KR20100067260A - Complex of antioxidant and hyaluronic acid having improved stability - Google Patents

Abstract

PURPOSE: An antioxidant-hyaluronic acid composite is provided to enhance stability of antioxidant and to block antioxidant deterioration. CONSTITUTION: An antioxidant-hyaluronic acid is manufactured by reacting antioxidant and hyaluronic acid under the presence of 3-dimethyl-aminopropyl-N-ethyl carbodimide. The antioxidant is retinol, coenzyme Q10, alpha-tocopherol, ascorbic acid, butylated hydroxyl toluene, genistein, propyl gallate, epigallocatechingallate, or gallocatechin gallate. The reaction is performed under the presence of acid. The acid is hydrochloric acid, acetic acid, sulfonic acid, phosphoric acid. A method for manufacturing the antioxidant-hyaluronic acid composite comprises: a step of reacting hyaluronic acid and adipic acid dihydrazide under the presence of 3-dimethyl-aminopropyl-N-ethyl carbodimide; and a step of reacting alpha-lipoic acid or glutathione under the presence of the product and 3-dimethyl-aminopropyl-N-ethyl carbodimide.

Description

Complex of antioxidant and hyaluronic acid having improved stability

The present invention relates to a complex of an antioxidant and hyaluronic acid, and more particularly, by preparing an antioxidant directly coupled to hyaluronic acid or binding adipic acid dihydrazide (ADH) as a linker. Regarding the complex, the antioxidant-hyaluronic acid complex not only effectively increases the stability of the antioxidant, but also has a strong moisturizing ability and can block the odor of the antioxidant, and thus is usefully applied to pharmaceutical compositions and cosmetic compositions. Can be.

Pharmaceutical compositions and / or cosmetic compositions include various antioxidants. The antioxidants include α-lipoic acid, coenzyme Q10, α-tocopherol, retinol, glutathione, ascorbic acid, butylated hydroxy toluene, genistein, cucetin, propyl gallate, epigallocatechin gallate Various compounds are known, such as late, gallocatechin gallate, silybin, diosmethin, camphorol, epicatechin, galangin and the like.

For example, glutathione plays an important role in the respiration of mammals and plant tissues. It protects red blood cells by reducing hydrogen peroxide, a harmful byproduct that occurs during various metabolic reactions, into water, and acts as a cofactor for various enzymes to regenerate immune cells. To get involved. In the elderly, 75 mg of glutathione per day has been reported to significantly increase the activity of immune cells. Coenzyme Q10 is a coenzyme that helps mitochondria create energy. It has been reported to have various pharmacological actions including antiviral, antibacterial and anticancer activity. In addition, tocopherol helps increase oxygen in blood, improves blood circulation, prevents arteriosclerosis, maintains elasticity of blood vessels and prevents blood coagulation, and has been reported to prevent and treat diseases such as cardiovascular diseases. .

On the other hand, when the antioxidants are present alone, there is a problem that they are easily denatured (e.g., reduced) due to low stability, especially in an aqueous medium, and depending on the antioxidants, an adverse smell, that is, a bad smell, occurs.

For example, α-lipoic acid, which is known to have various pharmacological effects such as enhancing the body's immune function, lowering blood sugar, and suppressing appetite, is easily reduced to produce dihydrolipoic acid, thereby causing an adverse smell. There is a problem that occurs. In order to solve this problem, encapsulation using liposomes has been proposed, but it is very difficult to encapsulate an excess of α-lipoic acid.

The inventors have conducted various studies to develop a method that can increase the stability of various antioxidants, including α-lipoic acid, glutathione, and the like, particularly in aqueous media. As a result, it was found that when the antioxidant is prepared in the form of a complex with hyaluronic acid, the obtained antioxidant-hyaluronic acid complex not only has excellent stability, but also fundamentally blocks the problem of antioxidant deodorization. In addition, since the antioxidant-hyaluronic acid complex may decompose and be present as an antioxidant and hyaluronic acid in a living environment (for example, a stomach having an acidic pH) or skin tissue, the antioxidant and hyaluronic acid complex may be elevated. It has been found that synergistic effects can be expected, for example, in pharmacological effects such as whitening, anti-aging, skin protection, wrinkle improvement, especially skin moisturizing.

