TREATMENT OF SIRTUIN (SIRT) RELATED DISEASES BY INHIBITION OF NATURAL ANTISENSE TRANSCRIPT TO A SIRTUIN (SIRT)

• US 20200131517A1
• Filed: 01/10/2020
• Published: 04/30/2020
• Est. Priority Date: 07/24/2009
• Status: Active Grant

• Current Assignee: CURNA, Inc. (Opko Health, Inc.)
• Original Assignee: CURNA, Inc. (Opko Health, Inc.)
• Inventors: COLLARD, Joseph, KHORKOVA SHERMAN, Olga
• Granted Patent: US 10,934,546 B2
• Time in Patent Office: Days
• 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 synthetic, modified single stranded oligonucleotide of 14 to 24 nucleotides in length comprising at least one modification wherein the at least one modification is selected from:

at least one modified sugar moiety at least one modified internucleotide linkage;

at least one modified nucleotide, and combinations thereof, wherein said oligonucleotide is an antisense compound which 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, or 1 to 373 of SEQ ID NO;

9, or 1 to 589 of SEQ ID NO;

12 or 1 to 4026 of SEQ ID NO;

14, thereby upregulating a function and/or expression of a Sirtuin (SIRT) polynucleotide having SEQ ID NOS;

1, 3 or 4 in vivo or in vitro as compared to a normal control.

2. The oligonucleotide of claim 1, wherein the at least one modification comprises an internucleotide linkage selected from the group consisting of:

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

3. The oligonucleotide of claim 1, wherein said oligonucleotide comprises at least one phosphorothioate internucleotide linkage.

4. The oligonucleotide of claim 1, wherein said oligonucleotide comprises a backbone of phosphorothioate internucleotide linkages.

5. The oligonucleotide of claim 1, wherein the olignucleotide comprises at least one modified nucleotide, said modified nucleotide selected from:

a peptide nucleic acid, a locked nucleic acid (LNA), analogue, derivative, and a combination thereof.

6. The oligonucleotide of claim 1, wherein the oligonucleotide comprises a plurality of modifications, wherein said modifications comprise modified nucleotides selected from:

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

7. The oligonucleotide of claim 1, wherein the oligonucleotide comprises a plurality of modifications, wherein said modifications comprise modified nucleotides selected from:

peptide nucleic acids, locked nucleic acids (LNA), analogues, derivatives, and a combination thereof.

8. The oligonucleotide of claim 1, wherein the oligonucleotide comprises 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 a combination thereof.

9. The oligonucleotide of claim 1, wherein the oligonucleotide comprises a plurality of modifications, wherein said modifications comprise modified sugar moieties 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 a combination thereof.

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

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

11. A pharmaceutical composition comprising one or more oligonucleotides according to claim 1 and a pharmaceutically acceptable excipient.

12. The pharmaceutical composition of claim 11, wherein the oligonucleotides have at least about 95% sequence identity as compared to any one of the nucleotide sequences set forth as SEQ ID NOS:

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

13. A single stranded, modified oligonucleotide having 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 1-156 of SEQ ID NO;

7 or nucleotides 1 to 593 of SEQ ID NO;

8 or 1 to 373 of SEQ ID NO;

9 or 1 to 159 of SEQ ID NO;

12 or 1 to 428 of SEQ ID NO;

13 or 1 to 4041 of SEQ ID NO;

14 and 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 wherein said antisense oligonucleotides upregulate the expression of SEQ ID NOS;

1, 3 or 4 in mammalian cells or tissues in vivo or in vitro.

17. A pharmaceutical composition comprising a single stranded modified oligonucleotide according to claims 13, 14 or 16 and a pharmaceutically acceptable excipient.

14. A single stranded, modified oligonucleotide of 14 to 24 nucleotides in length wherein said oligonucleotide is 100% complementary to and hybridizes to a polynucleotide comprising 12 to 24 nucleotides within nucleotides 296 to 660 of SEQ ID NO:

11, thereby upregulating a function and/or expression of a Sirtuin (SIRT) polynucleotide having SEQ ID NOS;

1, 3 or 4 in patient cells or tissues in vivo or in vitro.

15. A 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 nucleotides 1 to 35 and 55 to 660 of SEQ ID NO:

11 and upregulating the expression or function of a Sirtuin (SIRT) polynucleotide having SEQ ID NOS;

3 or 4 in mammalian cells or tissues in vivo or in vitro.

18. A pharmaceutical composition comprising a siRNA according to claims 15 or 16 and a pharmaceutically acceptable excipient.

16. A single-stranded, modified 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 which targets and hybridizes to a complementary target region of a natural antisense polynucleotide of the Sirtuin (SIRT) polynucleotide thereby upregulating a function and/or expression of a SIRT polynucleotide having SEQ ID NOS:

1, 3 or 4 and wherein the target region is selected from nucleotides 296 to 660 of SEQ ID NO;

11.