# US 9,433,699 B2 · Hydrogel comprising catechol group-coupled chitosan or polyamine and poloxamer comprising thiol group coupled to end thereof, preparation method thereof, and hemostat using same

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 9,433,699 B2
- Document ID: US9433699B2
- Status: Granted
- Kind: B2
- Application number: 14/239,538
- Filed: 2011-11-21
- Published: 2014-07-10
- Granted: 2016-09-06
- Earliest claimed: 2011-11-21
- Assignees: Innotherapy Inc.
- Applicants: Haeshin Lee; Moon Sue Lee; Ji Hyun Ryu
- Inventors: Haeshin Lee; Moon Sue Lee; Ji Hyun Ryu
- Web page: https://patentgrep.com/patent/US9433699B2

## CPC

- A61L 24/08: Polysaccharides {(A61L24/043 takes precedence)}
- A61L 24/0015: {Medicaments; Biocides}
- A61L 24/0031: {Hydrogels or hydrocolloids}
- A61L 24/0042: {Materials resorbable by the body}
- A61L 24/043: {Mixtures of macromolecular materials}
- A61K 9/06: Ointments; Bases therefor; {Other semi-solid forms, e.g. creams, sticks, gels (composition of ointments, creams or gels A61K47/00)}
- A61K 51/1213: {Semi-solid forms, gels, hydrogels, ointments, fats and waxes that are solid at room temperature}
- A61L 2400/04: Materials for stopping bleeding
- A61L: METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES (preservation of bodies or disinfecting characterised by the agents employed A01N; preserving, e.g. sterilising, food or foodstuffs A23; preparations for medical, dental or toiletry purposes A61K)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)

## Abstract

The present invention relates to an adhesive hydrogel composition containing catechol group-coupled chitosan and Pluronic comprising a thiol group coupled to the end thereof, and more specifically, to an adhesive composition which is safe in vivo and in vitro, is temperature sensitive, and has an excellent hemostatic effect and thus can be used as a bioadhesive, and a medical adhesive, an adhesion barrier and a surface adsorption inhibitor comprising the same.

## Claims

### Claim 1

A temperature sensitive hydrogel composition comprising: (i) a chitosan, covalently bound to catechol groups along its backbone; and (ii) a poloxamer containing terminal thiol groups, wherein each sulfur from said terminal thiol groups of the poloxamer is bound to a different catechol group, as respective sulfur-bound catechol groups, and wherein each of the sulfur-bound catechol group contains dihydroxy functional groups.

### Claim 2 (depends on 1)

The hydrogel composition of claim 1, wherein said chitosan, covalently bound to catechol groups along its backbone, has a molecular weight of 10,000 Da-1,000,000 Da.

### Claim 3 (depends on 1)

The hydrogel composition of claim 1, wherein said chitosan, covalently bound to catechol groups along its backbone, has a molecular weight of 50,000 Da-200,000 Da.

### Claim 4 (depends on 1)

The hydrogel composition of claim 1,
wherein said poloxamer comprises a block copolymer represented by the following formula 2, to which thiol groups have been coupled to the termini:
wherein a is an integer in a range of from 2 to 200; b is an integer in a range of from 10 to 100; and the ratio of a:b is in a range of from 1:9 to 8:2;
or a block copolymer represented by the following Formula 3, to which thiol groups have been coupled to the termini:
wherein a is an integer in a range of from 2 to 600; b is an integer in a range of from 10 to 150; and the ratio of a:b is in a range of from 1:9 to 9:1.

### Claim 5 (depends on 1)

The hydrogel composition of claim 1, wherein the hydrogel composition further comprises a therapeutic drug.

### Claim 6 (depends on 5)

The hydrogel composition of claim 5, wherein the therapeutic drug is one or more selected from the group consisting of human growth hormones, erythropoietin, interferon, insulin, interleukin, calcitonin, G-CSF, Angiopoietin, VEGF-Trap, monoclonal antibodies, antibody fragments, hemostatic proteins, antibiotics, analgesics, angina therapeutic agents, allergen extract, anti-histamines, and anti-allergic agents.

### Claim 7 (depends on 1)

A hemostat comprising the hydrogel composition of claim 1.

### Claim 8

A method for preparing a hydrogel, the method comprising:
(A) preparing a chitosan, covalently bound to catechol groups along its backbone;
(B) preparing a poloxamer containing terminal thiol groups;
(C) preparing a catechol-chitosan solution by dissolving the chitosan, covalently bound to catechol groups along its backbone, in a buffer;
(D) preparing a thiol-poloxamer solution by dissolving the poloxamer containing terminal thiol groups in a buffer;
(E) preparing an adhesive hydrogel solution by mixing the catechol-chitosan solution of step (C) with the thiol-poloxamer solution of step (D); and
(F) gelating the adhesive hydrogel solution of step (E).

