Cell-penetrating peptides

Patent metadata

Field	Value
Publication number	US-12472264-B2
Application number	US-201917266940-A
Country	US
Kind code	B2
Filing date	Aug 9, 2019
Priority date	Aug 9, 2018
Publication date	Nov 18, 2025
Grant date	Nov 18, 2025

Abstract

the present invention relates to peptides, in particular cell-penetrating peptides, having a first hydrophobic domain positioned at the C-terminus of the peptide and a second hydrophobic domain positioned at the N-terminus of the peptide, and to conjugates of such cell-penetrating peptides with a therapeutic molecule. The present invention further relates to use of such peptides or conjugates in methods of treatment or as a medicament, especially in the treatment of genetic disorders and in particular muscular dystrophies such as Duchenne muscular dystrophy.

First claim

The invention claimed is:

1 . A peptide having a total length of 40 amino acid residues or less, the peptide comprising:
one cationic domain, wherein the cationic domain comprises RBRXRBRXB (SEQ ID NO: 1), RBRXRBRXBRXRB (SEQ ID NO:2), RBRXRBRXBR (SEQ ID NO:3), RBRRXRBRXBRXRB (SEQ ID NO:4), RBRXRBRBRXRB (SEQ ID NO:5), RBRXRBRBRBRB (SEQ ID NO:6), RBRBRBRBRBRB (SEQ ID NO:7), RBRXRBRBRXR (SEQ ID NO:8), RBRRBRBRBRRB (SEQ ID NO:9), RBRRBRBRXBRXRB (SEQ ID NO:10), RBRRBRBRBBRXRB (SEQ ID NO:11), RBRRBRBRBBRBRB (SEQ ID NO:12), or any combination thereof; and two hydrophobic domains, wherein each hydrophobic domain independently comprises YQFLI (SEQ ID NO:13), FQILY (SEQ ID NO:14), ILFQY (SEQ ID NO:15), or any combination thereof; wherein one of the hydrophobic domains is positioned at the C-terminus of the peptide and one of the hydrophobic domains is positioned at the N-terminus of the peptide, and wherein the two hydrophobic domains flank the cationic domain, wherein the N-terminus of the peptide is optionally acetylated.

2 . A peptide according to claim 1 , wherein each hydrophobic domain has a length of 5 amino acids.

3 . The peptide according to claim 1 wherein the cationic domain has a length of between 9 to 20 amino acid residues.

4 . A peptide according to claim 1 , wherein the cationic core domain consists of RBRXRBRXB (SEQ ID NO:1), RBRXRBRXBRXRB (SEQ ID NO:2), RBRXRBRXBR (SEQ ID NO:3), RBRRXRBRXBRXRB (SEQ ID NO:4), RBRXRBRBRXRB (SEQ ID NO:5), RBRXRBRBRBRB (SEQ ID NO:6), RBRBRBRBRBRB (SEQ ID NO:7), RBRXRBRBRXR (SEQ ID NO: 8), RBRRBRBRBRRB (SEQ ID NO:9), RBRRBRBRXBRXRB (SEQ ID NO:10), RBRRBRBRBBRXRB (SEQ ID NO:11), or RBRRBRBRBBRBRB (SEQ ID NO:12), and the two hydrophobic arm domains each independently consist of YQFLI (SEQ ID NO:13), FQILY (SEQ ID NO:14), or ILFQY (SEQ ID NO:15).

5 . A peptide according to claim 1 , wherein the peptide consists of one of the following sequences: YQFLIRBRRXRBRXBRXRBYQFLI (SEQ ID NO:19), YQFLIRBRRBRBRBRRBYQFLI (SEQ ID NO:24), and YQFLIRBRRBRBRBBRXRBYQFLI (SEQ ID NO: 26), wherein the N-terminus of the peptide is optionally acetylated.

6 . A conjugate comprising the peptide of claim 1 covalently linked to a therapeutic molecule.

7 . The conjugate according to claim 6 further comprising a linker covalently linking the peptide, wherein the linker links the conjugate to the therapeutic molecule.

8 . The conjugate according to claim 6 , wherein the linker is selected from G, BC, XC, C, GGC, BBC, BXC, XBC, X, XX, B, BB, BX and XB.

9 . The conjugate according to claim 6 , wherein the therapeutic molecule is a nucleic acid, peptide nucleic acid, antisense oligonucleotide, short interfering RNA, micro RNA, peptide, cyclic peptide, protein, pharmaceutical or drug.

10 . A method for the treatment of DMD, the method comprising administering the conjugate of claim 6 to a subject in need thereof.

11 . A peptide according to claim 1 , wherein the peptide consists of one of the following sequences: YQFLIRBRXRBRXBYQFLI (SEQ ID NO:16), YQFLIRBRXRBRXBRXRBYQFLI (SEQ ID NO: 17), YQFLIRBRXRBRXBRYQFLI (SEQ ID NO:18), YQFLIRBRXRBRBRXRBYQFLI (SEQ ID NO: 20), YQFLIRBRXRBRBRBRBYQFLI (SEQ ID NO:21), YQFLIRBRBRBRBRBRBYQFLI (SEQ ID NO: 22), YQFLIRBRXRBRBRXRYQFLI (SEQ ID NO:23), YQFLIRBRRBRBRXBRXRBYQFLI (SEQ ID NO: 25), YQFLIRBRRBRBRBBRBRBYQFLI (SEQ ID NO:27), FQILYRBRRBRBRBBRBRBFQILY (SEQ ID NO:28), YQFLIRBRRXRBRXBRXRBFQILY (SEQ ID NO:29), wherein the N-terminus of the peptide is optionally acetylated.

