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US-12617827-B2 - Self-assembling peptide

US12617827B2US 12617827 B2US12617827 B2US 12617827B2US-12617827-B2

Abstract

An object is to provide a peptide gelling agent which gels under physiological conditions and which has a relatively short chain length, and a sustained-release gel based on the gelling agent. A hydrogelling self-assembling peptide is provided having one or two core peptides with an amino acid sequence of the formula: Xaa-Yaa-Zaa-Yaa-Xaa-Yaa-Zaa-Yaa-Xaa, wherein Xaa is independently Ile or Met, Yaa is independently Asp, Glu, Lys, or Arg, and Zaa is independently Ala or Gly. The full length of the amino acid sequence constituting the self-assembling peptide is 25 amino acids or less.

Inventors

  • Takahiro MURAOKA
  • Atsuya YAGUCHI
  • Itsuki AJIOKA
  • Go Watanabe

Assignees

  • NATIONAL UNIVERSITY CORPORATION TOKYO MEDICAL AND DENTAL UNIVERSITY
  • KANAGAWA INSTITUTE OF INDUSTRIAL SCIENCE AND TECHNOLOGY

Dates

Publication Date
20260505
Application Date
20210729
Priority Date
20200730

Claims (20)

  1. 1 . A hydrogelling self-assembling peptide selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, and SEQ ID NO: 45.
  2. 2 . A fusion peptide formed by linking a functional peptide to the self-assembling peptide of claim 1 .
  3. 3 . The fusion peptide of claim 2 , wherein said linking is mediated by covalent bonding or supramolecular interaction.
  4. 4 . The fusion peptide of claim 2 , wherein the functional peptide is laminin, VEGF, or N-cadherin.
  5. 5 . A gelling agent consisting of the self-assembling peptide of claim 1 or a fusion peptide formed by linking a functional peptide to said self-assembling peptide.
  6. 6 . A gelling composition comprising the gelling agent of claim 5 .
  7. 7 . The gelling composition of claim 6 , comprising two or more of the gelling agents.
  8. 8 . A method of gelling, comprising maintaining the gelling agent of claim 5 or a gelling composition comprising said gelling agent in a sol state, at a temperature not higher than the gelling temperature thereof in water or in an aqueous solution to thereby allow said gelling agent or said gelling composition to gel.
  9. 9 . The method of claim 8 , wherein said temperature at which said gelling agent or said gelling composition is maintained in water or in the aqueous solution is from 4 to 60° C.
  10. 10 . The method of claim 8 , wherein said gelling agent or said gelling composition comprises any one or more negative ions selected from the group consisting of a hydrogen carbonate ion, a carbonate ion, a citrate ion, a tartrate ion, and a sulfate ion.
  11. 11 . The method of claim 8 , wherein said gelling agent or said gelling composition in a gel state has a pH of from 6.0 to 8.0.
  12. 12 . The method of claim 8 , wherein the concentration of said gelling agent and/or said gelling composition is from 0.4% by weight to 10% by weight.
  13. 13 . An artificial extracellular matrix comprising the self-assembling peptide of claim 1 or a fusion peptide formed by linking a functional peptide to said self-assembling peptide, wherein the self-assembling peptide or the fusion peptide is gelled.
  14. 14 . A composition for preparing a sustained-release gel, comprising a gelling agent consisting of a first self-assembling peptide, and a functional molecule formed by linking a second self-assembling peptide and a functional moiety, wherein said first and second self-assembling peptides are selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, and SEQ ID NO: 45, wherein said functional moiety is a functional peptide and/or a low molecular weight compound.
  15. 15 . The composition for preparing a sustained-release gel of claim 14 , comprising two or more of the gelling agents.
  16. 16 . The composition for preparing a sustained-release gel of claim 14 , wherein said linking is mediated by covalent bonding or supramolecular interaction.
  17. 17 . The composition for preparing a sustained-release gel of claim 14 , wherein the functional peptide is selected from the group consisting of a vascular endothelial growth factor (VEGF), a fibroblast growth factor (FGF), and a hepatocyte growth factor (HGF).
  18. 18 . A sustained-release gel comprising the composition for preparing a sustained-release gel of claim 14 .
  19. 19 . A pharmaceutical composition comprising the sustained-release gel of claim 18 .
