Antisense antiviral compound and method for treating influenza viral infection

• US 9,394,323 B2
• Filed: 04/11/2014
• Issued: 07/19/2016
• Est. Priority Date: 11/13/2009
• Status: Active Grant

• Current Assignee: Sarepta Therapeutics, Inc.
• Original Assignee: Sarepta Therapeutics, Inc.
• Inventors: Iversen, Patrick L.
• Primary Examiner(s): Chong, Kimberly
• Application Number:
  US14/251,124
• Publication Number:
  US 20140303073A1
• Time in Patent Office: 830 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61K 31/165 having aromatic rings, e.g. colchicine, atenolol, progabide
• CPC Class Codes: A61K 31/43 Compounds containing 4-thia-1-azabicyclo [3.2.0] heptane ring systems, i.e. compounds containing a ring system of the formula , e.g. penicillins, penems
• CPC Class Codes: A61K 31/5377 not condensed and containing further heterocyclic rings, e.g. timolol
• CPC Class Codes: A61K 31/545 Compounds containing 5-thia-1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula: , e.g. cephalosporins, cefaclor, or cephalexine
• CPC Class Codes: A61K 31/7056 containing five-membered rings with nitrogen as a ring hetero atom
• CPC Class Codes: A61K 38/08 Peptides having 5 to 11 amino acids A61K38/043 - A61K38/046 take precedence
• CPC Class Codes: A61K 38/10 Peptides having 12 to 20 amino acids A61K38/043 - A61K38/046 take precedence
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/16 for influenza or rhinoviruses
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07F 9/65616 containing the ring system having three or more than three double bonds between ring members or between ring members and non-ring members, e.g. purine or analogs
• CPC Class Codes: C07K 7/06 having 5 to 11 amino acids
• CPC Class Codes: C07K 7/08 having 12 to 20 amino acids gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68
• 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 15/1131 against viruses
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/3181 Peptide nucleic acid, PNA
• 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 2310/3233 : Morpholino-type ring
• CPC Class Codes: C12N 2310/3513 : Protein; Peptide
• CPC Class Codes: C12N 2310/3521 : Methyl

Abstract

The present invention relates to antisense antiviral compounds and methods of their use and production in inhibition of growth of viruses of the Orthomyxoviridae family and in the treatment of a viral infection. The compounds are particularly useful in the treatment of influenza virus infection in a mammal. Exemplary antisense antiviral compounds are substantially uncharged, or partially positively charged, morpholino oligonucleotides having 1) a nuclease resistant backbone, 2) 12-40 nucleotide bases, and 3) a targeting sequence of at least 12 bases in length that hybridizes to a target region selected from the following: a) the 5′ or 3′ terminal 25 bases of the negative sense viral RNA segment of Influenzavirus A, Influenzavirus B and Influenzavirus C; b) the terminal 30 bases of the 5′ or 3′ terminus of the positive sense vcRNA; c) the 45 bases surrounding the AUG start codon of an influenza viral mRNA and; d) 50 bases surrounding the splice donor or acceptor sites of influenza mRNAs subject to alternative splicing.

Claims

1. An antiviral antisense oligonucleotide consisting of the following structure:

2. The antiviral antisense oligonucleotide of claim 1, wherein the antiviral antisense oligonucleotide is conjugated to an arginine-rich polypeptide that enhances the uptake of the compound into host cells.

3. The antiviral antisense oligonucleotide of claim 2, wherein the arginine-rich polypeptide is any one of SEQ ID NOS:

115-128.

28. The antiviral antisense oligonucleotide of claim 1, wherein the antiviral antisense oligonucleotide is linked to a polyethylene glycol moiety.

4. A pharmaceutical composition, comprising a pharmaceutically acceptable carrier, and an antiviral antisense oligonucleotide consisting of the following structure:

5. The pharmaceutical composition of claim 4, wherein the antiviral antisense oligonucleotide is conjugated to an arginine-rich polypeptide that enhances the uptake of the compound into host cells.

