Polynucleotide constructs
Inventors
Sakamuri, Sukumar • Bradshaw, Curt W. • Eltepu, Laxman • Meade, Bryan R. • Lam, Son
Assignees
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Publication Number
US-11981703-B2
Publication Date
2024-05-14
Expiration Date
Abstract
Disclosed are polynucleotide constructs having a strand linked to a moiety carrying one or more auxiliary moieties. Also disclosed are polynucleotide constructs interrupted with a sugar analogue, and polynucleotide constructs with stereochemical{circumflex over ( )} enriched phosphorothioates. The polynucleotide constructs may be provided as hybridized polynucleotide constructs. Also featured are methods of delivery a polynucleotide construct to a cell and methods of reducing the expression of a protein in a cell by contacting the cell with the disclosed polynucleotide construct or hybridized polynucleotide construct.
Core Innovation
The invention relates to polynucleotide constructs comprising a linker conjugated to a polynucleotide and to a plurality of targeting moieties. Prior to conjugation of the targeting moieties, the linker has the structure X2, and conjugation occurs through one or more alkyne moieties of X2. The targeting moieties comprise N-acetyl galactosamine (GalNAc), and the constructs couple multiple GalNAc targeting moieties to the polynucleotide construct.
The disclosed embodiments describe linker conjugation arrangements in which a single X2 linker is used for attaching multiple GalNAc targeting moieties to the polynucleotide construct. Dependent refinements specify click reaction conjugation and additional chemical features of the X2 linker, including a terminal phosphotriester. The targeting moieties are further described as named NAG moieties, including NAG26, NAG37, NAG38, and NAG39.
The disclosure also describes broader nucleic-acid architecture associated with the constructs, including passenger and guide strands and RISC loading. In the provided material, additional structural terminology includes internucleoside abasic spacers, modified nucleosides, stereochemically enriched phosphorothioates, and non-bioreversible internucleoside linkages, together with auxiliary moieties such as cell-penetrating peptides and endosomal escape moieties.
Claims Coverage
The consolidated claim coverage centers on one independent claim directed to a polynucleotide construct or salt thereof with an X2 linker conjugated to a polynucleotide and to a plurality of GalNAc targeting moieties through one or more alkyne moieties of X2. Dependent features refine the conjugation chemistry, linker composition, targeting-moiety selection, and nucleic-acid architecture; six inventive features are present.
X2 linker conjugated to multiple GalNAc targeting moieties
A polynucleotide construct or salt thereof comprising a linker conjugated to a polynucleotide and to a plurality of targeting moieties, wherein prior to conjugation the linker has the structure X2 and each targeting moiety comprises N-acetyl galactosamine (GalNAc).
Alkyne moiety conjugation
Conjugation of the targeting moieties to the linker occurs through one or more of the alkyne moieties of X2.
Click reaction conjugation
The X2 linker is conjugated to targeting moieties using a click reaction.
Terminal phosphotriester
The X2 linker has a terminal phosphotriester.
Exactly one X2 linker
The polynucleotide construct includes exactly one X2 linker.
Specified NAG targeting moieties and passenger strand length
Targeting moieties are selected from NAG26, NAG37, NAG38, and NAG39, and a passenger strand is 15 to 32 nucleotides long.
Overall, the claims define a polynucleotide construct with an X2 linker that attaches multiple GalNAc targeting moieties through alkyne moieties, with dependent refinements for click reaction conjugation, a terminal phosphotriester, exactly one X2 linker, selected NAG variants, and a passenger strand length constraint.
Stated Advantages
Reduced off-target effects.
Improved efficacy/extended duration of action.
Documented Applications
Targeting in the context of polynucleotide constructs with guide strands loaded into a RISC complex.
In vitro and in vivo target gene knockdown is supported by activity evidence, including AT3 and TTR target gene knockdown metrics.
Time-course in vivo mouse studies are supported by reported in vivo time courses.
Stereochemistry effects are supported by reported stereochemical results.
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