Treatment of vascular endothelial growth factor (VEGF) related diseases by inhibition of natural antisense transcript to VEGF

• US 8,927,511 B2
• Filed: 12/02/2009
• Issued: 01/06/2015
• 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): Chong, Kimberly
• Application Number:
  US13/132,997
• Publication Number:
  US 20110237650A1
• Time in Patent Office: 1,861 Days
• Field of Search: None
• US Class Current: 514/44
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• CPC Class Codes: A61K 31/712 Nucleic acids or oligonucleotides having modified sugars, i.e. other than ribose or 2'-deoxyribose
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/02 Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
• CPC Class Codes: A61P 11/06 Antiasthmatics
• 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 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 17/06 Antipsoriatics
• 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 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 25/16 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 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 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 37/06 : Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 37/08 : Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• 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 9/04 : 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: 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: C12N 2310/14 : interfering N.A.
• CPC Class Codes: C12N 2310/315 : Phosphorothioates
• CPC Class Codes: C12Q 1/6816 : characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6883 : for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 1/6886 : for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Q 2600/136 : Screening for pharmacological compounds
• CPC Class Codes: C12Q 2600/156 : Polymorphic or mutational markers

Abstract

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

Claims

1. A method of upregulating the function and/or expression of Vascular Endothelial Growth Factor (VEGF) polynucleotide in patient cells or tissues in vivo or in vitro comprising:

contacting said cells or tissues with an antisense single stranded oligonucleotide 10 to 30 nucleotides in length wherein said oligonucleotide specifically hybridizes to a natural antisense polynucleotide of the VEGF gene and has at least 50% sequence identity to a reverse complement of polynucleotide comprising 10 to 30 nucleotides within nucleotides 1 to 643 of SEQ ID NO;

2 or nucleotides 1 to 513 of SEQ ID NO;

3;

thereby upregulating the function and/or expression of the Vascular Endothelial Growth Factor (VEGF) polynucleotide in patient cells or tissues in vivo or in vitro.

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

4 to 9.

2. A method of upregulating the function and/or expression of an Vascular Endothelial Growth Factor (VEGF) polynucleotide in patient cells or tissues in vivo or in vitro comprising:

contacting said cells or tissues with an antisense single stranded oligonucleotide 10 to 30 nucleotides in length wherein said oligonucleotide specifically hybridizes to a natural antisense polynucleotide of the VEGF gene and has at least 90% sequence identity to a 10 to 30 nucleotide segment of a reverse complement of the natural antisense polynucleotide of the Vascular Endothelial Growth Factor (VEGF) polynucleotide;

thereby upregulating the function and/or expression of the Vascular Endothelial Growth Factor (VEGF) polynucleotide in patient cells or tissues in vivo or in vitro.

3. A method of upregulating the function and/or expression of an Vascular Endothelial Growth Factor (VEGF) polynucleotide in patient cells or tissues in vivo or in vitro comprising:

contacting said cells or tissues with an antisense single stranded modified oligonucleotide 10 to 30 nucleotides in length wherein said oligonucleotide specifically hybridizes to a natural antisense polynucleotide of the VEGF gene and has at least 90% sequence identity to a 10 to 30 nucleotide region of an RNA polynucleotide encoded by a Vascular Endothelial Growth Factor (VEGF) polynucleotide;

thereby upregulating the function and/or expression of the Vascular Endothelial Growth Factor (VEGF) polynucleotide in patient cells or tissues in vivo or in vitro.

4. A method of upregulating the function and/or expression of an Vascular Endothelial Growth Factor (VEGF) polynucleotide in patient cells or tissues in vivo or in vitro comprising:

contacting said cells or tissues with an antisense single stranded oligonucleotide of about 15 to 30 nucleotides in length that targets a complementary region of a natural antisense transcript of a Vascular Endothelial Growth Factor (VEGF) polynucleotide;

thereby upregulating the function and/or expression of an Vascular Endothelial Growth Factor (VEGF) polynucleotide in patient cells or tissues in vivo or in vitro.

5. The method of claim 4, wherein the expression and/or function of the Vascular Endothelial Growth Factor (VEGF) is increased in vivo or in vitro with respect to a control.

6. The method of claim 4, wherein the antisense oligonucleotide targets a natural antisense sequence of Vascular Endothelial Growth Factor (VEGF) polynucleotide comprising SEQ ID NO:

2 or 3.

