Treatment of tristetraproline (TTP) related diseases by inhibition of natural antisense transcript to TTP

• US 9,163,285 B2
• Filed: 12/24/2013
• Issued: 10/20/2015
• 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): Chong, Kimberly
• Application Number:
  US14/140,305
• Publication Number:
  US 20140155464A1
• Time in Patent Office: 665 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• 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 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 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 31/18 for HIV
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/04 Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• 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 2310/313 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: C12N 2310/3231 : having an additional ring, e.g. LNA, ENA
• CPC Class Codes: C12Q 1/6876 : Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
• 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 Tristetraproline (TTP), in particular, by targeting natural antisense polynucleotides of Tristetraproline (TTP). The invention also relates to the identification of these antisense oligonucleotides and their use in treating diseases and disorders associated with the expression of TTP.

Claims

1. A method of upregulating a function of and/or the expression of a Tristetraproline (TTP) polynucleotide 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 has at least 90% sequence identity to a reverse complement of a polynucleotide comprising 12 to 30 nucleotides within nucleotides 1 to 970 of SEQ ID NO;

3, 1 to 1117 of SEQ ID NO;

4 and 1 to 297 of SEQ ID NO;

5;

thereby upregulating a function of and/or the expression of the Tristetraproline (TTP) 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:

6 to 10.

2. A method of upregulating a function of and/or the expression of a Tristetraproline (TTP) polynucleotide 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 has at least 90% sequence identity to a reverse complement of a natural antisense of a Tristetraproline (TTP) polynucleotide;

thereby upregulating function of and/or the expression of the Tristetraproline (TTP) polynucleotide in patient cells or tissues in vim or in vitro.

3. A method of upregulating a function of and/or the expression of a Tristetraproline (TTP) polynucleotide 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 oligonucleotide has at least 90% sequence complementarily to a natural antisense polynucleotide of the Tristetraproline (TTP) polynucleotide;

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

4. A method of upregulating a function of and/or the expression of a Tristetraproline (TTP) polynucleotide in patient cells or tissues in vivo or in vitro comprising:

contacting said cells or tissues with at least one antisense oligonucleotide that specifically targets a region of a natural antisense polynucleotide of the Tristetraproline (TTP) polynucleotide;

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

5. The method of claim 4, wherein a function of and/or the expression of the Tristetraproline (TTP) is increased in vivo or in vitro with respect to a mock-transfected control.

6. The method of claim 4, wherein the ill least one antisense oligonucleotide targets a natural antisense sequence of a Tristetraproline (TTP) polynucleotide selected from SEQ ID NOS:

3, 4 or 5.

7. The method of claim 4, wherein the at least one antisense oligonucleotide targets a natural antisense polynucleotide antisense to coding and/or non-coding nucleic acid sequences of a Tristetraproline (TTP) polynucleotide.

8. The method of claim 4, 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), or an arabino-nucleic acid (FANA).

13. A method of upregulating a function of and/or the expression of a Tristetraproline (TTP) 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 non-overlapping region of a natural antisense polynucleotide of a Tristetraproline (TTP) polynucleotide, wherein said at least one siRNA oligonucleotide has at least 90% sequence complementarily to said natural antisense polynucleotide of the Tristetraproline (TTP) polynucleotide; and

, upregulating a function of and/or the expression of Tristetraproline (TTP) in mammalian cells or tissues in vivo or in vitro.

14. The method of claim 13, wherein said oligonucleotide has sequence complementarity to a sequence of at least about 19 nucleotides of the natural antisense polynucleotide of the Tristetraproline (TTP) polynucleotide.

15. A method of upregulating a function of and/or the expression of Tristetraproline (TTP) in mammalian cells or tissues in vivo or in vitro comprising:

contacting said cells or tissues with at least one antisense oligonucleotide of about 10 to 30 nucleotides in length specific for noncoding and/or coding sequences of a natural antisense strand of a Tristetraproline (TTP) polynucleotide wherein said at least one antisense oligonucleotide has at least 90% sequence identity to at least one nucleotide sequence set forth as SEQ. ID NO;

1, or an RNA transcribed from the TTP polynucleotide and, upregulating the function and/or expression of the Tristetraproline (TTP) in mammalian cells or tissues in vim or in vitro.

16. A method of preventing or treating a disease associated with at least one Tristetraproline (TTP) 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 Tristetraproline (TTP) polynucleotide selected from the group consisting of SEQ ID NOS;

3, 4 and 5 and upregulates expression of said at least one Tristetraproline (TTP) polynucleotide;

thereby treating the disease associated with the at least one Tristetraproline (TTP) polynucleotide and/or at least one encoded product thereof wherein the disease is selected from tristetraprolin-deficiency syndrome.