Treatment of sirtuin (SIRT) related diseases by inhibition of natural antisense transcript to a sirtuin (SIRT)

• US 9,089,588 B2
• Filed: 01/13/2011
• Issued: 07/28/2015
• Est. Priority Date: 05/03/2010
• Status: Active Grant

• Current Assignee: CURNA, Inc. (Opko Health, Inc.)
• Original Assignee: CURNA, Inc. (Opko Health, Inc.)
• Inventors: Collard, Joseph, Khorkova Sherman, Olga, Coito, Carlos, De Leon, Belinda
• Primary Examiner(s): Vivlemore, Tracy
• Assistant Examiner(s): POLIAKOVA-GEORGAN, EKATERINA
• Application Number:
  US13/695,801
• Publication Number:
  US 20130072421A1
• Time in Patent Office: 1,657 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• 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 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 13/00 Drugs for disorders of the urinary system
• CPC Class Codes: A61P 13/02 of urine or of the urinary tract, e.g. urine acidifiers
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/08 Antiseborrheics
• CPC Class Codes: A61P 17/18 Antioxidants, e.g. antiradicals preparations for protection against sunlight A61Q17/00
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 19/06 Antigout agents, e.g. antihyperuricemic or uricosuric agents
• CPC Class Codes: A61P 19/08 for bone diseases, e.g. rachitism, Paget's disease
• CPC Class Codes: A61P 19/10 for osteoporosis
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 21/02 Muscle relaxants, e.g. for tetanus or cramps
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/02 for peripheral neuropathies
• CPC Class Codes: A61P 25/14 : for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 25/16 : Anti-Parkinson drugs
• 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 25/32 : Alcohol-abuse
• CPC Class Codes: A61P 27/02 : Ophthalmic agents
• CPC Class Codes: A61P 27/12 : for cataracts
• 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 3/04 : Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/06 : Antihyperlipidemics
• CPC Class Codes: A61P 3/08 : for glucose homeostasis pancreatic hormones A61P5/48
• CPC Class Codes: A61P 3/10 : for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/18 : for HIV
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 35/02 : specific for leukemia
• CPC Class Codes: A61P 37/02 : Immunomodulators
• 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 7/00 : Drugs for disorders of the blood or the extracellular fluid
• 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 15/1137 : against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• 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 2330/10 : naturally occurring

Abstract

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

Claims

1. A method of upregulating function of and/or the expression of a Sirtuin (SIRT) gene/ polynucleotide selected from the group consisting of SIRT 3, 4, 5 and 7 in patient cells or tissues in vivo or in vitro comprising:

contacting said cells or tissues with at least one antisense oligonucleotide of 10-30 nucleotides in length that specifically targets a complementary region of a natural antisense oligonucleotide of the Sirtuin (SIRT) polynucleotide wherein said natural antisense oligonucleotide is selected from the group consisting of polynucleotides 1 to 726 of SEQ ID NO;

17, 1 to 320 of SEQ ID NO;

18, 1 to 616 of SEQ ID NO;

19, 1 to 492 of SEQ ID NO;

20, 1 to 705 of SEQ ID NO;

23 or 1 to 2714 of SEQ ID NO;

141 or 1 to 1757 of SEQ ID NO;

142 or 1 to 3647 of SEQ ID NO;

143; and

wherein said oligonucleotide is specifically hybridizable thereto thereby upregulating a function of and/or the expression of the Sirtuin (SIRT) polynucleotide in patient cells or tissues in vivo or in vitro.

2. The method of claim 1, wherein the at least one antisense oligonucleotide targets a natural antisense sequence of a Sirtuin (SIRT) polynucleotide selected from SEQ ID NO:

23.

3. The method of claim 1 wherein the at least one antisense oligonucleotide targets a natural antisense oligonucleotide antisense to coding regions of the Sirtuin (SIRT) polynucleotide.

4. The method of claim 1, wherein said cells or tissues are targeted with an antisense oligonucleotide that targets a natural antisense oligonucleotide having overlapping sequences with a Sirtuin (SIRT) RNA polynucleotide transcribed from the SIRT gene.

5. The method of any 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.

6. The method of claim 5, 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.

7. The method of claim 5, wherein the one or more modifications comprise at least one modified internucleoside linkage selected from:

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

8. The method of claim 5, 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.

9. The method of claim 1, wherein the at least one oligonucleotide comprises a sequence as set forth in any of SEQ ID NOS:

94-111.122 to 127 and 144-145.

10. A method of upregulating a function of and/or the expression of a Sirtuin (SIRT) polynucleotide selected from the group consisting of SIRT 2, 4, 5 and 7 in a discordant manner in mammalian cells or tissues in vivo or in vitro comprising:

contacting said cells or tissues with at least one single stranded oligonucleotide of 10 to 30 nucleotides in length or a short interfering RNA (siRNA) oligonucleotide 19 to 30 nucleotides in length, said at least one single stranded oligonucleotide or siRNA oligonucleotide being specific for a natural antisense polynucleotide of the Sirtuin (SIRT) polynucleotide, wherein said at least one single stranded or siRNA oligonucleotide has at least 80% sequence complementarity to at least about 10-19 nucleotides respectively of the natural antisense polynucleotide of the Sirtuin (SIRT) polynucleotide; and

, upregulating a function of and/or the expression of the Sirtuin (SIRT) in mammalian cells or tissues in vivo or in vitro.

11. The method of claim 10, wherein said oligonucleotide has at least 90% sequence complementarity.

12. A method of upregulating a function of and/or the expression of a Sirtuin (SIRT) polynucleotide selected from the group consisting of SIRT 2, 4, 5 and 7 in mammalian cells or tissues in vivo or in vitro comprising:

contacting said cells or tissues with at least one antisense oligonucleotide of about 15 to 30 nucleotides in length specific for noncoding and/or coding sequences of a natural antisense strand of the Sirtuin (SIRT) polynucleotide and wherein said oligonucleotide is specifically hybridizable thereto wherein said at least one antisense oligonucleotide has at least 90% sequence identity to at least one 15-30 nucleic acid sequence set forth within SEQ ID NOS;

3, 5, 6, 8 and 134-136 and 0.139 or an RNA transcribed from the SIRT 2, 4, 5 or 7 polynucleotide; and

, upregulating the function and/or expression of the Sirtuin (SIRT) in mammalian cells or tissues in vivo or in vitro.