Polymerized peptide and gel having collagen-like structure

Patent metadata

Field	Value
Publication number	US-10851152-B2
Application number	US-201615579286-A
Country	US
Kind code	B2
Filing date	Jun 23, 2016
Priority date	Jun 25, 2015
Publication date	Dec 1, 2020
Grant date	Dec 1, 2020

Abstract

The present invention is produced as a polymerized peptide prepared by oxidatively cross-linking collagen-like peptides having the triple-helical structure containing three peptide chains each containing multiple cysteine (Cys) residues. This polymerized peptide can be processed into a hydrogel and a thin membrane sheet before use, and is capable of imparting a particular physiological function such as cellular adhesion by incorporating functional amino acid sequences present on biological polymers. Further, the present invention provides a method for producing the abovementioned polymerized peptide and the like; and use applications employing the same, such as cell culturing, wound dressing, and compositions including regenerative medical materials and research materials.

First claim

The invention claimed is:

1. A gelling agent comprising at least one kind of a polymerized peptide that has, as a polymerization unit, a triple-chain peptide having a triple-helical structure, and is polymerized by oxidative cross-linking, wherein peptide chains composing the triple-chain peptide are identical to or different from one another, and each peptide chain has:
one or more triple helix forming peptidyl groups each having a repeating structure in which -(Xaa-Yaa-Gly)- as a basic unit is repeated at least 5 times; and
one or more cross-link forming peptidyl groups each comprising at least two cysteine (Cys) residues within 6 residues from each of an amino-terminus and a carboxy-terminus, and wherein
each of Xaa and Yaa is independently selected from a proline (Pro or P) residue, a hydroxyproline (Hyp or O) residue, an arginine (Arg or R) residue, a lysine (Lys or K) residue, a valine (Val or V) residue, a leucine (Leu or L) residue, an isoleucine (Ile or I) residue, a serine (Ser or S) residue, a threonine (Thr or T) residue, an alanine (Ala or A) residue, a glycine (Gly or G) residue, an N-isobutyl glycine residue, a phenylalanine (Phe or F) residue, a methionine (Met or M) residue, a glutamate (Glu or E) residue, an aspartate (Asp or D) residue, an asparagine (Asn or N) residue, a glutamine (Gln or Q) residue, a histidine (His or H) residue, a tryptophan (Trp or W) residue or a tyrosine (Tyr or Y) residue, the proline residue may be modified by an amino group or fluorine atom.

2. The gelling agent according to claim 1 , comprising at least one kind of the polymerized peptide polymerized with the peptide chains selected from the group consisting of the following (i) to (iii):
(i) a peptide chain comprising at least one of the triple helix forming peptidyl group (s) and at least one of the cross-link forming peptidyl group(s);
(ii) a peptide chain comprising at least one of the triple helix forming peptidyl group(s), at least one of the cross-link forming peptidyl group(s) and at least one peptidyl group(s) having a motif having a bioactivity; and
(iii) a peptide chain comprising at least one of the triple helix forming peptidyl group (s), at least one of the cross-link forming peptidyl group(s), and at least one peptidyl group(s) having an amino acid residue linking a biologically active motif to a side chain thereof via a linker.

3. The gelling agent according to claim 2 , comprising at least one kind of a copolymerized peptide produced by combining not less than two kinds of the biologically active motifs.

4. A method for producing the gelling agent as set forth in claim 1 , comprising:
dissolving into a solvent identical or different peptide chains each having a repeating structure in which (Xaa-Yaa-Gly) as a basic unit is repeated at least 5 times, comprising at least two cysteine (Cys) residues within 6 residues from each of an amino-terminus and a carboxy-terminus;
forming a triple-chain peptide having a triple-helical structure comprised of three of the peptides, by self-assembly; and
performing polymerization by oxidative cross-linking with the triple-chain peptide being a polymerization unit, wherein each of Xaa and Yaa is independently selected from a pro line (Pro or P) residue, a hydroxyproline (Hyp or O) residue, an arginine (Arg or R) residue, a lysine (Lys or K) residue, a valine (Val or V) residue, a leucine (Leu or L) residue, an isoleucine (lie or I) residue, a serine (Ser or S) residue, a threonine (Thr or T) residue, an alanine (Ala or A) residue, a glycine (Gly or G) residue, an N-isobuty glycine residue, a phenylalanine (Phe or F) residue, a methionine (Met or M) residue, a glutamate (Glu or E) residue, an aspartate (Asp or D) residue, an asparagine (Asn or N) residue, a glutamine (Gin or Q) residue, a histidine (His or H) residue, a tryptophan (Trp or W) residue or a tyrosine (Tyr or Y) residue, the proline residue may be modified by an amino group or fluorine atom.

5. A gel comprising at least one kind of the gelling agents as set forth in claim 1 .

6. The gel according to claim 5 , comprising a gelling agent prepared by combining at least two kinds of triple-chain peptides with the triple-chain peptide as set forth in claim 1 being a polymerization unit.

7. The gel according to claim 5 , further comprising at least one kind of a triple-chain peptide comprising no cysteine residue.

8. The gel according to claim 5 , wherein the gel is a hydrogel used as a base material for culturing selected cells.

9. A polymerized peptide thin membrane produced by drying the gel as set forth in claim 5 .

10. A regenerative medical material comprising the gel as set forth in claim 5 or the polymerized peptide thin membrane as set forth in claim 9 .

11. The regenerative medical material according to claim 10 , wherein the regenerative medical material is a composition for promoting wound healing.

Assignees

• Univ Waseda
• Kola Gen Pharma Inc

Inventors

• Koide Takaki
• ICHISE Shinichiro
• Takeuchi Shungo
• Nose Hiroshi

Classifications

• C07K14/00

  Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof · CPC title

• A61L31/145

  Hydrogels or hydrocolloids · CPC title

• A61L31/047

  Other specific proteins or polypeptides not covered by A61L31/044 - A61L31/046 · CPC title

• A61L26/008

  Hydrogels or hydrocolloids · CPC title

• A61L26/0047

  Specific proteins or polypeptides not covered by groups A61L26/0033 - A61L26/0042 · CPC title