6. The pharmaceutical composition of claim 5, wherein the arginine-rich polypeptide is any one of SEQ ID NOS:

115-128.

7. The pharmaceutical composition of any one of claims 4-6, further comprising an antibiotic.

8. The pharmaceutical composition of claim 7, wherein the antibiotic is bacteriostatic or bactericidal.

9. The pharmaceutical composition of claim 7, wherein the antibiotic is a beta-lactam.

10. The pharmaceutical composition of claim 7, wherein the antibiotic is selected from penicillin, amoxicillin, cephalosporin, chloramphenicol, or clindamycin.

11. The pharmaceutical composition of any one of claims 4-6, further comprising a second oligonucleotide targeted against an RNA molecule encoding CD200 or the CD200 receptor.

29. The pharmaceutical composition of claim 4, wherein the antiviral antisense oligonucleotide is linked to a polyethylene glycol moiety.

31. The pharmaceutical composition of claim 4, further comprising an additional influenza virus-targeted therapy.

32. The pharmaceutical composition of claim 31, wherein the additional influenza virus-targeted therapy is oseltamivir phosphate.

12. A method of reducing replication of an Influenza A virus, comprising contacting an Influenza A virus infected cell with an antiviral antisense oligonucleotide consisting of the following structure:

13. The method of claim 12, wherein the method comprises administering the antiviral antisense oligonucleotide to a subject having an Influenza A virus infected cell.

14. The method of claim 13, further comprising administering an antibiotic in combination with the antiviral antisense oligonucleotide when the subject has a secondary bacterial infection.

15. The method of claim 14, wherein the antibiotic is administered separately from the antiviral antisense oligonucleotide.

16. The method of claim 14, wherein the antibiotic is administered concurrently with the antiviral antisense oligonucleotide.

17. The method of claim 14, wherein the antibiotic is bacteriostatic or bactericidal.

18. The method of claim 14, wherein the antibiotic is a beta-lactam.

19. The method of claim 14, wherein the antibiotic is selected from penicillin, amoxicillin, cephalosporin, chloramphenicol, or clindamycin.

20. The method of claim 14, wherein the secondary bacterial infection is a Streptococcal infection.

21. The method of claim 20, wherein the Streptococcal infection is a Streptococcus pneuomoniae infection.

22. The method of claim 12, further comprising administering a second antisense oligonucleotide targeted against an RNA molecule encoding CD200 or the CD200 receptor separately or concurrently in combination with the antiviral antisense oligonucleotide.

23. The method of claim 22, wherein the second antisense oligonucleotide targeted against an RNA molecule encoding CD200 or the CD200 receptor is administered separately from the antiviral antisense oligonucleotide.

24. The method of claim 22, wherein the second antisense oligonucleotide targeted against an RNA molecule encoding CD200 or the CD200 receptor is administered concurrently with the antiviral antisense oligonucleotide.

25. The method of claim 14, further comprising administering an antisense oligonucleotide targeted against an RNA molecule encoding CD200 or the CD200 receptor, separately or concurrently in combination with the antiviral antisense oligonucleotide.

26. The method of claim 25, wherein the second antisense oligonucleotide targeted against an RNA molecule encoding CD200 or the CD200 receptor is administered separately from the antiviral antisense oligonucleotide.

27. The method of claim 25, wherein the second antisense oligonucleotide targeted against an RNA molecule encoding CD200 or the CD200 receptor is administered concurrently with the antiviral antisense oligonucleotide.

30. The method of claim 12, wherein the antiviral antisense oligonucleotide is linked to a polyethylene glycol moiety.

33. The method of claim 12, further comprising administering an additional influenza virus-targeted therapy separately or concurrently in combination with the antiviral antisense oligonucleotide.

34. The method of claim 33, wherein the additional influenza virus-targeted therapy is administered concurrently with the antiviral antisense oligonucleotide.

35. The method of claim 33, wherein the additional influenza virus-targeted therapy is administered separately from the antiviral antisense oligonucleotide.

36. The method of claim 33, wherein the additional influenza virus-targeted therapy is oseltamivir phosphate.