Enhanced targeting platform
Inventors
Saltzman, W. Mark • Tietjen, Gregory T. • Koide, Shohei • Albert, Claire • Pober, Jordan • Koide, Akiko • Bracaglia, Laura
Assignees
Yale University • New York University NYU
Publication Number
US-12162953-B2
Publication Date
2024-12-10
Expiration Date
2040-12-04
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Abstract
A platform technology provides particle and nucleic acid conjugates, and compositions thereof, with enhanced targeting to cells, tissues, organs. The particles and nucleic acids and other deliverables contain a synthetic binding protein such as a polypeptide monobody covalently conjugated to the surface of the particle or the nucleic acid, for linking a targeting agent to the particle's surface or the nucleic acid. The particles and nucleic acids and other deliverables optionally contain an antibody non-covalently conjugated to the binding protein, via an Fc domain of the antibody. The particles can include therapeutic agents, diagnostic agents, prophylactic agents, or a combination thereof, to be delivered to desired cells, tissues, and/or organs. The particles and nucleic acids and other deliverables can be used in a wide array of applications including, but not limited to, ex vivo perfusion of mammalian organs and in vivo disease treatment.
Core Innovation
The invention provides a platform technology comprising particles and nucleic acid conjugates that are functionalized with synthetic binding proteins, such as polypeptide monobodies, covalently conjugated to their surfaces. These synthetic binding proteins have a targeting agent binding site capable of binding to the Fc domain of an antibody, while being covalently linked to the particle or nucleic acid at a chemically defined location, commonly via a cysteine residue. This structure ensures enhanced spatial orientation and preserves the binding activity of the targeting agent on the deliverable surface.
A key problem addressed by this invention relates to prior methods of particle targeting that rely on direct chemical conjugation of targeting agents (such as antibodies) to particles using amine-coupling chemistries. Such methods result in random orientation of the targeting agents and lack adaptability, leading to compromised targeting efficacy and limited rapid design. The invention overcomes these issues by utilizing synthetic binding proteins as universal linkers, enabling modular, site-specific, and adaptable conjugation of a wide variety of targeting agents to particle surfaces without the need to re-engineer each targeting molecule.
The particles and nucleic acids so constructed can be loaded with therapeutic, diagnostic, or prophylactic agents and used for enhanced targeting to cells, tissues, or organs. The targeting agent, such as an antibody, can be bound non-covalently to the synthetic binding protein via the Fc domain, or the synthetic binding protein itself can serve as the targeting agent. The platform is compatible with various particle materials, including polymeric, liposomal, and inorganic types, and allows for site-specific delivery and improved adaptability for therapeutic or diagnostic interventions.
Claims Coverage
The patent claims coverage centers on five main inventive features involving functionalized particles with synthetic binding proteins, precise conjugation chemistry, adaptable use of monobody linkers, specific polypeptide sequences, and broad material/cargo compatibility.
Particles functionalized with monobodies having distinct binding and conjugation sites
The invention provides a composition comprised of a particle functionalized with one or more monobodies. Each monobody includes: - A first surface with a targeting agent binding site capable of binding to an Fc domain of an antibody. - A second surface comprising a chemical moiety used to conjugate the monobody to the particle's surface. The monobody's dual-surface design enables both site-specific particle attachment and modular antibody-based targeting.
Defined polypeptide sequences for monobody functionalization
The monobodies used for functionalization possess a polypeptide comprising the amino acid sequence of SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4, or a variant thereof with at least 90% sequence identity (with or without N- or C-terminal tags). This feature defines the precise molecular identity and scope of the synthetic binding protein component in the functional system.
Covalent linkage via specific chemical structures and defined amino acids
The conjugation of the monobody to the particle's surface is defined as a covalent linkage that comprises: - The structure —X—Ra—Y— (Formula I), with X and Y independently containing 3–90 atoms and Ra chosen from specific moieties such as a 3-thiopyrrolidine-2,5-dione, 3-aminopyrrolidine-2,5-dione, or other listed groups. - The chemical moiety on the monobody being an amino acid selected from cysteine, lysine, ornithine, arginine, serine, threonine, tyrosine, or combinations thereof, preferably a cysteine. This enables precise, site-specific, and reproducible attachment of the monobody.
Adaptability to various particle types and inclusion of additional agents
The inventive composition encompasses particles that can be polymeric particles, liposomes, inorganic particles, or expression vectors. These particles can comprise one or more therapeutic agents, diagnostic agents, prophylactic agents, or combinations thereof. The platform thus ensures compatibility across numerous particle types and applications.
Methods for delivering functionalized particles using antibody targeting
The invention covers methods for delivering the functionalized particles to a subject or to organs, tissues, or cells for transplantation. The methods include administering an effective amount of the composition and an antibody, either sequentially (with the antibody first) or together. Ex vivo application to organs is specifically claimed.
In summary, the claims protect the use of synthetic binding protein-functionalized particles enabling modular antibody targeting, with site-specific conjugation chemistries, using defined monobody polypeptide sequences, broadly applicable to various particle materials and cargos, and covering both compositions and methods for targeted delivery.
Stated Advantages
Provides significantly enhanced targeting efficiency of particles to desired cells, tissues, or organs compared to conventional methods.
Allows modular and adaptable conjugation, enabling rapid development of targeted particles using already available antibodies without re-engineering each targeting molecule.
Ensures proper orientation and display of targeting agents (such as antibodies), leading to improved binding and reduced off-target effects.
Permits use of a broad range of particle types and cargos, including therapeutic, diagnostic, and prophylactic agents.
Facilitates high binding affinity and robust targeting in diverse experimental and physiological conditions, including static cultures, flow conditions, and ex vivo organ perfusion.
Enables use of any nanoparticles presenting a thiol reactive group on the surface, thereby providing compatibility across different particle materials.
Streamlines formulation, as development of the targeted nanoparticles needs to be done only once for a given antibody isotype, with facile change of target or species.
Documented Applications
Targeted delivery of therapeutic, diagnostic, or prophylactic agents to cells, tissues, and organs in vivo.
Ex vivo perfusion of mammalian organs, such as in organ or tissue transplant settings.
Controlled release of active agents in a targeted manner to enhance treatment efficacy and reduce toxicity.
In vitro or ex vivo delivery of drugs or nucleic acids to primary or cultured cells for applications including cell therapy and immunomodulation.
Preparation of CAR-T cells and other cell manipulations via nucleic acid or drug delivery.
Transfection of cells with polynucleotides for gene therapy or disease treatment.
Targeted delivery to endothelial cells relevant for treating cancer, stroke, thrombosis, ischemia reperfusion injury, or improving organ transplant outcomes.
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