Treatment of 'IQ motif containing GTPase activating protein' (IQGAP) related diseases by inhibition of natural antisense transcript to IQGAP

• US 9,023,822 B2
• Filed: 08/25/2010
• Issued: 05/05/2015
• Est. Priority Date: 08/25/2009
• 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): Chong, Kimberly
• Application Number:
  US13/388,817
• Publication Number:
  US 20120142610A1
• Time in Patent Office: 1,714 Days
• Field of Search: None
• US Class Current: 514/44
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 15/10 for impotence
• 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 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/02 Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• 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 2310/11 Antisense
• 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 2310/3125 : Methylphosphonates
• 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 2310/316 : Phosphonothioates
• 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 2310/323 : modified ring structure
• CPC Class Codes: C12N 2310/3231 : having an additional ring, e.g. LNA, ENA

Abstract

The present invention relates to antisense oligonucleotides that modulate the expression of and/or function of ‘IQ motif containing GTPase activating protein’ (IQGAP), in particular, by targeting natural antisense polynucleotides of ‘IQ motif containing GTPase activating protein’ (IQGAP). The invention also relates to the identification of these antisense oligonucleotides and their use in treating diseases and disorders associated with the expression of IQGAP.

Claims

1. A method of upregulating a function of and/or the expression of an ‘

IQ motif containing GTPase activating protein’

(IQGAP) polynucleotide selected from SEQ ID NO;

1 or a variant thereof in patient cells or tissues in vivo or in vitro comprising;

contacting said cells or tissues with at least one single stranded modified antisense oligonucleotide of 12 to 25 nucleotides in length that specifically targets a complementary region of a natural antisense polynucleotide of the ‘

IQ motif containing GTPase activating protein’

(IQGAP) polynucleotide selected from SEQ ID NO;

2 ;

thereby upregulating a function of and/or the expression of the ‘

IQ motif containing GTPase activating protein’

(IQGAP) 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 ‘

IQ motif containing GTPase activating protein’

(IQGAP) is increased in vivo or in vitro with respect to a mock-transfected control.

3. The method of claim 1, wherein the at least one antisense oligonucleotide targets a natural antisense sequence of an ‘

IQ motif containing GTPase activating protein’

(IQGAP) polynucleotide having SEQ ID NO;

2.

4. 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 an ‘

IQ motif containing GTPase activating protein’

(IQGAP) RNA polynucleotide.

5. 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 an ‘

IQ motif containing GTPase activating protein’

(IQGAP) polynucleotide.

6. 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.

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 set forth as SEQ ID NO:

3.

11. A method of upregulating a function of and/or the expression of an ‘

IQ motif containing GTPase activating protein’

(IQGAP) 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, said at least one siRNA oligonucleotide being specific for a natural antisense polynucleotide of an ‘

IQ motif containing GTPase activating protein’

(IQGAP) polynucleotide selected from SEQ ID NO;

2, and, upregulating a function of and/or the expression of ‘

IQ motif containing GTPase activating protein’

(IQGAP) in mammalian cells or tissues in vivo or in vitro.

12. The method of claim 11, wherein said oligonucleotide has at least 90% sequence complementarity to the natural antisense polynucleotide of the ‘

IQ motif containing GTPase activating protein’

(IQGAP) polynucleotide.

13. A method of upregulating a function of and/or the expression of ‘

IQ motif containing GTPase activating protein 2’

(IQGAP2) in mammalian cells or tissues in vivo or in vitro comprising;

contacting said cells or tissues with at least one single stranded antisense oligonucleotide of about 10 to 30 nucleotides in length specific for noncoding and/or coding sequences of a natural antisense strand of said ‘

IQ motif containing GTPase activating protein 2’

(IQGAP2) polynucleotide selected from SEQ ID NO;

2 wherein said at least one antisense oligonucleotide has at least 80% sequence identity to 10 to 30 nucleotides of at least one nucleic acid sequence set forth as SEQ ID NO;

1 or an RNA transcribed from said IQGAP2 gene; and

, upregulating the function and/or expression of the ‘

IQ motif containing GTPase activating protein 2’

(IQGAP2) in mammalian cells or tissues in vivo or in vitro.

14. A method of upregulating the expression of ‘

IQ motif containing GTPase activating protein’

(IQGAP) polynucleotide having SEQ ID NO;

1 and/or at least one encoded product thereof in a patient in need of treatment thereof, comprising;

administering to a patient a therapeutically effective dose of at least one single stranded modified antisense oligonucleotide of 12-25 nucleotides in length that specifically binds to a complementary region of a natural antisense polynucleotide of said at least one ‘

IQ motif containing GTPase activating protein’

(IQGAP) polynucleotide selected from SEQ ID NO;

2 and upregulates expression of said at least one ‘

IQ motif containing GTPase activating protein’

(IQGAP) polynucleotide;

thereby preventing or treating the disease associated with the at least one ‘

IQ motif containing GTPase activating protein’

(IQGAP) polynucleotide and/or at least one encoded product thereof.

15. The method of claim 14, wherein said upregulation treats cancer, neoplastic transformation, Non small cell lung carcinoma (NSCLCs), liposarcoma, gastric cancer or glioma.