7. The method of claim 4, wherein the antisense oligonucleotide targets a nucleic acid sequence antisense to the coding nucleic acid sequences of Vascular Endothelial Growth Factor (VEGF) RNA polynucleotide.

8. The method of claim 4, wherein the antisense oligonucleotide targets the natural antisense transcript having overlapping and/or non-overlapping sequences with the Vascular Endothelial Growth Factor (VEGF) RNA polynucleotide.

9. The method of claim 4, wherein the antisense oligonucleotides comprise one or more modifications comprising:

a modified sugar moiety, a modified internucleoside linkage, a modified nucleotide and/or combinations thereof.

10. The method of claim 9, wherein the modified sugar moiety comprises a 2′

-O-methoxyethyl modified sugar moiety, a 2′

-methoxy modified sugar moiety, a 2′

-O-alkyl modified sugar moiety, or a bicyclic sugar moiety.

11. The method of claim 9, wherein the modified internucleoside linkages comprise a phosphorothioate, alkylphosphonate, phosphorodithioate, alkylphosphonothioate, phosphoramidate, carbamate, carbonate, phosphate triester, acetamidate, carboxymethyl ester, and/or combinations thereof.

12. The method of claim 9, wherein the oligonucleotides optionally having at least one modified nucleotides comprising, peptide nucleic acids, locked nucleic acid (LNA) molecules, arabino-nucleic acid (FANA), peptide nucleic acids (PNAs), analogues or derivatives thereof.

14. A method of upregulating an Vascular Endothelial Growth Factor (VEGF) gene expression and/or function in mammalian cells or tissues in vivo or in vitro comprising:

contacting said cells or tissues with a short interfering RNA (siRNA) oligonucleotide 19 to 30 nucleotides in length specific for a natural antisense polynucleotide of an Vascular Endothelial Growth Factor (VEGF) polynucleotide wherein said oligonucleotide has at least 80% sequence identity to about nineteen consecutive nucleic acids of the antisense RNA nucleic acid molecule transcribed from the Vascular Endothelial Growth Factor (VEGF) polynucleotide; and

, upregulating Vascular Endothelial Growth Factor (VEGF) gene expression and/or function in mammalian cells or tissues in vivo or in vitro.

15. The method of claim 14, wherein said oligonucleotide has at least 90% sequence identity to about nineteen consecutive nucleic acids of the antisense nucleic acid molecule of the Vascular Endothelial Growth Factor (VEGF) polynucleotide.

16. A method of upregulating Vascular Endothelial Growth Factor (VEGF) gene expression and/or function in mammalian cells or tissues in vivo or in vitro comprising:

contacting said cells or tissues with an antisense oligonucleotide of about 10 to 30 nucleotides in length specific for noncoding or coding and noncoding sequences of a natural antisense strand of a polynucleotide encoding an Vascular Endothelial Growth Factor (VEGF) molecule wherein the antisense oligonucleotide has at least 50% sequence identity to at least one 10 to 30 polynucleotide region within the nucleic acid sequences set forth as SEQ ID NO;

1 or an RNA polynucleotide encoded by the VEGF gene or has at least 50% complementarity to SEQ ID NO;

17; and

, upregulating Vascular Endothelial Growth Factor (VEGF) gene expression and/or function in mammalian cells or tissues in vivo or in vitro.

17. A method of preventing or treating diseases associated with Vascular Endothelial Growth Factor (VEGF) polynucleotides and/or encoded products thereof, comprising:

administering to a patient a therapeutically effective dose of at least one antisense oligonucleotide that binds to a natural antisense sequence of Vascular Endothelial Growth Factor (VEGF) polynucleotide and modulates expression of said Vascular Endothelial Growth Factor (VEGF) polynucleotides;

thereby preventing or treating diseases associated with Vascular Endothelial Growth Factor (VEGF) polynucleotides and/or encoded products thereof.

18. The method of claim 17, wherein a disease associated with Vascular Endothelial Growth Factor (VEGF) polynucleotides comprise:

DiGeorge syndrome, low VEGF and neuropilin-1 expression;

atherosclerosis;

nerve injury, brain injury and Neurodegenerative disorders selected from Alzheimer'"'"'s disease, or amyotrophic lateral sclerosis;

or ulceration.