EP-4739698-A1 - SELF-HEALING ANTI-MICROBIAL PEPTIDE HYDROGELATORS

Abstract

A hydrogelling peptide having the sequence X1-WTW–Link–WTW-X2 (eg. SWTW–Link– WTWK.) X1 and X2 are independently selected from any amino acid, for example Leucine (L), Isoleucine (I), Valine (V), Alanine (A), Methionine (M), Phenylalanine (F), Tryptophan (W), Proline (P), Glycine (G), Serine (S), Asparagine (N), Glutamine (Q), Threonine (T), Cysteine (C), Tyrosine (Y), Aspartic acid (D), Glutamic acid (E), Lysine (K), Arginine (R) or Histidine (H), where Link is a peptide motif selected to provide the hydrogelling peptide with hydrogelling properties. Link can be any number (e.g. 4-8) amino acid residues, for example, Link is a four amino acid residue of the following formula -aa1-GN-aa2- where aa1 and aa2 are independently selected from E, K, V or Q, (e.g. X1-WTWQGNVWTW-X2). Also the resultant hydrogels and methods of controlling mechanical properties (for example stiffness) of hydrogels and controlling basal-out: apical-out generation of organoids.

Inventors

• NGUYEN, Ashley Khanh Mai
• KILLIAN, Kristopher Alan

Assignees

• NewSouth Innovations Pty Limited

Dates

Publication Date

20260513

Application Date

20240705

Claims (20)

1. 1. A hydrogelling peptide having the sequence SWTW-Link-WTWK, where Link is a peptide motif selected to provide the hydrogelling peptide with hydrogelling properties.
2. 2. A hydrogelling peptide according to claim 1 wherein Link is a peptide motif having any number of amino acid residues.
3. 3. A hydrogelling peptide according to claim 2 wherein the peptide motif has 4, 5, 6, 7 or 8 amino acid residues.
4. 4 A hydrogelling peptide according to claim 1 where Link is a four amino acid residue of the following formula -aa1 -GN-aa2- where aa1 and aa2 are independently selected from E, K, V or Q.
5. 5. A hydrogelling peptide having the sequence X1 -WTW-Link-WTW-X2, wherein Link is a peptide motif selected to provide the hydrogelling peptide with hydrogelling properties and where X1 and X2 are independently selected from any amino acid, for example Leucine (L), Isoleucine (I), Valine (V), Alanine (A), Methionine (M), Phenylalanine (F), Tryptophan (W), Proline (P), Glycine (G), Serine (S), Asparagine (N), Glutamine (Q), Threonine (T), Cysteine (C), Tyrosine (Y), Aspartic acid (D), Glutamic acid (E), Lysine (K), Arginine (R) or Histidine (H).
6. 6. A hydrogelling peptide according to claim 5 wherein Link is a peptide motif having any number of amino acid residues.
7. 7. A hydrogelling peptide according to claim 6 wherein the peptide motif has 4, 5, 6, 7 or 8 amino acid residues.
8. 8 A hydrogelling peptide according to claim 5 where Link is a four amino acid residue of the following formula -aa1 -GN-aa2- where aa1 and aa2 are independently selected from E, K, V or Q.
9. 9. A hydrogelling peptide having the sequence SWTWQGNVWTWK.
10. 10. A hydrogelling peptide of the structure X1 -WTWQGNVWTW-X2, where X1 and X2 are independently selected from any amino acid, for example Leucine (L), Isoleucine (I), Valine (V), Alanine (A), Methionine (M), Phenylalanine (F), Tryptophan (W), Proline (P), Glycine (G), Serine (S), Asparagine (N), Glutamine (Q), Threonine (T), Cysteine (C), Tyrosine (Y), Aspartic acid (D), Glutamic acid (E), Lysine (K), Arginine (R) or Histidine (H).
11. 11. A hydrogelling peptide having the sequence selected from the group consisting of: Sub1- SWTWQGNVWTWK-Sub2, Sub1 -SWTWQGNVWTWK and SWTWQGNVWTWK-Sub2 wherein Sub1 and Sub2 are independently variable substituents.
12. 12. A hydrogelling peptide according to claim 11 wherein Sub1 and/or Sub2 is a peptide substituent which contains the laminin motif -lle-Lys-Val-Ala-Val (IKVAV).
13. 13. A hydrogelling peptide according to claim 11 wherein Sub1 and/or Sub2 substituent is a peptide substituent which contains motif selected from one or more of YIGSR, GRGDSC or GFOGER.
14. 14. A hydrogelling peptide according to claim 11 wherein Sub1 and/or Sub2 is selected from one or more of a small molecule; a protein; an enzyme; a carbohydrate (for example a polysaccharide); a nucleic acid; a synthetic polymer; a natural polymer; a metal nanoparticle; a metal oxide nanoparticle; a lipid nanoparticle, a quantum dot, or a combination thereof.
15. 15. A hydrogelling peptide having the sequence Sub1 -SWTW-Link-WTWK-Sub2, where Link is a peptide motif selected to provide the hydrogelling peptide with hydrogelling properties and Sub1 and Sub2 are independently variable substituents.
16. 16. A hydrogelling peptide according to claim 15 wherein Link is a peptide motif having any number of amino acid residues.
17. 17. A hydrogelling peptide according to claim 16 wherein the peptide motif has 4, 5, 6, 7 or 8 amino acid residues.
18. 18 A hydrogelling peptide according to claim 15 where Link is a four amino acid residue of the following formula -aa1 -GN-aa2- where aa1 and aa2 are independently selected from E, K, V or Q.
19. 19. A hydrogelling peptide according to any one of claims 15 to 18 wherein Sub1 and/or Sub2 is a peptide substituent which contains the laminin motif -lle-Lys-Val-Ala-Val (IKVAV).
20. 20. A hydrogelling peptide according to any one of claims 15 to18 wherein Sub1 and/or Sub2 substituent is a peptide substituent which contains motif selected from one or more of YIGSR, GRGDSC or GFOGER.