Accordingly, an object of the present invention is to provide a complex of an antioxidant and hyaluronic acid.

According to one aspect of the invention, retinol, coenzyme Q10, α-tocopherol, ascorbic acid, butylated hydroxy toluene, genistein, cucetin, propyl gallate, epigallocatechin gallate In the presence of 3-dimethyl-aminopropyl-N-ethyl carbodiimide, at least one antioxidant and hyaluronic acid selected from the group consisting of gallocatechin gallate, silibine, diosmethine, camphorol, epicatechin, and galangin, An antioxidant-hyaluronic acid complex obtained by a manufacturing method comprising the step of reacting is provided.

More preferably, the antioxidant is retinol, coenzyme Q10, or α-tocopherol. The reaction may preferably be carried out in the presence of an acid as a catalyst, the acid including hydrochloric acid, acetic acid, sulfonic acid, sulfuric acid, phosphoric acid and the like. Further, after performing the reaction, the precipitate separated from the reaction mixture is dispersed in water and filtered through a semipermeable membrane having a molecular weight cut-off of 2,000 to 10,000, more preferably molecular weight cut-off of about 2 000, followed by drying. It may further comprise.

According to another aspect of the invention, (a) reacting hyaluronic acid with adipic acid dihydrazide in the presence of 3-dimethyl-aminopropyl-N-ethyl carbodiimide; And (b) reacting α-lipoic acid or glutathione with the product obtained in step (a) in the presence of 3-dimethyl-aminopropyl-N-ethyl carbodiimide in the presence of an antioxidant-hyaluronic acid complex. do.

The reaction of step (b) may preferably be carried out in the presence of an acid as a catalyst, the acid comprising hydrochloric acid, acetic acid, sulfonic acid, sulfuric acid, phosphoric acid and the like. In addition, each reaction product of step (a) and step (b) is subjected to the reaction of step (a) and step (b), respectively, and then the precipitate separated from the reaction mixture is dispersed in water and the molecular weight cut-off is 2,000 to It can be separated by filtration through a semipermeable membrane of 10,000, more preferably molecular weight cut-off of about 2 000, followed by drying.

The antioxidant-hyaluronic acid complex according to the present invention effectively increases the stability of the antioxidant in the aqueous medium, thereby minimizing the alteration caused by the environment such as temperature, light, oxygen and water even if stored in the aqueous composition for a long time. In particular, when an antioxidant such as α-lipoic acid is prepared in a complex with hyaluronic acid according to the present invention, it is possible to fundamentally block the generation of inverse smells due to degeneration (eg, reduction) of the antioxidant, that is, the problem of malodor. . In addition, since the antioxidant-hyaluronic acid complex according to the present invention may exist as an antioxidant and hyaluronic acid in an in vivo environment (for example, a stomach having an acidic pH) or skin tissue, a synergy effect with an antioxidant, For example, synergistic effects can be expected in pharmacological effects such as whitening, anti-aging, skin protection, wrinkle improvement, skin moisturizing, especially skin moisturizing. Therefore, the antioxidant-hyaluronic acid complex according to the present invention can be usefully applied to pharmaceutical compositions and cosmetic compositions.

The present invention relates to retinol, coenzyme Q10, α-tocopherol, ascorbic acid, butylated hydroxy toluene, genistein, cucetin, propyl gallate, epigallocatechin gallate, gallocatechin gallate And reacting hyaluronic acid with at least one antioxidant selected from the group consisting of silicine, diosmethine, camphorol, epicatechin, and galangin, in the presence of 3-dimethyl-aminopropyl-N-ethyl carbodiimide. It provides an antioxidant-hyaluronic acid complex obtained by the preparation method. Preferably, the antioxidant may be retinol, coenzyme Q10, or α-tocopherol.

The hyaluronic acid used in the preparation of the antioxidant-hyaluronic acid complex of the present invention is a non-toxic, biodegradable, bio-compatible material, and has a very strong moisture retention ability (for example, attracts about 6000 times the weight of the hyaluronic acid itself. Pulling ability). In addition, hyaluronic acid promotes the transport of nutrition and minerals from the outside of the cell, and induces immune cells in the wound tissue during the initial inflammatory reaction, thereby promoting wound healing, and the result of enzymatic decomposition of hyaluronic acid and free radicals It reduces the toxicity of free radicals and inhibits free radical removal and migration of free radicals in the skin. Hyaluronic acid has a structure in which glycosaminoglycan units are linked via oxygen (see Formula 1), and may exist in various forms depending on the degree of polymerization of the glycosaminoglycan units. The weight average molecular weight of the hyaluronic acid used in the preparation of the antioxidant-hyaluronic acid complex according to the present invention is not particularly limited. Preferably, the weight average molecular weight may be 200 kDa to 600 kDa, more preferably about 300 kDa.

The reaction of the antioxidant with hyaluronic acid is carried out using a coupling agent, i.e. 3-dimethyl-aminopropyl-N-ethyl carbodiimide. The antioxidant-hyaluronic acid complex obtained from the reaction is bound through an ester bond with the antioxidant via a carboxylic acid of hyaluronic acid (see Formula 2).

The coupling reaction may be preferably carried out in the presence of an acid as a catalyst, and the acid may include hydrochloric acid, acetic acid, sulfonic acid, sulfuric acid, phosphoric acid, or the like, preferably acetic acid. In addition, the coupling reaction may be carried out at a range of 20 ° C. to about 50 ° C., more preferably at room temperature (about 25 ° C.), to prepare an antioxidant-hyaluronic acid complex.

The equivalent ratio of the antioxidant to hyaluronic acid is used in the range of 0.5 to 1.5 equivalents, more preferably 0.9 to 1 equivalents, relative to 1 equivalent of hyaluronic acid, leaving no unreacted antioxidant (for example, α-lipoic acid). Can be easily purified.

As described above, the antioxidant-hyaluronic acid complex obtained by the coupling reaction has auto-emulsifying properties by having water solubility and weak fat solubility at the same time, and therefore, it is preferable to separate by dialysis using a semipermeable membrane. That is, after performing the coupling reaction, the precipitate separated from the reaction mixture is dispersed in water and filtered through a semipermeable membrane having a molecular weight cut-off of 2,000 to 10,000, more preferably molecular weight cut-off of about 2 000, thereby removing the unreacted material. After removal, it is preferable to separate the antioxidant-hyaluronic acid complex by drying the filtrate obtained. The drying may be carried out by ordinary vacuum drying, freeze drying, and the like, and more preferably by freeze drying.

The antioxidant-hyaluronic acid complex of the present invention can be obtained by reacting an antioxidant with hyaluronic acid through a linker. That is, in the present invention, an antioxidant such as α-lipoic acid and glutathione is connected to hyaluronic acid through a linker having a hydrazide moiety on both sides, preferably an adipic acid dihydrazide. Obtained antioxidant-hyaluronic acid complex. Specifically, the present invention comprises the steps of (a) reacting hyaluronic acid and adipic acid dihydrazide in the presence of 3-dimethyl-aminopropyl-N-ethyl carbodiimide; And (b) reacting α-lipoic acid or glutathione with the product obtained in step (a) in the presence of 3-dimethyl-aminopropyl-N-ethyl carbodiimide in the presence of an antioxidant-hyaluronic acid complex. do.

Antioxidants that can be applied to the linker-mediated complex formation include various antioxidants, and can be, for example, α-lipoic acid and / or glutathione.

The product obtained by reacting the hyaluronic acid with the adipic acid dihydrazide in the presence of 3-dimethyl-aminopropyl-N-ethyl carbodiimide (that is, the product of step (a)) is represented by the following formula (3). Adipic acid dihydrazide has a structure in which an adipic acid dihydrazide is bound through an ester bond. In addition, the antioxidant-hyaluronic acid complex of the product of step (b) has the structure of formula (4).

The reaction of hyaluronic acid and adipic acid dihydrazide in step (a) is carried out using a coupling agent, i.e. 3-dimethyl-aminopropyl-N-ethyl carbodiimide. The reaction can be carried out according to known methods (eg, Key Engineering Matrials (2007) p 252-258) using a mixed solvent of water and ethanol. The equivalent ratio of hyaluronic acid and adipic acid dihydrazide may be in the range of 0.5 to 0.6 equivalents, more preferably about 0.5 equivalents, of adipic acid dihydrazide per 1 equivalent of hyaluronic acid. The reaction product obtained from the reaction is dispersed in water and the precipitate separated from the reaction mixture is filtered through a semipermeable membrane having a molecular weight cut-off of 2,000 to 10,000, more preferably a semipermeable membrane of molecular weight cut-off of about 2,000 to remove unreacted material. The filtrate can be separated by drying the obtained filtrate. The drying may be carried out by ordinary vacuum drying, lyophilization and the like, more preferably by lyophilization.

The reaction of the antioxidant with the product obtained in step (a) [ie step (b)] is carried out using a coupling agent, i.e. 3-dimethyl-aminopropyl-N-ethyl carbodiimide. The coupling reaction may be preferably carried out in the presence of an acid as a catalyst, and the acid may include hydrochloric acid, acetic acid, sulfonic acid, sulfuric acid, phosphoric acid, or the like, preferably acetic acid. In addition, the coupling reaction is carried out at 20 ° C. to about 50 ° C., more preferably at room temperature (about 25 ° C.). The equivalent ratio of the product and the antioxidant obtained in step (a) is unreacted using an antioxidant in the range of 0.9 to 1.5 equivalents, more preferably 0.9 to 1 equivalent, based on 1 equivalent of the product obtained in step (a). It can be easily purified because it can leave no antioxidants.

The antioxidant-hyaluronic acid complex (specifically, antioxidant-linker-chitosan complex) obtained as described above has water-soluble and weak fat solubility at the same time, and thus has an auto-emulsifying property, and thus is separated by dialysis using a semipermeable membrane. It is preferable. That is, after performing the coupling reaction, the precipitate separated from the reaction mixture is dispersed in water and filtered through a semipermeable membrane having a molecular weight cut-off of 2,000 to 10,000, more preferably molecular weight cut-off of about 2 000, thereby removing the unreacted material. After removal, it is preferable to separate the antioxidant-hyaluronic acid complex by drying the filtrate obtained. The drying may be carried out by ordinary vacuum drying, lyophilization and the like, more preferably by lyophilization.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, these examples are intended to illustrate the invention and should not be construed as limiting the invention.

Example 1 Preparation of Complex of α-Lipoic Acid and Hyaluronic Acid

Step (a): prepared by hyaluronic acid-adipic acid dihydrazide (HA-ADH)

5.00 g (12.21 mmol) of hyaluronic acid having a molecular weight of 35,000 Da and 2.13 g (12.21 mmol) of adipic acid dihydrazide were added to 80 ml of 50% ethanol while stirring, and adjusted to pH 4.8 with 1M HCl. 2.81 g (14.65 mmol) of 3-dimethyl-aminopropyl-N-ethyl carbodiimide were slowly added over 30 minutes, adjusted to pH 7.0 with 1M NaOH, and 0.30 g (2.44 mmol) of dimethylaminopyridine was added to the reaction mixture. After stirring at room temperature for 9 hours. 500 ml of tetrahydrofuran was added to the reaction mixture, and the precipitate was separated by filtration. The obtained precipitate was completely dissolved in 300 ml of water, and then the Milipore dialysis tube, M.W. Unreacted material was removed by semipermeable membrane filtration for 2 days with cut-off 2,000. The obtained filtrate was lyophilized to obtain 5.46 g of HA-ADH (yield 79%).

¹ H NMR 400 MHz, DMSO-d ₆ , ppm; 2.02 (hyaluronic acid, -COCH ₃ , 3H, s), 2.19-2.17 (ADH, 4H, q), 1.59-1.55 (ADH, 4H, m)

Step (b) Reaction of α-Lipoic Acid with HA-ADH

5.00 g (8.84 mmol) of HA-ADH prepared in step (a) and 1.86 g (9.72 mmol) of 3-dimethyl-aminopropyl-N-ethyl carbodiimide were dissolved in 250 ml of a 1% acetic acid aqueous solution. To the obtained solution, a solution in which 1.83 g (8.84 mmol) of α-lipoic acid was dissolved in 30 ml of tetrahydrofuran was added, and then reacted at room temperature for 72 hours. 500 ml of tetrahydrofuran was added to the reaction mixture, and the precipitate was separated by filtration. The obtained precipitate was completely dissolved in 300 ml of water, and then the Milipore dialysis tube, M.W. Unreacted material was removed by semipermeable membrane filtration for 2 days with cut-off 2,000. The obtained filtrate was lyophilized to obtain 4.95 g (yield 74%) of a complex of α-lipoic acid and hyaluronic acid.

¹ H NMR 400 MHz, DMSO-d ₆ , ppm; 2.03 (hyaluronic acid, -COCH ₃ , 3H, s), 2.18-2.16 (ADH, 4H, q), 1.58-1.54 (ADH, 4H, m), 2.51-2.60 (α-lipoic acid, -SCH-, -SCH ₂ -, 3H, m) 1.55-2.18 (α-lipoic acid, -CH _2- , 8H, m)

Example 2 Preparation of Complexes of Glutathione and Hyaluronic Acid

The reaction was carried out in the same manner as in Example 1, except that 2.72 g (8.84 mmol) of glutathione was used instead of α-lipoic acid to obtain 5.76 g (yield 76%) of a complex of glutathione and hyaluronic acid.

¹ H NMR 400 MHz, DMSO-d ₆ , ppm; 2.03 (hyaluronic acid, -COCH ₃ , 3H, s), 2.19-2.15 (ADH, 4H, q), 1.59-1.54 (ADH, 4H, m), 4.08 (glutathione, -NHCO CH ₂ NH-, 2H, t) 3.51 (glutathione, -CH ₂ CH ₂ CHNH _2- , 2 H, t) 3.19-2.94 (glutathione, -CH CH ₂ SH, 2H, m) 2.19 (gluthathione, -CO CH ₂ CH _2- , 2H, t)

Example 3 Preparation of Complex of Retinol and Hyaluronic Acid

The reaction was carried out in the same manner as in Step (b) of Example 1, except that 2.53 g (8.84 mmol) of retinol was used instead of HA-ADH to obtain 4.75 g (yield 79%) of the retinol and hyaluronic acid complex. .

¹ H NMR 400 MHz, DMSO-d ₆ , ppm; 2.10 (hyaluronic acid, -COCH ₃ , 3H, s), 6.48-5.61 (retionol, -C = CH- , 6H, s) 4.68 (retinol, -OCH _2- , 2H, s), 1.89-1.52 (retinol ring, -CH _2- , 6H, m) 1.22 (retinol ring, -CH ₃ , 6H, s)

Example 4 Preparation of a Complex of Reduced Coenzyme Q10 and Hyaluronic Acid

The reaction was carried out in the same manner as in step (b) of Example 1, except that 5.29 g (8.84 mmol) of reduced coenzyme Q10 was used instead of HA-ADH, and 5.55 g of reduced coenzyme Q10 and hyaluronic acid complex (yield 65) %) Was obtained.

¹ H NMR 400 MHz, DMSO-d ₆ , ppm; 2.09 (hyaluronic acid, -COCH ₃ , 3H, s), 5.25 (coenzyme Q ₁₀ , -C = CH- , 3H, s), 3.49 (coenzyme Q ₁₀ , -OCH ₃ , 6H, s), 2.13-2.64 (coenzyme Q ₁₀ , -CH _2- , 10H, m), 1.93 (coenzyme Q ₁₀ ring, -CH ₃ , 3H) 1.72-1.75 (coenzyme Q ₁₀ , -CH ₃ , 15H, s)

Example 5 Preparation of Complex of Tocopherol and Hyaluronic Acid

The reaction was carried out in the same manner as in Step (b) of Example 1, except that 3.81 g (8.84 mmol) of tocopherol was used instead of HA-ADH, thereby obtaining 5.18 g (yield 71%) of tocopherol and hyaluronic acid complex.

