Treatment of tumor necrosis factor receptor 2 (TNFR2) related diseases by inhibition of natural antisense transcript to TNFR2

• US 8,859,515 B2
• Filed: 06/24/2010
• Issued: 10/14/2014
• Est. Priority Date: 06/24/2009
• Status: Active Grant

• Current Assignee: CURNA, Inc. (Opko Health, Inc.)
• Original Assignee: CURNA, Inc. (Opko Health, Inc.)
• Inventors: Collard, Joseph, Khorkova Sherman, Olga
• Primary Examiner(s): McGarry, Sean
• Application Number:
  US13/379,728
• Publication Number:
  US 20120094934A1
• Time in Patent Office: 1,573 Days
• Field of Search: None
• US Class Current: 514/44.A
• CPC Class Codes: A61K 31/711 Natural deoxyribonucleic acids, i.e. containing only 2'-deoxyriboses attached to adenine, guanine, cytosine or thymine and having 3'-5' phosphodiester links
• 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 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• 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 33/06 Antimalarials
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 39/06 Free radical scavengers or antioxidants
• 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 15/1138 against receptors or cell surface proteins
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/111 : spanning the whole gene, or a large part of it
• CPC Class Codes: C12N 2310/113 : targeting other non-coding nucleic acids, e.g. antagomirs
• CPC Class Codes: C12N 2320/30 : Special therapeutic applications

Abstract

The present invention relates to antisense oligonucleotides that modulate the expression of and/or function of Tumor Necrosis Factor Receptor 2 (TNFR2), in particular, by targeting natural antisense polynucleotides of Tumor Necrosis Factor Receptor 2 (TNFR2). The invention also relates to the identification of these antisense oligonucleotides and their use in treating diseases and disorders associated with the expression of TNFR2.

Claims

1. A method of upregulating a function of and/or the expression of a Tumor Necrosis Factor Receptor 2 (TNFR2) polynucleotide having SEQ ID NO:

1 in human patient cells or tissues in vivo or in vitro comprising;

contacting said cells or tissues with at least one antisense oligonucleotide 16 to 26 nucleotides in length wherein said at least one oligonueleotide has at least 80% sequence identity to a reverse complement of a polynueleotide comprising 10 to 30 consecutive nucleotides within nucleotides 1 to 413 of SEQ ID NO;

2 and 1 to 413 of SEQ ID NO;

3;

thereby upregulating a function of and/or the expression of the Tumor Necrosis Factor Receptor 2 (TNFR2) polynucleotide in patient cells or tissues in vivo or in vitro.

12. The method of claim 1, wherein the at least one oliuonucleotide comprises at least one oligonucleotide sequences set forth as SEQ ID NOS:

4 to 9.

2. A method of upregulating a function of and/or the expression of a Tumor Necrosis Factor Receptor 2 (TNFR2) polynucleotide having SEQ ID NO:

1 in human patient cells or tissues in vivo or in vitro comprising;

contacting said cells or tissues with at least one antisense oligonucleotide 16 to 26 nucleotides in length wherein said at least one oligonucleotide is specific for a natural antisense polynucleotide of the TNFR2 polynucleotide and has at least 80% sequence identity to a 16 to 26 nucleotide region of as reverse complement of said natural antisense of a Tumor Necrosis Factor Receptor 2 (TNFR2) polynueleotide;

thereby upregulating a function of and/or the expression of the Tumor Necrosis Factor Receptor 2 (TNFR2) polynucleotide in patient cells or tissues in vivo or in vitro.

3. A method of upregulating a function of and/or the expression of a Tumor Necrosis Factor Receptor 2 (TNFR2) polynucleotide having SEQ ID NO:

1 in human patient cells or tissues in vivo or in vitro comprising;

contacting said cells or tissues with at least one antisense oligonucleotide of 18 to 24 nucleotides in length that specifically targets a complementary region of a natural antisense oligonucleotide of the Tumor Necrosis Factor Receptor 2 (TNFR2) polynucleotide;

thereby upregulating a function of and/or the expression of the Tumor Necrosis Factor Receptor 2 (TNFR2) polynucleotide in patient cells or tissues in vivo or in vitro.

4. The method of claim 3, wherein a function of and/or the expression of the Tumor Necrosis Factor Receptor 2 (TNFR2) is increased in vivo or in vitro with respect to a control.

5. The method of claim 3, wherein the at least one antisense oligonuceleotide targets a natural antisense sequence of a Tumor Necrosis Factor Receptor 2 (TNFR2) polynucleotide selected from SEQ ID NOS:

2 or 3.

