Compositions and methods for inhibiting expression of transthyretin
Inventors
Sah, Dinah Wen-Yee • Hinkle, Gregory • Alvarez, Rene • Milstein, Stuart • Chen, Qingmin
Assignees
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Publication Number
US-12227744-B2
Publication Date
2025-02-18
Expiration Date
Abstract
The invention relates to a double-stranded ribonucleic acid (dsRNA) targeting a transthyretin (TTR) gene, and methods of using the dsRNA to inhibit expression of TTR.
Core Innovation
The invention relates to double-stranded RNA (dsRNA) duplexes designed to inhibit transthyretin (TTR) expression. The dsRNA comprises a sense strand and an antisense strand forming a double stranded region, and the disclosure provides sequence-specific constraints tied to TTR targeting, including specified SEQ ID NOs and chemically modified strand variants.
The pharmaceutical composition includes dsRNA together with a lipid formulation comprising a cationic lipid, a non-cationic lipid, and a polyethyleneglycol (PEG)-lipid. The disclosure also includes chemically modified human TTR dsRNA candidates, including 2′-O-methyl substitutions and dTdT 3′ extensions, as well as dsRNA design aspects using transcript sources and off-target prediction scoring to select candidate duplex oligos.
The disclosure further addresses TTR gene expression inhibition through TTR gene mRNA transcript degradation in the context of TTR-mediated diseases. It includes evaluation of selected duplexes for knockdown performance, immunostimulation assessment, and in vivo efficacy, with results reported across human, rat, transgenic mouse, non-human primate, and mutant V30M models using LNP/SNALP formulations.
Claims Coverage
The claim set covers two inventive features centered on TTR-targeting dsRNA: a sequence-length/sequence-content constrained pharmaceutical composition and a lipid formulation with defined lipid classes. Dependent refinements specify particular SEQ ID pairings, sequence constraints, and dsRNA chemical modification requirements, together with a method for inhibiting TTR gene expression in a cell.
TTR-inhibiting dsRNA composition with defined strand lengths and antisense sequence constraints
A pharmaceutical composition comprising dsRNA for inhibiting expression of transthyretin (TTR), where the dsRNA comprises a sense strand and an antisense strand forming a double stranded region, each strand is 19, 20, 21, 22, 23, or 24 nucleotides in length, and the antisense strand comprises 15 or more contiguous nucleotides of SEQ ID NO: 170, SEQ ID NO:450, SEQ ID NO:730, or SEQ ID NO: 1010.
Lipid formulation combining cationic lipid, non-cationic lipid, and PEG-lipid with the dsRNA
The pharmaceutical composition of the dsRNA includes a lipid formulation comprising a cationic lipid, a non-cationic lipid, and a polyethyleneglycol (PEG)-lipid.
Sense-strand sequence constraint using specified SEQ ID NOs
The sense strand contains at least 15 contiguous nucleotides matching one of SEQ ID NO:169, SEQ ID NO:449, SEQ ID NO:729, or SEQ ID NO:1009.
Specific sense/antisense SEQ ID pairing
The sense strand is SEQ ID NO:449 and the antisense strand is SEQ ID NO:450.
Antisense sequence constraint together with dsRNA chemical modification requirement
The antisense strand includes at least 15 contiguous nucleotides of SEQ ID NO: 170 and the dsRNA contains at least one modified nucleotide.
Cell-contacting and time-maintenance for degrading TTR gene mRNA transcripts
A method for inhibiting TTR gene expression in a cell by contacting the cell with the pharmaceutical composition and maintaining the treated cell long enough to degrade the TTR gene mRNA transcript.
Overall, the claims cover a TTR-targeting dsRNA pharmaceutical composition constrained by defined strand length and specified contiguous antisense and/or sense sequence identities, formulated with a lipid mixture comprising a cationic lipid, a non-cationic lipid, and a PEG-lipid. The disclosure also includes a cell-treatment method centered on maintaining contact long enough for degradation of TTR gene mRNA transcripts to inhibit TTR gene expression.
Stated Advantages
Inhibits expression of transthyretin (TTR).
Degrades the TTR gene mRNA transcript to inhibit TTR gene expression.
Dose-dependent reduction of liver TTR mRNA in in vivo models.
Reduction of plasma TTR protein where measured.
Lead TTR dsRNA duplexes show dose-response IC50 values in the pM range.
No significant TNF-α/IFN-α induction by human PBMC in the immunostimulation assessment.
Documented Applications
Treating TTR-mediated diseases by targeting TTR gene expression inhibition, including TTR amyloidosis (e.g., FAP/FAC/CNS amyloidosis and related forms described in the disclosure).
Patient identification and diagnosis in the context of TTR-mediated disease (e.g., family history, DNA test, biopsy/amyloid plaques as described).
TTR inhibition using human, rat, and TTR mutant (V30M) dsRNA duplexes, including evaluations in transgenic mice and non-human primates, using LNP/SNALP formulations.
In vitro TTR knockdown assessment in HepG2, Hep3B, and a rat cell line.
Immunostimulation assessment using human PBMC to evaluate TNF-α/IFN-α induction.
A brief human treatment description is included.
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