Coronavirus iRNA compositions and methods of use thereof
Inventors
Akinc, Akin • McIninch, James D. • Anglero-Rodriguez, Yesseinia • Schlegel, Mark K. • Hebner, Christy M. • LEMPP, Florian A.
Assignees
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Publication Number
US-11208660-B1
Publication Date
2021-12-28
Expiration Date
Abstract
The present invention relates to RNAi agents, e.g., dsRNA agents, targeting the coronavirus genome. The invention also relates to methods of using such RNAi agents to inhibit expression of a coronavirus genome and to methods of treating or preventing a coronavirus-associated disease in a subject.
Core Innovation
The invention relates to a composition comprising a first double stranded RNAi agent and a second double stranded RNAi agent for inhibiting expression of a coronavirus genome in a cell. Each RNAi agent comprises a sense strand and an antisense strand forming a double stranded region, and the composition is defined by two RNAi agents rather than a single agent.
For the first double stranded RNAi agent, the antisense strand comprises at least 15 contiguous nucleotides differing by no more than three nucleotides from the nucleotide sequence 5′-GUUUGAAAAGCAACAUUGUUAGU-3′ (SEQ ID NO: 6). For the second double stranded RNAi agent, the antisense strand comprises at least 15 contiguous nucleotides differing by no more than three nucleotides from the nucleotide sequence 5′-UGACAUUUUAGACUGUACAGUGG-3′ (SEQ ID NO: 8). The corresponding sense strands form the double stranded regions with the antisense strands.
The disclosed embodiments also include specific sense and antisense strand nucleotide sequences and defined nucleotide chemistries and linkages. The nucleotide definitions include 2′-O-methyl, 2′-fluoro, phosphorothioate linkage, 2′-O-hexadecyl-adenosine-3′-phosphate, and vinyl-phosphonate, and carbohydrate conjugate structures including triantennary GalNAc and RNAi agent attachment linker embodiments are described.
Claims Coverage
The consolidated content identifies two independent claims. Across these claims, the inventive features focus on two double stranded RNAi agents for inhibiting expression of a coronavirus genome in a cell, with defined sense/antisense strand structures and either antisense sequence constraints or explicit modified nucleotide sequences.
Two double stranded RNAi agents for coronavirus inhibition with defined sense/antisense double stranded regions
A composition comprising a first double stranded RNAi agent for inhibiting expression of a coronavirus genome in a cell and a second double stranded RNAi agent for inhibiting expression of a coronavirus genome in a cell, wherein the first double stranded RNAi agent comprises a first sense strand and a first antisense strand forming a first double stranded region and the second double stranded RNAi agent comprises a second sense strand and a second antisense strand forming a second double stranded region.
Antisense sequence constrained by limited divergence from specified SEQ ID sequences
The first antisense strand comprises at least 15 contiguous nucleotides differing by no more than three nucleotides from the nucleotide sequence 5′-GUUUGAAAAGCAACAUUGUUAGU-3′ (SEQ ID NO: 6); and the second antisense strand comprises at least 15 contiguous nucleotides differing by no more than three nucleotides from the nucleotide sequence 5′-UGACAUUUUAGACUGUACAGUGG-3′ (SEQ ID NO: 8).
Defined modified sense/antisense sequences with chemical entities and linkages
A composition comprising a first double stranded RNAi agent and a second double stranded RNAi agent, wherein the first sense strand comprises the nucleotide sequence 5′-usasaca(Ahd)UfgUfUJfGfcuuuucaasasa-3′ (SEQ ID NO: 9) and the first antisense strand comprises the nucleotide sequence 5′-VPusUfsuugAfaaagcaaCfaUfuguuasgsu-3′ (SEQ ID NO: 10); and the second sense strand comprises the nucleotide sequence 5′-ascsugu(Ahd)CfaGfUfCfuaaaauguscsa-3′ (SEQ ID NO: 11) and the second antisense strand comprises the nucleotide sequence 5′-VPusGfsacaUfuuuagacUfgUfacagusgsg-3′ (SEQ ID NO: 12), wherein a, g, c and u are 2′-O-methyl A, G, C, and U; Af, Gf, Cf and Uf are 2′-fluoro A, G, C and U; s is a phosphorothioate linkage; (Ahd) is 2′-O-hexadecyl-adenosine-3′-phosphate; and VP is vinyl-phosphonate.
Across the independent claims, the core coverage is for a composition that uses two double stranded RNAi agents to inhibit expression of a coronavirus genome in a cell. The claims define each agent’s sense/antisense strand architecture and enforce either antisense sequence constraints relative to SEQ ID NO: 6 and SEQ ID NO: 8 or specify particular fully defined modified sense/antisense sequences with defined chemical modifications and linkages.
Stated Advantages
Inhibiting expression of a coronavirus genome in a cell.
Inhibition of SARS-CoV-2 expression/infection as reported in the disclosed evaluations.
Reduction of off-target gene silencing through thermally destabilizing modifications in the antisense seed region.
Documented Applications
A composition for inhibiting expression of a coronavirus genome in a cell.
A pharmaceutical composition for inhibiting expression of a coronavirus genome in a cell, including a pharmaceutically acceptable carrier.
In vitro message inhibition using siRNA/dsRNA duplexes evaluated in Cos-7 cells.
In vitro SARS-CoV-2 replication screening using RT-qPCR and in-cell ELISA (IFA).
Resistance-selection assessment for duplexes using duplex EC50/IFA metrics and selection of resistance-associated changes.
Animal evaluation with intranasal delivery in hamsters, including reported inhibition of SARS-CoV-2 expression/infection and comparison with subcutaneous delivery.
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