Potent LNA oligonucleotides for the inhibition of HIF-1a expression

• US 7,939,507 B2
• Filed: 06/03/2009
• Issued: 05/10/2011
• Est. Priority Date: 11/09/2004
• Status: Active Grant

• Current Assignee: Roche Innovation Center Copenhagen A/S (Roche Holding AG)
• Original Assignee: Enzon Pharmaceuticals Inc. (Icahn Associates Holding LLC), Santaris Pharma A/S (Roche Holding AG)
• Inventors: Thrue, Charlotte Albaek, Hansen, Henrik Frydenlund, Westergaard, Majken, Rasmussen, Frank Winther
• Primary Examiner(s): ZARA, JANE J
• Application Number:
  US12/477,261
• Publication Number:
  US 20090286859A1
• Time in Patent Office: 706 Days
• Field of Search: 435/6, 435/91.1, 435/91.31, 435/455, 514/44, 536/23.1, 536/24.5
• US Class Current: 514/44.00R
• 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 11/06 Antiasthmatics
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 17/08 Antiseborrheics
• CPC Class Codes: A61P 17/12 Keratolytics, e.g. wart or anti-corn preparations
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 27/00 Drugs for disorders of the senses
• 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 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/08 Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• 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 2310/3231 having an additional ring, e.g. LNA, ENA
• CPC Class Codes: C12N 2310/3341 5-Methylcytosine
• CPC Class Codes: C12N 2310/341 Gapmers, i.e. of the type ===---===
• CPC Class Codes: C12N 2310/345 : having at least two different backbone modifications
• CPC Class Codes: C12N 2310/346 : having a combination of backbone and sugar modifications

Abstract

The present disclosure relates to an LNA oligonucleotide consisting of a sequence selected from the group consisting of 5′-(T_x)G_xG_xc_sa_sa_sg_sc_sa_st_sc_sc_sT_xG_xT-3′ and 5′-(G_x)T_xT_xa_sc_st_sg_sc_sc_st_st_sc_sT_xT_xA-3′, wherein capital letters designate a beta-D-oxy-LNA nucleotide analogue, small letters designate a 2-deoxynucleotide, underline designates either a beta-D-oxy-LNA nucleotide analogue or a 2-deoxynucleotide, subscript “s” designates a phosphorothioate link between neighbouring nucleotides/LNA nucleotide analogues, and subscript “x” designates either a phosphorothioate link or a phosphorodiester link between neighbouring nucleotides/LNA nucleotide analogues, and wherein the sequence is optionally extended by up to five 2-deoxynucleotide units. The LNA oligonucleotides are useful for modulating the expression of hypoxia-inducible factor-1a (HIF-1a), e.g. in the treatment of cancer diseases, inhibiting angiogenesis, inducing apoptosis, preventing cellular proliferation, or treating an angiogenic disease, e.g. diabetic retinopathy, macular degeneration (ARMD), psoriasis, rheumatoid arthritis and other inflammatory diseases.

Claims

1. A method for treating cancer, said method comprising administering a therapeutically effective amount of an LNA oligonucleotide consisting of the sequence:

5′

-T_sG_sG_sc_sa_sa_sg_sc_sa_st_sc_sc_sT_sG_sT_sa-3′

(SEQ ID NO.

     1)

wherein capital letters designate a beta-D-oxy-LNA nucleotide analogue, lower case letters designate a 2-deoxynucleotide, and subscript “

s”

designates a phosphorothioate link between neighboring nucleotides/LNA nucleotide analogues to a patient in need thereof.

2. The method according to claim 1, wherein said cancer is a solid tumor.

3. The method according to claim 1, wherein the cancer is selected from the group consisting of multiple myeloma, renal cancer, cervical cancer, colon cancer, brain cancer, and breast cancer.

4. A method of inducing apoptosis in a cell comprising contacting the cell with an effective amount of an LNA oligonucleotide consisting of the sequence:

5′

-T_sG_sG_sc_sa_sa_sg_sc_sa_st_sc_sc_sT_sG_sT_sa-3′

(SEQ ID NO.

     1)

wherein capital letters designate a beta-D-oxy-LNA nucleotide analogue, lower case letters designate a 2-deoxynucleotide, and subscript “

s”

designates a phosphorothioate link between neighboring nucleotides/LNA nucleotide analogues.

5. A method of preventing proliferation of a cell comprising contacting the cell with an effective amount of an LNA oligonucleotide consisting of the sequence:

5′

-T_sG_sG_sc_sa_sa_sg_sc_sa_st_sc_sc_sT_sG_sT_sa-3′

(SEQ ID NO.

     1)

wherein capital letters designate a beta-D-oxy-LNA nucleotide analogue, lower case letters designate a 2-deoxynucleotide, and subscript “

s”

designates a phosphorothioate link between neighboring nucleotides/LNA nucleotide analogues.