Ultrasound-sensitive peptide particles for spatially resolved molecule delivery

Patent metadata

Field	Value
Publication number	US-12064514-B2
Application number	US-201816634679-A
Country	US
Kind code	B2
Filing date	Jul 30, 2018
Priority date	Jul 28, 2017
Publication date	Aug 20, 2024
Grant date	Aug 20, 2024

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.

First claim

The invention claimed is:

1. An amphiphilic peptide represented by Formula (III):
HB-CL-HP-NH2  (III)

wherein HB is a fluorinated hydrophobic block consisting of three to five consecutively connected pentafluorinated hydrophobic amino acid residues;
wherein CL is an amino acid sequence consisting of two to 10 amino acid residues, at least two of which are cross-linking cysteine residues;
wherein HP is a hydrophilic amino acid sequence, and
wherein said amphiphilic peptide consists of 8 to 30 total amino acid residues.

2. The amphiphilic peptide according to claim 1 , wherein HB consists of three, four or five consecutively connected pentafluoro-phenylalanine residues, and is located at the N-terminal end of the peptide sequence.

3. The amphiphilic peptide according to claim 1 , wherein HP comprises lysine, glycine, arginine, aspartic acid, or any combination thereof.

4. The amphiphilic peptide according to claim 1 , wherein HP comprises the sequence KGRGD (SEQ ID NO: 35), where K is lysine, G is glycine, R is arginine, and D is aspartic acid.

5. The amphiphilic peptide according to claim 1 , wherein CL comprises GGGCCGG (SEQ ID NO: 46), where G is glycine and C is cysteine.

6. The amphiphilic peptide according to claim 1 , wherein said hydrophilic amino acid sequence of HP comprises a targeting motif.

7. The amphiphilic peptide according to claim 1 , wherein said hydrophilic amino acid sequence comprises a conserved targeting motif selected from the group consisting of: HGK, RGD, KAR, RSR, KAA, RGRR(SEQ ID NO:1), RGRRS (SEQ ID NO:2), YQLDV (SEQ ID NO:3), EYQ, RPM, PSP, VGVA (SEQ ID NO:4), NGR, CRKRLDRNC (SEQ ID NO:43), EFEEFEIDEEEK (SEQ ID NO:44), and DFEEIPEEYLQ (SEQ ID NO:45).

8. The amphiphilic peptide according to claim 1 , wherein said hydrophilic amino acid sequence comprises a hydrophilic amino acid sequence selected from the group consisting of: GHGKHKNK (SEQ ID NO:5), CRGDKGPDC (SEQ ID NO:6), CKGAKAR (SEQ ID NO:7), CRVSRQNKC (SEQ ID NO:8), CGGERGKSC (SEQ ID NO:9), CRSRKG (SEQ ID NO:10), CKAAKN (SEQ ID NO:11), CRGRRST (SEQ ID NO:12), CRGRRT (SEQ ID NO:50), CEYQLDVE (SEQ ID NO:13), TVRTSAD (SEQ ID NO:14), PIEDRPM (SEQ ID NO:15), ALRDRPM (SEQ ID NO:16), PEKFRPM (SEQ ID NO:17), IKVGKLQ (SEQ ID NO:18), SVSVGMKPSPRP (SEQ ID NO:19), VPEQRPM (SEQ ID NO:20), CAKIDPELC (SEQ ID NO:21), CSNIDARAC (SEQ ID NO:22), RLQLKL (SEQ ID NO:23), PMMRQRPM (SEQ ID NO:24), AKATCPA (SEQ ID NO:25), QPPMEYS (SEQ ID NO:26), SISSLTD (SEQ ID NO:27), FRVGVADV (SEQ ID NO:28), CNGRCVSGCAGRC (SEQ ID NO:29), NWGDRIL (SEQ ID NO:30), CVSNPRWKC (SEQ ID NO:31), CDCRGDCFC (SEQ ID NO:32), YSAYPDSVPMMS (SEQ ID NO:33), and PLASRPM (SEQ ID NO:34).

9. The amphiphilic peptide according to claim 1 , wherein said hydrophilic amino acid sequence comprises a hydrophilic amino acid sequence selected from the group consisting of: KGRGD (SEQ ID NO:35), RGDS (SEQ ID NO:36), GRGD (SEQ ID NO:37), GRGDS (SEQ ID NO:38), GRGDSP (SEQ ID NO:39), GRGDSPK (SEQ ID NO:40), GRGDNP (SEQ ID NO:41), and GRGDTP (SEQ ID NO:42).

10. The amphiphilic peptide according to claim 1 , wherein said amphiphilic peptide comprises an amphiphilic peptide represented by Formula (IV) or Formula (V):
F F F F F F GGGCCGGKGRGD (IV) (SEQ ID NO: 47), F F F F F F F F GGGCCGGKGRGD-NH2 (V) (SEQ ID NO:49), wherein F F is pentafluoro-phenylalanine, G is glycine, C is cysteine, K is lysine, G is glycine, R is arginine, and D is aspartic acid.

11. The amphiphilic peptide according to claim 1 , wherein said amino acid sequence of CL consists of two to 10 amino acid residues and said hydrophilic amino acid sequence of HP consists of 3 to 15 hydrophilic amino acids;
wherein said amphiphilic peptide consists of 10 to 30 total amino acid residues.

12. The amphiphilic peptide according to claim 1 , wherein the amphiphilic peptide has a molecular weight in the range of about 2000-5000 daltons, wherein the amphiphilic peptide includes at least eight amino acid residues, and a total number of no more than 30 amino acid residues, wherein at least two of the amino acid residues are directly connected consecutively by peptide bonds without any intervening amino acid residues.

Assignees

• Penn State Res Found
• Us Health
• The Us Secretary Department Of Health And Human Services Office Of Technology Transfer National Inst

Inventors

• Schneider Joel P
• Medina Scott H

Classifications

• C07K2319/01

  containing a localisation/targetting motif · CPC title

• C07K7/06

  having 5 to 11 amino acids · CPC title

• B82Y5/00

  Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery · CPC title

• A61K9/0009

  involving or responsive to electricity, magnetism or acoustic waves; Galenical aspects of sonophoresis, iontophoresis, electroporation or electroosmosis · CPC title

• A61K47/65

  Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers · CPC title