Ultrasound-sensitive peptide particles for spatially resolved molecule delivery
Inventors
Schneider, Joel P. • Medina, Scott H.
Assignees
Penn State Research Foundation • Office of Technology Transfer
Publication Number
US-12064514-B2
Publication Date
2024-08-20
Expiration Date
2038-07-30
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Abstract
The present invention is directed to peptisomes, including nanopeptisomes, which have a perfluorocarbon liquid core containing a perfluorocarbon liquid and a cargo, such as a therapeutically active agent, dispersed in the perfluorocarbon. liquid, and a plurality of amphiphilic peptide molecules surrounding the perfluorocarbon liquid core, wherein the amphiphilic peptide is represented by Formula (I) HB-CL-HP wherein HB is a fluorinated hydrophobic block, such as a fluorinated hydrophobic amino acid sequence, CL is a cross-linking motif, and HP is a hydrophilic amino acid sequence. The present invention is also directed to methods of use of the amphiphilic peptides and peptisomes, such as nanopeptisomes, to deliver a cargo, such as a therapeutically active agent, to a cell, Q wherein the cell may be in vitro, ex vivo, or in vivo.
Core Innovation
The invention relates to peptisomes, including nanopeptisomes, which are assemblies comprising a perfluorocarbon liquid core containing a therapeutically active agent or cargo dispersed therein, surrounded by a plurality of amphiphilic peptides. These amphiphilic peptides have a defined structure represented by Formula (I): HB-CL-HP, where HB is a fluorinated hydrophobic block, CL is a cross-linking motif, and HP is a hydrophilic amino acid sequence. The amphiphilic peptides assemble at the surface of the perfluorocarbon liquid core to form peptisomes, which are capable of ultrasound-triggered delivery of cargo into cells.
The problem addressed by the invention is the challenge in delivering protein and nucleic acid-based therapeutic agents effectively into disease tissues, particularly intracellularly. Existing biocarriers rely on endocytic transport leading to degradation in endosomal compartments and require harsh formulation methods, often damaging biological cargo. There is a need for a delivery vehicle capable of bypassing the endocytic pathway, achieving direct cytoplasmic delivery with spatial and temporal control, and preserving cargo activity. The invention provides such a solution through the design of amphiphilic peptides that assemble into ultrasound-activated peptisomes enabling direct intracellular delivery, avoiding endosomal uptake and degradation.
Claims Coverage
The patent contains one independent claim directed to an amphiphilic peptide with specific structural features enabling assembly into nanopeptisomes and ultrasound-triggered cargo delivery. The main inventive features relate to the peptide composition, structure, and ultrasound-activated delivery mechanism.
Amphiphilic peptide structure and composition
An amphiphilic peptide represented by Formula (III): HB-CL-HP-NH2, where HB is a fluorinated hydrophobic block comprising three to five consecutively connected pentafluorinated hydrophobic amino acid residues, CL is an amino acid sequence with two to ten residues including at least two cross-linking cysteine residues, HP is a hydrophilic amino acid sequence, and the peptide consists of 8 to 30 total amino acid residues.
Fluorinated hydrophobic block specifics
The HB consists of three, four, or five consecutively connected pentafluoro-phenylalanine residues located at the N-terminal end of the peptide.
Hydrophilic amino acid sequence features
The HP includes lysine, glycine, arginine, aspartic acid or any combination thereof, and can comprise the sequence KGRGD (SEQ ID NO: 35). The HP can include a conserved targeting motif selected from a defined group of peptide sequences known for targeting various cells or tissues.
Cross-linking motif characteristics
The CL comprises the sequence GGGCCGG (SEQ ID NO: 46), containing two or more cysteines enabling disulfide cross-linking between amphiphilic peptides for particle stabilization.
Assembly capability and ultrasound activation
The amphiphilic peptide is capable of assembling at the surface of a perfluorocarbon liquid to form a nanopeptisome, which upon ultrasound application causes phase transition of the perfluorocarbon liquid core, leading to particle rupture and direct intracellular delivery of cargo.
The claims cover an amphiphilic peptide with defined fluorinated hydrophobic, cross-linking, and hydrophilic domains that assemble into nanopeptisomes with perfluorocarbon liquid cores. These particles enable ultrasound-triggered delivery of therapeutic cargo directly into target cells, providing a novel drug delivery system addressing limitations of prior biocarriers.
Stated Advantages
Ultrasound-mediated delivery yields spatially and temporally controlled cargo release into the cytoplasm, avoiding endosomal degradation.
The amphiphilic peptides can be chemically synthesized in high yield and purity using standard solid-phase techniques despite high fluorination.
The assembly method is mild and avoids harsh synthetic conditions that could degrade biologics.
The size of peptisomes can be precisely controlled for various applications, including imaging and therapy.
Nanopeptisomes remain stable under physiological conditions and storage, enabling practical use.
The system accommodates a broad range of cargo including proteins, peptides, nucleic acids, small molecules, and imaging agents.
Peptisomes can be targeted to different cells or tissues via hydrophilic sequence motifs for enhanced specificity.
Peptisomes provide theranostic capabilities combining therapeutic delivery and ultrasound imaging guidance.
Documented Applications
Direct intracellular delivery of therapeutic agents including proteins, peptides, nucleic acids, small molecule drugs, gene editing tools into target cells in vitro, ex vivo, and in vivo.
Treatment of cancers such as lung cancer, pancreatic cancer, hepatic cancer, and colorectal cancer by targeted cargo delivery using ultrasound activation.
Imaging and targeted treatment of cardiovascular diseases including atherosclerotic plaques and blood clots using targeted peptisomes and ultrasound imaging modalities.
Treatment of bacterial infections by delivering antibacterial agents via peptisomes targeted to bacteria, activated by ultrasound.
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