Treatment of discs large homolog (DLG) related diseases by inhibition of natural antisense transcript to DLG

• US 9,902,958 B2
• Filed: 06/07/2016
• Issued: 02/27/2018
• Est. Priority Date: 07/14/2010
• Status: Expired due to Fees

• 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): Gibbs, Terra C
• Application Number:
  US15/175,701
• Publication Number:
  US 20160281088A1
• Time in Patent Office: 630 Days
• Field of Search: 435 61, 435 911, 435325, 435375
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• 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 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• 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 25/18 Antipsychotics, i.e. neuroleptics; Drugs for mania or schizophrenia
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• 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 2310/111 spanning the whole gene, or a large part of it
• 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 2330/10 naturally occurring
• CPC Class Codes: C12Q 1/6813 Hybridisation assays

Abstract

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

Claims

1. A method of identifying and selecting at least one modified antisense oligonucleotide specific for a target natural antisense transcript of a Discs large homolog 1 (DLG 1) gene wherein the function of and/or expression of said DLG1 gene is modulated by said at least one modified antisense oligonucleotide comprising:

identifying at least one modified oligonucleotide of 10 to 30 nucleotides in length which are at least 90% complementary to a 10 to 30 nucleotide region of a natural antisense polynucleotide of said DLG 1 gene;

measuring the thermal melting point of a hybrid of said modified antisense oligonucleotide and said natural antisense polynucleotide of said DLG-1 gene under stringent hybridization conditions to identify and select at least one modified antisense oligonucleotide having a high thermal melting point; and

measuring the modulation activity of said modified antisense oligonucleotide identified as having said high thermal melting point; and

identifying and selecting at least one modified antisense oligonucleotide having modulation activity and binding specificity for said natural antisense polynucleotide wherein the at least one modified antisense oligonucleotide comprises one or more modifications selected from;

at least one modified sugar moiety, at least one modified intemucleoside linkage, at least one modified nucleotide, and combinations thereof.

2. The method according to claim 1 wherein said at least one modified antisense oligonucleotide has at least 80% sequence identity to a reverse complement of a polynucleotide comprising 10 to 30 consecutive nucleotides within the natural antisense polynucleotide having nucleotides 1 to 1706 of SEQ ID NO:

2.

3. The method of claim 1, wherein a function of and/or the expression of the Discs large homolog 1 (DLG 1) is increased in vivo or in vitro with respect to a control.

4. The method of claim 1, wherein the at least one antisense oligonucleotide targets a natural antisense polynucleotide of a Discs large homolog 1 (DLG 1) polynucleotide having SEQ ID NO:

2.

5. The method of claim 1, wherein the at least one antisense oligonucleotide targets a natural antisense polynucleotide having coding and/or non-coding nucleic acid sequences of a Discs large homolog 1 (DLG 1) polynucleotide.

6. The method of claim 1, wherein the at least one modified antisense oligonucleotide comprises one or more modifications selected from:

at least one modified sugar moiety, at least one modified intemucleoside linkage or at least one modified nucleotide.

7. The method of claim 6, wherein the one or more modifications comprise at least one modified sugar moiety selected from:

a 2′

-0-methoxyethyl modified sugar moiety, a 2′

-methoxy modified sugar moiety, a 2′

-0-alkyl modified sugar moiety, a bicyclic sugar moiety, and combinations thereof.

8. The method of claim 6, wherein the one or more modifications comprise at least one modified intenucleoside linkage selected from:

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

9. The method of claim 6, 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.

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

3 to 12.

11. The method according to claim 1 wherein the antisense oligonucleotide is a siRNA having 19-30 nucleotides.