Modulation of HIF-1 beta expression

• US 7,618,947 B2
• Filed: 08/25/2005
• Issued: 11/17/2009
• Est. Priority Date: 08/25/2004
• Status: Expired due to Fees

• Current Assignee: ISIS Pharmaceuticals Incorporated
• Original Assignee: ISIS Pharmaceuticals Incorporated
• Inventors: Henry, Scott W., Marcusson, Eric G., Dobie, Kenneth W., Kim, Youngsoo
• Primary Examiner(s): MCGARRY, SEAN
• Application Number:
  US11/213,593
• Publication Number:
  US 20060252720A1
• Time in Patent Office: 1,545 Days
• Field of Search: None
• US Class Current: 514/44.00R
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• 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 2310/11 Antisense
• 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

Abstract

Compounds, compositions and methods are provided for modulating the expression of HIF1-beta. The compositions comprise oligonucleotides, targeted to nucleic acid encoding HIF1-beta. Methods of using these compounds for modulation of HIF1-beta expression and for diagnosis and treatment of diseases and conditions associated with expression of HIF1-beta are provided.

Claims

1. A compound comprising a modified oligonucleotide consisting of 13 to 30 linked nucleosides, wherein said modified oligonucleotide has a nucleobase sequence comprising at least 8 consecutive nucleobases complementary to nucleotides 1200-1239 of SEQ ID NO:

4, wherein said nucleobase sequence of said modified oligonucleotide is at least 90% complementary to SEQ ID NO;

4 as measured over the entirety of said nucleobase sequence of said modified oligonucleotide, wherein said compound inhibits expression of human HIF-1beta.

2. A composition comprising the compound of claim 1, or a salt thereof, and a pharmaceutically acceptable carrier or diluent.

3. A method of inhibiting expression of human HIF-1 beta in cells or tissues, comprising contacting said cells or tissues with the compound of claim 1, such that expression of human HIF-1 beta is inhibited.

4. A method of inhibiting expression of a HIF-1 beta regulated gene in a cell or tissue comprising contacting said cells or tissues with the compound of claim 1, such that expression of the HIF-1 beta regulated gene is inhibited.

5. The method of claim 4, wherein the HIF-1 beta regulated gene is selected from the group consisting of VEGF, GLUT-1, PGK- 1, PAI-1 and Epo.

6. A method of treating an animal having a disease or condition associated with HIF-1 beta comprising administrating to said animal a therapeutically or prophylactically effective amount of the composition of claim 2 so that expression of HIF-1 beta is inhibited.

7. The method of claim 6, wherein the disease or condition is a hyperproliferative disorder.

8. The method of claim 7, wherein the hyperproliferative disorder is cancer.

9. The method of claim 7, wherein the hyperproliferative disorder is an angiogenic disorder.

10. The method of claim 9, wherein the angiogenic disorder is an ocular disorder.

11. The method of claim 10, wherein the ocular disorder is selected from the group consisting of macular degeneration, diabetic retinopathy, macular edema and retinopathy of prematurity.

12. A method of inhibiting aberrant angiogenesis in an animal, comprising administering to said animal the compound of claim 1 such that aberrant angiogenesis is inhibited.

13. The compound of claim 1, wherein said modified oligonucleotide has a nucleobase sequence comprising at least 8 consecutive nucleobases of a nucleobase sequence selected from the group consisting of SEQ ID NOs:

56, 57, 58, 59, and 60.

14. The compound of claim 1, wherein said modified oligonucleotide has a nucleobase sequence comprising at least 8 consecutive nucleobases of the nucleobase sequence of SEQ ID NO:

60.

15. The compound of claim 1, wherein said modified oligonucleotide has a nucleobase sequence comprising the nucleobase sequence of SEQ ID NO:

60.

16. The compound of claim 1, consisting of a single-stranded modified oligonucleotide.

17. The compound of claim 16, wherein the nucleobase sequence of the modified oligonucleotide is 100% complementary to SEQ ID NO:

4.

18. The compound of claim 16, wherein at least one internucleoside linkage is a modified internucleoside linkage.

19. The compound of claim 18, wherein each internucleoside linkage is a phosphorothioate internucleoside linkage.

20. The compound of claim 16, wherein at least one nucleoside comprises a modified sugar.

21. The compound of claim 20, wherein at least one modified sugar is a bicyclic sugar.

22. The compound of claim 20, wherein at least one modified sugar comprises a 2′

-O-methoxyethyl.

23. The compound of claim 20, wherein at least one modified sugar comprises a 4′

-(CH₂)_n—

O-2′

bridge, wherein n is 1 or 2.

24. The compound of claim 16, wherein at least one nucleoside comprises a modified nucleobase.

25. The compound of claim 24, wherein the modified nucleobase is a 5-methylcytosine.

26. The compound of claim 1, wherein the modified oligonucleotide comprises:

a gap segment consisting of linked deoxynucleosides;

a 5′

wing segment consisting of linked nucleosides;

a 3′

wing segment consisting of linked nucleosides;

wherein the gap segment is positioned between the 5′

wing segment and the 3′

wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.

27. The compound of claim 26, wherein the modified oligonucleotide comprises:

a gap segment consisting often linked deoxynucleosides;

a 5′

wing segment consisting of five linked nucleosides;

a 3′

wing segment consisting of five linked nucleosides;

wherein the gap segment is positioned between the 5′

wing segment and the 3′

wing segment, wherein each nucleoside of each wing segment comprises a 2′

-O-methoxyethyl sugar; and

wherein each internucleoside linkage of said modified oligonucleotide is a phosphorothioate linkage.

28. The compound of claim 16, wherein the modified oligonucleotide consists of 20 linked nucleosides.

29. The composition of claim 2, consisting of a single-stranded oligonucleotide.

30. The composition of claim 2, wherein the modified oligonucleotide consists of 20 linked nucleosides.

31. The compound of claim 15, wherein the modified oligonucleotide comprises:

a gap segment consisting often linked deoxynucleosides;

a 5′

wing segment consisting of five linked nucleosides;

a 3′

wing segment consisting of five linked nucleosides;

wherein the gap segment is positioned between the 5′

wing segment and the 3′

wing segment, wherein each nucleoside of each wing segment comprises a 2′

-O-methoxyethyl sugar; and

wherein each internucleoside linkage of said modified oligonucleotide is a phosphorothioate linkage.

32. The compound of claim 31, wherein the modified oligonucleotide consists of 20 linked nucleosides.