Method for sulfating glycosaminoglycan

Patent metadata

Field	Value
Publication number	US-10259889-B2
Application number	US-201615563196-A
Country	US
Kind code	B2
Filing date	Mar 31, 2016
Priority date	Mar 31, 2015
Publication date	Apr 16, 2019
Grant date	Apr 16, 2019

Abstract

An object of the present invention is provide a method for sulfating a glycosaminoglycan in a solution of a non-organic solvent. In the present invention, sulfation reaction of a glycosaminoglycan is performed with a sulfating agent in a strongly basic solution of a non-organic solvent. In the present invention, pH of the strongly basic solution is preferably set to be 11.5 or higher. According to the present invention, for example, a glycosaminoglycan having heparin-like anticoagulant activity can be produced from N-acetylheparosan through one-pot procedure. In one embodiment, a sulfated glycosaminoglycan produced by the method of the present invention has a unique disaccharide composition and is expected to be a novel useful material.

First claim

The invention claimed is:

1. A method for sulfating a glycosaminoglycan, the method comprising performing sulfation reaction in a strongly basic solution under coexistence of a glycosaminoglycan with a sulfating agent, wherein the pH of the strongly basic solution is set to be 11.5 or higher.

2. The method according to claim 1 , wherein the glycosaminoglycan is selected from among the following glycosaminoglycans (A) to (D):
(A) a glycosaminoglycan having a hexuronic acid residue;
(B) a glycosaminoglycan prepared through addition or elimination of a substituent or a functional group to or from the glycosaminoglycan (A);
(C) a glycosaminoglycan prepared through deacetylation of the glycosaminoglycan (A); and
(D) a glycosaminoglycan prepared through alkylation of the glycosaminoglycan (A).

3. A method for producing a sulfated glycosaminoglycan, the method comprising performing sulfation reaction in a strongly basic solution under coexistence of a glycosaminoglycan with a sulfating agent, wherein the pH of the strongly basic solution is set to be 11.5 or higher.

4. The method according to claim 3 , the method further comprising a step of performing deacetylation reaction of the glycosaminoglycan.

5. The method according to claim 3 , the method further comprising a step of performing alkylation reaction of the glycosaminoglycan.

6. The method according to claim 3 , wherein the sulfated glycosaminoglycan has heparin-like anticoagulant activity.

7. The method according to claim 3 , wherein the sulfated glycosaminoglycan is sulfated heparosan having heparin-like anticoagulant activity.

8. The method according to claim 3 , wherein the sulfated glycosaminoglycan is chondroitin sulfate containing 3 mol % or higher of a disaccharide having a structure represented by the following formula as the composition ratio of disaccharide units:
[HexA(2S)1-3GalN1-4]
(where “HexA” represents a hexuronic acid residue; “GalN” represents a galactosamine residue; “1-3” represents a 1-3 glycosidic bond; “1-4” represents a 1-4 glycosidic bond; and “2S” represents a 2-O-sulfate group, respectively).

9. The method according to claim 3 , wherein the sulfated glycosaminoglycan is chondroitin sulfate containing a disaccharide having a structure represented by the following formula (b) over 3 times as many as a disaccharide having a structure represented by the following formula (a):
[HexA1-3GalN(4S)1-4]  (a)

[HexA(2S)1-3GalN1-4]  (b)

(where “HexA” represents a hexuronic acid residue; “GalN” represents a galactosamine residue; “1-3” represents a 1-3 glycosidic bond; “1-4” represents a 1-4 glycosidic bond; “4S” represents a 4-O-sulfate group; and “2S” represents a 2-O-sulfate group, respectively).

10. The method according to claim 3 , wherein the sulfated glycosaminoglycan is chondroitin sulfate containing a disaccharide having a structure represented by the following formula (d) over 0.1 times as many as a disaccharide having a structure represented by the following formula (c):
[HexA1-3GalN(6S)1-4]  (c)

[HexA(2S)1-3GalN1-4]  (d)

(where “HexA” represents a hexuronic acid residue; “GalN” represents a galactosamine residue; “1-3” represents a 1-3 glycosidic bond; “1-4” represents a 1-4 glycosidic bond; “6S” represents a 6-O-sulfate group; and “2S” represents a 2-O-sulfate group, respectively).

11. The method according to claim 3 , wherein the sulfated glycosaminoglycan is chondroitin sulfate having 25 mol % or higher of a disaccharide having a structure represented by the following formula as the composition ratio of disaccharide units:
[HexA(2S)1-3GalN(6S)1-4]
(where “HexA” represents a hexuronic acid residue; “GalN” represents a galactosamine residue; “1-3” represents a 1-3 glycosidic bond; “1-4” represents a 1-4 glycosidic bond; “2S” represents a 2-O-sulfate group; and “6S” represents a 6-O-sulfate group, respectively).

12. The method according to claim 3 , wherein the sulfated glycosaminoglycan is sulfated hyaluronic acid containing a disaccharide having a structure represented by the following formula as the disaccharide composition:
[HexA1-3GlcN(4S)1-4]
(where “HexA” represents a hexuronic acid residue; “GlcN” represents a glucosamine residue; “1-3” represents a 1-3 glycosidic bond; “1-4” represents a 1-4 glycosidic bond; and “4S” represents a 4-O-sulfate group, respectively).

13. The method according to claim 3 , wherein the sulfated glycosaminoglycan is sulfated hyaluronic acid having the following characteristics (A) and (B):
(A) a molecular weight of 2,000,000 Da or higher, and
(B) a sulfur content of 2 mass % or higher.

Assignees

• Seikagaku Kogyo Co Ltd

Inventors

• Minamisawa Toshikazu
• Asami Nagamasa
• Fujita Hiroshi
• Suzuki Kiyoshi

Classifications

• C08B37/00

  Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof (cellulose D21; {microbiological processes C12P}) · CPC title

• A61P7/02

  Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors · CPC title

• C08B37/0072

  Hyaluronic acid, i.e. HA or hyaluronan; Derivatives thereof, e.g. crosslinked hyaluronic acid (hylan) or hyaluronates · CPC title

• C08B37/0063

  Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan · CPC title

• C08B37/0075 Primary

  Heparin; Heparan sulfate; Derivatives thereof, e.g. heparosan; Purification or extraction methods thereof · CPC title