Antisense modulation of protein expression

• US 20030176385A1
• Filed: 11/27/2002
• Published: 09/18/2003
• Est. Priority Date: 02/15/2000
• Status: Abandoned Application

• Current Assignee: CuraGen Corporation (Celldex Therapeutics Incorporated)
• Original Assignee: CuraGen Corporation (Celldex Therapeutics Incorporated)
• Inventors: Huang, Chunli, Ju, Jingfang, Taillon, Bruce E., Zhong, Haihong, Simons, Jan Fredrik, Smithson, Glennda, Peyman, John A., Millet, Isabelle, Chant, John S.
• Application Number:
  US10/305,810
• Publication Number:
  US 20030176385A1
• Time in Patent Office: Days
• US Class Current: 514/44
• 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 14/47 from mammals
• CPC Class Codes: C07K 14/475 Growth factors; Growth regulators
• CPC Class Codes: C07K 14/5421 IL-8
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C07K 14/82 Translation products from oncogenes
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 15/1135 against oncogenes or tumor suppressor genes
• CPC Class Codes: C12N 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 15/1138 against receptors or cell surface proteins
• CPC Class Codes: C12N 9/1051 Hexosyltransferases (2.4.1)
• CPC Class Codes: C12N 9/1211 Thymidine kinase (2.7.1.21)
• CPC Class Codes: C12Y 204/0115 N-Acetyllactosaminide beta-1,6-N-acetylglucosaminyl-transferase (2.4.1.150)

Abstract

Antisense compounds, compositions and methods are provided for modulating the expression of H-Ras, WNT-7B, acetylglucosaminyltransferase, voltage-gated K channel, IL-8, ion transport, Map3K8 and Thymidine kinase. The compositions comprise antisense compounds, particularly antisense oligonucleotides, targeted to nucleic acids encoding H-Ras, WNT-7B, acetylglucosaminyltransferase, voltage-gated K channel, IL-8, ion transport, Map3K8 and Thymidine kinase. Methods of using these compounds for modulation of H-Ras, WNT-7B, acetylglucosaminyltransferase, voltage-gated K channel, IL-8, ion transport, Map3K8 and Thymidine kinase.expression and for treatment of diseases associated with expression of H-Ras, WNT-7B, acetylglucosaminyltransferase, voltage-gated K channel, 1L-8, ion transport, Map3KS and Thymidine kinase are provided.

Claims

2. An oligonucleotide 10-50 nucleotides in length comprising at least 10 contiguous nucleotides of the nucleic acid sequence selected from the group consisting of SEQ ID NO:

7-11, wherein said oligonucleotide inhibits the expression of WNT-7B.

38. A method of inhibiting the expression of WNT-7B in a cell, comprising contacting said cell with one or more of the oligonucleotides of claim 2.

50. A method of inhibiting cell proliferation, comprising contacting a cell with one or more of the oligonucleotides of claim 2.

3. An oligonucleotide comprising the nucleic acid sequence of selected from the group consisting of SEQ ID NO:

7-11.

4. An oligonucleotide 20-50 nucleotides in length targeted to nucleotides 224-366, nucleotides 761-841, or 1062-1142 of SEQ ID NO:

2, wherein said oligonucleotide specifically hybridizes with one of said regions and inhibits expression of N-acetylglucoaminyltransferase.

22. The oligonucleotide of claim 4, wherein said oligonucleotide comprises at least one modified internucleoside linkage.

23. The oligonucleotide of claim 4, wherein said oligonucleotide comprises at least one modified sugar moiety.

24. The oligonucleotide of claim 4, wherein said oligonucleotide comprises at least one modified nucleotide.

39. A method of inhibiting the expression of N-acetylglucoaminyltransferase in a cell, comprising contacting said cell with the oligonucleotide of claim 4.

5. An oligonucleotide 10-50 nucleotides in length comprising at least 10 contiguous nucleotides of the nucleic acid sequence selected from the group consisting of SEQ ID NO:

12-16, wherein said oligonucleotide inhibits the expression of N-acetylglucoaminyltransferase.

1. An oligonucleotide 20-50 nucleotides in length targeted to nucleotides 1-109 or nucleotides 434-513 of SEQ ID NO:

1, wherein said oligonucleotide specifically hybridizes with one of said regions and inhibits expression of WNT-7B.

19. The oligonucleotide of claim 1, wherein said oligonucleotide comprises at least one modified internucleoside linkage.

20. The oligonucleotide of claim 1, wherein said oligonucleotide comprises at least one modified sugar moiety.

21. The oligonucleotide of claim 1, wherein said oligonucleotide comprises at least one modified nucleotide.

37. A method of inhibiting the expression of WNT-7B in a cell, comprising contacting said cell with the oligonucleotide of claim 1.

40. A method of inhibiting the expression of N-acetylglucoaminyltransferase in a cell, comprising contacting said cell with one or more of the oligonucleotides of claim 5.

49. A method of inhibiting cell proliferation, comprising contacting a cell with the oligonucleotide of claim 1

51. The method of claim 49, wherein said oligonucleotide is present at a concentration of between 50 mM and 400 mM.

52. The method of claim 49, wherein said oligonucleotide comprises the nucleotide sequence selected from the group consisting of SEQ ID NO:

7-11.

53. A method of inhibiting cell proliferation, comprising contacting a cell with the oligonucleotide of claim 4.

54. The method of claim 53, wherein said oligonucleotide is present at a concentration of between SOmM and 400 mM

55. The method of claim 53, wherein said oligonucleotide comprises the nucleotide sequence selected from the group consisting of SEQ ID NO:

12-16.

58. The method of claim 53, wherein said oligonucleotide is present at a concentration of at least 400 mM.

6. An oligonucleotide comprising the nucleic acid sequence of selected from the group consisting of SEQ ID NO:

12-16.

7. An oligonucleotide 20-50 nucleotides in length targeted to nucleotides 1-121, nucleotides 1226-1201 or 1185-1953 of SEQ ID NO:

3, wherein said oligonucleotide specifically hybridizes with one of said regions and inhibits expression of voltage gated K channel.

25. The oligonucleotide of claim 7, wherein said oligonucleotide comprises at least one modified internucleoside linkage.

26. The oligonucleotide of claim 7, wherein said oligonucleotide comprises at least one modified sugar moiety.

27. The oligonucleotide of claim 7, wherein said oligonucleotide comprises at least one modified nucleotide.

41. A method of inhibiting the expression of voltage gated K channel in a cell, comprising contacting said cell with the oligonucleotide of claim 7.

8. An oligonucleotide 10-50 nucleotides in length comprising at least 10 contiguous nucleotides of the nucleic acid sequence selected from the group consisting of SEQ ID NO:

17-21, wherein said oligonucleotide inhibits the expression of voltage gated K channel.

42. A method of inhibiting the expression of voltage gated K channel in a cell, comprising contacting said cell with one or more of the oligonucleotides of claim 8.

9. An oligonucleotide comprising the nucleic acid sequence of selected from the group consisting of SEQ ID NO:

17-21.

10. An oligonucleotide 20-50 nucleotides in length targeted to nucleotides 1-91, nucleotides 77-157, nucleotides 902-982 or nucleotides 1541-1621 of SEQ ID NO:

4, wherein said oligonucleotide specifically hybridizes with one of said regions and inhibits expression of ion transport.

28. The oligonucleotide of claim 10, wherein said oligonucleotide comprises at least one modified internucleoside linkage.

29. The oligonucleotide of claim 10, wherein said oligonucleotide comprises at least one modified sugar moiety.

30. The oligonucleotide of claim 10, wherein said oligonucleotide comprises at least one modified nucleotide.

43. A method of inhibiting the expression of ion transport in a cell, comprising contacting said cell with the oligonucleotide of claim 10.

11. An oligonucleotide 10-50 nucleotides in length comprising at least 10 contiguous nucleotides of the nucleic acid sequence selected from the group consisting of SEQ ID NO:

23-27, wherein said oligonucleotide inhibits the expression of ion transport.

44. A method of inhibiting the expression of ion transport in a cell, comprising contacting said cell with one or more of the oligonucleotides of claim 11.

12. An oligonucleotide comprising the nucleic acid sequence of selected from the group consisting of SEQ ID NO:

23-27.

13. An oligonucleotide 20-50 nucleotides in length targeted to nucleotides 63-162 nucleotides 197-246, nucleotides 1037-1186 or nucleotides 1447-1526 of SEQ ID NO:

5, wherein said oligonucleotide specifically hybridizes with one of said regions and inhibits expression of Map3K8.

31. The oligonucleotide of claim 13, wherein said oligonucleotide comprises at least one modified internucleoside linkage.

32. The oligonucleotide of claim 13, wherein said oligonucleotide comprises at least one modified sugar moiety.

33. The oligonucleotide of claim 13, wherein said oligonucleotide comprises at least one modified nucleotide.

45. A method of inhibiting the expression of Map3K8 in a cell, comprising contacting said cell with the oligonucleotide of claim 13.

14. An oligonucleotide 10-50 nucleotides in length comprising at least 10 contiguous nucleotides of the nucleic acid sequence selected from the group consisting of SEQ ID NO:

28-32, wherein said oligonucleotide inhibits the expression of Map3K8.

46. A method of inhibiting the expression of Map3K8 in a cell, comprising contacting said cell with one or more of the oligonucleotides of claim 14.

15. An oligonucleotide comprising the nucleic acid sequence of selected from the group consisting of SEQ ID NO:

28-32.

16. An oligonucleotide 20-50 nucleotides in length targeted to nucleotides 15-116 nucleotides 132-211, nucleotides 629-708 or nucleotides 1286-1165 of SEQ ID NO:

6, wherein said oligonucleotide specifically hybridizes with one of said regions and inhibits expression of thymidine kinase.

34. The oligonucleotide of claim 16, wherein said oligonucleotide comprises at least one modified internucleoside linkage.

35. The oligonucleotide of claim 16, wherein said oligonucleotide comprises at least one modified sugar moiety.

36. The oligonucleotide of claim 16, wherein said oligonucleotide comprises at least one modified nucleotide.

47. A method of inhibiting the expression of thymidine kinase in a cell, comprising contacting said cell with the oligonucleotide of claim 16.

56. A method of increasing the production of Il-1b in a cell comprising contacting a cell with the oligonucleotide of claim 16.

17. An oligonucleotide 10-50 nucleotides in length comprising at least 10 contiguous nucleotides of the nucleic acid sequence selected from the group consisting of SEQ ID NO:

33-37, wherein said oligonucleotide inhibits the expression of thymidine kinase.

48. A method of inhibiting the expression of thymidine kinase in a cell, comprising contacting said cell with one or more of the oligonucleotides of claim 17.

57. A method of increasing the production of Il-1b in a cell, comprising contacting a cell with one or more of the oligonucleotides of claim 17.

18. An oligonucleotide comprising the nucleic acid sequence of selected from the group consisting of SEQ ID NO:

33-37.

53-1. A method of inhibiting cell proliferation, comprising contacting a cell with one or more of the oligonucleotides of claim 5.