Antisense antiviral compound and method for treating picornavirus infection

• US 20070066556A1
• Filed: 09/08/2006
• Published: 03/22/2007
• Est. Priority Date: 09/08/2005
• Status: Active Grant

• Current Assignee: Sarepta Therapeutics, Inc.
• Original Assignee: Sarepta Therapeutics, Inc.
• Inventors: Stein, David, Weller, Dwight, Bestwick, Richard, Iversen, Patrick
• Granted Patent: US 8,524,676 B2
• Time in Patent Office: Days
• US Class Current: 514/44.A
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: C12N 15/1131 against viruses
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/314 Phosphoramidates
• CPC Class Codes: C12N 2310/3233 Morpholino-type ring
• CPC Class Codes: C12N 2770/32311 Enterovirus
• CPC Class Codes: C12N 2770/32711 Rhinovirus
• 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 invention provides antisense antiviral compounds and methods of their use and production in inhibition of growth of viruses of the Picornaviridae family and in the treatment of a viral infection. The compounds are particularly useful in the treatment of Enterovirus and/or Rhinovirus infection in a mammal. The antisense antiviral compounds are substantially uncharged, morpholino oligonucleotides have a sequence of 12-40 subunits, including at least 12 subunits having a targeting sequence that is complementary to a region associated with viral RNA sequences within a 32 nucleotide region of the viral 5′ untranslated region identified by SEQ ID NO:7.

Claims

1. A method of inhibiting viral infection in mammalian cells by an Enterovirus or Rhinovirus in the Picornaviridae family, comprising

(a) exposing the cells to an antisense oligonucleotide compound characterized by:

(i) a substantially uncharged, nuclease-resistant backbone,

(ii) capable of uptake by mammalian host cells,

(iii) containing between 12-40 nucleotide bases, and

(iv) having a targeting sequence of at least 12 subunits complementary to SEQ ID NO;

7 in the positive-sense strand of the virus, and

(b) by said exposing, forming a heteroduplex structure (i) composed of the virus'"'"' positive sense strand and the oligonucleotide compound, and (ii) characterized by a Tm of dissociation of at least 45°

C.

2. The method of claim 1, wherein said oligonucleotide compound to which the host cells are exposed is composed of morpholino subunits and phosphorus-containing intersubunit linkages joining a morpholino nitrogen of one subunit to a 5′

exocyclic carbon of an adjacent subunit.

3. The method of claim 2, wherein the morpholino subunits in the oligonucleotide compound to which the host cells are exposed is administered to the subject are joined by phosphorodiamidate linkages having the structure:

4. The method of claim 2, in which at least 2 and no more than half of the total number of intersubunit linkages are positively charged at physiological pH.

5. The method of claim 2, wherein said morpholino subunits are joined by phosphorodiamidate linkages, in accordance with the structure:

6. The method of claim 1, wherein the oligonucleotide compound to which the cells are exposed has a sequence contained in SEQ ID NO:

10.

7. The method of claim 6, wherein said sequence of the oligonucleotide compound is selected from the group consisting of SEQ ID NOS:

11, 12, and 13.

8. The method of claim 1, wherein the compound to which the cells are exposed is conjugated to an arginine-rich polypeptide effective to promote uptake of the compound into infected host cells.

9. The method of claim 8, wherein the arginine rich peptide has one of the sequences identified as SEQ ID NOS:

14-19.

10. The method of claim 1, for use in treating a mammalian subject infected by an Enterovirus or Rhinovirus in the Picornaviridae family, wherein said exposing includes administering to the subject, a pharmaceutically effective amount of the oligonucleotide compound, and which further includes continuing said administering until a significant reduction in viral infection or the symptoms thereof is observed.

11. The method of claim 10, which further includes administering a second anti-viral compound to the subject.

12. The method of claim 1, for use in treating a mammalian subject at risk of infection by an Enterovirus or Rhinovirus in the Picornaviridae family, wherein said exposing includes administering to the subject, an amount of the oligonucleotide compound effective to inhibit infection of subject host cells by the virus.

21. The compound of claim 1, in combination with another anti-viral compound.

13. An antisense oligonucleotide compound for use in inhibiting viral infection in mammalian cells by an Enterovirus or Rhinovirus in the Picornaviridae family, characterized by:

(i) a substantially uncharged, nuclease-resistant backbone,

(ii) capable of uptake by mammalian host cells,

(iii) containing between 12-40 nucleotide bases,

(iv) having a targeting sequence of at least 12 subunits contained in SEQ ID NO;

10; and

(v) capable of binding to the virus'"'"' positive sense strand to form a heteroduplex structure having by a Tm of dissociation of at least 45°

C.

14. The compound of claim 13, which is substantially uncharged and composed of morpholino subunits and phosphorus-containing intersubunit linkages joining a morpholino nitrogen of one subunit to a 5′

exocyclic carbon of an adjacent subunit.

15. The compound of claim 14, wherein said morpholino subunits are joined by phosphorodiamidate linkages having the structure:

16. The compound of claim 14, in which at least 2 and no more than half of the total number of intersubunit linkages are positively charged at physiological pH.

17. The compound of claim 16, wherein said morpholino subunits are joined by phosphorodiamidate linkages, in accordance with the structure:

18. The compound of claim 13, wherein said sequence of the oligonucleotide compound is selected from the group consisting of SEQ ID NO:

11, SEQ ID NO;

12, and SEQ ID NO;

13.

19. The compound of claim 13, which is conjugated to an arginine-rich polypeptide effective to promote uptake of the compound into infected host cells.

20. The compound of claim 19, Wherein the arginine rich peptide has one of the sequences identified as SEQ ID NOS:

14-19.