12 . A peptide according to claim 1 , wherein the peptide consists of the amino acid sequence YQFLIRBRXRBRXBYQFLI (SEQ ID NO:16), wherein the N-terminus of the peptide is optionally acetylated.

13 . A peptide according to claim 1 , wherein the peptide consists of the amino acid sequence YQFLIRBRXRBRXBRXRBYQFLI (SEQ ID NO:17), wherein the N-terminus of the peptide is optionally acetylated.

14 . A peptide according to claim 1 , wherein the peptide consists of the amino acid sequence YQFLIRBRXRBRXBRYQFLI (SEQ ID NO:18), wherein the N-terminus of the peptide is optionally acetylated.

15 . A peptide according to claim 1 , wherein the peptide consists of the amino acid sequence YQFLIRBRRXRBRXBRXRBYQFLI (SEQ ID NO:19), wherein the N-terminus of the peptide is optionally acetylated.

16 . A peptide according to claim 1 , wherein the peptide consists of the amino acid sequence YQFLIRBRXRBRBRXRBYQFLI (SEQ ID NO:20), wherein the N-terminus of the peptide is optionally acetylated.

17 . A peptide according to claim 1 , wherein the peptide consists of the amino acid sequence YQFLIRBRXRBRBRBRBYQFLI (SEQ ID NO:21), wherein the N-terminus of the peptide is optionally acetylated.

18 . A peptide according to claim 1 , wherein the peptide consists of the amino acid sequence YQFLIRBRBRBRBRBRBYQFLI (SEQ ID NO:22), wherein the N-terminus of the peptide is optionally acetylated.

19 . A peptide according to claim 1 , wherein the peptide consists of the amino acid sequence YQFLIRBRXRBRBRXRYQFLI (SEQ ID NO:23), wherein the N-terminus of the peptide is optionally acetylated.

20 . A peptide according to claim 1 , wherein the peptide consists of the amino acid sequence YQFLIRBRRBRBRBRRBYQFLI (SEQ ID NO:24), wherein the N-terminus of the peptide is optionally acetylated.

21 . A peptide according to claim 1 , wherein the peptide consists of the amino acid sequence YQFLIRBRRBRBRXBRXRBYQFLI (SEQ ID NO:25), wherein the N-terminus of the peptide is optionally acetylated.

22 . A peptide according to claim 1 , wherein the peptide consists of the amino acid sequence YQFLIRBRRBRBRBBRXRBYQFLI (SEQ ID NO:26), wherein the N-terminus of the peptide is optionally acetylated.

23 . A peptide according to claim 1 , wherein the peptide consists of the amino acid sequence YQFLIRBRRBRBRBBRBRBYQFLI (SEQ ID NO:27), wherein the N-terminus of the peptide is optionally acetylated.

24 . A peptide according to claim 1 , wherein the peptide consists of the amino acid sequence FQILYRBRRBRBRBBRBRBFQILY (SEQ ID NO:28), wherein the N-terminus of the peptide is optionally acetylated.

25 . A peptide according to claim 1 , wherein the peptide consists of the amino acid sequence YQFLIRBRRXRBRXBRXRBFQILY (SEQ ID NO:29), wherein the N-terminus of the peptide is optionally acetylated.

26 . The conjugate according to claim 6 , wherein the therapeutic molecule is an antisense oligonucleotide.

27 . The conjugate according to claim 26 further comprising a linker covalently linking the peptide, wherein the linker links the conjugate to the therapeutic molecule.

28 . The conjugate according to claim 27 , wherein the linker is selected from G, BC, XC, C, GGC, BBC, BXC, XBC, X, XX, B, BB, BX and XB.

29 . The conjugate according to claim 28 , wherein the peptide consists of an amino acid sequence selected from: YQFLIRBRXRBRXBYQFLI (SEQ ID NO:16), YQFLIRBRXRBRXBRXRBYQFLI (SEQ ID NO: 17), YQFLIRBRXRBRXBRYQFLI (SEQ ID NO:18), YQFLIRBRRXRBRXBRXRBYQFLI (SEQ ID NO: 19), YQFLIRBRXRBRBRXRBYQFLI (SEQ ID NO:20), YQFLIRBRXRBRBRBRBYQFLI (SEQ ID NO: 21), YQFLIRBRBRBRBRBRBYQFLI (SEQ ID NO:22), YQFLIRBRXRBRBRXRYQFLI (SEQ ID NO: 23), YQFLIRBRRBRBRBRRBYQFLI (SEQ ID NO:24), YQFLIRBRRBRBRXBRXRBYQFLI (SEQ ID NO: 25), YQFLIRBRRBRBRBBRXRBYQFLI (SEQ ID NO:26), YQFLIRBRRBRBRBBRBRBYQFLI (SEQ ID NO:27), FQILYRBRRBRBRBBRBRBFQILY (SEQ ID NO:28), and YQFLIRBRRXRBRXBRXRBFQILY (SEQ ID NO:29), wherei

Assignees

• Univ Oxford Innovation Ltd
• Res & Innovation Uk

Inventors

• Wood Matthew
• El Andaloussi Samir
• Mcclorey Graham
• Manzano Raquel
• Gait Michael J
• Godfrey Caroline
• Arzumanov Andrey
• O'Donovan Liz
• Raz Richard

Classifications

• C12N2320/32

  Special delivery means, e.g. tissue-specific · CPC title

• C12N2310/3513

  Protein; Peptide · CPC title

• C12N2310/11

  Antisense · CPC title

• C12N15/113

  Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)} · CPC title

• C07K2319/10

  containing a tag for extracellular membrane crossing, e.g. TAT or VP22 · CPC title