Viral nanoparticle multimers
Inventors
Steinmetz, Nicole F. • Wen, Amy M.
Assignees
Case Western Reserve University
Publication Number
US-12359175-B2
Publication Date
2025-07-15
Expiration Date
2033-12-27
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Abstract
Virus particle multimers and methods of making and using such virus particle multimer are described. Virus particle multimers are constructed by preparing a plurality of asymmetrically functionalized virus particles bearing one or more functional groups and contacting the asymmetrically functionalized virus particles with a first linker molecule that reacts with the functional groups to form a virus particle multimer that includes a plurality of asymmetrically functionalized virus particles connected by the linker molecule. The asymmetrically functionalized virus particles are typically prepared by attaching the virus particles to a support surface to allow asymmetrical functionalization to be introduced.
Core Innovation
The invention describes virus particle multimers constructed by preparing a plurality of asymmetrically functionalized virus particles bearing one or more functional groups and contacting these virus particles with a first linker molecule that reacts with the functional groups to form a virus particle multimer. These multimers include a plurality of asymmetrically functionalized virus particles connected by the linker molecule. The asymmetrically functionalized virus particles are typically prepared by attaching the virus particles to a support surface to facilitate asymmetrical functionalization.
The technical problem addressed is the challenge of creating mesoscale nanostructured materials with high precision manufacturing technologies that allow robust large-scale assembly and spatial control at the nanometer level. Prior bottom-up techniques often result in symmetric nanoparticle assemblies limiting hierarchical assembly possibilities. There is also a need to eliminate confounding variables like surface charge to better understand nanoparticle interactions with cells and to prepare assemblies that control size, shape, and spatial organization of targeting molecules for improved delivery and imaging applications.
The invention provides a novel method of producing nanoparticle clusters and chains of varying aspect ratio from identical, asymmetrically functionalized virus particles. This symmetry-breaking approach enables higher spatial control over nanoparticle interconnection, allowing the formation of versatile modular nanoparticle assemblies via reaction with polymers containing appropriate functionalities. The method is demonstrated with plant virus particles, especially cowpea mosaic virus, to prepare dimers and larger multimers, including multifunctional assemblies displaying targeting molecules and fluorescent dyes with controlled spatial distribution. These assemblies show enhanced cell uptake and highlight the influences of shape and targeting molecule organization on biological interaction.
Claims Coverage
The patent includes 2 independent claims covering the composition of plant virus particle multimers and methods of delivering cargo molecules using these multimers.
Asymmetrically functionalized plant virus particle multimers prepared by support surface attachment
A plant virus particle multimer comprising a plurality of asymmetrically functionalized plant virus particles bearing one or more functional groups disposed asymmetrically, connected by a linker molecule, formed by attaching plant virus particles to a support surface for asymmetrical functionalization and releasing them, with the preparation done at physiological pH. The asymmetric functional groups inhibit further cluster formation and aggregation.
Method of delivering cargo molecules to a cell using asymmetrically functionalized plant virus particle multimers
A method of delivering a cargo molecule to a cell by contacting the cell with a plant virus particle multimer, prepared by forming asymmetrically functionalized plant virus particles attached to a support surface to introduce asymmetrical functionalization and releasing them, then reacting with linker molecules to form multimers, where asymmetrically disposed functional groups inhibit additional aggregation, and the step of preparing is performed at physiological pH.
The claims focus on compositions of asymmetrically functionalized plant virus particle multimers prepared via support surface asymmetrical functionalization and methods using these multimers for delivery of cargo molecules including imaging and antitumor agents, emphasizing inhibition of aggregation and preparation at physiological pH.
Stated Advantages
Virus particle multimers exhibit significantly more uptake by cells compared to single virus particles, especially when bearing targeting molecules such as RGD peptides.
The asymmetrical functionalization prevents uncontrolled aggregation and allows for precise spatial control in nanoparticle assembly.
The method is modular and versatile, suitable for a wide selection of nanoparticles and allows formation of complex three-dimensional nanoparticle assemblies.
Maintaining identical particle composition ensures consistent charge and surface properties, eliminating confounding variables in cellular interaction studies.
The assemblies show enhanced pharmacokinetic and flow properties and reduced clearance by macrophages compared to single particles.
Documented Applications
Use of virus particle multimers for delivering cargo molecules such as imaging agents or therapeutic (antitumor) agents to cells.
Imaging of tissues in vivo using multimers loaded with imaging agents detectable by fluorescent imaging, MRI, PET, CT, or optical imaging.
Treatment and imaging of cancer cells, including various tumors such as colon, brain, breast, fibrosarcoma, and squamous carcinoma.
Targeting inflamed tissue such as atherosclerotic blood vessels for imaging and treatment.
Use in ex vivo applications for imaging or killing cancer cells in tissues or organs maintained viable outside the body.
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