Modulation of HIF1-beta expression

• US 7,799,764 B2
• Filed: 10/13/2009
• Issued: 09/21/2010
• Est. Priority Date: 08/25/2004
• Status: Expired due to Fees

• Current Assignee: ISIS Pharmaceuticals Incorporated (Ionis Pharmaceuticals, Inc.)
• Original Assignee: ISIS Pharmaceuticals Incorporated (Ionis Pharmaceuticals, Inc.)
• Inventors: Marcusson, Eric G., Kim, Youngsoo, Henry, Scott W., Dobie, Kenneth
• Primary Examiner(s): MCGARRY, SEAN
• Application Number:
  US12/578,468
• Publication Number:
  US 20100113573A1
• Time in Patent Office: 343 Days
• Field of Search: None
• US Class Current: 514/44.00A
• 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 12 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8 contiguous nucleobases of the nucleobase sequence recited in SEQ ID NO:

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

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

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

3. The compound of claim 2, wherein the nucleobase sequence of the modified oligonucleotide is 100% complementary to SEQ ID NO:

4 as measured over the entirety of said modified oligonucleotide.

4. The compound of claim 2, wherein at least one internucleoside linkage is a modified internucleoside linkage.

5. The compound of claim 4, wherein each internucleoside linkage is a phosphorothioate internucleoside linkage.

6. The compound of claim 2, wherein at least one nucleoside comprises a modified sugar.

7. The compound of claim 6, wherein at least one modified sugar is a bicyclic sugar.

8. The compound of claim 6, wherein at least one modified sugar comprises a 2′

-O-methoxyethyl or a 4′

-(CH₂)_n—

O-2′

bridge, wherein n is 1 or 2.

9. The compound of claim 2, wherein at least one nucleoside comprises a modified nucleobase.

10. The compound of claim 9, wherein the modified nucleobase is a 5-methylcytosine.

11. 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.

12. The compound of claim 11, wherein the modified oligonucleotide comprises:

a gap segment consisting of ten 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;

wherein each cytosine in said modified oligonucleotide is a 5-methylcytosine, and wherein each internucleoside linkage of said modified oligonucleotide is a phosphorothioate linkage.

13. The compound of claim 2, wherein the modified oligonucleotide consists of 20 linked nucleosides.

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

15. The composition of claim 14, wherein said compound consists of a single-stranded oligonucleotide.

16. The composition of claim 14, wherein the modified oligonucleotide consists of 20 linked nucleosides.

17. A method of inhibiting HIF-1 beta comprising administering to an animal an effective amount of the compound of claim 1.

18. The method of claim 17, wherein the animal is a human.

19. The method of claim 18, wherein administering the compound treats a hyperproliferative disorder.

20. The method of claim 19, wherein the hyperproliferative disorder is cancer.

21. The method of claim 19, wherein the hyperproliferative disorder is an angiogenic disorder.

22. The method of claim 21, wherein the angiogenic disorder is an ocular disorder.

23. The method of claim 22, wherein the ocular disorder is selected from the group consisting of macular degeneration, diabetic retinopathy, macular edema and retinopathy of prematurity.

24. The method of claim 17, wherein said administration comprises subcutaneous or intravitreous injection.

25. A method comprising identifying a human with a hyperproliferative disorder and administering to the human a therapeutically effective amount of the compound of claim 1.

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

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

28. The method of claim 27, wherein the HIF-1 beta regulated gene is selected from the group consisting of VEGF, GLUT-1, PGK-1, PAI-1 and Epo.

29. The compound of claim 2, wherein the nucleobase sequence of the modified oligonucleotide is at least 95% complementary to SEQ ID NO:

4 as measured over the entirety of said modified oligonucleotide.

30. The compound of claim 12, wherein said nucleobase sequence comprises SEQ ID NO:

30.

31. The compound of claim 12, wherein said modified oligonucleotide consists of 20 linked nucleosides having a nucleobase sequence consisting of SEQ ID NO:

30.