Compositions and methods for inhibiting gene expression of hepatitis B virus

• US 8,809,293 B2
• Filed: 06/28/2012
• Issued: 08/19/2014
• Est. Priority Date: 06/30/2011
• Status: Expired

• Current Assignee: Arrowhead Pharmaceuticals, Inc.
• Original Assignee: Arrowhead Madison Inc (Arrowhead Pharmaceuticals, Inc.)
• Inventors: Deckert, Jochen, Hossbach, Markus, John, Matthias, Chin, Daniel
• Primary Examiner(s): GIBBS, TERRA C
• Application Number:
  US13/535,454
• Publication Number:
  US 20130005793A1
• Time in Patent Office: 782 Days
• Field of Search: 514/44.A, 536/24.5
• US Class Current: 514/44.A
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• 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 31/12 Antivirals
• CPC Class Codes: A61P 31/20 for DNA viruses
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C12N 15/1131 against viruses
• 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 2310/335 Modified T or U
• CPC Class Codes: C12N 2310/351 Conjugate
• CPC Class Codes: C12N 2310/3521 Methyl

Abstract

The invention relates to a double-stranded ribonucleic acid (dsRNA) for inhibiting the expression of a Hepatitis B Virus gene. The invention also relates to a pharmaceutical composition comprising the dsRNA or nucleic acid molecules or vectors encoding the same together with a pharmaceutically acceptable carrier; methods for treating diseases caused by Hepatitis B Virus infection using said pharmaceutical composition; and methods for inhibiting the expression of a Hepatitis B Virus gene in a cell.

Claims

1. A double-stranded ribonucleic acid molecule capable of inhibiting the expression of a Hepatitis B Virus gene in vitro wherein said double-stranded ribonucleic acid molecule comprises a sense strand comprising in order nucleotides 1-19 of SEQ ID 2, or 6 and an antisense strand at least partially complementary to the sense strand, wherein said sense strand and said antisense strand are each less than 30 nucleotides in length.

2. The double-stranded ribonucleic acid molecule of claim 1, wherein said antisense strand comprises in order nucleotides 1-19 of SEQ ID 158 or 163.

3. The double-stranded ribonucleic acid molecule of claim 2, wherein said double-stranded ribonucleic acid molecule comprises sequence pairs selected from the group consisting of SEQ ID NOs:

2/158 and 6/163.

4. The double-stranded ribonucleic acid molecule of claim 1, wherein the sense strand or the antisense strand further comprises a 3′

overhang of 1-5 nucleotides in length.

5. The double-stranded ribonucleic acid molecule of claim 4, wherein the 3′

overhang of the antisense strand comprises uracil or nucleotides which are complementary to the pregenomic RNA and/or the mRNA encoding the protein necessary for replication or pathogenesis of Hepatitis B Virus.

6. The double-stranded ribonucleic acid molecule of claim 4 wherein the 3′

overhang of the sense strand comprises uracil or nucleotides which are identical to the pregenomic RNA and/or the mRNA encoding the protein necessary for replication or pathogenesis of Hepatitis B Virus.

7. The double-stranded ribonucleic acid molecule of claim 1, wherein said double-stranded ribonucleic acid molecule comprises at least one modified nucleotide selected from the group consisting of:

2′

-O-methyl modified nucleotide, nucleotide comprising a 5′

-phosphorothioate group, terminal nucleotide linked to a cholesteryl derivative, terminal nucleotide linked to a dodecanoic acid bisdecylamide group, 2′

-deoxy- 2′

-fluoro modified nucleotide, 2-deoxy-nucleotide, locked nucleotide, abasic nucleotide, deoxythymidine, inverted deoxythymidine, 2′

-amino-modified nucleotide, 2′

-alkyl-modified nucleotide, morpholino nucleotide, phosphoramidate, and non-natural base comprising nucleotide.

8. The double-stranded ribonucleic acid molecule of claim 7, wherein said double-stranded ribonucleic acid molecule contains a 2′

-O-methyl modified nucleotide, a nucleotide comprising a 5′

-phosphomthioate group, and a deoxythymidine.

9. The double-stranded ribonucleic acid molecule of claim 8, wherein said sense strand or said antisense strand comprises an overhang of 1-2 deoxythymidines.

10. The double-stranded ribonucleic acid molecule of claim 9, wherein said double-stranded ribonucleic acid molecule comprises the sequence pairs selected from the group consisting of SEQ ID NOs:

322/486 and 327/491.

11. The double-stranded ribonucleic acid molecule of claim 1 wherein said double-stranded ribonucleic acid molecule comprises a nucleic acid sequence encoding said sense strand or said antisense strand.

12. A pharmaceutical composition comprising:

a) first and second double-stranded ribonucleic acid molecules each as defined in claim 1;

b) at least one nucleic acid sequence encoding sense strands or antisense strands comprising first and second double-stranded ribonucleic acid molecules each as defined in claim 1;

or,

c) a cell, tissue or non-human organism comprising first and second double-stranded ribonucleic acid molecules each as defined in claim 1.

13. The pharmaceutical composition of claim 12 wherein:

a) said first double stranded ribonucleic acid has a sense strand comprising in order nucleotides 1-19 of SEQ ID 2 and said second double-stranded ribonucleic acid has a sense strand comprising in order nucleotides 1-19 of SEQ ID 4 or 6 or

b) said first double stranded ribonucleic acid has a sense strand comprising in order nucleotides 1-19 of SEQ ID 6 and said second double-stranded ribonucleic acid has a sense strand comprising in order nucleotides 1-19 of SEQ ID 2 or 7.

14. The pharmaceutical composition of claim 12 further comprising a pharmaceutically acceptable carrier, stabilizer and/or diluent.

15. A method for inhibiting the expression of Hepatitis B Virus gene in a cell, a tissue, or an organism comprising:

a) introducing into the cell, tissue, or organism the double-stranded ribonucleic acid molecule as defined in claim 1; and

b) maintaining the cell, tissue or organism produced in step a) for a time sufficient to obtain degradation of the mRNA transcript of a Hepatitis B Virus gene, thereby inhibiting expression of a Hepatitis B Virus gene in the cell.

16. The method of claim 15 wherein inhibiting expression of Hepatitis B Virus gene in an organism treats or manages a pathological condition or disease caused by infection with the Hepatitis B Virus.

17. The method claim 16 wherein the pathological condition and disease caused by the infection with the Hepatitis B Virus is selected from the group consisting of:

chronic liver disorder, inflammation, fibrotic condition, and proliferative disorder.