Antisense antiviral compounds and methods for treating a filovirus infection
View Patent ↗The invention provides antisense antiviral compounds and methods of their use and production in inhibition of growth of viruses of the Filoviridae family, and in the treatment of a viral infection. The compounds and methods relate to the treatment of viral infections in mammals including primates by Ebola and Marburg viruses. The antisense antiviral compounds are morpholino oligonucleotides having: a) a nuclease resistant backbone, b) 15-40 nucleotide bases, and c) a targeting sequence of at least 15 bases in length that hybridizes to a target region selected from the following: i) the AUG start site region of VP35, as exemplified by SEQ ID NOS:67-71 or ii) the AUG start site region of VP24, as exemplified by SEQ ID NOS:72-76.
1. A method of treating Filovirus infection in a subject, comprising:
administering to a subject a therapeutically effective amount of an antiviral composition of an antisense oligomer, wherein the oligomer comprises:
morpholino subunits linked by phosphorous-containing intersubunit linkages which join a morpholino nitrogen of one subunit to a 5′ exocyclic carbon of an adjacent subunit, wherein at least 2 and no more than half of the total number of intersubunit linkages are positively charged at physiological pH; and
a targeting sequence which forms a heteroduplex with a target sequence of the AUG start-site region of a positive-strand mRNA for Filovirus viral protein 35 (VP35);
wherein the antisense oligomer inhibits virus production.
2. The method of claim 1 , wherein the targeting sequence forms a heteroduplex with the AUG start-site region defined by SEQ ID NO:1.
3. The method of claim 2 , wherein the targeting sequence is complementary to at least 12 contiguous bases of the sequence of SEQ ID NO:1.
4. The method of claim 2 , wherein the oligomer has between 12-40 subunits.
5. The method of claim 2 , wherein the heteroduplex has a Tm of dissociation of at least 45° C.
6. The method of any one of claims 1 to 5 , wherein the morpholino subunits are joined by 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 selected from alkyl alkoxy; thioalkoxy; —NR 2 , wherein each R is independently H or lower alkyl; or 1-piperizino.
7. The method of claim 1 , wherein the Filovirus is ebola virus.
8. The method of claim 7 , wherein the ebola virus is viral strain ebola Zaire.