up to 50 nucleotide units in length which specifically binds mRNA encoding human protein kinase C, wherein said antisense oligonucleotide comprises a nucleotide sequence selected from the group consisting of SEQ ID NO:

21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 54, 55, 56, 57, 58, 59, 60, 61, 62, 67, 74, 75, 87, 93, 106, 107, 109, 110, 113, 115, 116, 117, 118, and 121.

2. The antisense oligonucleotide of claim 1 wherein at least one of the intersugar linkages between nucleotide units of the antisense oligonucleotide is a phosphorothioate.

3. The antisense oligonucleotide of claim 1 wherein at least one of the nucleotides comprises a modification on the 2'"'"' position of the sugar.

4. The antisense oligonucleotide of claim 3 wherein the modification is a 2'"'"'-O-alkyl or 2'"'"'-fluoro modification.

5. The antisense oligonucleotide of claim 3 wherein the modification is a 2'"'"'-O-methyl or 2'"'"'-O-propyl modification.

6. The antisense oligonucleotide of claim 1 which is a chimeric antisense oligonucleotide.

7. The antisense oligonucleotide of claim 6 which comprises a deoxy region flanked by 2'"'"'-O-alkyl or 2'"'"'-fluoro regions.

8. The antisense oligonucleotide of claim 1, wherein said nucleotide sequence is selected from the group consisting of SEQ ID NO:

21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, and 34.

9. The antisense oligonucleotide of claim 1, wherein said nucleotide sequence is selected from the group consisting of SEQ ID NO:

35, 36, 37, 38, and 39.

10. The antisense oligonucleotide of claim 1, wherein said nucleotide sequence is selected from the group consisting of SEQ ID NO:

40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 54, 55, 56, 57, 58, 59, 60, 61, and 62.

11. The antisense oligonucleotide of claim 1, wherein said nucleotide sequence is selected from the group consisting of SEQ ID NO:

67, 74, and 75.

12. The antisense oligonucleotide of claim 1, wherein said nucleotide sequence is selected from the group consisting of SEQ ID NO:

87, and 93.

13. The antisense oligonucleotide of claim 1, wherein said nucleotide sequence is selected from the group consisting of SEQ ID NO:

106, 107, 109, 110, 113, 115, 116, 117, 118, and 121.

14. A composition comprising the antisense oligonucleotide of claim 1 and a pharmaceutically acceptable carrier or diluent.

15. An antisense oligonucleotide which specifically binds mRNA encoding human protein kinase C, wherein said antisense oligonucleotide consists of a nucleotide sequence selected from the group consisting of SEQ ID NO:

21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 54, 55, 56, 57, 58, 59, 60, 61, 62, 67, 74, 75, 87, 93, 106, 107, 109, 110, 113, 115, 116, 117, 118, and 121.

16. The antisense oligonucleotide of claim 15 wherein at least one of the intersugar linkages between nucleotide units of the antisense oligonucleotide is a phosphorothioate.

17. The antisense oligonucleotide of claim 15 wherein at least one of the nucleotides comprises a modification on the 2'"'"' position of the sugar.

18. The antisense oligonucleotide of claim 17 wherein the modification is a 2'"'"'-O-alkyl or 2'"'"'-fluoro modification.

19. The antisense oligonucleotide of claim 17 wherein the modification is a 2'"'"'-O-methyl or 2'"'"'-O-propyl modification.

20. The antisense oligonucleotide of claim 15 which is a chimeric antisense oligonucleotide.

21. The antisense oligonucleotide of claim 20 which comprises a deoxy region flanked by 2'"'"'-O-alkyl or 2'"'"'-fluoro regions.

22. The antisense oligonucleotide of claim 15 consisting of SEQ ID NO:

21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33 or 34.

23. The antisense oligonucleotide of claim 15 consisting of SEQ ID NO:

35, 36, 37, 38 or 39.

24. The antisense oligonucleotide of claim 15 consisting of SEQ ID NO:

40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 54, 55, 56, 57, 58, 59, 60, 61 or 62.

25. The antisense oligonucleotide of claim 15 consisting of SEQ ID NO:

67, 74 or 75.

26. The antisense oligonucleotide of claim 15 consisting of SEQ ID NO:

87 or 93.

27. The antisense oligonucleotide of claim 15 consisting of SEQ ID NO:

106, 107, 109, 110, 113, 115, 116, 117, 118 or 121.

28. A composition comprising the antisense oligonucleotide of claim 15 and a pharmaceutically acceptable carrier or diluent.