Antisense antiviral compound and method for treating arenavirus infection
View Patent ↗The invention provides antisense antiviral compounds and methods of their use and production in inhibition of growth of viruses of the Arenaviridae family and in the treatment of a viral infection. The compounds are particularly useful in the treatment of Arenavirus 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 19 nucleotide region of the 5′-terminal regions of the viral RNA, viral complementary RNA and/or mRNA identified by SEQ ID NO:1.
1. A method of inhibiting viral infection in mammalian cells by an Arenavirus in the Arenaviridae family, comprising
(a) exposing the cells to an antisense oligonucleotide compound 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, and 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 contiguous subunits complementary to SEQ ID NO:1, and
(v) conjugated to the oligonucleotide, an arginine-rich polypeptide effective to promote uptake of the compound into infected host cells, and
(b) by said exposing, forming a heteroduplex structure (i) composed of the virus' vRNA or vcRNA 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 the morpholino subunits in the oligonucleotide compound to which the host cells are exposed are joined by phosphorodiamidate linkages having the structure:
where 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 alkyl amino, including dialkylamino.
3. The method of claim 1 , wherein the oligonucleotide compound to which the cells are exposed has a sequence comprising SEQ ID NO:5.
4. The method of claim 1 , wherein the oligonucleotide compound to which the cells are exposed has a sequence comprising SEQ ID NO:2.
5. The method of claim 1 , for use in treating a mammalian subject infected by an Arenavirus in the Arenaviridae 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.
6. The method of claim 5 , which further includes administering a second anti-viral compound to the subject.
7. The method of claim 1 , for use in treating a mammalian subject at risk of infection by an Arenavirus in the Arenaviridae 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.
8. The method of claim 1 , wherein the arginine rich peptide has the sequence identified as SEQ ID NO: 8.