Treatment of filaggrin (FLG) related diseases by modulation of FLG expression and activity

• US 10,113,166 B2
• Filed: 11/25/2014
• Issued: 10/30/2018
• Est. Priority Date: 09/25/2009
• 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): Poliakova-Georgantas, Ekaterina
• Application Number:
  US14/552,726
• Publication Number:
  US 20150152417A1
• Time in Patent Office: 1,435 Days
• Field of Search: None
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/135 having aromatic rings , e.g. ketamine, nortriptyline methadone A61K31/137
• CPC Class Codes: A61K 31/325 Carbamic acids; Thiocarbamic acids; Anhydrides or salts thereof thiurams A61K31/145
• CPC Class Codes: A61K 31/357 having two or more oxygen atoms in the same ring, e.g. crown ethers, guanadrel
• CPC Class Codes: A61K 31/4406 only substituted in position 3, e.g. zimeldine nicotinic acid A61K31/455
• CPC Class Codes: A61K 31/4439 containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole nicotine A61K31/465
• CPC Class Codes: A61K 31/444 containing a six-membered ring with nitrogen as a ring heteroatom, e.g. amrinone
• CPC Class Codes: A61K 31/5415 ortho- or peri-condensed with carbocyclic ring systems, e.g. phenothiazine, chlorpromazine, piroxicam
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 11/06 Antiasthmatics
• 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 17/04 Antipruritics
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 17/16 Emollients or protectives, e.g. against radiation
• CPC Class Codes: A61P 17/18 Antioxidants, e.g. antiradicals preparations for protection against sunlight A61Q17/00
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• 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 29/00 : Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 31/22 : for herpes viruses
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 37/00 : Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 : Immunomodulators
• CPC Class Codes: A61P 37/08 : Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• 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 2310/11 : Antisense
• CPC Class Codes: C12N 2310/14 : interfering N.A.
• CPC Class Codes: C12N 2310/315 : Phosphorothioates
• CPC Class Codes: C12N 2310/3181 : Peptide nucleic acid, PNA
• CPC Class Codes: C12N 2310/321 : 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 : 2'-R Modification
• CPC Class Codes: C12N 2310/3231 : having an additional ring, e.g. LNA, ENA

Abstract

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

Claims

1. A method of upregulating a function of and/or the expression of a Filaggrin (FLG) 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 of 15 to 25 nucleotides in length or a siRNA oligonucleotide of 19 to 25 nucleotides in length wherein said at least one is 100% complementary with and specifically hybridizes to a 15 to 25 nucleotide region or, in the case of siRNA, to a 19 to 25 nucleotide non-overlapping sequence of SEQ ID NO;

2;

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

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

3, 5, and 9, 10, 12 and 13.

2. A method of upregulating a function of and/or the expression of a Filaggrin (FLG) 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 of 15 to 21 nucleotides in length that is 100% complementary with, targets and specifically hybridizes to a region of a natural antisense RNA polynucleotide of the Filaggrin (FLG) polynucleotide having sequence identity to SEQ ID NO;

2;

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

3. The method of claim 2, wherein a function of and/or the expression of the Filaggrin (FLG) 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 Filaggrin (FLG) polynucleotide selected from SEQ ID NO:

2 and has a length of between 18-21 nucleotides.

5. The method of claim 2, wherein the at least one antisense oligonucleotide targets a natural antisense polynucleotide comprising non-coding nucleic acid sequences.

6. The method of claim 2, wherein the at least one antisense oligonucleotide targets a natural antisense polynucleotide having overlapping sequences with a Filaggrin (FLG) mRNA 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 and combinations thereof.

12. A method of upregulating a function of and/or the expression of a Filaggrin (FLG) 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 23 nucleotides in length, said at least one siRNA oligonucleotide is 100% complementary with and specific for a natural antisense polynucleotide of a Filaggrin (FLG) polynucleotide selected from SEQ ID NO;

2, wherein said at least one siRNA oligonucleotide specifically hybridizes to a non-overlapping, complementary region of said natural antisense polynucleotide of the Filaggrin (FLG) polynucleotide; and

, upregulates a function of and/or the expression of Filaggrin (FLG) in mammalian cells or tissues in vivo or in vitro.

13. A method of treating a disease associated with at least one Filaggrin (FLG) polynucleotide and/or at least one encoded product thereof having SEQ ID NO:

1, comprising;

administering to a patient a therapeutically effective dose of at least one (i) single-stranded antisense oligonucleotide of 15 to 25 nucleotides in length that specifically hybridizes to a natural antisense polynucleotide of said at least one Filaggrin (FLG) polynucleotide or (ii) a siRNA of 19 to 25 nucleotides in length that specifically hybridizes to a non-overlapping region of said at least one Filaggrin (FLG) polynucleotide and upregulates expression of said at least one Filaggrin (FLG) polynucleotide;

thereby treating the disease associated with the at least one Filaggrin (FLG) polynucleotide and/or at least one encoded product thereof wherein said disease associated with the at least one Filaggrin (FLG) polynucleotide is selected from;

psoriasis, atopic dermatitis (AD), ichthyosis vulgaris or eczema.

14. A method of treating a disease or disorder in a subject according to claim 13, the method comprises administering to the subject a composition further comprising one or more FLG modulating molecules selected from the group of Pioglitazone, Lomerizine, Phenprobamate, Benidipine, bupropion, Piroxicam, Topiramate, isradipine, Nicorandil, Piribedil, Oxaprozin, Glycopyrrolate, Granisetron, Memantine, Nimodipine and Amlodipine and a pharmaceutically acceptable carrier.

15. The method of claim 14, wherein the compound is selected from the group of Pioglitazone, Lomerizine, Bupropion, Phenprobamate and Benidipine.