Method for treating enterovirus or rhinovirus infection using antisense antiviral compounds
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.
1. A method of treating viral infection of mammalian cells by an Enterovirus or Rhinovirus , comprising exposing host cells to an effective amount of an antiviral compound that comprises a morpholino antisense oligonucleotide of between 15 and 25 morpholino subunits having a substantially uncharged, nuclease-resistant backbone containing phosphorodiamidate intersubunit linkages joining a morpholino nitrogen of one subunit to a 5′ exocyclic carbon of an adjacent subunit, wherein the morpholino antisense oligonucleotide comprises a targeting sequence of at least 15 nucleotide bases complementary to SEQ ID NO:7 and, conjugated to the morpholino antisense oligonucleotide, an arginine-rich polypeptide effective to promote uptake of the antiviral compound into the host cells, wherein the arginine-rich polypeptide has the sequence of SEQ ID NO:15.
2. The method of claim 1 , wherein the morpholino antisense oligonucleotide forms a heteroduplex with its target sequence, wherein the heteroduplex has a T m of at least 45° C.
3. The method of claim 1 , wherein the morpholino antisense oligonucleotide contains an intersubunit linkages in accordance with the structure:
wherein Y 1 =O, Z=O, Pj is a purine or pyrimidine base-pairing moiety effective to bind, by base-specific hydrogen bonding, to a base in a polynucleotide, and X is alkyl, alkoxy, thioalkoxy, amino, or —NR 2 , where each R is independently hydrogen or methyl.
4. The method of claim 1 , wherein the morpholino antisense oligonucleotide contains an intersubunit linkages in accordance with the structure:
wherein Y 1 =O, Z=O, Pj is a purine or pyrimidine base-pairing moiety effective to bind, by base-specific hydrogen bonding, to a base in a polynucleotide, and X is alkyl, alkoxy, thioalkoxy, amino, —NR 2 , where each R is independently hydrogen or methyl, or 1-piperazine, wherein at least 2 and no more than half of the total number of intersubunit linkages are 1-piperazine.
5. The method of claim 1 , wherein the oligonucleotide has at least 15 nucleotide bases contained in SEQ ID NO:10.
6. The method of claim 5 , wherein the oligonucleotide is selected from the group consisting of SEQ ID NOS:11, 12, and 13.
7. The method of claim 1 , wherein the exposing is by administering to a subject a therapeutically effective amount of the antiviral compound.
8. The method of claim 7 , further comprising administering a second antiviral compound to the subject.