Treatment of sex hormone binding globulin (SHBG) related diseases by inhibition of natural antisense transcript to SHBG

• US 9,200,277 B2
• Filed: 01/11/2011
• Issued: 12/01/2015
• Est. Priority Date: 01/11/2010
• 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): ZARA, JANE J
• Application Number:
  US13/520,503
• Publication Number:
  US 20120277290A1
• Time in Patent Office: 1,785 Days
• Field of Search: 435/6, 435/91.1, 435/91.31, 435/455, 435/6.1, 435/375, 514/44, 536/23.1, 536/24.5
• US Class Current: 1/1
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 15/08 for gonadal disorders or for enhancing fertility, e.g. inducers of ovulation or of spermatogenesis
• CPC Class Codes: A61P 17/14 for baldness or alopecia
• CPC Class Codes: A61P 19/10 for osteoporosis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/14 for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 25/18 Antipsychotics, i.e. neuroleptics; Drugs for mania or schizophrenia
• CPC Class Codes: A61P 25/20 Hypnotics; Sedatives
• CPC Class Codes: A61P 25/22 Anxiolytics
• CPC Class Codes: A61P 25/24 Antidepressants
• 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 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• 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: 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 2310/113 : targeting other non-coding nucleic acids, e.g. antagomirs
• CPC Class Codes: C12N 2310/14 : interfering N.A.
• CPC Class Codes: C12N 2310/315 : Phosphorothioates
• CPC Class Codes: Y10T 436/143333 : Saccharide [e.g., DNA, etc.]

Abstract

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

Claims

1. A method of upregulating a function of and/or the expression of a Sex Hormone Binding Globulin (SHBG) polynucleotide having SEQ ID NO:

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

contacting said cells or tissues with at least one antisense oligonucleotide 12 to 30 nucleotides in length wherein said at least one oligonucleotide is specific for a natural antisense polynucleotide of the SHBG gene having SEQ ID NOS;

2 or 3 and has at least 80% sequence identity to a reverse complement of a polynucleotide comprising 12 to 30 consecutive nucleotides within nucleotides 1 to 3016 of SEQ ID NO;

2 or nucleotides 1 to 1609 of SEQ ID NO;

3;

thereby upregulating a function of and/or the expression of the Sex Hormone Binding Globulin (SHBG) polynucleotide in patient cells or tissues in vivo or in vitro.

12. 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 17.

2. A method of upregulating a function of and/or the expression of a Sex Hormone Binding Globulin (SHBG) polynucleotide having SEQ ID NO:

1 in patient cells or tissues in vim or in vitro comprising;

contacting said cells or tissues with at least one antisense oligonucleotide 15 to 30 nucleotides in length wherein said oligonucleotide is specific for a natural antisense polynucleotide of the SHBG gene having SEQ ID NOS;

2 or 3 and has at least 80% sequence identity to a 15 to 30 nucleotide region of an RNA polynucleotide transcribed from the SHBG gene or a polynucleotide derived therefrom;

thereby upregulating a function of and/or the expression of the Sex Hormone Binding Globulin (SHBG) polynucleotide in patient cells or tissues in vivo or in vitro.

3. A method of upregulating a function of and/or the expression of a Sex Hormone Binding Globulin (SHBG) polynucleotide having SEQ ID NO:

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

contacting said cells or tissues with at least one antisense oligonucleotide of 15-30 nucleotides in length that specifically targets a 15 to 30 nucleotide region of a natural antisense polynucleotide of the Sex Hormone Binding Globulin (SHBG) polynucleotide wherein said natural antisense polynucleotide comprises SEQ ID NOS;

2 or 3;

thereby upregulating a function of and/or the expression of the Sex Hormone Binding Globulin (SHBG) polynucleotide in patient cells or tissues in vivo or in vitro.

4. The method of claim 3, wherein a function of and/or the expression of the Sex Hormone Binding Globulin (SHBG) is increased in vivo or in vitro with respect to a control.

5. The method of claim 3, wherein the at least one antisense oligonucleotide targets a natural antisense sequence of a Sex Hormone Binding Globulin (SHBG) polynucleotide selected from SEQ ID NOS:

2.

6. The method of claim 3, wherein the at least one antisense oligonucleotide targets a natural antisense polynucleotide antisense to coding and/or non-coding nucleic acid sequences of a Sex Hormone Binding Globulin (SHBG) RNA polynucleotide.

7. The method of claim 3, wherein the at least one antisense oligonucleotide targets a natural antisense polynucleotide having overlapping and/or non-overlapping sequences with a Sex Hormone Binding Globulin (SHBG) RNA polynucleotide.

8. The method of claim 3, 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.

9. The method of claim 8, 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.

10. The method of claim 8, 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.

11. The method of claim 8, 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.

13. A method of upregulating a function of and/or the expression of a Sex Hormone Binding Globulin (SHBG) gene encoding SEQ ID NO:

1 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 a Sex Hormone Binding Globulin (SHBG) polynucleotide selected from SEQ ID NOS;

2 or 3; and

, upregulating a function of and/or the expression of Sex Hormone Binding Globulin (SHBG) in mammalian cells or tissues in vivo or in vitro.

14. A method of upregulating a function of and/or the expression of Sex Hormone Binding Globulin (SHBG) having SEQ ID NO:

1 in mammalian cells or tissues in vivo or in vitro comprising;

contacting said cells or tissues with at least one antisense oligonucleotide of about 20 to 30 nucleotides in length specific for a natural antisense strand of a Sex Hormone Binding Globulin (SHBG) polynucleotide wherein said natural antisense strand comprises SEQ ID NOS;

2 or 3 and wherein said at least one antisense oligonucleotide has at least 80% sequence identity to a 20 to 30 nucleotide region of at least one nucleic acid sequence set forth as SEQ ID NOS;

1 or a polynucleotide encoded by the SHBG gene; and

, upregulating the function and/or expression of the Sex Hormone Binding Globulin (SHBG) in mammalian cells or tissues in vivo or in vitro.