Hyaluronic acid derivative and use thereof

Abstract

The present invention provides a hydrogel platform using, as a substrate, hyaluronic acid (HA) conjugated to a pyrogallol (PG) moiety. The HA-PG conjugate of the present invention can be rapidly crosslinked by two different methods, in each of which an oxidizing agent is used or a pH is adjusted. The hydrogel of the present invention can not only have excellent biocompatibility, but also can have efficiently controlled physical characteristics such as a crosslinking rate, elasticity, and adhesive strength, depending on each crosslinking method. On the basis of such excellent stability and functionality, the hydrogel of the present invention can be used in various fields including drug delivery, biopharmaceutical materials such as a wound healing agent or anti-adhesive agent, medicines, and cosmetic products.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application is a divisional application of U.S. application Ser. No. 16 / 483,146 filed on Aug. 2, 2019, which is a U.S. national phase application of International Application No. PCT / KR2018 / 001473 filed on Feb. 2, 2018, which claims priority to Korean Application No. 10-2017-0014856 filed on Feb. 2, 2017 and Korean Application No. 10-2017-0014855 filed on Feb. 2, 2017. The applications are incorporated herein by reference.TECHNICAL FIELD

[0002] The present invention relates to a hydrogel based on a hyaluronic acid derivative modified with a gallol group, and a use thereof.BACKGROUND ART

[0003] Interest in functional biomaterials is increasing as markets for medical and biotech industries, cosmetics industry, and the like are rapidly expanding. In particular, development of biocompatible materials using natural polymers for which stability is ensured, rather than chemically synthesized polymers which may cause toxicity or side effe...

Examples

preparation examples

Preparation Example 1

[0104]Preparation of Hyaluronic Acid Derivative Modified with Gallol Group

[0105]As illustrated in FIG. 1, a hyaluronic acid derivative modified with a gallol group according to the present invention was prepared. Specifically, hyaluronic acid (MW 200K, Lifecore Biomedical, LLC, IL, USA) was completely dissolved in water (TDW), and 1 equivalent of N-hydroxysuccinimide (NHS, Sigma-Aldrich, Inc., St. Louis, Mo., USA) and 1.5 equivalent of 1-(3-dimethylaminopropyl)-3-ehtylcarbodiimide hydrochloride (EDC, Thermo Scientific, Rockford, Ill., USA) were added thereto. After 30 minutes, 1 equivalent of 5′-hydroxydopamine (Sigma-Aldrich, Inc.) was added thereto as a PG moiety and the resultant was allowed to react at pH 4 to 4.5 for 24 hours. Reactants having two different molar ratios were used in the preparation of HA-PG conjugates (HA:EDC:NHS:PG=1:1.5:1:1 or 2:1.5:1:1). Then, EDC, NHS, and 5′-hydroxydopamine were removed by dialysis based on PBS and water (Cellu / Sep (tr...

preparation example 2

[0106]Preparation of Hyaluronic Acid Hydrogel

[0107]The hyaluronic acid derivatives of Preparation Example 1 were crosslinked to prepare a hyaluronic acid hydrogel, in which each crosslinking method using sodium periodate (NaIO4) as an oxidizing agent or sodium hydroxide (NaOH) as a pH adjusting agent (pH 8 to 9) was employed. Specifically, the hyaluronic acid derivatives were dissolved in PBS (1% (w / v), 2% (w / v)), and then crosslinking was allowed to proceed while performing mixing with 4.5 mg / ml of NaIO4 or 0.08 M NaOH at a volumetric ratio of 1.5:1 to 4:1 relative to the hyaluronic acid derivative solution. As illustrated in FIG. 3, through each of these crosslinking methods, a light brown-colored hyaluronic acid hydrogel or a blue-colored hyaluronic acid hydrogel was prepared.

[0108]In order to specifically identify crosslinking of the hyaluronic acid hydrogel, analysis with ultraviolet-visible spectroscopy (UV-vis) was performed. In a case of using NaIO4, as illustrated in FIG. 4...

example 1

[0109]Changes in Physical Properties of Hyaluronic Acid Hydrogel Depending on Crosslinking Method

[0110]In the present example, changes in physical properties of hyaluronic acid hydrogels, depending on difference in crosslinking method in Preparation Example 2, were compared. On the other hand, in the hydrogel preparation step, a molar concentration ratio of hyaluronic acid to 5′-hydroxydopamine (0.5×(HA:EDC:NHS:5′-hydroxydopamine=2:1.5:1:1), lx (HA:EDC:NHS:5′-hydroxydopamine=1:1.5:1:1)) could be used to adjust a rate of being substituted with a gallol group to 4% to 5% (0.5×) or 8% to 9% (1×) (not shown), and changes in physical properties of hydrogels depending on a degree of substitution of 5′-hydroxydopamine were also compared. Specifically, hydrogel formation and changes in elastic modulus over time were compared depending on crosslinking methods; and elasticity, adhesive strength, swelling, and degradation patterns depending on the crosslinking method and the degree of substitu...

