Peptide/β-1,3-glucan complex and production method thereof and pharmaceutical composition containing peptide/β-1,3-glucan complex

Patent metadata

Field	Value
Publication number	US-10195270-B2
Application number	US-201415116306-A
Country	US
Kind code	B2
Filing date	Dec 25, 2014
Priority date	Feb 6, 2014
Publication date	Feb 5, 2019
Grant date	Feb 5, 2019

Abstract

This peptide/β-1,3-glucan complex includes β-1,3-glucan and a peptide/polynucleotide conjugate in which an antigenic peptide is bonded covalently to a polynucleotide or a derivative thereof. The polynucleotide or derivative thereof of the peptide/polynucleotide conjugate bonds via a hydrogen bond with β-1,3-glucan, forming a complex having a triple helix structure including a single molecular chain of the polynucleotide or derivative thereof and two molecular chains of the β-1,3-glucan. Alternatively, the side chain of the β-1,3-glucan and the antigenic peptide are bonded covalently formed by either a cycloaddition reaction between an alkyne and an azide derivative, or a reaction between a maleimide group or a vinyl sulfone group and a thiol group.

First claim

The invention claimed is:

1. A peptide/β-1,3-glucan complex, comprising:
a polysaccharide having a β-1,3-glucan backbone; and
a peptide/polynucleotide conjugate in which an antigenic peptide is bonded covalently to a polynucleotide or derivative thereof, wherein
the polynucleotide or derivative thereof of the peptide/polynucleotide bonds through hydrogen bonding with the polysaccharide having the β-1,3-glucan backbone, and forms a complex having a triple helix structure consisting of one molecular chain of the polynucleotide or derivative thereof and two molecular chains of the polysaccharide having the β-1,3-glucan backbone.

2. The peptide/β-1,3-glucan complex according to claim 1 , wherein the polysaccharide having the β-1,3-glucan backbone is schizophyllan, lentinan, scleroglucan or curdlan.

3. The peptide/β-1,3-glucan complex according to claim 1 , wherein the polynucleotide or derivative thereof is poly-deoxyadenosine.

4. The peptide/β-1,3-glucan complex according to claim 1 , wherein
the polynucleotide or derivative thereof is a polynucleotide derivative in which at least a portion of phosphodiester bonds of DNA or RNA is substituted with a phosphorothioate group.

5. The peptide/β-1,3-glucan complex according to claim 4 , wherein
in the polynucleotide derivative in which the at least the portion of phosphodiester bonds of DNA or RNA is substituted with a phosphorothioate group, at least 50% of the phosphodiester bonds are substituted with the phosphorothioate group.

6. The peptide/β-1,3-glucan complex according to claim 1 , wherein
the polynucleotide or derivative thereof and the antigenic peptide included in the peptide/polynucleotide conjugate are bonded through covalent bonding formed by:
a cycloaddition reaction between an alkyne and an azide derivative,
a reaction between a maleimide group and a thiol group, or
a reaction between a thiol group of a thiol-modified nucleic acid and a thiol group of a peptide C-terminal cysteine residue.

7. The peptide/β-1,3-glucan complex according to claim 6 , wherein the polysaccharide having the β-1,3-glucan backbone is schizophyllan, lentinan, scleroglucan or curdlan.

8. The peptide/β-1,3-glucan complex according to claim 1 , wherein
the peptide/β-1,3-glucan complex further comprises a polynucleotide or derivative thereof having a partial base sequence having immunostimulatory activity;
the polynucleotide or derivative thereof and the polysaccharide having the β-1,3-glucan backbone are bonded by hydrogen bonding to form a complex having a triple helix structure consisting of one molecular chain of the polynucleotide or derivative thereof and two molecular chains of the polysaccharide having the β-1,3-glucan backbone.

9. The peptide/β-1,3-glucan complex according to claim 8 , wherein a portion of the polynucleotide or derivative thereof forming the complex having the triple helix structure is poly-deoxyadenosine.

10. The peptide/β-1,3-glucan complex according to claim 8 , wherein the portion of the polynucleotide or derivative thereof forming the complex having the triple helix structure is a polynucleotide derivative in which at least a portion of phosphodiester bonds of DNA or RNA is substituted with a phosphorothioate group.

11. The peptide/β-1,3-glucan complex according to claim 10 , wherein
in the polynucleotide derivative in which the portion of phosphodiester bonds of DNA or RNA is substituted with a phosphorothioate group, at least 50% of the phosphodiester bonds are substituted with the phosphorothioate group.

12. A pharmaceutical composition comprising:
a polynucleotide or derivative thereof having a partial base sequence having immunostimulatory activity, and
the peptide/β-1,3-glucan complex of claim 1 .

13. The pharmaceutical composition according to claim 12 , wherein
the polynucleotide or derivative thereof and the polysaccharide having the β-1,3-glucan backbone are bonded by hydrogen bonding to form a complex having a triple helix structure consisting of one molecular chain of the polynucleotide or derivative thereof and two molecular chains of the polysaccharide having the β-1,3-glucan backbone.

14. The pharmaceutical composition according to claim 13 , wherein
the polysaccharide having the β-1,3-glucan backbone included in the polynucleotide/β-1,3-glucan complex having a triple helix structure is schizophyllan, lentinan, scleroglucan, or curdlan.

15. A pharmaceutical composition comprising the peptide/β-1,3-glucan complex according to claim 8 .

16. The peptide/β-1,3-glucan complex according to claim 6 , wherein the polysaccharide having the β-1,3-glucan backbone is schizophyllan, lentinan, scleroglucan or curdlan.

Assignees

• Japan Science & Tech Agency

Inventors

• Sakurai Kazuo
• Mochizuki Shin-Ichi
• Morishita Hiromi

Classifications

• A61P37/04

  Immunostimulants · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61P31/12

  Antivirals · CPC title

• A61P31/04

  Antibacterial agents · CPC title

• C07K19/00

  Hybrid peptides {, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes} · CPC title