Methods and compositions for the specific inhibition of Complement Component 5(C5) by double-stranded RNA
Inventors
Brown, Bob D. • Dudek, Henryk T.
Assignees
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Publication Number
US-11773390-B2
Publication Date
2023-10-03
Expiration Date
Abstract
This invention relates to compounds, compositions, and methods useful for reducing C5 target RNA and protein levels via use of dsRNAs, e.g., Dicer substrate siRNA (DsiRNA) agents.
Core Innovation
The invention concerns nucleic acid molecules, including single-stranded and double stranded nucleic acids (dsNA), that comprise oligonucleotide strands complementary to a target C5 mRNA sequence set forth as SEQ ID NO: 1262. The nucleic acids are configured to reduce C5 target mRNA expression when introduced into a mammalian cell, and the nucleic acids comprise a modified nucleotide.
The dsNA embodiments specify RNA strands of defined length, with complementarity to SEQ ID NO: 1262 across at least 20 consecutive nucleotides of the second strand or at least 21 consecutive nucleotides of the oligonucleotide strand. The disclosure further includes structural and positional constraints such as duplex region length, overhangs, blunt ends, tetraloop and discontinuity features, and defined terminal nucleotide and ribonucleotide positioning.
The disclosure also encompasses processing-oriented features, including mammalian Ago2 cleavage at a site between positions 9 and 10 of the C5 mRNA sequence set forth as SEQ ID NO: 1262, and dsNA embodiments that are sufficiently highly modified to substantially prevent dicer cleavage while optionally yielding 19-23 nucleotide strand length dsNAs capable of reducing C5 mRNA expression. The overall problem being solved is reducing C5 target mRNA expression in mammalian cells using modified nucleic acids complementary to SEQ ID NO: 1262.
Claims Coverage
The independent claims cover three main inventive features: modified oligonucleotide strands complementary to SEQ ID NO: 1262, dsNA with defined RNA strand lengths and complementarity to SEQ ID NO: 1262, and composition or hybridization-complex alternatives with additional structural and processing constraints. Across the independent claims, the coverage is anchored to reducing C5 target mRNA expression in mammalian cells with modified nucleotides.
Modified oligonucleotide complementary to SEQ ID NO: 1262
A nucleic acid comprising an oligonucleotide strand of up to 80 nucleotides in length, wherein the oligonucleotide strand is complementary to a target C5 mRNA sequence set forth as SEQ ID NO: 1262 along at least 21 consecutive nucleotides of the oligonucleotide strand length to reduce C5 target mRNA expression when introduced into a mammalian cell, wherein the nucleic acid comprises a modified nucleotide.
Double stranded nucleic acid with defined RNA strand lengths
A double stranded nucleic acid (dsNA) comprising first and second nucleic acid strands comprising RNA, wherein the first strand is 22-66 nucleotides in length and the second strand is 20-66 nucleotides in length, wherein the second strand is complementary to a target C5 mRNA sequence set forth as SEQ ID NO: 1262 along at least 20 consecutive nucleotides of the second oligonucleotide strand length to reduce C5 target mRNA expression when introduced into a mammalian cell, wherein the dsNA comprises a modified nucleotide.
Composition with structural and processing constraints
A composition selected from the group consisting of dsNA and nucleic acid alternatives targeting SEQ ID NO: 1262, including dsNA with specified strand-length ranges and modified nucleotide; an alternative in which mammalian Ago2 cleaves the mRNA between positions 9 and 10 of SEQ ID NO: 1262; nucleic acids with end structures including blunt end, 3′ overhang, and 5′ overhang; alternatives with tetraloop and discontinuity features; alternatives sufficiently highly modified to substantially prevent dicer cleavage and optionally yield shorter dsNAs; and in vivo and in vitro hybridization complexes comprising an exogenous nucleic acid and a duplex region of at least 21 consecutive nucleotides along SEQ ID NO: 1262.
Collectively, the independent claims cover modified nucleic acids and dsNA that target C5 mRNA sequence SEQ ID NO: 1262 to reduce C5 target mRNA expression in mammalian cells, with specified complementarity lengths and, in some alternatives, structural and processing constraints including mammalian Ago2 cleavage positioning and resistance to dicer cleavage.
Stated Advantages
Reduce C5 target mRNA expression when introduced into a mammalian cell.
In some alternatives, include a mammalian Ago2 cleavage site between positions 9 and 10 of the C5 mRNA sequence set forth as SEQ ID NO: 1262.
In some alternatives, substantially prevent dicer cleavage of the dsNA while optionally yielding shorter dsNAs capable of reducing C5 mRNA expression.
Predicted knockdown efficacy greater than 50% is stated for the listed human anti-C5 DsiRNAs.
Documented Applications
Treatment of paroxysmal nocturnal hemoglobinuria (PNH) or atypical hemolytic uremic syndrome (aHUS) by administering the dsNA to a subject.
Use in mammalian cells to reduce C5 target mRNA expression.
DsiRNA/dsRNA nucleic acid targeting of a C5 mRNA set, including Selected Human Anti-C5 “Blunt/Blunt” DsiRNAs with sequence listings.
Use in vivo and in vitro hybridization complexes present within a cell comprising an exogenous nucleic acid and a target C5 mRNA sequence set forth as SEQ ID NO: 1262.
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