Treatment of erythropoietin (EPO) related diseases by inhibition of natural antisense transcript to EPO

• US 8,921,329 B2
• Filed: 12/03/2009
• Issued: 12/30/2014
• Est. Priority Date: 12/04/2008
• Status: Active Grant

• Current Assignee: CURNA, Inc. (Opko Health, Inc.)
• Original Assignee: CURNA, Inc. (Opko Health, Inc.)
• Inventors: Collard, Joseph, Sherman, Olga Khorkova
• Primary Examiner(s): WHITEMAN, BRIAN A
• Application Number:
  US13/133,025
• Publication Number:
  US 20110237651A1
• Time in Patent Office: 1,853 Days
• Field of Search: 514/44.A
• US Class Current: 514/44.A
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• 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 13/12 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 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• 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 25/08 Antiepileptics; Anticonvulsants
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/18 Antipsychotics, i.e. neuroleptics; Drugs for mania or schizophrenia
• CPC Class Codes: A61P 25/22 Anxiolytics
• CPC Class Codes: A61P 25/24 Antidepressants
• 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 27/06 Antiglaucoma agents or miotics
• 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 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• 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 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 7/06 : Antianaemics
• 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 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: A61P 9/12 : Antihypertensives
• CPC Class Codes: C07K 14/505 : Erythropoietin [EPO]
• 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 15/1136 : against growth factors, growth regulators, cytokines, lymphokines or hormones
• 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: C12Q 1/6883 : for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/136 : Screening for pharmacological compounds
• CPC Class Codes: C12Q 2600/158 : Expression markers
• CPC Class Codes: C12Q 2600/178 : miRNA, siRNA or ncRNA
• CPC Class Codes: Y10T 436/143333 : Saccharide [e.g., DNA, etc.]

Abstract

Oligonucleotide compounds modulate expression and/or function of Erythropoietin (EPO) polynucleotides and encoded products thereof. Methods for treating diseases associated with Erythropoietin (EPO) comprise administering one or more oligonucleotide compounds designed to inhibit the EPO natural antisense transcript to patients.

Claims

1. A method of upregulating a function of and/or the expression of an Erythropoietin (EPO) polynucleotide in patient cells or tissues in vivo or in vitro comprising:

contacting said cells or tissues with at least one single stranded antisense oligonucleotide 10 to 30 nucleotides in length or a double stranded antisense oligonucleotide 19 to 30 nucleotides in length wherein said oligonucleotide has at least 80% sequence identity to a reverse complement of a polynucleotide comprising 10 to 30 or 19 to 30 nucleotides respectively within nucleotides 1 to 156 of a natural antisense oligonucleotide having SEQ ID NO;

3;

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

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

4 to 6.

3. A method of upregulating a function of and/or the expression of an Erythropoictin (EPO) polynucleotide having SEQ ID NO:

1 or 2 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 10-30 nucleotides in length or a double stranded antisense oligonucleotide of 19-30 nucleotides in length that targets a region of a natural antisense polynucleotide of the Erythropoietin (EPO) polynucleotide wherein said natural antisense polynucleotide comprises SEQ ID NO;

3;

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

4. The method of claim 3 wherein the expression and/or function of the Erythropoietin (EPO) is increased in vivo or in vitro with respect to a control.

5. The method of claim 3, wherein the at least one antisense oligonucleotide targets a region of the natural antisense oligonucleotide of the Erythropoietin (EPO) polynucleotide wherein said natural antisense sequence is selected from SEQ ID NO:

3.

6. The method of claim 3, wherein the at least one antisense oligonucleotide targets a nucleic acid sequence of said natural antisense oligonucleotide that is complementary to coding nucleic acid sequences of an Erythropoietin (EPO) polynucleotide.

7. The method of claim 3, wherein the at least one antisense oligonucleotide targets a natural antisense oligonucleotide having overlapping sequences with said Erythropoietin (EPO) polynucleotide.

8. The method of claim 3, 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 nuclecotide, 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 internucleoside linkage selected from:

a phosphorothioate, 2′

-O-methoxyethyl (MOE), 2′

-fluoro, alkylphosphonate, phosphorodithioate, 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), analogues, derivatives, and combinations thereof.

12. The method of claim 3 wherein the natural antisense transcript is coding or non-coding.

13. A method of upregulating a function of and/or the expression of an Erythropoictin (EPO) 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, said at least one siRNA oligonucleotide being specific for a region of a natural antisense polynucleotide of an Erythropoictin (EPO) polynucleotide, wherein said ea-leas-eone natural antisense polynucleotide of the Erythropoietin (EPO) polynucleotide comprises SEQ ID NO;

3; and

, upregulating Erythropoietin (EPO) gene expression and/or function in mammalian cells or tissues in vivo or in vitro.

14. The method of claim 13, wherein said oligonucleotide has at least 80% sequence identity to a sequence of at least about five consecutive nucleic acids that is complementary to the antisense and/or sense nucleic acid molecule of the Erythropoietin (EPO) polynucleotide.

15. A method of upregulating an Erythropoietin (EPO) gene expression and/or function 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 an EPO polynucleotide wherein said natural antisense strand comprises SEQ ID NO;

3 and wherein the at least one antisense oligonucleotide has at least 80% sequence identity to at least one nucleic acid sequence encoded by SEQ ID NO;

2, and, upregulating Erythropoietin (EPO) gene expression and/or function in mammalian cells or tissues in vivo or in vitro.

16. A method of treating a disease or disorder associated with at least one Erythropoietin (EPO) polynucleotide having SEQ ID NOS:

1 or 2 and/or at least one encoded product thereof wherein said disease or disorder associated with the at least one Erythropoictin (EPO) polynucleotide is selected from;

a disease, disorder, or state of hematologic irregularity, a blood disorder characterized by low or defective red blood cell production or an anemia, comprising;

administering to a patient a therapeutically effective dose of at least one antisense oligonucleotide of 10 to 30 nucleotides in length that binds to a natural antisense sequence of said at least one Erythropoietin (EPO) polynucleotide which comprises SEQ ID NO;

3 and upregulates expression of said at least one Erythropoietin (EPO) polynucleotide;

thereby treating the disease or disorder associated with the at least one Erythropoietin (EPO) polynucleotide and/or at least one encoded product thereof.