Modular functional peptides for the intracellular delivery of nanoparticles
Inventors
Delehanty, James B. • Medintz, Igor L. • Mattoussi, Hedi M. • Deschamps, Jeffrey R. • Dawson, Glyn • Dawson, Philip E. • Blanco-Canosa, Juan Bautista • Boeneman, Kelly
Assignees
University of Chicago • US Department of Navy
Publication Number
US-8409858-B2
Publication Date
2013-04-02
Expiration Date
2030-10-20
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Abstract
Described are peptides for delivery of a nanoparticle to the cytosol, the peptide comprising: (a) a nanoparticle association domain; (b) a proline-rich spacer domain; (c) an uptake domain; and (d) a vesicle escape domain comprising a non-hydrolyzable lipid moiety, wherein the spacer domain is between the nanoparticle association domain and the uptake and vesicle escape domains, and wherein the peptide, when attached to an extracellular nanoparticle, is effective to induce uptake of the nanoparticle by a cell and delivery of the nanoparticle to the cytosol of the cell. Also described are methods of delivery of a nanoparticle to the cytosol of a cell, the method comprising providing to a cell a nanoparticle attached to such a peptide. Exemplary nanoparticles include quantum dots.
Core Innovation
The invention describes modular peptides designed for the intracellular delivery of nanoparticles, such as quantum dots, into the cytosol of cells. These peptides comprise four distinct functional domains arranged in a specific order: a nanoparticle association domain, a spacer domain, an uptake domain, and a vesicle escape domain containing a non-hydrolyzable lipid moiety or perfluoro group. When attached to an extracellular nanoparticle, these peptides effectively induce cellular uptake and facilitate delivery of the nanoparticle into the cytosol, overcoming typical cellular barriers.
The problem being addressed stems from the limitations of existing nanoparticle intracellular delivery methods. Traditional passive and facilitated delivery approaches result predominantly in nanoparticle entrapment within endolysosomal vesicles, necessitating methods to liberate nanoparticles into the cytosol, which are often toxic or invasive. Active delivery methods, such as electroporation and microinjection, while delivering nanoparticles directly to the cytosol, are highly invasive and reduce cell viability. Therefore, there is a need for a delivery system that is efficient, non-toxic, minimally invasive, and capable of mediating both cellular uptake and subsequent endosomal escape of nanoparticles.
Claims Coverage
The claims cover several inventive features related to modular peptides for nanoparticle delivery and associated methods, with multiple independent claims detailing these features.
Peptide for cytosolic delivery of nanoparticles with ordered multi-domain structure
A peptide comprising, in order, a nanoparticle association domain, a spacer domain, an uptake domain, and a vesicle escape domain containing a non-hydrolyzable lipid moiety or perfluoro group, effective to induce cellular uptake and cytosolic delivery of the attached nanoparticle.
Spacer domain consisting of proline residues
The spacer domain specifically consists of from six to fifteen proline residues to serve as a rigid or extended linker between the nanoparticle association and uptake/escape domains.
Vesicle escape domain comprising a non-hydrolyzable lipid moiety
The vesicle escape domain contains a non-hydrolyzable lipid moiety, such as palmitoyldiamoniproprionyl, which facilitates escape from endocytic vesicles into the cytosol.
Nanoparticle association domain comprising polyhistidine
The nanoparticle association domain includes polyhistidine sequences enabling noncovalent metal ion coordination with nanoparticles for assembly.
Modular peptide sequences and specific compositions
Peptides comprising particular sequences including polyhistidine, 8-10 proline residues in the spacer, the uptake domain with sequence VKIKK (SEQ ID No: 7), and the vesicle escape domain with non-hydrolyzable palmitic acid, e.g., peptides of SEQ ID No: 3 or 4.
Method of nanoparticle delivery to cytosol using attached peptides
Methods of delivering nanoparticles to the cytosol of a cell by providing nanoparticles attached to the described modular peptides and allowing cellular uptake, where the nanoparticles can be quantum dots or other nano-scale materials.
The claims collectively define peptides with a modular domain structure enabling nanoparticle attachment, cellular uptake, and vesicle escape via a non-hydrolyzable lipid moiety, along with compositions of such peptides attached to nanoparticles and methods for their cytosolic delivery.
Stated Advantages
Efficient cellular uptake of nanoparticles mediated by peptides with modular functional domains.
Facilitation of endosomal escape of nanoparticles, enabling delivery into the cytosol rather than sequestration in endolysosomal vesicles.
Minimized cytotoxicity relative to other delivery methods such as electroporation, polymers, or chemical agents.
Modular design allows for iterative development and adaptation of peptide domains to optimize uptake and escape.
Documented Applications
Intracellular labeling and visualization of cellular structures using quantum dots.
Use in drug delivery and drug monitoring by delivering nanoparticles carrying therapeutic agents.
Cellular tagging for sorting and cataloging of cells.
In vivo tissue and cellular labeling for biological imaging.
Development of biological labels, biosensors, and probes combining nanoparticles and peptides.
Potential extension to delivery of proteins, carbohydrates, contrast agents, nucleic acids, and gene therapy agents.
Interested in licensing this patent?