Compositions containing nucleic acid nanoparticles with modular functionality
Inventors
Rushworth, James Luke • Foot, George William • Rosell, Anna Perdrix • Brzosko, Zuzanna Aleksandra
Assignees
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Publication Number
US-12472267-B2
Publication Date
2025-11-18
Expiration Date
Abstract
The invention provides compositions containing cargo molecules attached to elements that improve the function of the cargo molecules in the body of a subject. The compositions are useful for therapeutic and diagnostic purposes. Furthermore, the invention outlines ways in which these compositions can be produced; the core molecule can be functionalized, via bioorthogonal click chemistry, in such a way as to impart modular characteristics. This functionalization simultaneously allows for loading of biologically relevant cargo and provides stabilization to the overall structure of the molecule.
Core Innovation
The invention relates to a composition comprising an RNA oligonucleotide covalently linked to one or more cargo molecules. The cargo molecules are an siRNA duplex molecule or an ASO, and the cargo molecules are covalently linked to the RNA via an orthogonal click chemistry reaction between a modified nucleotide at a terminal end of the RNA and a modified nucleotide at a terminal end of the siRNA duplex or ASO.
The disclosed chemistry includes stimulus-responsive and cleavable linker concepts and covalent stabilization approaches for forming the conjugate. The composition further supports orthogonal terminal-end conjugation using CuAAC and SPAAC, together with additional orthogonal chemistries such as IEDDA and SuFEx/SPANC, and terminal nucleotide modifications including DBCO, tetrazine/TCO, azide, and norbornene/alkyne derivatives, as well as PEG linker variants.
The invention additionally describes multi-cargo architectures in which an oligonucleotide is linked to a first cargo molecule and a second cargo molecule is linked to the first, with optional further cargo molecules linked to the first and/or second. In some embodiments, the composition is provided as a nucleic acid nanoparticle tertiary structure with three or more junctions formed by at least two oligonucleotide strands interacting through hydrogen bonding and/or base-stacking interactions.
Claims Coverage
The provided claim coverage centers on covalently linked RNA–cargo compositions using orthogonal click chemistry between terminal-modified nucleotides. The inventive features also include multi-cargo connectivity, cleavable or dicer-substrate and extended spacer regions, and nucleic acid nanoparticle tertiary structure constraints; across the provided scope, the key inventive features number four.
Orthogonal terminal-end RNA-to-siRNA/ASO covalent click conjugation
An oligonucleotide that is RNA covalently linked to one or more cargo molecules, wherein the cargo molecules are an siRNA duplex molecule or an ASO, and the cargo molecules are covalently linked to the oligonucleotide via an orthogonal click chemistry reaction between a modified nucleotide at a terminal end of the RNA and a modified nucleotide at a terminal end of the siRNA duplex molecule or ASO.
Selected orthogonal click chemistry reaction from enumerated types
The orthogonal click chemistry reaction is selected from a specified group of named click/ligation reaction types.
Multi-cargo architecture connected via first and second cargo linkage
The oligonucleotide is attached to a first cargo molecule, with a second cargo molecule linked to the first, and additional cargo molecules are linked to the first or second cargo molecule.
Cleavable dicer substrate or extended nucleic acid spacer region linking cargo
At least one of the first and second cargo molecules is linked to a third cargo molecule via a dicer substrate or an extended nucleic acid spacer region that can be cleaved.
Nucleic acid nanoparticle tertiary structure with hydrogen bonding/base-stacking junctions
A nucleic acid nanoparticle composition in which the nanoparticle is a tertiary structure with three or more junctions formed by at least two oligonucleotide strands that interact with other strands via hydrogen bonding and/or base-stacking interactions.
Overall, the claim set as provided centers on RNA oligonucleotide covalent conjugates to siRNA duplex or ASO cargo molecules using orthogonal terminal-end click chemistry between modified terminal nucleotides. Refinements cover selected orthogonal click/ligation reaction types, multi-cargo architectures, cleavable dicer-substrate or extended spacer regions, and nucleic acid nanoparticle tertiary structures defined by junction count and strand interactions.
Stated Advantages
Enables covalent linking of RNA to siRNA duplex or ASO cargo molecules via orthogonal click chemistry between terminal modified nucleotides.
Provides specific orthogonal terminal-end conjugation chemistries and linker moieties for constructing the RNA–cargo conjugates.
Supports multi-cargo architectures in which additional cargo molecules can be linked through first and second cargo connectivity.
Incorporates cleavable or dicer-substrate/extended spacer regions that can be cleaved for cargo linkage.
Allows formation of nucleic acid nanoparticle tertiary structures with junctions formed by strand interactions via hydrogen bonding and/or base-stacking.
Documented Applications
Gene knockdown/biological activity is described for conjugated siRNA constructs, including PLK1 mRNA knockdown with a reported reduction range.
Receptor targeting and internalization-related modalities are described conceptually for conjugates, including ASGPR targeting using GalNAc, and endosomal/escape-related contexts.
Multi-cargo conjugate use is described conceptually for combining RNA/siRNA cargo with other cargo types, including receptor-binding ligands and other payload categories.
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