Oligomeric compounds for the modulation of HIF-1A expression

• US 8,785,617 B2
• Filed: 12/21/2012
• Issued: 07/22/2014
• Est. Priority Date: 04/05/2002
• Status: Expired due to Fees

• Current Assignee: Roche Innovation Center Copenhagen A/S (Roche Holding AG)
• Original Assignee: Santaris Pharma A/S (Roche Holding AG)
• Inventors: Thrue, Charlotte Albaek, Hog, Anja Molhart, Kristjansen, Paul E. G.
• Primary Examiner(s): Chong, Kimberly
• Application Number:
  US13/725,540
• Publication Number:
  US 20130116309A1
• Time in Patent Office: 578 Days
• Field of Search: None
• US Class Current: 536/24.5
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/70 Carbohydrates; Sugars; Derivatives thereof sorbitol A61K31/047
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• 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: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/04 specific for metastasis
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• 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 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/3231 having an additional ring, e.g. LNA, ENA
• CPC Class Codes: C12N 2310/331 Universal or degenerate base
• CPC Class Codes: C12N 2310/334 : Modified C
• CPC Class Codes: C12N 2310/3341 : 5-Methylcytosine
• CPC Class Codes: C12N 2310/335 : Modified T or U
• CPC Class Codes: C12N 2310/341 : Gapmers, i.e. of the type ===---===

Abstract

Oligonucleotides directed against the hypoxia-inducible factor-1α (HIF-1α) gene are provided for modulating the expression of HIF-1α. The compositions comprise oligonucleotides, particularly antisense oligonucleotides, targeted to nucleic acids encoding the HIF-1α. Methods of using these compounds for modulation of HIF-1α expression and for the treatment of diseases associated with the hypoxia-inducible factor-1α are provided. Examples of diseases are cancer and pre-eclampsia. The oligonucleotides may be composed of deoxyribonucleosides, a nucleic acid analog, or Locked Nucleic Acid (LNA) or a combination thereof.

Claims

1. A compound of 16 to 50 nucleotides or nucleotide analogues, comprising SEQ ID NO:

3, wherein adjacent nucleotides or nucleotide analogues are covalently linked by an internucleotide linkage, wherein the compound comprises at least one nucleotide analogue, and wherein the compound modulates HIF-1α

expression.

2. The compound according to claim 1 which inhibits the expression of HIF-1α

in a cell or tissue.

3. The compound according to claim 1, which is an antisense oligonucleotide.

4. The compound according to claim 3, wherein the nucleotide analogue is a Locked Nucleic Acid (LNA) unit.

5. The compound according to claim 4, wherein the LNA unit has the structure:

6. The compound according to claim 4, wherein the LNA unit has a structure of any of the following formulas:

7. The compound according to claim 5, wherein the internucleotide linkage is selected from the group consisting of —

O—

P(O)₂—

O—

, —

O—

P(O,S)—

O—

, —

O—

P(S)₂—

O—

, —

S—

P(O)₂—

O—

, —

S—

P(O,S)—

O—

, —

S—

P(S)₂—

O—

, —

O—

P(O)₂—

S—

, —

O—

P(O,S)—

S—

, —

S—

P(O)₂—

S—

, —

O—

PO(R^H)—

O—

, O—

PO(OCH₃)—

O—

, —

O—

PO(NR^H)—

O—

, —

O—

PO(OCH₂CH₂S—

R)—

O—

, —

O—

PO(BH₃)—

O—

, —

O—

PO(NHR^H)—

O—

, —

O—

P(O)₂—

HR^H—

, —

NR^H—

P(O)₂—

O—

, and —

NR″

—

CO—

O—

, where R^H is selected form hydrogen and C_1-4-alkyl.

8. The compound of claim 7, wherein at least one internucleotide linkage is a phosphorothioate linkage.

9. The compound according to claim 5, wherein at least one nucleobase is a modified nucleobase independently selected from the group consisting of 5-methylcytosine, isocytosine, pseudoisocytosine, 5-bromouracil, 5-propynyluracil, 6-aminopurine, 2-aminopurine, inosine, diaminopurine, and 2-chloro-6-aminopurine.

10. The compound according to claim 4, wherein each of the LNA units is independently selected from β

-D-oxy-LNA, β

-D-thio-LNA, β

-D-amino-LNA, α

-L-oxy-LNA, α

-L-thio-LNA and α

-L-amino-LNA.

11. The compound of claim 3, wherein the antisense oligonucleotide is a chimeric oligonucleotide.

12. The compound of claim 11, wherein the chimeric oligonucleotide is a gapmer, a headmer, or a tailmer.

13. The compound according to claim 4, which comprises at least four LNA units.

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

15. A pharmaceutical composition comprising the compound of claim 1 in the form of a pharmaceutically acceptable salt.

16. A pharmaceutical composition comprising the compound of claim 1, which is in the form of a conjugate.

17. A pharmaceutical composition comprising the compound of claim 1, which is in the form of a pro-drug.

18. A compound represented by the formula:

T_sG_sG_sT_sg_sa_sg_sg_sc_st_sg_st_sC_sC_sG_sA (SEQ ID NO.

     3) wherein uppercase letters denote oxy-LNA nucleotides, lowercase letters denote deoxynucleotides and the subscript “

s”

denotes a phosphorothioate linkage.

19. A compound selected from: