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 Ebola virus AUG start site region of VP24; ii) the Ebola virus AUG start site region of VP35; iii) the Marburg virus AUG start site region of VP24; or iv) the Marburg virus AUG start site region of NP.
1. A method of treating Ebola virus infection in a subject, comprising:
administering to a subject a therapeutically effective amount of an antisense oligomer
of 12-40 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 comprising
a targeting sequence which forms a heteroduplex with a target sequence of the AUG start-site region of a positive-strand mRNA for Ebola viral protein 24 (VP24);
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:2.
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:2.
4. The method of claim 2 , wherein the oligomer has 15-25 subunits.
5. The method of claim 2 , wherein the heteroduplex has a Tm of dissociation of at least 45° C.
6. The method of claim 1 , 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-piperazino.
7. The method of claim 1 , wherein the Ebola virus is viral strain Ebola Zaire.
8. The method of claim 1 , which further includes administering to the subject a therapeutically effective amount of a second antisense oligomer having the same structure recited in claim 1 , but having a targeting sequence which forms a heteroduplex with a target sequence of the AUG start-site region of a positive-strand mRNA for Ebola viral protein 35 (VP35).
9. The method of claim 8 , wherein the Ebola viral protein 35 targeting sequence forms a heteroduplex with the AUG start-site region defined by SEQ ID NO: 1.
10. The method of claim 8 , wherein the Ebola viral protein 35 targeting sequence is complementary to at least 12 contiguous bases of the sequence of SEQ ID NO:1.
11. The method of claim 1 , wherein the targeting sequence comprises the sequence of SEQ ID NO:34.
12. The method of claim 11 , wherein the targeting sequence consists of the sequence of SEQ ID NO:34.
13. The method of claim 8 , wherein the VP24 targeting sequence consists of the sequence of SEQ ID NO:34, and the VP35 targeting sequence of the second oligomer consists of the sequence of SEQ ID NO:22.
14. The method of claim 13 , wherein the antisense oligomers have between 2-8 piperazine-containing intersubunit linkages.
15. The method of claim 13 , wherein the oligomer targeted against VP24 and the oligomer targeted against VP35 are administered sequentially.
16. The method of claim 13 , wherein the oligomer targeted against VP24 and the oligomer targeted against VP35 are administered concurrently.