Treatment of RNASE H1 related diseases by inhibition of natural antisense transcript to RNASE H1

• US 8,946,182 B2
• Filed: 01/25/2011
• Issued: 02/03/2015
• Est. Priority Date: 01/25/2010
• 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): WHITEMAN, BRIAN A
• Application Number:
  US13/575,206
• Publication Number:
  US 20120295959A1
• Time in Patent Office: 1,470 Days
• Field of Search: 514/44.A
• US Class Current: 514/44.A
• 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 31/12 Antivirals
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 31/20 for DNA viruses
• CPC Class Codes: A61P 33/02 Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis
• 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: C12N 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• 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: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 2'-R Modification
• CPC Class Codes: C12N 9/22 Ribonucleases RNAses, DNAses catalytic nucleic acids C12N15/113
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The present invention relates to antisense oligonucleotides that modulate the expression of and/or function of RNase H1, in particular, by targeting natural antisense polynucleotides of RNase H1. The invention also relates to the identification of these antisense oligonucleotides and their use in treating diseases and disorders associated with the expression of RNASE H1.

Claims

1. A method of increasing the expression of an RNase H1 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 15 to 30 nucleotides in length wherein said at least one oligonucleotide is specific for a natural antisense polynucleotide of the RNase H1 gene and has at least 90% sequence identity to a reverse complement of a polynucleotide comprising 15 to 30 consecutive nucleotides within nucleotides 1 to 437 of SEQ ID NO;

2 or nucleotides 1 to 278 of SEQ ID NO;

3 or nucleotides 1 to 436 of SEQ ID NO;

4 or nucleotides 1 to 1140 of SEQ ID NO;

5;

thereby increasing the expression of the RNase H1 polynucleotide in patient cells or tissues in vivo or in vitro and used in conjunction with antisense oligonucleotides targeted against other targets to increase their activity.

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 13.

2. A method of increasing the expression of an RNase H1 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 15 to 30 nucleotides in length wherein said at least one oligonucleotide is specific for a natural antisense polynucleotide of the RNase H1 gene and has at least 90% sequence identity to a 15 to 30 nucleotide region of a reverse complement of a natural antisense of an RNase H1 polynucleotide;

thereby increasing the expression of the RNase H1 polynucleotide in patient cells or tissues in vivo or in vitro.

3. A method of upregulating a function of and/or the expression of an RNase H1 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 to 30 nucleotides in length that specifically targets a region of a natural antisense polynucleotide of the RNase H1 polynucleotide selected from SEQ ID NOS;

2-5;

thereby upregulating a function of and/or the expression of the RNase H1 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 RNase H1 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 an RNase H1 polynucleotide selected from the group consisting of SEQ ID NO:

4.

6. The method of claim 3, wherein the at least one antisense oligonucleotide targets a natural antisense polynucleotide that is antisense to coding and/or non-coding sequences of an RNA polynucleotide transcribed by the RNase H1 gene.

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 RNase H1 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 RNase H1 having SEQ ID Na 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 18 to 24 nucleotides in length specific for a natural antisense strand of an RNase H1 polynucleotide selected from SEQ ID NOS;

2-5 wherein said at least one antisense oligonucleotide has at least 80% sequence identity to 18-24 nucleotides within at least one nucleic acid sequence set forth as SEQ ID NOS;

1 or a sense RNA polynucleotide encoded by the RNase gene; and

, upregulating the function and/or expression of the RNase H1 in mammalian cells or tissues in vivo or in vitro.

14. A method of increasing the activity of a first oligonucleotide of 18 to 24 nucleotides in length targeted to a natural antisense polynucleotide of a targeted gene to increase the levels of a therapeutic protein of said targeted gene comprising the combination of said first oligonucleotide with a second oligonucleotide of 18 to 24 nucleotides in length specifically targeted to a natural antisense polynucleotide of an RNASE H1 gene selected from SEQ ID NOS:

2-5 to upregulate the expression of RNASE H1 thereby increasing the activity of said first oligonucleotide.