Treatment of LCAT gene related diseases by inhibition of a natural antisense transcript to LCAT

• US 9,957,503 B2
• Filed: 07/31/2015
• Issued: 05/01/2018
• Est. Priority Date: 05/06/2009
• 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): Schnizer, Richard A
• Application Number:
  US14/815,007
• Publication Number:
  US 20160024506A1
• Time in Patent Office: 1,005 Days
• Field of Search: None
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• 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 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 13/10 of the bladder
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• 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 25/28 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 25/30 for treating abuse or dependence
• CPC Class Codes: A61P 27/06 Antiglaucoma agents or miotics
• 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 31/00 : Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 : Antibacterial agents
• CPC Class Codes: A61P 33/00 : Antiparasitic agents
• 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 37/06 : 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 5/14 : of the thyroid hormones, e.g. T3, T4
• 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: 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 15/1137 : against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 15/1138 : against receptors or cell surface proteins
• CPC Class Codes: C12N 2310/11 : Antisense
• CPC Class Codes: C12N 2310/14 : interfering N.A.
• CPC Class Codes: C12N 2310/313 : Phosphorodithioates
• CPC Class Codes: C12N 2310/314 : Phosphoramidates
• CPC Class Codes: C12N 2310/315 : Phosphorothioates
• CPC Class Codes: C12N 2310/321 : 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 : 2'-R Modification
• CPC Class Codes: C12Y 203/01043 : Phosphatidylcholine-sterol O-acyltransferase (2.3.1.43), i.e. lecithin-cholesterol acyltransferase or LCAT

Abstract

The present invention relates to antisense oligonucleotides that modulate the expression of and/or function of a Lipid transport and metabolism gene, in particular, by targeting natural antisense polynucleotide of a Lipid transport and metabolism gene. The invention also relates to the identification of these antisense oligonucleotides and their use in treating diseases and disorders associated with the expression of a Lipid transport and metabolism genes.

Claims

1. A method of upregulating a function of and/or the expression of a lecithin-cholesterol acetyltransferase (LCAT) gene in mammalian cells or tissues in vivo or in vitro comprising:

contacting said cells or tissues with at least one modified antisense oligonucleotide of 12 to 30 nucleotides in length that targets, is 100% complementary with and specifically hybridizes to a complementary region within nucleotides 1 to 1550 of a natural antisense polynucleotide of the LCAT gene wherein said natural antisense polynucleotide consists of SEQ ID NO;

10, thereby upregulating a function of and/or the expression of the LCAT gene in mammalian cells or tissues in vivo or in vitro.

2. The method of claim 1, wherein expression of the LCAT gene is increased in cells in vitro relative to cells of a mock-transfected control.

3. The method of claim 1, wherein the at least one modified antisense oligonucleotide comprises at least one modification selected from the group consisting of at least one modified sugar moiety, at least one modified internucleoside linkage, at least one modified nucleotide, and combinations thereof.

4. The method of claim 1, wherein the complementary region of the natural antisense polynucleotide is complementary to a sequence within a coding region of a LCAT RNA.

5. The method of claim 1, wherein the 12-30 nucleotides of the at least one modified antisense oligonucleotide are 100% complementary to an equal length sequence of the natural antisense that is complementary to both coding and non-coding sequences of a LCAT RNA.

6. The method of claim 1, wherein the at least one modified antisense oligonucleotide is 12 to 24 nucleotides in length and 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.

7. The method of claim 6, 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.

8. The method of claim 6, 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.

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), or an arabino-nucleic acid (FANA).

10. The method of claim 1, wherein the at least one oligonucleotide comprises at least one oligonucleotide sequence selected from the group consisting of SEQ ID NOS:

41, 42, 52, 54, 55, 56, and 58.

11. A method of upregulating a function of and/or the expression of a LCAT 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 30 nucleotides in length, wherein said at least one siRNA oligonucleotide comprises an oligonucleotide strand that is sufficiently complementary to a portion of SEQ ID NO;

10 to specifically hybridize under physiological conditions to a natural antisense transcripts that is the RNA version of SEQ ID NO;

10, wherein the portion of the natural antisense transcript to which the siRNA oligonucleotide strand can hybridize is not complementary to LCAT mRNA.

12. A method of treating a disease associated with an LCAT gene in a patient in need of treatment thereof, comprising:

administering to said patient a therapeutically effective dose of at least one modified antisense oligonucleotide of 10 to 30 nucleotides in length that binds and specifically hybridizes to a complementary region of a natural antisense sequence of said LCAT gene consisting of SEQ ID NO;

10 and upregulates expression of said LCAT gene thereby treating the disease associated with the LCAT gene, wherein the disease associated with the LCAT gene is Fish Eye disease.