### Claim 9 (depends on 8)

The method of claim 8, wherein step (A) of preparing the chitosan, covalently bound to catechol groups along its backbone, comprises the steps of:
(a) preparing a chitosan solution by adding 0.2-5 parts by weight of chitosan to 100 parts by weight of distilled water;
(b) preparing a hydrocaffeic acid solution by adding 1-5 parts by weight of hydrocaffeic acid and 2-10 parts by weight of 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC) to 100 parts by weight of distilled water;
(c) preparing a mixture of chitosan, covalently bound to catechol groups along its backbone, by mixing the chitosan solution of step (a) with the hydrocaffeic acid solution of step (b) at a volume ratio of 1:1 to 3:1 and stirring the mixture; and
(d) purifying the chitosan, covalently bound to catechol groups along its backbone, by dialyzing the mixture of chitosan, covalently bound to catechol groups along its backbone, of step (c) through a dialysis membrane in a buffer for 6-48 hours, and then drying purified chitosan, covalently bound to catechol groups along its backbone.

### Claim 10 (depends on 8)

The method of claim 8, wherein step (B) of preparing the poloxamer containing terminal thiol groups comprising the steps of:
(1) preparing a poloxamer solution by adding 5-15 parts by weight of poloxamer to 100 parts by weight of a non-polar organic solvent and 0.1-1 part by volume of triethylamine to 100 parts by volume of the non-polar organic solvent;
(2) preparing a nitrophenyl chloroformate solution by adding 5-20 parts by weight of nitrophenyl chloroformate to 100 parts by weight of a non-polar organic solvent;
(3) preparing a poloxamer mixture solution by mixing the poloxamer solution of step (1) with the nitrophenyl chloroformate solution of step (2) at a volume ratio of 5:1 to 10:1, and stirring the mixture;
(4) preparing a poloxamer mixture by precipitating and drying the poloxamer mixture solution of step (3);
(5) preparing a mixture of poloxamer containing terminal thiol groups by adding 5-20 parts by weight of the poloxamer mixture of step (4) and 0.2-5 parts by weight of cysteamine to 100 parts by weight of a mixture of a non-polar organic solvent and a polar solvent, and stirring the mixture;
(6) preparing a block copolymer mixture containing terminal thiol groups by precipitating and drying the mixture of poloxamer containing a thiol group of step (5); and
(7) purifying the block copolymer containing terminal thiol groups by dialyzing the block copolymer mixture containing terminal thiol groups of step (6) through a dialysis membrane in distilled water for 12-120 hours, and then drying to produce a purified block copolymer containing terminal thiol groups.

### Claim 11 (depends on 8)

The method of claim 8, wherein the catechol-chitosan solution of step (C) is prepared by adding 0.1-10 wt % of the chitosan, covalently bound to catechol groups along its backbone, to the buffer.

### Claim 12 (depends on 8)

The method of claim 8, wherein the thiol-poloxamer solution of step (D) is prepared by adding 3-50 wt % of the poloxamer containing terminal thiol groups, to 100 wt % of the buffer.

### Claim 13 (depends on 8)

The method of claim 8, wherein the adhesive hydrogel solution of step (E) is prepared by mixing the catechol-chitosan solution with the thiol-poloxamer solution at a volume ratio of 1:3 to 3:1.

### Claim 14 (depends on 8)

The method of claim 8, wherein step (F) is carried out at a temperature of 4 to 37° C.

### Claim 15 (depends on 9)

The method of claim 9, wherein each of the catechol groups in step (A) is represented by the following formula 1:
wherein the chitosan, covalently bound to the catechol groups along its backbone, has a molecular weight of 10 kDa-1,000 kDa.

### Claim 16 (depends on 10)

The method of claim 10, wherein the poloxamer containing terminal thiol groups of step (B) comprises a block copolymer represented by the following formula 2, to which terminal thiol groups have been coupled at the termini:
wherein a is an integer in a range of from 2 to 200; b is an integer in a range of from 10 to 100; and the ratio of a:b is in a range of from 1:9 to 8:2; and
or a block copolymer represented by the following Formula 3, to which terminal thiol groups have been coupled to the termini:
wherein a is an integer in a range of from 2 to 600; b is an integer in a range of from 10 to 150; and the ratio of a:b is in a range of from 1:9 to 9:1.