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

• US 10,696,966 B2
• Filed: 10/26/2015
• Issued: 06/30/2020
• 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:
  US14/922,954
• Publication Number:
  US 20160046942A1
• Time in Patent Office: 1,709 Days
• Field of Search: 435 61, 435 611, 435 9131, 435455, 435458, 435375, 435 911, 514 44, 536 231, 536 245, 536 241, 536 243, 536 2433
• 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 Globulia (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 synthetic, modified single-stranded oligonucleotide of 20 to 21 nucleotides in length comprising at least one modification wherein the at least one modification is selected from:

at least one modified sugar moiety;

at least one modified internucleotide linkage;

at least one modified nucleotide, and combinations thereof;

wherein said oligonucleotide is an antisense compound which is 100% complementary to and specifically hybridizes to a natural antisense polynucleotide of a Sex Hormone Binding Globulin (SHBG) gene selected from SEQ ID NO;

2 or to a target region from nucleotides 1 to 831 of SEQ ID NO;

3 and upregulates the function and/or expression of a Sex Hormone Binding Globulin (SHBG) gene in vivo or in vitro as compared to a normal control.

2. The oligonucleotide of claim 1, wherein the at least one modification comprises an internucleotide linkage selected from the group consisting of:

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

3. The oligonucleotide of claim 1, wherein said oligonucleotide comprises at least one phosphorothioate internucleotide linkage.

4. The oligonucleotide of claim 1, wherein said oligonucleotide comprises a backbone of phosphorothioate internucleotide linkages.

5. The oligonucleotide of claim 1, wherein the oligonucleotide comprises at least one modified nucleotide, said modified nucleotide selected from:

a peptide nucleic acid, a locked nucleic acid (LNA), analogue, derivative, and a combination thereof.

6. The oligonucleotide of claim 1, wherein the oligonucleotide comprises a plurality of modifications, wherein said modifications comprise modified nucleotides selected from:

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

7. The oligonucleotide of claim 1, wherein the oligonucleotide comprises a plurality of modifications, wherein said modifications comprise modified nucleotides selected from:

peptide nucleic acids, locked nucleic acids (LNA), analogues, derivatives, and a combination thereof.

8. The oligonucleotide of claim 1, wherein the oligonucleotide comprises 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 a combination thereof.

9. The oligonucleotide of claim 1, wherein the oligonucleotide comprises a plurality of modifications, wherein said modifications comprise modified sugar moieties 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 a combination thereof.

10. The oligonucleotide of claim 1, wherein said oligonucleotide hybridizes to nucleotides 1 to 831 of the natural antisense polynucleotide having SEQ ID NO:

3 and upregulates expression and/or function of at least one Sex Hormone Binding Globulin (SHBG) polynucleotide in vivo or in vitro, as compared to a mock-transfected control.

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

8, 9, 13, 15, and 17.

12. A composition comprising one or more oligonucleotides according to claim 1 and a pharmaceutically acceptable excipient.

14. The composition of claim 12, wherein the oligonucleotide comprises any one of a nucleotide sequence selected from the group consisting of SEQ ID NOS:

8, 9, 13, 16 and 17.

15. The composition of claim 14, wherein the oligonucleotides set forth as SEQ ID NOS:

8, 9, 13, 16 and 17 comprise one or more modifications or substitutions.

16. The composition of claim 15, wherein the one or more modifications are selected from:

phosphorothioate, methylphosphonate, peptide nucleic acid, locked nucleic acid (LNA) molecules, and combinations thereof.

13. A pharmaceutical composition comprising an oligonucleotide selected from the group consisting of SEQ ID NOS:

8, 9, 13, 15, and 17 and a pharmaceutically acceptable excipient.

17. A method of preventing or treating a disease associated with at least one Sex Hormone Binding Globulin (SHBG) polynucleotide and/or at least one encoded product thereof, comprising:

administering to a patient a therapeutically effective dose of at least one antisense oligonucleotide of 10 to 30 nucleotides in length that binds and specifically hybridizes to a natural antisense sequence of said at least one Sex Hormone Binding Globulin (SHBG) polynucleotide and upregulates expression of said at least one Sex Hormone Binding Globulin (SHBG) polynucleotide;

thereby preventing or treating the disease associated with the at least one Sex Hormone Binding Globulin (SHBG) polynucleotide and/or at least one encoded product thereof.

18. The method of claim 17, wherein a disease associated with the at least one Sex Hormone Binding Globulin (SHBG) polynucleotide is:

a disease or disorder associated with abnormal function and/or expression of SHBG, a neurological disease or disorder, a psychological disease or disorder (e.g., disorders of mood and affect, a memory dysfunction disease or disorder, an amnestic disease or disorder, a motor and tic disorder, substance abuse disease or disorder, a psychotic disease or disorder, an anxiety disease or disorder etc.), a psychotic disease or disorder (e.g., schizophrenia, schizofreniform disorder, schizoaffective disorder, and delusional disorder, anxiety disorders such as panic disorder, phobias (including agoraphobia), an obsessive-compulsive disorder, posttraumatic stress disorder etc.), a fertility disease or disorder, hirsutism, a disease or disorder associated with spermatogenesis, a disease or disorder associated with semen quality, ovarian failure, a mood disorder (e.g., minor and major depression, dysthymic disorder, bipolar disorders and the like), motor and tic disorder (e.g., Tourette'"'"'s disorder, chronic motor or vocal tic disorder, transient tic disorder, and stereotypic movement disorder), Asperger syndrome, a substance abuse disorder (e.g., substance dependence, substance abuse and the sequalae of substance abuse/dependence, such as substance-induced psychological disorders, substance withdrawal and substance-induced dementia, amnestic disorders etc.), a metabolic disease or disorder (e.g., hypothyroidism, a disorder associated with chronic stress, symptoms associated with spinal cord injury such as decreased pulmonary function, increased colonic transit time and the like), a chronic stress-related disease or disorder (e.g., Chronic stress-related conditions include fibromyalgia, chronic fatigue syndrome, hypothalamic-pituitary axis dysregulation, chronic sleep deprivation, and conditions associated with elevated levels of interleukin 6 (IL-6)), a sleep disease or disorder, alopecia, steroid dependent cancer (e.g., including prostate cancer and breast cancer, such as estrogen receptor positive (ER+)), a sexual disease or disorder or dysfunction, polycystic ovary syndrome, osteoporosis, hyperinsulinemia, glucose intolerance, insulin resistance, a cardiovascular disease or disorder and diabetes.

19. A method of identifying and selecting at least one oligonucleotide for in vivo administration to treat a disease associated with the at least one Sex Hormone Binding Globulin (SHBG) polynucleotide comprising:

selecting a target SHBG natural antisense polynucleotide of said at least one Sex Hormone Binding Globulin (SHBG) polynucleotide;

identifying at least one oligonucleotide of 10 to 30 nucleotides in length comprising at least ten consecutive nucleotides which are complementary to the selected target natural antisense polynucleotide measuring the thermal melting point of a hybrid of said antisense oligonucleotide and the target polynucleotide under stringent hybridization conditions; and

/or determining the expression level of the mRNA of the SHBG polynucleotide by said hybridization of said antisense oligonucleotide to said natural antisense polynucleotide relative to a control and selecting at least one oligonucleotide for in vivo administration based on the thermal melting point and/or expression level of said mRNA.

20. A modified oligonucleotide 19 nucleotides in length having SEQ ID NO:

6.