Compounds are substantially uncharged morpholino oligonucleotides having 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
Inventors
Neuman, Benjamin • Stein, David A. • Buchmeier, Michael • Iversen, Patrick L.
Assignees
Scripps Research Institute • Sarepta Therapeutics Inc • US Department of Health and Human Services
Publication Number
US-7582615-B2
Publication Date
2009-09-01
Expiration Date
2027-03-07
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Abstract
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.
Core Innovation
The invention provides antisense antiviral compounds and methods for inhibiting the growth of viruses of the Arenaviridae family and treating viral infections caused by such viruses. The compounds are substantially uncharged morpholino oligonucleotides with sequences of 12-40 subunits, including at least 12 subunits targeting a sequence complementary to a region associated with viral RNA sequences within a 19 nucleotide region at the 5′-terminal regions of the viral RNA, viral complementary RNA, and/or mRNA identified by SEQ ID NO:1.
The antisense compounds are characterized by a substantially uncharged, nuclease-resistant backbone, capable of uptake by mammalian host cells, and form heteroduplex structures with the viral RNA strands having a Tm of dissociation of at least 45° C. These compounds can be conjugated to arginine-rich peptides to enhance cellular uptake and may be used therapeutically in mammals for inhibiting Arenavirus infections.
The technical problem addressed is the need for more effective antiviral therapies against the Arenaviridae family, which includes viruses causing significant human diseases such as Lassa fever and Argentine Hemorrhagic Fever. Existing treatments, such as ribavirin, have limited effectiveness and penetration, especially in certain tissues like the cerebral spinal fluid. Current vaccines are limited, and some Arenaviruses pose bioterrorism threats, underscoring the demand for improved antiviral compounds capable of targeting conserved viral RNA sequences to inhibit viral replication efficiently.
Claims Coverage
The patent includes 8 claims with one independent claim detailing a method of inhibiting Arenavirus infection using a specific antisense oligonucleotide compound. The inventive features focus on the composition and characteristics of the antisense compound and its application in treating or inhibiting Arenavirus infection.
Use of morpholino subunit antisense oligonucleotide with uncharged nuclease-resistant backbone
The method employs an antisense oligonucleotide compound made of morpholino subunits linked by phosphorus-containing intersubunit linkages connecting a morpholino nitrogen to a 5′ exocyclic carbon, characterized by a substantially uncharged, nuclease-resistant backbone.
Oligonucleotide compound with specific size and targeting sequence
The oligonucleotide compound contains between 12 and 40 nucleotide bases and has a targeting sequence of at least 12 contiguous subunits complementary to SEQ ID NO:1, which corresponds to the 5′ terminal region of Arenavirus RNA.
Conjugation of the oligonucleotide to an arginine-rich polypeptide
The antisense oligonucleotide compound is conjugated to an arginine-rich polypeptide that promotes uptake of the compound into infected host cells, enhancing cellular entry and efficacy.
Formation of heteroduplex with viral RNA strands with defined melting temperature
Upon exposing mammalian cells to the compound, a heteroduplex structure is formed between the antisense oligonucleotide and viral RNA or viral complementary RNA strands, characterized by a melting temperature (Tm) of dissociation of at least 45° C., indicating stable binding.
The independent claim covers a method of inhibiting Arenavirus infection in mammalian cells using a morpholino-based antisense oligonucleotide compound with specific biochemical and targeting features, conjugated to an arginine-rich peptide to enhance uptake, which forms stable heteroduplexes with conserved viral RNA sequences. Dependent claims further specify the chemical structure and sequences of the oligonucleotide and applications in treatment.
Stated Advantages
The antisense compounds target a highly conserved 5′-terminal sequence of Arenavirus RNA, allowing broad-spectrum inhibition across multiple Arenavirus species.
The compounds form stable duplexes with viral RNA having a melting temperature of at least 45°C, ensuring effective and specific binding.
Conjugation to arginine-rich peptides enhances cellular uptake, improving the delivery of antisense compounds into infected host cells.
The antisense morpholino oligonucleotides possess nuclease resistance, increasing their stability and persistence within biological environments.
The method offers potential for effective antiviral therapy against Arenaviruses where existing treatments are limited or only moderately effective.
Documented Applications
Treatment of Arenavirus infections in mammals, including humans, by administering antisense morpholino oligonucleotides targeting viral RNA.
Prophylactic treatment of mammals at risk of Arenavirus infection to inhibit viral entry and replication.
Use of antisense compounds conjugated to arginine-rich polypeptides to promote uptake and antiviral activity within infected host cells.
Combination therapy where antisense compounds are administered along with other antiviral agents.
Identification of viral infections by administering targeted antisense oligomers and detecting antisense-RNA heteroduplexes in body fluids as diagnostic indicators.
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