6. The method of claim 3, wherein the at least one antisense oligonucleotide targets a natural antisense oligonucleotidc antisense to coding and/or coding and non-coding nucleic acid sequences of a Tumor Necrosis Factor Receptor 2 (TNFR2) polynucleotide.

7. The method of claim 3, wherein the at least one antisense oligonucleotide targets a natural antisense oligonucleotide having overlapping and/or non-overlapping sequences with a Tumor Necrosis Factor Receptor 2 (INFR2) polynucleotide.

8. The method of claim 3, wherein the at least one antisense oligonuceleotide comprises one or more modifications selected from:

at least one modified sugar moiety, at least one modified internclueoside linkage, at least one modified nucleotide, and combinations thereof.

9. The method of claim 8, wherein the one or more modifications comprise at least one modified sugar moiety selected from:

a 2′

-O-methoxyethyl modified sugar moiety, a 2′

-methoxy modified sugar moiety, a 2′

-O-alkyl modified sugar moiety, a bicyclic sugar moiety, and combinations thereof.

10. The method of claim 8, wherein the one or more modifications comprise at least one modified intemucleoside linkage selected from:

a phosphorothioate alkylphosphonate, phosphorodithioate,l alkylphosphonothioate, phosphoramidate, carbamate, carbonate, phosphate triester, acetamidate, carboxymethyl ester, and combinations thereof.

11. The method of claim 8, wherein the one or more modifications comprise at least one modified nucleotide selected from:

a peptide nucleic acid (PNA), a locked nucleic acid (LNA), an arabino-nucleic acid (FANA), an analogue, a derivative, and combinations thereof.

13. A method of modulating a function of and/or the expression of a Tumor Necrosis Factor Receptor 2 (TNFR2) polyttucleotide having SEQ ID NO:

1 in human cells or tissues in vivo or in vitro comprising;

contacting said cells or tissues with at least one short interferina, RNA (siRNA) oligonucleotide 19 to 26 nucleotides in length, said at least one siRNA oligonucleotide being specific for a natural antisense polynucleotide of a Tumor Necrosis Factor Receptor 2 (TNFR2) polynucleotide, wherein said at least one siRNA oligonucleotide has at least 80% sequence identity to a complementary sequence of at least about 19 consecutive nucleic acids of the sense nucleic acid molecule of the Tumor Necrosis Factor Receptor 2 (TNFR2) polynucleotide; and

, upregulating a function of and/or the expression of Tumor Necrosis Factor Receptor 2 (TNFR2) in mammalian cells or tissues in vivo or in vitro.

14. The method of claim 13, wherein said oligonucleotide has at least 90% sequence identity to a sequence of at least about 19 consecutive nucleic acids of the sense nucleic acid molecule of the Tumor Necrosis Factor Receptor 2 (TNFR2) polynuceleotide.

15. A method of upregulating a function of and/or the expression of Tumor Necrosis Factor Receptor 2 (TNFR2) in mammalian cells or tissues in vivo or in vitro comprising:

contacting said cells or tissues with at least one antisense oligonucleotide of about 10 to 30 nucleotides in length specific for noncoding and/or coding sequences of a natural antisense strand of a Tumor Necrosis Factor Receptor 2 (TNFR2) polynucleotide wherein said at least one antisense oligonucleotide has at least 50% sequence identity to at least one nucleic acid sequence of about 10 to 30 nucleotides in length set forth within SEQ ID NO;

1 or a polynucleotide encoded by the TNFR2 gene; and

, upregulating the function and/or expression of the Tumor Necrosis Factor Receptor 2 (TNFR2) in mammalian cells or tissues in vivo or in vitro.

16. A method of preventing or treating, a disease or condition associated with at least one Tumor Necrosis Factor Receptor 2 (INFR2) polynucleotide having SEQ ID NO:

1, comprising;

administering to a human patient a therapeutically effective dose of at least one modified antisense oligonucleotide of 16 to 26 nucleotides in length that binds to a natural antisense sequence of said at least one Tumor Necrosis Factor Receptor 2 (TNFR2) polynucleotide and upregulates expression of said at least one Tumor Necrosis Factor Receptor 2 (TNFR2) polynucleotide;

thereby preventing or treating the disease associated with the at least one Tumor Necrosis Factor Receptor 2 (TNFR2) polynucleotide and/or at least one encoded product thereof Wherein said disease or condition is selected from inflammation or rheumatoid arthritis or an autoimmune disease or disorder or Crohn'"'"'s disease or ischemia.