Description

SELF-HEALING ANTI-MICROBIAL PEPTIDE HYDROGELATORS Technical Field The invention relates to peptide hydrogelators which mimic natural extracellular matrices while having other desirable properties such as the uniformity of starting material, ease of synthesis, biodegradability, low cytotoxicity, characteristics of yield-stress fluids, and self-healing behavior. Background Natural extracellular matrices are composed of a meshwork of multiple proteins with an interconnected and hierarchical structure that are ideal for guiding tissue assembly. These are predominantly Matrigel or collagen materials. While natural hydrogel materials are the gold standard for 3D cell culture, these materials are derived from animals and are hampered by poor uniformity and batch -to-batch variability. The in vitro use of these matrices also gives rise to concerns of immunogenicity. Thus, the discovery of synthetic alternatives remains a principal goal for cell biologists and tissue engineers. In pursuit of this, hydrogels comprised of self-assembling synthetic peptides have attracted broad interest due to the uniformity of starting material, ease of synthesis, biodegradability, and low cytotoxicity. These synthetic hydrogelators have been designed to form entangled networks of peptide nanofibers that mimic the structural characteristics of native matrices, including mesh size, pore size, and nanofiber architecture. One approach to forming new peptide hydrogelators is via functionalization of ultra-short hydrophobic peptides (2-5 residues), with large N-terminal capping groups that favor selfassembly through pi-stacking interactions. Another approach is through peptide amphiphiles, which are another class of gelators typically formed from longer peptide (16 residues) with alternating charged amino acids that selfassemble through electrostatic interactions. Still further alternative approaches involve rational design of peptide sequences that imitate naturally occurring secondary protein structures, such as the alpha helix or beta sheet. However, despite extensive research into supramolecular peptide assembly, the discovery of new hydrogelators is most often driven by serendipity or permutation of pre-existing gelator sequences. There exists a need for new forms of peptide hydrogelator which can conveniently mimic native extracellular matrices. It is an object of the present invention to overcome or ameliorate at least one of the disadvantages of the prior art, or to provide a useful alternative. Any discussion of the prior art throughout the specification should in no way be considered as an admission that such prior art is widely known or forms part of common general knowledge in the field. Summary of the Invention According to a first aspect, the invention provides a peptide having a sequence SWTWQGNVWTWK. In another aspect, the invention also provides peptides which have a sequence selected from the group consisting of: Sub1-SWTWQGNVWTWK-Sub2, Sub1 -SWTWQGNVWTWK and SWTWQGNVWTWK-Sub2 wherein Sub1 and Sub2 are independently variable peptide substituents. The peptide may, for example comprise the laminin motif at a terminal position-lle-Lys-Val- Ala-Val (IKVAV), i.e., SWTWQGNVWTWKIKVAV Other adhesion sequences could be added at the terminal positions, for example other laminin derived sequences (YIGSR), adhesive fibronectin (RGD) type peptides such as GRGDSC, and other adhesive peptides of the GFOGER type. The terminal modifications need not be restricted to amino acid sequences, but can include other types of species such as small molecules; proteins; enzymes; carbohydrates (for example polysaccharides); nucleic acids; synthetic polymers; natural polymers; metal nanoparticles; metal oxide nanoparticles; lipid nanoparticles; quantum dots; and combinations thereof. The invention also provides a hydrogel comprising a peptide according to any one of the preceding aspects and water. The peptide may be present in an amount of at least 0.1% w/v, or it may be present in an amount of at least 3% w/v. Typically, the concentration range for cell culture applications is around 0.5% w/v. In some embodiments, the peptide is present in an amount up to 5.0% or even up to 10.0% by weight. The balance is generally water but other agents (for example pH adjusters, tonicity adjusters, colourants, bioactive agents) may all be added depending on the intended purpose of the hydrogel. The hydrogel may contain other components depending upon the intended purpose, for example, it may contain one or more of small molecules, proteins, enzymes, carbohydrates (such as polysaccharides), hydrogel forming polymers (such as poly(ethylene glycol) for example. The hydrogel may comprise a peptide having the sequence SWTWQGNVWTWK and a peptide having the sequence SWTWQGNVWTWKIKVAV. These may be present in any desired total amount with respect to the water and they may be present in any desired ratio. For example, the ratio of SWTWQGNVWTWK: SWTWQGNVWTWKIKVAV maybe