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

• US 10,563,202 B2
• Filed: 05/01/2015
• Issued: 02/18/2020
• Est. Priority Date: 07/24/2009
• Status: Active Grant

• Current Assignee: CURNA, Inc. (Opko Health, Inc.)
• Original Assignee: Gurna Incorporated
• Inventors: Collard, Joseph, Khorkova Sherman, Olga
• Primary Examiner(s): Poliakova-Georgantas, Ekaterina
• Application Number:
  US14/701,998
• Publication Number:
  US 20150267204A1
• Time in Patent Office: 1,754 Days
• Field of Search: None
• 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 13/08 of the prostate
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• 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 19/08 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 21/04 for myasthenia gravis
• 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 25/04 Centrally acting analgesics, e.g. opioids
• 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 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 3/10 : for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/10 : Antimycotics
• 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 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 9/02 : Non-specific cardiovascular stimulants, e.g. drugs for syncope, antihypotensives
• CPC Class Codes: A61P 9/04 : Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: A61P 9/06 : Antiarrhythmics
• 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 15/1137 : against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 2310/11 : Antisense
• CPC Class Codes: C12N 2310/14 : interfering N.A.
• CPC Class Codes: C12N 2310/315 : Phosphorothioates
• CPC Class Codes: C12N 2320/30 : Special therapeutic applications
• CPC Class Codes: C12Y 305/01098 : Histone deacetylase (3.5.1.98), i.e. sirtuin deacetylase

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 a function of and/or the expression of a Sirtuin (SIRT) polynucleotide having SEQ ID NOS:

1, 3 or 4 in patient cells or tissues in vivo or in vitro comprising;

contacting said cells or tissues with at least one single-stranded antisense oligonucleotide 14 to 24 nucleotides in length wherein said at least one oligonucleotide is 100% complementary and hybridizes to a complementary polynucleotide comprising 12 to 24 nucleotides within nucleotides 1 to 1028 of SEQ ID NO;

5 or nucleotides 1 to 352 and 373 to 429 of SEQ ID NO;

6, or nucleotides 1 to 156 of SEQ ID NO;

7 or nucleotides 269 to 593 of SEQ ID NO;

8, 1 to 373 of SEQ ID NO;

9, 1 to 589 of SEQ ID NO;

12, and 1 to 4026 of SEQ ID NO;

14;

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 a function of and/or the expression of the Sirtuin (SIRT) is increased in vivo or in vitro with respect to a control wherein the control is selected from a mock transfected control.

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

4. The method of claim 1, wherein the at least one antisense oligonucleotide targets a natural antisense polynucleotide having overlapping sequences with a Sirtuin (SIRT) polynucleotide.

5. 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.

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, alkylphosphonothioate, 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) and combinations thereof.

9. A method of upregulating a function of and/or the expression of a Sirtuin (SIRT) polynucleotide having SEQ ID NOS:

1, 3 or 4 in mammalian cells or tissues in vivo or in vitro comprising;

contacting said cells or tissues with at least one oligonucleotide wherein the at least one oligonucleotide consists of one oligonucleotide sequence selected from SEQ ID NOs;

15, 29-37, 41-51, 56-60, 73-78, 81, 82, 85-90 and 92-94.

10. A method of upregulating a function of and/or the expression of a Sirtuin (SIRT) polynucleotide having SEQ ID NO:

1, 3 or 4 in mammalian cells or tissues in vivo or in vitro comprising;

contacting said cells or tissues with at least one single stranded antisense oligonucleotide of about 10 to 24 nucleotides in length having 100% complementarity to noncoding and/or coding sequences of a natural antisense strand of a Sirtuin (SIRT) polynucleotide selected from nucleotides 1 to 411 and 423 to 436 and 456 to 1028 of SEQ ID NO;

5 or nucleotides 1 to 429 of SEQ ID NO;

6, or nucleotides 1 to 156 of SEQ ID NO;

7 or nucleotides 1 to 593 of SEQ ID NO;

8, 1 to 373 of SEQ ID NO;

9, 1 to 589 of SEQ ID NO;

12, 1 to 428 of SEQ ID NO;

13 and 1 to 4041 of SEQ ID NO;

14 wherein said at least one antisense oligonucleotide has sequence identity to at least one nucleic acid sequence of 10 to 24 nucleotides in length set forth within SEQ ID NOS;

1, 3 or 4; and

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

11. A method of upregulating a function of and/or the expression of a Sirtuin (SIRT) polynucleotide having SEQ ID NOS:

1, 3 or 4 in patient cells or tissues in vivo or in vitro comprising;

contacting said cells or tissues with at least one single-stranded antisense oligonucleotide 14 to 24 nucleotides in length wherein said at least one oligonucleotide is 100% complementary and hybridizes to a complementary polynucleotide comprising 12 to 24 nucleotides within nucleotides 296 to 660 of SEQ ID NO;

11;

thereby upregulating a function of and/or the expression of the Sirtuin (SIRT) polynucleotide in patient cells or tissues in vivo or in vitro.

12. A method of upregulating a function of and/or the expression of a Sirtuin (SIRT) polynucleotide having SEQ ID NOS:

1, 3 or 4 in patient cells or tissues in vivo or in vitro comprising;

contacting said cells or tissues with at least one single-stranded antisense oligonucleotide of 14 to 24 nucleotides in length or at least one siRNA oligonucleotide of 20-27 nucleotides in length having 100% complementarity with and that targets and hybridizes to a complementary target region of a natural antisense polynucleotide of the Sirtuin (SIRT) polynucleotide;

thereby upregulating a function of and/or the expression of the Sirtuin (SIRT) polynucleotide in patient cells or tissues in vivo or in vitro wherein the SIRT natural antisense polynucleotides are selected from nucleotides 296 to 660 of SEQ ID NO;

11.

13. A method of upregulating a function of and/or the expression of a Sirtuin (SIRT) polynucleotide having SEQ ID NOS:

1, 3 or 4 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 of 20 to 27 nucleotides in length having 100% complementarity with a target region of a natural antisense polynucleotide of a Sirtuin (SIRT) polynucleotide selected from 1 to 35 and 55 to 660 of SEQ ID NO;

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