  20. 20 . A method for producing a sustained-release gel, comprising: mixing the composition for preparing a sustained-release gel of claim 14 , with water or an aqueous solution; and maintaining the mixture obtained after said mixing step, at a temperature not higher than the gelling temperature, to thereby allow the mixture to gel.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application is a 371 of PCT/JP2021/028175, filed Jul. 29, 2021, which claims the benefit of Japanese Patent Application No. 2020-128805, filed Jul. 30, 2020 and Japanese Patent Application No. 2020-146597, filed Sep. 1, 2020. REFERENCE TO A SEQUENCE LISTING SUBMITTED ELECTRONICALLY VIA EFS-WEB The content of the electronically submitted sequence listing, file name: 522-1226_SequenceListing.txt; size: 20,051 bytes; and date of creation: Jan. 6, 2026 filed herewith, is incorporated herein by reference in its entirety. TECHNICAL FIELD The present invention relates to a self-assembling peptide, a gelling agent, a gelling composition comprising a gelling agent as an active ingredient, a composition for preparing a sustained-release gel, a sustained-release gel, and a method for producing a sustained-release gel. BACKGROUND ART Peptides (peptide gelling agents) capable of forming hydrogels have been used as biomaterials excellent in biocompatibility and biodegradability, for cell and tissue culturing, and have been recently applied to the fields of regenerative medicine. For example, applications as scaffold materials (artificial extracellular matrices, cell scaffold materials, and the like) for adhesion with cells, organs, and the like in vivo or in vitro, and as regenerative medicine materials for tissue regeneration are expected. For example, gelatin and collagen extracted from animal cells, and matrigels derived from mouse sarcoma cells have been widely used as peptide gelling agents conventionally utilized, for cell and tissue culturing. However, animal-derived peptide gelling agents are not necessarily uniform in molecular weight distribution and component composition, and have differences in quality of each production lot. Additionally, in the case of introduction in vivo, the risk of causing allergy or unknown infection due to trace components or contaminations incorporated into extracts cannot also be avoided (Non-Patent Literatures 1 to 5). Development of chemically synthesized low-molecular peptide gelling agents (synthetic low-molecular peptide gelling agents) as potent materials capable of overcoming the above problems of animal-derived peptide gelling agents is ongoing. Synthetic low-molecular peptide gelling agents can be synthesized as peptides having particular amino acid sequences, at high purities by solution synthesis or solid-phase synthesis methods. Therefore, there are advantages of stable quality of each production lot and of extremely reduced contaminations. Known synthetic low-molecular peptide gelling agents include (i) an amphiphilic peptide in which one or two amino acids and an aromatic site (cinnamoyl group, Fmoc group, and the like) are linked, (ii) an amphiphilic peptide in which a long alkyl chain (palmitoyl group or the like) is linked to a terminus of a β-sheet forming peptide, and (iii) an amphiphilic peptide in which a hydrophilic amino acid and a hydrophobic amino acid are alternately linked (Non-Patent Literatures 6 to 7). The synthetic low-molecular peptide gelling agent corresponding to above (iii) does not comprise a non-natural backbone and can be composed only of natural amino acids, and thus is considered to be suitable particularly for clinical applications (Non-Patent Literatures 8 to 10). However, synthetic low-molecular peptide gelling agents developed so far have problems of poor gelling in physiological conditions, and of high production cost due to large chain length. For providing a particular function to a peptide gelling agent, it is necessary to combine the peptide gelling agent with a biologically functional molecule such as a protein or to allow sustained release of the combined biologically functional molecule from the gel over a long period of time. For forming complexes with maintaining the activity of the biologically functional molecule, it is necessary to allow for gelling in physiological temperature and physiological pH conditions in which the biologically functional molecule does not denature. For example, (RADA)4 peptides (SEQ ID NO: 19) as representative examples of (iii) described above gel in acidic conditions, but do not gel in physiological pH conditions. In acidic conditions, most of biologically functional molecules such as proteins are denatured, and the activity is reduced or lost. Therefore, it is difficult to form complexes of a biologically functional molecule such as a protein with maintaining the activity in hydrogels formed by (RADA)4 peptides (SEQ ID NO: 19), and availability as a material for regenerative medicine is limited. Furthermore, technologies for allowing sustained release of a complexed biologically functional molecule from a gel over a long period of time are not established. Accordingly, there are needs for a synthetic low-molecular peptide gelling agent which can allow for hydrogel formation in physiological conditions and which has a relatively short chain length, and a