Treatment of uncoupling protein 2 (UCP2) related diseases by inhibition of natural antisense transcript to UCP2

• US 9,068,183 B2
• Filed: 12/23/2010
• Issued: 06/30/2015
• Est. Priority Date: 12/23/2009
• 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): Vivlemore, Tracy
• Assistant Examiner(s): POLIAKOVA-GEORGA, EKATERINA
• Application Number:
  US13/518,226
• Publication Number:
  US 20120329727A1
• Time in Patent Office: 1,650 Days
• US Class Current: 1/1
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• 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 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 Immunomodulators
• 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 15/1138 against receptors or cell surface proteins
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/113 targeting other non-coding nucleic acids, e.g. antagomirs
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/31 of the backbone
• CPC Class Codes: C12N 2310/312 : Phosphonates
• CPC Class Codes: C12N 2310/313 : Phosphorodithioates
• CPC Class Codes: C12N 2310/314 : Phosphoramidates
• CPC Class Codes: C12N 2310/315 : Phosphorothioates
• CPC Class Codes: C12N 2310/316 : Phosphonothioates
• CPC Class Codes: C12N 2310/3181 : Peptide nucleic acid, PNA
• CPC Class Codes: C12N 2310/32 : of the sugar
• CPC Class Codes: C12N 2310/321 : 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 : 2'-R Modification
• CPC Class Codes: C12N 2310/3231 : having an additional ring, e.g. LNA, ENA

Abstract

The present invention relates to antisense oligonucleotides that modulate the expression of and/or function of Uncoupling Protein 2 (UCP2), in particular, by targeting natural antisense polynucleotides of Uncoupling Protein 2 (UCP2). The invention also relates to the identification of these antisense oligonucleotides and their use in treating diseases and disorders associated with the expression of UCP2.

Claims

1. A method of upregulating a function of and/or the expression of a Uncoupling Protein 2 (UCP2) polynucleotide in patient cells or tissues in vivo or in vitro, said patient having a UCP2 associated disease or disorder, comprising:

contacting said cells or tissues with at least one antisense oligonucleotide of 12-30 nucleotides in length that specifically targets and specifically hybridizes to a region of a natural antisense oligonucleotide of the Uncoupling Protein 2 (UCP2) polynucleotide selected from SEQ ID NOS;

2 or 3;

thereby upregulating a function of and/or the expression of the Uncoupling Protein 2 (UCP2) polynucleotide in patient cells or tissues in vivo or in vitro.

2. The method of claim 1, wherein a function of and/or the expression of the Uncoupling Protein 2 (UCP2) is increased in vivo or in vitro with respect to a mock-transfected control.

3. The method of claim 1, wherein the at least one antisense oligonucleotide targets a natural antisense sequence of a Uncoupling Protein 2 (UCP2) polynucleotide selected from SEQ ID NO 3.

4. The method of claim 1, wherein the at least one antisense oligonucleotide targets a natural antisense oligonucleotide antisense to nucleic acid sequences of the RNA transcribed from the Uncoupling Protein 2 (UCP2) gene.

5. The method of claim 1, wherein the at least one antisense oligonucleotide targets a natural antisense oligonucleotide having overlapping sequences with the RNA transcribed from the Uncoupling Protein 2 (UCP2) gene.

6. The method of claim 1, wherein the at least one antisense oligonucleotide comprises one or more modifications selected from:

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

7. The method of claim 6, 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.

8. The method of claim 6, wherein the one or more modifications comprise at least one modified internucleoside linkage selected from:

a phosphorothioate, 2′

-Omethoxyethyl (MOE), 2-fluoro, alkylphosphonate, phosphorodithioate, alkylphosphonothioate, phosphoramidate, carbamate, carbonate, phosphate triester, acetamidate, carboxymethyl ester, and combinations thereof.

9. The method of claim 6, 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.

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

4 5, 11 and 13.

11. A method of upregulating a function of and/or the expression of a Uncoupling Protein 2 (UCP2) gene in mammalian cells or tissues in vivo or in vitro comprising:

contacting said cells or tissues with at least one short interfering RNA (siRNA) oligonucleotide 19 to 30 nucleotides in length, wherein said at least one siRNA oligonucleotide specifically hybridizes to a region of a natural antisense polynucleotide of a Uncoupling Protein 2 (UCP2) polynucleotide selected front SEQ ID NOS;

2 or 3, and upregulates a function of and/or the expression of Uncoupling Protein 2 (UCP2) in mammalian cells or tissues in vivo or in vitro.

12. A method of upregulating a function of and/or the expression of Uncoupling Protein 2 (UCP2) in mammalian cells or tissues in vivo or in vitro comprising:

contacting said cells or tissues with at least one single stranded modified antisense oligonucleotide of about 20 to 30 nucleotides in length that specifically hybridizes to a natural antisense strand of a Uncoupling Protein 2 (UCP2) polynucleotide selected from SEQ ID NOS;

2 or 3 wherein said at least one antisense oligonucleotide has at least 90% sequence identity to at least one nucleic acid sequence of about 20 to 30 nucleotides set forth within SEQ ID NO;

1 or a polynucleotide encoded by the UCP2 gene; and

, upregulating the function and/or expression of the Uncoupling Protein 2 (UCP2) in mammalian cells or tissues in vivo or in vitro.