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application is a divisional application of U.S. application Ser. No. 16 / 483,146 filed on Aug. 2, 2019, which is a U.S. national phase application of International Application No. PCT / KR2018 / 001473 filed on Feb. 2, 2018, which claims priority to Korean Application No. 10-2017-0014856 filed on Feb. 2, 2017 and Korean Application No. 10-2017-0014855 filed on Feb. 2, 2017. The applications are incorporated herein by reference.TECHNICAL FIELD

[0002] The present invention relates to a hydrogel based on a hyaluronic acid derivative modified with a gallol group, and a use thereof.BACKGROUND ART

[0003] Interest in functional biomaterials is increasing as markets for medical and biotech industries, cosmetics industry, and the like are rapidly expanding. In particular, development of biocompatible materials using natural polymers for which stability is ensured, rather than chemically synthesized polymers which may cause toxicity or side effe...

Examples

preparation examples

Preparation Example 1

[0104]Preparation of Hyaluronic Acid Derivative Modified with Gallol Group

[0105]As illustrated in FIG. 1, a hyaluronic acid derivative modified with a gallol group according to the present invention was prepared. Specifically, hyaluronic acid (MW 200K, Lifecore Biomedical, LLC, IL, USA) was completely dissolved in water (TDW), and 1 equivalent of N-hydroxysuccinimide (NHS, Sigma-Aldrich, Inc., St. Louis, Mo., USA) and 1.5 equivalent of 1-(3-dimethylaminopropyl)-3-ehtylcarbodiimide hydrochloride (EDC, Thermo Scientific, Rockford, Ill., USA) were added thereto. After 30 minutes, 1 equivalent of 5′-hydroxydopamine (Sigma-Aldrich, Inc.) was added thereto as a PG moiety and the resultant was allowed to react at pH 4 to 4.5 for 24 hours. Reactants having two different molar ratios were used in the preparation of HA-PG conjugates (HA:EDC:NHS:PG=1:1.5:1:1 or 2:1.5:1:1). Then, EDC, NHS, and 5′-hydroxydopamine were removed by dialysis based on PBS and water (Cellu / Sep (tr...

preparation example 2

[0106]Preparation of Hyaluronic Acid Hydrogel

[0107]The hyaluronic acid derivatives of Preparation Example 1 were crosslinked to prepare a hyaluronic acid hydrogel, in which each crosslinking method using sodium periodate (NaIO4) as an oxidizing agent or sodium hydroxide (NaOH) as a pH adjusting agent (pH 8 to 9) was employed. Specifically, the hyaluronic acid derivatives were dissolved in PBS (1% (w / v), 2% (w / v)), and then crosslinking was allowed to proceed while performing mixing with 4.5 mg / ml of NaIO4 or 0.08 M NaOH at a volumetric ratio of 1.5:1 to 4:1 relative to the hyaluronic acid derivative solution. As illustrated in FIG. 3, through each of these crosslinking methods, a light brown-colored hyaluronic acid hydrogel or a blue-colored hyaluronic acid hydrogel was prepared.

[0108]In order to specifically identify crosslinking of the hyaluronic acid hydrogel, analysis with ultraviolet-visible spectroscopy (UV-vis) was performed. In a case of using NaIO4, as illustrated in FIG. 4...

example 1

[0109]Changes in Physical Properties of Hyaluronic Acid Hydrogel Depending on Crosslinking Method

[0110]In the present example, changes in physical properties of hyaluronic acid hydrogels, depending on difference in crosslinking method in Preparation Example 2, were compared. On the other hand, in the hydrogel preparation step, a molar concentration ratio of hyaluronic acid to 5′-hydroxydopamine (0.5×(HA:EDC:NHS:5′-hydroxydopamine=2:1.5:1:1), lx (HA:EDC:NHS:5′-hydroxydopamine=1:1.5:1:1)) could be used to adjust a rate of being substituted with a gallol group to 4% to 5% (0.5×) or 8% to 9% (1×) (not shown), and changes in physical properties of hydrogels depending on a degree of substitution of 5′-hydroxydopamine were also compared. Specifically, hydrogel formation and changes in elastic modulus over time were compared depending on crosslinking methods; and elasticity, adhesive strength, swelling, and degradation patterns depending on the crosslinking method and the degree of substitu...

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