Factor XII (hageman factor) (F12), kallikrein B, plasma (fletcher factor) 1 (KLKB1), and kininogen 1 (KNG1) iRNA compositions and methods of use thereof
Inventors
Akinc, Akin • Hinkle, Gregory • Maier, Martin A. • Butler, James • Liu, Jingxuan
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-12338439-B2
Publication Date
2025-06-24
Expiration Date
Abstract
The present invention relates to RNAi agents, e.g., double stranded RNAi agents, targeting the Kallikrein B, Plasma (Fletcher Factor) 1 (KLKB1) gene, the Factor XII (Hageman Factor (F12) gene, or the Kininogen 1 (KNG1) gene, and methods of using such RNAi agents to inhibit expression of a KLKB1 gene, an F12 gene, and/or a KNG1 gene, and methods of treating subjects having an hereditary angioedema (HAE) and/or a contact activation pathway-associated disorder.
Core Innovation
The invention provides a double stranded ribonucleic acid (dsRNA) agent, or a salt thereof, for inhibiting expression of Factor XII (Hageman Factor) (F12). The dsRNA agent comprises a sense strand and an antisense strand forming a double stranded region, where the sense strand comprises at least 19 contiguous nucleotides from nucleotides 433-460 of the nucleotide sequence of SEQ ID NO:9 and the antisense strand comprises at least 19 contiguous nucleotides from corresponding nucleotides of the nucleotide sequence of SEQ ID NO:10.
All nucleotides of the sense strand and all nucleotides of the antisense strand comprise a nucleotide modification, and the sense strand is conjugated to a ligand attached at the 3′-terminus. The disclosed framework further includes phosphorothioate internucleotide linkages and methylphosphonate internucleotide linkages, phosphates at 5′/3′ ends or mimics, and carbohydrate ligands, including N-acetylgalactosamine (GalNAc) derivatives with monovalent and branched multivalent linker embodiments.
The disclosure further describes motifs comprising three identical nucleotide modifications on three consecutive nucleotides at or near the Dicer cleavage site, including embodiments with 2′-fluoro (2′-F) and 2′-O-methyl (2′-OMe) on “YYY” motifs, along with strand length, overhang, terminal capping, and end-group modification concepts. It also includes in vivo gene-silencing application for contact activation pathway genes including KLKB1, F12, and KNG1, including through degradation of mRNA transcripts after contacting cells and maintaining the cells long enough.
Claims Coverage
The claim coverage centers on dsRNA agents for inhibiting Factor XII (F12) expression, with multiple inventive features directed to defined sequence-derived dsRNA, complete nucleotide modification, and 3′-terminal ligand conjugation to the sense strand. Additional dependent features include specific nucleotide modification types, phosphorothioate and methylphosphonate linkage features, strand length, GalNAc derivative ligands, and a cell-based mRNA degradation method.
Defined dsRNA duplex targeting Factor XII (F12)
A double stranded ribonucleic acid (dsRNA) agent, or a salt thereof, for inhibiting expression of Factor XII (Hageman Factor) (F12), comprising a sense strand and an antisense strand forming a double stranded region.
Sequence-defined contiguous 19+ nucleotide regions from SEQ ID NO:9 and SEQ ID NO:10
The sense strand comprises at least 19 contiguous nucleotides from nucleotides 433-460 of SEQ ID NO:9 and the antisense strand comprises at least 19 contiguous nucleotides from the corresponding nucleotides of SEQ ID NO:10.
All-nucleotide modification across both strands
All of the nucleotides of the sense strand and all of the nucleotides of the antisense strand comprise a nucleotide modification.
3′-terminal ligand conjugation on the sense strand
The sense strand is conjugated to a ligand attached at the 3′-terminus.
Selectable nucleotide modification types
At least one nucleotide modification is selected from a group of specific nucleotide modification types, including 2′-fluoro (2′-F) and 2′-O-methyl (2′-OMe) embodiments described on consecutive motifs.
Phosphorothioate and methylphosphonate linkage features
The dsRNA agent includes phosphorothioate internucleotide linkages and methylphosphonate internucleotide linkages, with a cited 6-8 phosphorothioate internucleotide linkage constraint.
Strand length constraint (19-25 nucleotides per strand)
Each strand is independently 19-25 nucleotides long.
GalNAc derivative ligand
The ligand is an N-acetylgalactosamine (GalNAc) derivative, including monovalent and branched multivalent linker embodiments.
Cell inhibition via dsRNA contacting and mRNA transcript degradation
A method to inhibit expression of a contact activation pathway gene in a cell by contacting the cell with the dsRNA agent (or a salt thereof) and maintaining the cell long enough to degrade the gene’s mRNA transcript.
Overall, the claim coverage is anchored on a sequence-defined, fully nucleotide-modified dsRNA agent for Factor XII (F12) with a 3′-terminal ligand on the sense strand, and it further narrows to specific modification, linkage, strand length, and GalNAc derivative features, together with a cell-based method for mRNA transcript degradation.
Stated Advantages
Inhibiting expression of Factor XII (F12).
Decreasing bradykinin levels and/or decreasing F12 activity.
Treating or preventing symptoms of thrombophilia and hereditary angioedema (HAE).
Decreasing mRNA remaining for the targeted gene in in vitro screening.
Dose-response activity is reported via IC50 values for the screened siRNA duplexes.
Dose-dependent reduction of F12 mRNA/plasma protein.
Normalization of vascular permeability in mouse HAE models.
Non-human primate durability of F12 protein reduction.
Enhanced dsRNA activity is stated in connection with motifs comprising three identical nucleotide modifications on three consecutive nucleotides at or near the Dicer cleavage site.
Degrades a contact activation pathway gene mRNA transcript in a cell after contacting and maintaining the cell.
Provides therapeutic and prophylactic use for contact activation pathway-associated diseases such as hereditary angioedema (HAE), thrombophilia, and prekallikrein deficiency.
Documented Applications
Treating or preventing symptoms of thrombophilia and hereditary angioedema (HAE) using dsRNA to inhibit contact activation pathway gene expression, including KLKB1, F12, and/or KNG1.
Inhibiting expression of a contact activation pathway gene in a cell by contacting the cell with the dsRNA agent and degrading the gene’s mRNA transcript.
In vitro screening of siRNA duplexes in Hep3b cells and primary mouse hepatocytes, with mRNA remaining measured for targeted genes.
In vitro screening for KNG1 (kininogen 1) siRNA duplexes, with dose-response IC50 values reported.
Embodiments involving vectors and cells comprising RNAi, and use in connection with pharmaceutical compositions.
F12 silencing in Hep3b single-dose F12 silencing screen.
In vivo efficacy in mouse HAE models, including reduction of F12 mRNA/plasma protein and normalization of vascular permeability.
Durability of F12 protein reduction in non-human primates.
Primary mouse hepatocyte screening.
Potency effect studies related to 5′ antisense vinylphosphate modification.
In vivo gene-silencing application to contact activation pathway genes KLKB1, F12, and KNG1, including inhibition of gene expression via degradation of the mRNA transcript after contacting cells with the dsRNA agent and maintaining the cells long enough.
Therapeutic and prophylactic treatment of contact activation pathway-associated diseases including hereditary angioedema (HAE).
Therapeutic and prophylactic treatment of contact activation pathway-associated diseases including thrombophilia.
Therapeutic and prophylactic treatment of contact activation pathway-associated diseases including prekallikrein deficiency.
Interested in licensing this patent?