Treatment of delta-like 1 homolog (DLK1) related diseases by inhibition of natural antisense transcript to DLK1

• US 9,708,604 B2
• Filed: 03/16/2010
• Issued: 07/18/2017
• Est. Priority Date: 03/17/2009
• Status: Active Grant

• Current Assignee: CURNA, Inc. (Opko Health, Inc.)
• Original Assignee: CURNA, Inc. (Opko Health, Inc.)
• Inventors: Collard, Joseph, Khorkova Sherman, Olga, Coito, Carlos
• Primary Examiner(s): McGarry, Sean
• Application Number:
  US13/256,710
• Publication Number:
  US 20120010156A1
• Time in Patent Office: 2,681 Days
• Field of Search: None
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• 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 3/00 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 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/00 Drugs for disorders of the endocrine system
• CPC Class Codes: A61P 5/06 of the anterior pituitary hormones, e.g. TSH, ACTH, FSH, LH, PRL, GH
• 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: 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 15/1138 against receptors or cell surface proteins
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/313 Phosphorodithioates
• CPC Class Codes: C12N 2310/314 Phosphoramidates
• CPC Class Codes: C12N 2310/315 : Phosphorothioates

Abstract

The present invention relates to antisense oligonucleotides that modulate the expression of and/or function of Delta-like (1) homolog (DLK1), in particular, by targeting natural antisense polynucleotides of Delta-like (1) homolog (DLK1). The invention also relates to the identification of these antisense oligonucleotides and their use in treating diseases and disorders associated with the expression of (DLK1).

Claims

1. A method of upregulating a function of and/or the expression of a Delta-like 1 homolog (DLK1) polynucleotide having SEQ ID NOS:

1 or 2 in human patient cells or tissues in vivo or in vitro comprising;

contacting said cells or tissues with at least one antisense oligonucleotide of 12 to 30 nucleotides in length that specifically targets a region of a natural antisense oligonucleotide of the Delta-like 1 homolog (DLK1) polynucleotide selected from SEQ ID NO;

3;

thereby upregulating a function of and/or the expression of the Delta-like 1 homolog (DLK1) 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 Delta-like 1 homolog (DLK1) is increased in vivo or in vitro with respect to a control.

3. The method of claim 1, wherein the at least one antisense oligonucleotide targets a natural antisense polynucleotide antisense to coding and/or non-coding nucleic acid sequences of a Delta-like 1 homolog (DLK1) polynucleotide.

4. The method of claim 1, wherein the at least one antisense oligonucleotide targets a natural antisense polynucleotide having overlapping and/or non-overlapping sequences with a Delta-like 1 homolog (DLK1) 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), an analogue, a derivative, and combinations thereof.

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

4 to 5.

10. A method of upregulating a function of and/or the expression of a Delta-like 1 homolog (DLK1) polynucleotide having SEQ ID NOS:

1 or 2 in human 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 Delta-like 1 homolog (DLK1) polynucleotide having SEQ ID NO;

3, wherein said at least one siRNA oligonucleotide has at least 80% sequence identity to at least about 19 consecutive nucleic acids of the RNA transcribed from the Delta-like 1 homolog (DLK1) polynucleotide; and

, upregulating a function of and/or the expression of Delta-like 1 homolog (DLK1) in mammalian cells or tissues in vivo or in vitro.

11. The method of claim 10, wherein said oligonucleotide has at least 90% sequence identity to said RNA transcribed from the Delta-like 1 homolog (DLK1) polynucleotide.

12. A method of upregulating a function of and/or the expression of Delta-like 1 homolog (DLK1) polynucleotide having SEQ ID NOS:

1 or 2 in human cells or tissues in vivo or in vitro comprising;

contacting said cells or tissues with at least one antisense oligonucleotide of about 15 to 30 nucleotides in length specific for a natural antisense strand of a Delta-like 1 homolog (DLK1) polynucleotide having SEQ ID NO;

3 wherein said at least one antisense oligonucleotide has at least 80% sequence identity to a 15 to 30 nucleotide region of at least one nucleic acid sequence set forth as SEQ ID NOS;

1 or an RNA transcribed from the DLK1 gene; and

, upregulating the function and/or expression of the Delta-like 1 homolog (DLK1) in mammalian cells or tissues in vivo or in vitro.