Modulation of hepatitis B virus (HBV) expression

• US 9,034,841 B2
• Filed: 12/20/2013
• Issued: 05/19/2015
• Est. Priority Date: 04/21/2011
• Status: Active Grant

• Current Assignee: Glaxo Group Limited (Glaxosmithkline Holdings Ltd.)
• Original Assignee: ISIS Pharmaceuticals Incorporated
• Inventors: Swayze, Eric E., Freier, Susan M., McCaleb, Michael L.
• Primary Examiner(s): VIVLEMORE, TRACY ANN
• Application Number:
  US14/137,111
• Publication Number:
  US 20140107184A1
• Time in Patent Office: 515 Days
• Field of Search: None
• US Class Current: 514/44.A
• 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 31/14 for RNA viruses
• CPC Class Codes: A61P 31/20 for DNA viruses
• 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: C12N 15/1131 against viruses
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/313 Phosphorodithioates
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/3231 having an additional ring, e.g. LNA, ENA
• CPC Class Codes: C12N 2310/3341 5-Methylcytosine
• CPC Class Codes: C12N 2310/341 Gapmers, i.e. of the type ===---===
• CPC Class Codes: C12N 2310/344 Position-specific modifications, e.g. on every purine, at the 3'-end
• CPC Class Codes: C12N 2310/346 having a combination of backbone and sugar modifications
• CPC Class Codes: C12N 2310/3525 MOE, methoxyethoxy
• CPC Class Codes: C12N 2320/30 Special therapeutic applications

Abstract

Disclosed herein are antisense compounds and methods for decreasing HBV mRNA, DNA and protein expression. Such methods, compounds, and compositions are useful to treat, prevent, or ameliorate HBV-related diseases, disorders or conditions.

Claims

1. A compound comprising a single-stranded modified oligonucleotide consisting of the sequence recited in any one of SEQ ID NOs:

17, 50, 722, 1327, 1340, and 1379, wherein the modified oligonucleotide is 100% complementary over its entire length with a nucleic acid encoding hepatitis B virus (HBV) and comprises;

a gap segment consisting of linked deoxynucleosides;

a 5′

wing segment consisting of linked nucleosides; and

a 3′

wing segment consisting of linked nucleosides;

wherein the gap segment is positioned between the 5′

wing segment and the 3′

wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.

2. The compound of claim 1, wherein the modified oligonucleotide comprises at least one modified internucleoside linkage.

3. The compound of claim 2, wherein each internucleoside linkage is a phosphorothioate internucleoside linkage.

4. The compound of claim 1, wherein the modified sugar is a bicyclic sugar.

5. The compound of claim 4, wherein the bicyclic sugar comprises a 4′

-CH(CH3)-O-2′

group.

6. The compound of claim 1, wherein the at least one modified sugar comprises a 2′

-O-methoxyethyl group.

7. The compound of claim 1, wherein at least one nucleoside comprises a modified nucleobase.

8. The compound of claim 7, wherein the modified nucleobase is a 5-methylcytosine.

9. The compound of claim 1, wherein the oligonucleotide consists of 17 linked nucleosides having a nucleobase sequence consisting of the sequence recited in SEQ ID NO:

17, and wherein the oligonucleotide comprises;

a gap segment consisting of 10 linked deoxynucleosides;

a 5′

wing segment consisting of 3 linked nucleosides; and

a 3′

wing segment consisting of 4 linked nucleosides;

wherein the gap segment is positioned between the 5′

wing segment and the 3′

wing segment, wherein each nucleoside of each wing segment comprises a 2′

-O-methoxyethyl sugar, wherein each internucleoside linkage of the modified oligonucleotide is a phosphorothioate linkage, and wherein each cytosine of the modified oligonucleotide is a 5-methylcytosine.

10. The compound of claim 1, wherein the oligonucleotide consists of 20 linked nucleosides having a nucleobase sequence consisting of the sequence recited in SEQ ID NO:

50, and wherein the oligonucleotide comprises;

a gap segment consisting of 10 linked deoxynucleosides;

a 5′

wing segment consisting of 5 linked nucleosides; and

a 3′

wing segment consisting of 5 linked nucleosides;

wherein the gap segment is positioned between the 5′

wing segment and the 3′

wing segment, wherein each nucleoside of each wing segment comprises a 2′

-O-methoxyethyl sugar, wherein each internucleoside linkage of the modified oligonucleotide is a phosphorothioate linkage, and wherein each cytosine of the modified oligonucleotide is a 5-methylcytosine.

