Peptide hydrogels and use thereof
Inventors
Schneider, Joel • Walsh, Scott • Miller, Stephen • Yamada, Yuji • Durum, Scott • Andrews, Caroline • Li, Wenqing • Hixon, Julie • Tau, Steven
Assignees
US Department of Health and Human Services
Publication Number
US-11661439-B2
Publication Date
2023-05-30
Expiration Date
2037-12-17
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Abstract
This disclosure provides novel anionic amphiphilic β-hairpin peptides that self-assemble under appropriate conditions to form a reversible gel-sol hydrogel that can be used, for example, to readily deliver protein therapeutics and cells by injection to a target location in a subject.
Core Innovation
This disclosure provides novel anionic amphiphilic β-hairpin peptides that self-assemble under appropriate conditions to form reversible gel-sol hydrogels. These hydrogels can be used to readily deliver protein therapeutics and cells by injection to a target location in a subject. The peptides form an amphiphilic β-hairpin conformation in aqueous solution at physiological ionic strength and pH, resulting in fibrillar networks and hydrogels with shear-thin recovery properties enabling syringe delivery and subsequent gel reforming.
The problem being solved relates to current injectable hydrogel systems used for local delivery of protein therapeutics that have limitations in biocompatibility, predictably controlling protein release, and require individual hydrogel optimization for each protein. Additionally, prior synthetic hydrogels for cell encapsulation and 3D culturing lacked support for cell adhesion and proliferation, restricting their utility for delivering cell-based therapeutics.
The invention addresses these by providing amphiphilic β-hairpin peptides that form hydrogels exhibiting tunable protein release in vivo and supporting 3D cell culture matrices free of non-human components. These hydrogels promote cell growth and proliferation, maintain encapsulated cells within target tissue after injection, and are cytocompatible and biocompatible, showing minimal immune response on implantation.
Claims Coverage
The patent includes multiple independent claims covering isolated peptides, peptide hydrogels formed therefrom, peptides fused to integrin binding peptides, hydrogels comprising therapeutic proteins or cells, and methods of administration and culturing. The main inventive features involve the peptide sequence design, amphiphilic β-hairpin conformation, reversible hydrogel formation, incorporation of heterologous proteins or cells, and integrin-binding fusion for cell culture and delivery.
Isolated peptide with defined amino acid sequence forming amphiphilic β-hairpin conformation
An isolated peptide comprising or consisting of an amino acid sequence defined by specific patterns of L, I, T, V residues and negatively charged residues (D, E, S) arranged to form an amphiphilic β-hairpin in aqueous solution at physiological pH and ionic strength, with the ability to fold and self-assemble into fibrillar networks.
Peptide hydrogel exhibiting reversible gel-sol and sol-gel phase transitions
A peptide hydrogel formed from the isolated peptide that undergoes a gel-sol phase transition upon application of shear stress and a sol-gel transition upon removal of shear, having a storage modulus greater than 40 Pascal, forming in physiological conditions with 0.25-4.0% w/v peptide, 10 to 400 mM NaCl, and pH 7 to 9.
Peptide hydrogel incorporating heterologous proteins with tunable retention
A peptide hydrogel comprising at least one heterologous protein dispersed within, where the protein can be a therapeutic protein such as a cytokine (e.g., IL-7), and may be fused to a peptide tag with net positive or negative charge to modulate retention within the anionic hydrogel matrix.
Isolated peptide fused to integrin binding peptide for cell culture and delivery
An isolated peptide fused to an integrin binding peptide (e.g., RGDV) optionally via a heterologous peptide linker, forming an amphiphilic β-hairpin conformation and capable of self-assembly into a hydrogel that supports mammalian cell encapsulation, proliferation, and retention via integrin-mediated cell binding.
Peptide hydrogel comprising mammalian cells expressing integrins for retention and delivery
A peptide hydrogel formed with the integrin binding peptide-fused peptide that includes mammalian cells, such as CAR T cells, dispersed within, where cell-surface integrins bind the peptide ligand to increase retention of cells in the hydrogel after administration.
Methods and syringes for administrating peptide hydrogels
Methods of administering heterologous proteins or cells encapsulated in the peptide hydrogels to subjects, for instance by injection of syringes pre-loaded with the peptide hydrogels containing therapeutic proteins or cells to a target site.
The inventive features encompass the design of amphiphilic β-hairpin peptides forming reversible hydrogels capable of encapsulating therapeutic proteins and integrin-binding peptides for cell adhesion and proliferation. These hydrogels have tunable mechanical and release properties, enabling delivery of proteins and cells with improved biocompatibility and retention in vivo.
Stated Advantages
The hydrogels provide tunable protein release in vivo without requiring redesign of the hydrogel for each protein.
Hydrogels support 3D cell culture matrices free of non-human media components and promote cell growth and proliferation in vitro.
Hydrogels are cytocompatible and biocompatible, eliciting minimal lymphocytic inflammatory response in vivo.
Shear-thinning and rapid recovery properties allow easy delivery by syringe injection and localized gel reforming.
Increased retention of cells and proteins at the target location enhances therapeutic efficacy.
Documented Applications
Local delivery of encapsulated protein therapeutics to tissues to reduce systemic toxicity and improve therapeutic outcomes.
Controlled, tunable release of therapeutic proteins such as cytokines including IL-7 for immune modulation.
Encapsulation, 3D culture, and expansion of mammalian cells, including anchorage-dependent cells like fibroblasts, in chemically defined hydrogels.
Delivery of therapeutic cells, such as CAR T cells, by injection in hydrogels containing integrin binding peptides to enhance cell retention at target sites.
Syringe-based injectable hydrogels for localized administration of cells or proteins in vivo.
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