OLIGONUCLEOTIDE COMPOUND AND METHOD FOR TREATING NIDOVIRUS INFECTIONS

• US 20090012280A1
• Filed: 04/25/2008
• Published: 01/08/2009
• Est. Priority Date: 12/24/2003
• Status: Active Grant

• Current Assignee: Sarepta Therapeutics, Inc.
• Original Assignee: Sarepta Therapeutics, Inc.
• Inventors: Bestwick, Richard K., Buchmejer, Michael, Iversen, Patrick L., Weller, Dwight D., Stein, David A., Neuman, Benjamin
• Granted Patent: US 8,759,307 B2
• Time in Patent Office: Days
• US Class Current: 536/23.100
• CPC Class Codes: C07H 21/02 with ribosyl as saccharide radical
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C12N 15/1131 against viruses
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/3233 Morpholino-type ring
• CPC Class Codes: C12N 2310/3513 Protein; Peptide
• CPC Class Codes: C12N 2770/20022 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Abstract

A method and oligonucleotide compound for inhibiting replication of a nidovirus in virus-infected animal cells are disclosed. The compound (i) has a nuclease-resistant backbone, (ii) is capable of uptake by the infected cells, (iii) contains between 8-25 nucleotide bases, and (iv) has a sequence capable of disrupting base pairing between the transcriptional regulatory sequences in the 5′ leader region of the positive-strand viral genome and negative-strand 3′ subgenomic region. In practicing the method, infected cells are exposed to the compound in an amount effective to inhibit viral replication.

Claims

1. An oligonucleotide compound, which is composed of morpholino subunits and uncharged, phosphorus-containing intersubunit linkages joining a morpholino nitrogen of one subunit to a 5′

exocyclic carbon of an adjacent subunit, for use in inhibiting replication of a nidovirus in host cells, characterized by;

(i) a nuclease-resistant backbone,

(ii) capable of uptake by virus-infected host cells,

(iii) containing between 8-25 nucleotide bases,

(iv) having a sequence that is complementary to at least 12 contiguous bases contained in SEQ ID NO;

1; and

(v) capable of forming with SEQ ID NO;

1, a heteroduplex structure characterized by a T_m of dissociation of at least 45°

C.

2. The compound of claim 1, wherein the morpholino subunits are joined by phosphorodiamidate linkages, in accordance with the structure:

3. The compound 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.

4. The compound of claim 3, wherein said morpholino subunits are joined by phosphorodiamidate linkages, in accordance with the structure:

5. The compound of claim 1, which contains a sequence selected from the group consisting of SEQ ID NOS:

20 and 21.

6. The compound of claim 1, which further includes an Arg-rich peptide conjugated to the oligonucleotide.

7. The compound of claim 6, wherein the peptide is SEQ ID NO:

47.

8. An oligonucleotide compound, which is composed of morpholino subunits and uncharged, phosphorus-containing intersubunit linkages joining a morpholino nitrogen of one subunit to a 5′

exocyclic carbon of an adjacent subunit, for use in inhibiting replication of a nidovirus in host cells, characterized by;

(i) a nuclease-resistant backbone,

(ii) capable of uptake by virus-infected host cells,

(iii) containing between 8-25 nucleotide bases,

(iv) having a sequence that is complementary to at least 12 contiguous bases contained in SEQ ID NO;

3; and

(v) capable of forming with SEQ ID NO;

3, a heteroduplex structure characterized by a T_m of dissociation of at least 45°

C.

9. The compound of claim 8, wherein the morpholino subunits are joined by phosphorodiamidate linkages, in accordance with the structure:

10. The compound of claim 9, in which at least 2 and no more than half of the total number of intersubunit linkages are positively charged at physiological pH.

11. The compound of claim 10, wherein said morpholino subunits are joined by phosphorodiamidate linkages, in accordance with the structure:

12. The compound of claim 8, which contains a sequence selected from the group consisting of SEQ ID NOS:

22 and 23.

13. The compound of claim 8, which further includes an Arg-rich peptide conjugated to the oligonucleotide.

14. The compound of claim 13, wherein the peptide is SEQ ID NO:

47.