11. The compound of claim 1, wherein the oligonucleotide consists of 20 linked nucleosides having a nucleobase sequence consisting of the sequence recited in SEQ ID NO:

722, and wherein the oligonucleotide comprises;

a gap segment consisting of 10 linked deoxynucleosides;

a 5′

wing segment consisting of 6 linked nucleosides; and

a 3′

wing segment consisting of 4 linked nucleosides;

wherein the gap segment is positioned between the 5′

wing segment and the 3′

wing segment, wherein each nucleoside of each wing segment comprises a 2′

-O-methoxyethyl sugar, wherein each internucleoside linkage of the modified oligonucleotide is a phosphorothioate linkage, and wherein each cytosine of the modified oligonucleotide is a 5-methylcytosine.

12. The compound of claim 1, wherein the oligonucleotide consists of 16 linked nucleosides having a nucleobase sequence consisting of the sequence recited in SEQ ID NO:

1327, and wherein the oligonucleotide comprises;

a gap segment consisting of 10 linked deoxynucleosides;

a 5′

wing segment consisting of 2 linked nucleosides; and

a 3′

wing segment consisting of 4 linked nucleosides;

wherein the gap segment is positioned between the 5′

wing segment and the 3′

wing segment;

wherein the 2 linked nucleosides of the 5′

wing segment comprise a 2′

-O-methoxyethyl sugar and a constrained ethyl sugar in the 5′

to 3′

direction;

wherein the 4 linked nucleosides of the 3′

wing segment comprise a constrained ethyl sugar, a 2′

-O-methoxyethyl sugar, a constrained ethyl sugar, and a 2′

-O-methoxyethyl sugar in the 5′

to 3′

direction;

wherein each internucleoside linkage of the modified oligonucleotide is a phosphorothioate linkage; and

wherein each cytosine of the modified oligonucleotide is a 5-methylcytosine.

13. The compound of claim 1, wherein the oligonucleotide consists of 16 linked nucleosides having a nucleobase sequence consisting of the sequence recited in SEQ ID NO:

1340, and wherein the oligonucleotide comprises;

a gap segment consisting of 10 linked deoxynucleosides;

a 5′

wing segment consisting of 3 linked nucleosides; and

a 3′

wing segment consisting of 3 linked nucleosides;

wherein the gap segment is positioned between the 5′

wing segment and the 3′

wing segment;

wherein the 3 linked nucleosides of the 5′

wing segment comprise a 2′

-O-methoxyethyl sugar, a constrained ethyl sugar, and a constrained ethyl sugar in the 5′

to 3′

direction;

wherein the 3 linked nucleosides of the 3′

wing segment comprise a constrained ethyl sugar, a constrained ethyl sugar, and a 2′

-O-methoxyethyl sugar in the 5′

to 3′

direction;

wherein each internucleoside linkage of the modified oligonucleotide is a phosphorothioate linkage; and

wherein each cytosine of the modified oligonucleotide is a 5-methylcytosine.

14. The compound of claim 1, wherein the oligonucleotide consists of 18 linked nucleosides having a nucleobase sequence consisting of the sequence recited in SEQ ID NO:

1379, and wherein the oligonucleotide comprises;

a gap segment consisting of 8 linked deoxynucleosides;

a 5′

wing segment consisting of 5 linked nucleosides; and

a 3′

wing segment consisting of 5 linked nucleosides;

wherein the gap segment is positioned between the 5′

wing segment and the 3′

wing segment;

wherein the 5 linked nucleosides of the 5′

wing segment comprise a constrained ethyl sugar, a deoxy sugar, a constrained ethyl sugar, a deoxy sugar, and a constrained ethyl sugar in the 5′

to 3′

direction;

wherein each of the 5 linked nucleosides of the 3′

wing segment comprises a 2′

-O-methoxyethyl sugar;

wherein each internucleoside linkage of the modified oligonucleotide is a phosphorothioate linkage; and

wherein each cytosine of the modified oligonucleotide is a 5-methylcytosine.

15. A method of preventing, treating, ameliorating, or slowing progression of a HBV-related disease, disorder or condition in an animal comprising administering to the animal the compound of claim 1, thereby preventing, treating, ameliorating, or slowing progression of a HBV-related disease, disorder or condition in the animal.

16. The method of claim 15, wherein the disease, disorder or condition is jaundice, liver inflammation, liver fibrosis, inflammation, liver cirrhosis, liver failure, liver cancer, diffuse hepatocellular inflammatory disease, hemophagocytic syndrome, serum hepatitis, HBV viremia, or liver disease-related transplantation.

17. The method of claim 16, wherein administering the compound of claim 1 reduces HBV antigen levels in the animal.

18. The method of claim 17, wherein HBsAg levels are reduced.

19. The method of claim 17, wherein HBeAg levels are reduced.

20. The method of claim 15, wherein administering the compound of claim 1 induces HBsAG seroconversion in the animal.