Antisense oligonucleotides against human protein kinase C

• US 5,885,970 A
• Filed: 06/07/1995
• Issued: 03/23/1999
• Est. Priority Date: 03/16/1992
• Status: Expired due to Term

• Current Assignee: ISIS Pharmaceuticals Incorporated
• Original Assignee: ISIS Pharmaceuticals Incorporated
• Inventors: Bennett, C. Frank, Dean, Nicholas
• Primary Examiner(s): LeGuyader, John L.
• Assistant Examiner(s): SHIBUYA, MARK LANCE
• Application Number:
  US08/488,177
• Time in Patent Office: 1,385 Days
• Field of Search: 514/44, 536/23.5, 536/24.1, 536/24.5, 536/6, 536/91.1, 536/69.1, 536/172.3, 536/354, 536/366, 536/368, 536/325, 536/371, 536/375, 435/91.1, 435/91.2, 435/91.4, 435/91.41, 435/91.5, 435/240.2, 935/2, 935/5, 935/6, 935/8, 935/10
• US Class Current: 514/44.00A
• 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: 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. Methods of treating animals suffering from disease amenable to therapeutic intervention by modulating protein kinase C expression with an oligonucleotide targeted to PKC are disclosed.

Claims

1. A method of treating a hyperproliferative condition associated with expression of human protein kinase C comprising administering 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:

1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 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, 52, 53, 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, whereby expression of human protein kinase C is inhibited and said hyperproliferative condition is ameliorated.

2. The method of claim 1 wherein said hyperproliferative condition is cancer.

3. The method of claim 2 wherein said hyperproliferative condition is colorectal cancer, breast cancer, bladder cancer, lung cancer or brain cancer.

4. The method of claim 3 wherein said brain cancer is glioblastoma multiforme.

5. The method of claim 1 wherein at least one of the intersugar linkage between nucleotide units of the antisense oligonucleotide is a phosphorothioate.

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

7. The method of claim 6 wherein the modification is a 2'"'"'-O-alkyl or 2'"'"'-fluoro modification.

8. The method of claim 1 wherein the antisense oligonucleotide is a chimeric antisense oligonucleotide.

9. The method of claim 8 wherein the antisense oligonucleotide comprises a deoxy region flanked by regions of nucleotide units having 2'"'"'-O-alkyl or 2'"'"'-fluoro modifications.

10. The method of claim 1 wherein said antisense oligonucleotide is in a pharmaceutically acceptable carrier or diluent.

11. The method of claim 10 wherein said carrier or duluent comprises a cationic lipid.

12. The method of claim 1 wherein said antisense oligonucleotide comprises SEQ ID NO:

1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 52 or 53.

13. The method of claim 1 wherein said antisense oligonucleotide comprises SEQ ID NO:

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

14. The method of claim 1 wherein said antisense oligonucleotide comprises SEQ ID NO:

35, 36, 37, 38 or 39.

15. The method of claim 1 wherein said antisense oligonucleotide comprises 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.

16. The method of claim 1 wherein said antisense oligonucleotide comprises SEQ ID NO:

67, 74 or 75.

17. The method of claim 1 wherein said antisense oligonucleotide comprises SEQ ID NO:

87 or 93.

18. The method of claim 1 wherein said antisense oligonucleotide comprises SEQ ID NO:

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

19. A method of treating a hyperproliferative condition associated with human protein kinase C comprising administering an antisense oligonucleotide up to 50 nucleotide units in length and comprising SEQ ID NO:

2, 3, or 5, whereby expression of human protein kinase C is inhibited and said hyperproliferative condition is ameliorated.

20. A method of treating a hyperproliferative condition associated with expression of human protein kinase C comprising administering 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:

1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 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, 52, 53, 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, whereby expression of human protein kinase C is inhibited and said hyperproliferative condition is ameliorated.

21. The method of claim 20 wherein said hyperproliferative condition is cancer.

22. The method of claim 21 wherein said hyperproliferative condition is colorectal cancer, breast cancer, bladder cancer, lung cancer or brain cancer.

23. The method of claim 22 wherein said brain cancer is glioblastoma multiforme.

24. The method of claim 20 wherein at least one of the intersugar linkages between nucleotide units of antisense oligonucleotide is a phosphorothioate.

25. The method of claim 20 wherein at least one of the nucleotides of the antisense oligonucleotide comprises a modification on the 2'"'"' position of the sugar.

26. The method of claim 25 wherein the modification is a 2'"'"'-O-alkyl or 2'"'"'-fluoro modification.

27. The method of claim 20 wherein the antisense oligonucleotide is a chimeric oligonucleotide.

28. The method of claim 27 wherein the antisense oligonucleotide comprises a deoxy region flanked by regions of nucleotide units having 2'"'"'-O-alkyl or 2'"'"'-fluoro modifications.

29. The method of claim 20 wherein said antisense oligonucleotide is in a pharmaceutically acceptable carrier or diluent.

30. The method of claim 29 wherein said carrier or diluent comprises a cationic lipid.

31. The method of claim 20 wherein said antisense oligonucleotide consists of SEQ ID NO:

1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 52 or 53.

32. The method of claim 20 wherein said antisense oligonucleotide consists of SEQ ID NO:

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

33. The method of claim 20 wherein said antisense oligonucleotide consists of SEQ ID NO:

35, 36, 37, 38 or 39.

34. The method of claim 20 wherein said antisense oligonucleotide consists 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.

35. The method of claim 20 wherein said antisense oligonucleotide consists of SEQ ID NO:

67, 74 or 75.

36. The method of claim 20 wherein said antisense oligonucleotide consists of SEQ ID NO:

87 or 93.

37. The method of claim 20 wherein said antisense oligonucleotide consists of SEQ ID NO:

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

38. A method of treating a hyperproliferative condition associated with human protein kinase C comprising administering an antisense oligonucleotide consisting of SEQ ID NO:

2, 3, or 5, whereby expression of human protein kinase C is inhibited and said hyperproliferative condition is ameliorated.

39. A method of treating a hyperproliferative condition associated with expression of human protein kinase C comprising administering an antisense oligonucleotide up to 50 nucleotide units in length which specifically binds mRNA encoding human protein kinase C, wherein said oligonucleotide comprises SEQ ID NO:

2, whereby expression of human protein kinase C is inhibited and said hyperproliferative condition is ameliorated.

40. A method of treating a hyperproliferative condition associated with expression of human protein kinase C comprising administering an antisense oligonucleotide up to 50 nucleotide units in length which specifically binds mRNA encoding human protein kinase C, wherein said oligonucleotide comprises SEQ ID NO:

3, whereby expression of human protein kinase C is inhibited and said hyperproliferative condition is ameliorated.

41. A method of treating a hyperproliferative condition associated with expression of human protein kinase C comprising administering an antisense oligonucleotide up to 50 nucleotide units in length which specifically binds mRNA encoding human protein kinase C, wherein said oligonucleotide comprises SEQ ID NO:

5, whereby expression of human protein kinase C is inhibited and said hyperproliferative condition is ameliorated.