RNase L activators and antisense oligonucleotides effective to treat RSV infections

• US 5,998,602 A
• Filed: 02/14/1997
• Issued: 12/07/1999
• Est. Priority Date: 02/15/1996
• Status: Expired due to Fees

• Current Assignee: Cleveland Clinic Foundation, Health and Human Services The Government of The United Sates of America As Represented By
• Original Assignee: Cleveland Clinic Foundation
• Inventors: Li, Guiying, Xiao, Wei, Torrence, Paul F., Silverman, Robert Hugh, Cirino, Nick Mario
• Primary Examiner(s): Guzo, David
• Assistant Examiner(s): LARSON, THOMAS GEORGE
• Application Number:
  US08/801,898
• Time in Patent Office: 1,026 Days
• Field of Search: 435/6, 435/199, 435/91.1, 435/325, 435/375, 514/44, 536/24.3, 536/24.5, 935/34
• US Class Current: 536/24.5
• 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: A61K 47/549 Sugars, nucleosides, nucleotides or nucleic acids
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/14 for RNA viruses
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C12N 15/1131 against viruses
• CPC Class Codes: C12N 2310/31 of the backbone
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/3521 Methyl

Abstract

The invention concerns a compounds and methods for treating infection with Respiratory Syncytial Virus. The compounds comprise an antisense portion, which is complementary to a normally single stranded portion of the RSV antigenomic strand (the mRNA strand), a linker and a oligonucleotide activator of RNase L, a ubiquitous non-specific RNase. The method comprised forming a complex of an activated RNase L and the antisense molecule. The application teaches methods of determining which portions of the RSV antigenomic strand are normally single-stranded. The application teaches that an antisense oligonucleotide having the sequence of residues 8281-8299 of the RSV genome is particularly useful to practice the invention and provides in vitro results superior to those obtainable with the conventional drug of choice, ribavirin.

Claims

1. A chimeric molecule comprising:

(a) an antisense oligonucleotide in which the oligonucleotide is complementary to a portion of the antigenomic RNA strand of a strain of a Respiratory Syncytial Virus (RSV) spanning residues 617 to 663 of SEQ ID NO;

23;

residues 718 to 772 of SEQ ID NO;

23;

residues 2490 to 2530 of SEQ ID NO;

23, residues 3212 to 3247 of SEQ ID NO;

23;

residues 5240 to 5288 of SEQ ID NO;

23 or residues 8251 to 8299 of SEQ ID NO;

23 of the RSV strain A2 genome; and

(b) an oligonucleotide activator of RNase L attached to the antisense oligonucleotide by a linker.

2. The chimeric molecule of claim 1 wherein said chimeric molecule further comprises a modification to the 3'"'"' terminus of the oligonucleotide which protects the oligonucleotide from exonucleases.

3. The chimeric molecule of claim 1 which further comprises additional adenylate molecules attached to the 3'"'"' terminus of the oligonucleotide.

4. The chimeric molecule of claim 1 in which the oligonucleotide activator is selected from the group consisting of sp5'"'"'A2'"'"'(p5'"'"'A2'"'"')₂ --O--, sp5'"'"'A2'"'"'(p5'"'"'A2'"'"')₃ --O--, p5'"'"'A2'"'"'(p5'"'"'A2'"'"')₂ --O--, and p5'"'"'A2'"'"'(p5'"'"'A2'"'"')₃ --O--.

5. The chimeric molecule of claim 1 in which the first end is the 5'"'"' terminus, and the 3'"'"' terminal hydroxyl of the antisense oligonucleotide is blocked by a blocker selected from the group consisting of a -p3'"'"'N5'"'"' nucleotide, a p-O-alkylamine, a p-O-hydroxyalkylamine, a sp-O-alkylamine, a sp-O-hydroxyalkylamine, ethyl and methyl.

6. The chimeric molecule of claim 1 in which the antisense oligonucleotide is selected from the group consisting of residues 8251-8270 (SEQ ID NO. 21), 8261-8279 (SEQ ID NO. 20) and 8281-8299 (SEQ ID NO. 22), numbered in the 5'"'"'→

3'"'"' direction.

7. The chimeric molecule of claim 1 in which the antisense oligonucleotide contains one or more phospho-moieties selected from the group consisting of phosphorothioate, methylphosphonate and methylphosphonothioate.