Treatment of sialidase 4 (NEU4) related diseases by inhibition of natural antisense transcript to NEU4

• US 8,993,533 B2
• Filed: 10/05/2011
• Issued: 03/31/2015
• Est. Priority Date: 10/06/2010
• Status: Active Grant

• Current Assignee: CURNA, Inc. (Opko Health, Inc.)
• Original Assignee: CURNA, Inc. (Opko Health, Inc.)
• Inventors: Collard, Joseph, Khorkova Sherman, Olga
• Primary Examiner(s): Chong, Kimberly
• Application Number:
  US13/879,100
• Publication Number:
  US 20130245095A1
• Time in Patent Office: 1,273 Days
• US Class Current: 514/44
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• 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 17/06 Antipsoriatics
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• 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 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• 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 9/10 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 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• 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 2310/315 : Phosphorothioates
• CPC Class Codes: C12N 9/2402 : hydrolysing O- and S- glycosyl compounds (3.2.1)
• CPC Class Codes: C12Y 302/01018 : Exo-alpha-sialidase (3.2.1.18), i.e. trans-sialidase

Abstract

The present invention relates to antisense oligonucleotides that modulate the expression of and/or function of Sialidase 4 (NEU4), in particular, by targeting natural antisense polynucleotides of Sialidase 4 (NEU4). The invention also relates to the identification of these antisense oligonucleotides and their use in treating diseases and disorders associated with the expression of NEU4.

Claims

1. A method of upregulating a function of and/or the expression of a Sialidase 4 (NEU4) polynucleotide selected from SEQ ID No:

1 in a biological system comprising;

contacting said system with at least one antisense oligonucleotide 10 to 30 nucleotides in length wherein said at least one oligonucleotide specifically hybridizes to a natural antisense polynucleotide of the Neu4 polynucleotide and has at least 80% sequence identity to a 10 to 30 nucleotide region of a reverse complement of a natural antisense polynucleotide of the Sialidase 4 (NEU4) polynucleotide selected from nucleotides 1 to 2119 of SEQ ID NO;

2 or 1245 of SEQ ID NO;

3 or 1 to 490 of SEQ ID NO;

4 or 1 to 527 of SEQ ID NO;

5 or 1 to 946 of SEQ ID NO;

6 or 1 to 703 of SEQ ID NO 7;

thereby upregulating a function of and/or the expression of the Sialidase 4 (NEU4) polynucleotide.

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

8 to 11.

2. A method of upregulating a function of and/or the expression of a Sialidase 4 (NEU4) polynucleotide having SEQ ID NO:

1 in a biological system comprising;

contacting said system with at least one antisense oligonucleotide of 10 to 30 nucleotides in length that targets and specifically hybridizes to a region of a natural antisense polynucleotide of the Sialidase 4 (NEU4) polynucleotide wherein said natural antisense polynucleotide is selected from SEQ ID NOS;

2-7;

thereby upregulating a function of and/or the expression of the Sialidase 4 (NEU4) polynucleotide.

3. The method of claim 2, wherein a function of and/or the expression of the Sialidase 4 (NEU4) is increased in vivo or in vitro with respect to a mock-transfected control.

4. The method of claim 2, wherein the at least one antisense oligonucleotide targets a natural antisense sequence of a Sialidase 4 (NEU4) polynucleotide selected from SEQ ID NOS:

2-6.

5. The method of claim 2, wherein the at least one antisense oligonucleotide targets a natural antisense polynucleotide antisense to coding nucleic acid sequence of a Sialidase 4 (NEU4) polynucleotide.

6. The method of claim 2, wherein the at least one antisense oligonucleotide targets a natural antisense polynucleotide having overlapping sequences with a Sialidase 4 (NEU4) RNA polynucleotide.

7. The method of claim 2, 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.

8. The method of claim 7, 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.

9. The method of claim 7, 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.

10. The method of claim 7, 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.

12. A method of modulating a function of and/or the expression of a Sialidase 4 (NEU4) gene having SEQ ID NO:

1 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 natural antisense polynucleotide of a Sialidase 4 (NEU4) polynucleotide selected fromSEQ ID NOS;

27, and, upregulating a function of and/or the expression of Sialidase 4 (NEU4) in mammalian cells or tissues in vivo or in vitro.

13. A method of upregulating a function of and/or the expression of Sialidase 4 (NEU4) polynucleotide having SEQ ID NO:

1 in mammalian cells or tissues in vivo or in vitro comprising;

contacting said cells or tissues with at least one antisense oligonucleotide of about 14 to 30 nucleotides in length specific for noncoding and/or coding sequences of a sense and/or natural antisense strand of a Sialidase 4 (NEU4) polynucleotide having SEQ ID NOS;

2-7 wherein said at least one antisense oligonucleotide has at least 80% sequence identity to a 14 to 30 nucleotide region of at least one nucleic acid sequence set forth as SEQ ID NOS;

1 or an RNA transcribed from the Nue4 polynucleotide; and

, upregulating the function and/or expression of the Sialidase 4 (NEU4) in mammalian cells or tissues in vivo or in vitro.

14. A method of treating a disease associated with at least one Sialidase 4 (NEU4) polynucleotide having SEQ ID NO:

1 and/or at least one encoded product thereof, comprising;

administering to a patient a therapeutically effective dose of at least one antisense oligonucleotide that binds to a natural antisense sequence of said at least one Sialidase 4 (NEU4) polynucleotide selected from SEQ id NOS;

2-7 and upregulates expression of said at least one Sialidase 4 (NEU4) polynucleotide;

thereby treating the disease associated with the at least one Sialidase 4 (NEU4) polynucleotide and/or at least one encoded product thereof and wherein the disease or disorder is selected from Tay Sachs disease or a disease or disorder associated with impaired glycoprotein desialylation.