Use of single dendritic wedge cell penetrating peptides to facilitate cellular delivery of nanoparticles and nanoparticles carrying cargos
Inventors
Medintz, Igor L. • Delehanty, James B. • Breger, Joyce • Muttenthaler, Markus • Dawson, Philip E.
Assignees
Publication Number
US-10183080-B2
Publication Date
2019-01-22
Expiration Date
2037-09-01
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Abstract
Nanoparticles (and optionally a cargo such as a drug) can be delivered to cells by attaching just a single dendritic peptide to the nanoparticle. The dendritic peptide includes a polyhisitidine motif and a hinge and a spacer connecting the polyhistidine to a lysine-based dendritic wedge displaying at least two copies of a cell-penetrating peptide motif.
Core Innovation
The invention relates to facilitating cellular delivery of nanoparticles by attaching a single dendritic peptide to the nanoparticle. This dendritic peptide includes a polyhistidine motif, a hinge, and a spacer connecting the polyhistidine to a lysine-based dendritic wedge that displays at least two copies of the cell-penetrating peptide motif RRRRRRRRRFG (SEQ ID No: 2). Efficient intracellular delivery is achieved with just one dendritic peptide having multivalent cell-penetrating peptide (CPP) branches.
The problem being addressed is that conventional nanoparticle-mediated drug delivery typically requires decorating the nanoparticle surface with multiple copies (10 to 20 or more) of CPPs to achieve efficient cellular uptake. This high avidity requirement consumes significant nanoparticle surface area and cargo carrying capacity. Therefore, there is a need to improve nanoparticle delivery by reducing the number of CPP copies required while maintaining or improving uptake efficiency, thus increasing the available surface area for therapeutic cargo attachment.
The invention synthesizes modular, branched dendritic peptides that display increasing numbers of polyarginine CPP motifs (e.g., 1×, 2×, 4×, 8×, 16×) using oxime ligation to attach polyarginine branches to a lysine-based dendritic scaffold. This dendritic wedge peptide strategy enhances nanoparticle cellular uptake, allowing uptake to be achieved with a single peptide displaying as few as two polyarginine branches. This innovation frees more nanoparticle surface area for cargo attachment and enables delivery of large cargoes (over 600 kDa protein) with just one dendritic peptide per nanoparticle after short incubation times.
Claims Coverage
The claims include three independent claims that focus on methods of delivering a nanoparticle to a cell using a single dendritic peptide bound to the nanoparticle. These claims specify structural features of the dendritic peptide and the conditions of nanoparticle binding and cellular entry.
Use of a single dendritic peptide for nanoparticle delivery
A method of delivering a nanoparticle to a cell by contacting a living cell with a nanoparticle bound to a single dendritic peptide, resulting in nanoparticle entry into the cell.
Dendritic peptide structural features
The dendritic peptide comprises a polyhistidine (or hexahistidine) motif; a hinge; a spacer connecting the polyhistidine to a lysine-based dendritic wedge displaying at least two copies of the peptide sequence RRRRRRRRRFG (SEQ ID No: 2). In some embodiments, the spacer includes six prolines and two glycines.
Nanoparticle free from other cell-penetrating peptides
In one method, the nanoparticle is free of any other cell-penetrating peptides aside from the single dendritic peptide bound to it.
Co-delivery of cargo with the nanoparticle
The nanoparticle may also be bound to a cargo which is delivered to the cell along with the nanoparticle.
The independent claims cover methods of delivering nanoparticles to cells using a single dendritic peptide characterized by a polyhistidine motif and a dendritic lysine wedge displaying multiple copies of a specific polyarginine peptide. The claims encompass structural features of the dendritic peptide, delivery of nanoparticles free from other CPPs, and co-delivery of cargo, illustrating the innovation’s focus on efficient cellular delivery with minimal peptide decoration.
Stated Advantages
Ability to achieve nanoparticle cellular uptake with a single dendritic peptide displaying as few as two polyarginine branches after short incubation (30 minutes).
Increased nanoparticle cargo carrying capacity by reducing the number of cell-penetrating peptides needed on the nanoparticle surface.
Capability to deliver large protein cargo (>600 kDa) per nanoparticle with only one dendritic peptide and 1 hour incubation.
Simplistic, efficient synthetic assembly strategy producing monodisperse peptide-like dendrimers.
Spacer region extends dendrimer away from nanoparticle surface ensuring polyarginine availability for cell surface binding.
Biocompatibility and minimal cytotoxicity at effective concentrations and exposure times.
High affinity binding to nanoparticle surfaces due to hexahistidine motif and enhanced binding to cell membranes through localized arginine residues.
Modular design allowing synthesis with different chemical handles and applicability to a variety of nanoparticle materials.
Documented Applications
Facilitating cellular delivery of nanoparticles such as semiconductor quantum dots (QDs) for therapeutic or diagnostic applications.
Delivery of nanoparticles carrying cargos including peptides, drugs (e.g., doxorubicin-labeled peptides), and large proteins (e.g., mCherry fluorescent protein) into cells.
Delivery systems applicable to nanomaterials with surface ligands that enable coordination with dendritic peptides, including hard metals, polymers, and other nano-scale materials.
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