Oligonucleotides for enhanced modulation of protein kinase C expression

• US 6,117,847 A
• Filed: 06/15/1998
• Issued: 09/12/2000
• Est. Priority Date: 03/16/1992
• Status: Expired due to Fees

• Current Assignee: ISIS Pharmaceuticals Incorporated
• Original Assignee: ISIS Pharmaceuticals Incorporated
• Inventors: Dean, Nicholas M., Bennett, C. Frank
• Primary Examiner(s): Schwartzman, Robert A.
• Assistant Examiner(s): LARSON, THOMAS GEORGE
• Application Number:
  US09/094,714
• Time in Patent Office: 820 Days
• Field of Search: 435/6, 435/375, 514/44, 536/24.31, 536/24.55
• 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: 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 2310/3341 5-Methylcytosine
• 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 2310/3525 MOE, methoxyethoxy
• CPC Class Codes: C12N 2310/3527 Other alkyl chain
• 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

Abstract

Compositions and methods are provided for modulating the expression of protein kinase C. Oligonucleotides are provided which are targeted to nucleic acids encoding PKC. The oligonucleotides are from 5 to 50 nucleotides in length and in one referred embodiment are from 12 to 18 nucleotides in length. The oligonucleotides may be chimeric oligonucleotides and in a preferred embodiment comprise at least one 2'"'"'-O-methoxyethyl modification. Pharmaceutical compositions comprising the oligonucleotides of the invention are also provided. Methods of inhibiting protein kinase C expression and methods of treating conditions associated with expression of protein kinase C using oligonucleotides of the invention are disclosed.

Claims

1. An antisense oligonucleotide up to 50 nucleotides in length which is targeted to a nucleic acid encoding a human protein kinase C and which modulates expression of the human protein kinase C, wherein said oligonucleotide comprises SEQ ID NO:

2, 3, 18, 19, 20, 23, 25 or 26.

2. The antisense oligonucleotide of claim 1 which is up to 20 nucleotides in length.

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

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

9. A pharmaceutical composition comprising a therapeutically effective amount of the oligonucleotide of claim 1.

12. A method of inhibiting human protein kinase C expression in cells comprising contacting the cells with the oligonucleotide of claim 1.

15. The method of claim 12 wherein the cells are cancer cells.

18. A method of treating a condition associated with expression of protein kinase C comprising administering to a human, or a cell, a tissue, or a bodily fluid thereof, a therapeutically effective amount of the oligonucleotide of claim 1.

21. The method of claim 18 wherein said condition is an inflammatory or hyperproliferative disorder.

24. The method of claim 21 wherein the condition is cancer or psoriasis.

5. An antisense oligonucleotide consisting of SEQ ID NO:

18.

6. The antisense oligonucleotide of claim 5 wherein each intersugar linkage is a phosphorothioate linkage and wherein the first eight nucleotides at the 5'"'"' end of the oligonucleotide are 2'"'"'-deoxynucleotides and the remaining seven nucleotides are 2'"'"'-O--CH₂ CH₂ OCH₃ nucleotides, and wherein each 2'"'"'-O--CH₂ CH₂ OCH₃ cytidine nucleotide is a 5-methyl cytidine.

10. A pharmaceutical composition comprising a therapeutically effective amount of the oligonucleotide of claim 5.

13. A method of inhibiting human protein kinase C expression in cells comprising contacting the cells with the oligonucleotide of claim 5.

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

19. A method of treating a condition associated with expression of protein kinase C comprising administering to a human, or cells, tissues, or a bodily fluid thereof, a therapeutically effective amount of the oligonucleotide of claim 5.

22. The method of claim 19 wherein said condition is an inflammatory or hyperproliferative disorder.

25. The method of claim 22 wherein the condition is cancer or psoriasis.

7. An antisense oligonucleotide consisting of SEQ ID NO:

25.

8. The antisense oligonucleotide of claim 7 wherein each intersugar linkage is a phosphorothioate linkage and wherein the first eight nucleotides at the 5'"'"' end of the oligonucleotide are 2'"'"'-deoxynucleotides and the remaining seven nucleotides are 2'"'"'-O--CH₂ CH₂ OCH₃ nucleotides, and wherein each 2'"'"'-O--CH₂ CH₂ OCH₃ cytidine nucleotide is a 5-methyl cytidine.

11. A pharmaceutical composition comprising a therapeutically effective amount of the oligonucleotide of claim 7.

14. A method of inhibiting human protein kinase C expression in cells comprising contacting the cells with the oligonucleotide of claim 7.

17. The method of claim 14 wherein the cells are cancer cells.

20. A method of treating a condition associated with expression of protein kinase C comprising administering to a human, or cells, tissues, or a bodily fluid thereof, a therapeutically effective amount of the oligonucleotide of claim 7.

23. The method of claim 20 wherein said condition is an inflammatory or hyperproliferative disorder.

26. The method of claim 23 wherein the condition is cancer or psoriasis.

27. An oligonucleotide consisting of SEQ ID NO:

40.

28. The oligonucleotide of claim 27 wherein at least one of the nucleotides contains a modification on the 2'"'"' position of the sugar.

29. The oligonucleotide of claim 27 which is a chimeric oligonucleotide.

30. A pharmaceutical composition comprising a therapeutically effective amount of the oligonucleotide of claim 27.

31. A method of inhibiting human protein kinase C expression in cells comprising contacting the cells with the oligonucleotide of claim 27.

32. The method of claim 31 wherein the cells are cancer cells.

33. A method of treating a condition associated with expression of protein kinase C comprising administering to a human, or a cell, a tissue, or a bodily fluid thereof, a therapeutically effective amount of the oligonucleotide of claim 27.

34. The method of claim 33 wherein said condition is an inflammatory or hyperproliferative disorder.

35. The method of claim 34 wherein the condition is cancer or psoriasis.