¹ H NMR 400 MHz, DMSO-d ₆ , ppm; 2.06 (hyaluronic acid, -COCH ₃ , 3H, s), 2.63-2.07 (tocopherol ring, -CH ₂ CH _2- , 4H, m), 2.35 (tocopherol ring, -CH ₃ , 9H, s) 1.78 (tocopherol, 1H , s) 1.64-1.27 (tocopherol, -CH _2- , 18H, m) 1.53-1.04 (tocopherol, -CH ₃ , 15H, s)

Test example. Properties, chemical stability, and sensory tests

3.0 g of the complex of the antioxidant and hyaluronic acid prepared in Examples 1 to 5 were completely dissolved in 1,000 ml of distilled water to prepare a clear solution, and stirred at 40 ° C. for 30 minutes to prepare a micelle solution. 10 ml of the obtained aqueous solution was taken and stored at 45 ° C. for 0 weeks, 1 week, 2 weeks, 3 weeks, and 4 weeks, whereby a change in properties, a residual content of antioxidant, a sensory test, and a moisturizing effect experiment were performed.

The change in appearance was observed with the naked eye, and the moisturizing effect experiment was commissioned by the skin clinical research institute [Derma Pro Co., Ltd.] and the transepidermal water loss (TEWL) was measured after 0 and 28 days.

Residual content of the antioxidant was inverted by measuring the content (measurement content) of the antioxidant eluted from the antioxidant-hyaluronic acid complex into the aqueous solution. In the high performance liquid chromatography, each sample was taken in a 10 ml-volume flask, each 0.1 g of the sample, adjusted to 10 ml with chloroform, vortexed for 1 minute, and centrifuged at 10000 rpm for 10 minutes. It was measured by high performance liquid chromatography. The high performance liquid chromatography conditions are as follows: column-ACE 5-C18 (4.6 * 150mm, 5㎛), Guard Column-ACE 5-C18 (2.1 * 12.5mm, 5㎛), mobile phase Mixed solution of acetonitrile and 0.1% aqueous sulfuric acid solution (80:20), detector wavelength-UV 224 nm, flow rate-1 ml / min, injection amount-2 μl.

Sensory tests were performed as follows: Each sample was measured and averaged by measuring the intensity of odor of 10 women in their late 20s, the intensity of odor was set as shown in Table 1 below.

Bad smell Explanation 0 Odorless One Detection 2 Normal odor 3 Strong odor 4 Extreme odor 5 Unbearable odor

As described above, the results of the property change, the residual content of the antioxidant, the sensory test and the moisturizing effect test are shown in Table 2 below.

Complex
(Example)
Storage period
Change of appearance
Antioxidant Content (%) Sensory test
(directly
Sensory method) TEWL (g / m2 / h) Measurement content
(%) Remaining content
(%) 0 days 28 days Difference Example 1 Week 0 none 0 100 1.3 23.2 18.3 4.9 1 week none 0.33 99.77 1.6 2 weeks none 0.62 99.38 2.1 3 weeks none 1.10 98.90 2.7 4 Weeks Slightly cloudy 1.75 98.25 2.6 Example 2 Week 0 none 0 100 1.3 23.6 18.8 4.8 1 week none 0.26 99.74 1.2 2 weeks none 0.45 99.55 1.7 3 weeks Slightly cloudy 0.62 99.38 2.4 4 Weeks Slightly cloudy 0.93 99.07 2.3 Example 3 Week 0 none 0 100 0.7 24.1 19.0 5.1 1 week none 0.12 99.88 0.6 2 weeks none 0.36 99.64 1.2 3 weeks none 0.78 99.22 2.1 4 Weeks Slightly cloudy 1.21 98.79 2.2 Example 4 Week 0 none 0 100 0.6 23.5 19.3 4.2 1 week none 0.24 99.76 1.7 2 weeks none 0.39 99.61 1.5 3 weeks none 0.73 99.27 1.7 4 Weeks Slightly cloudy 1.32 98.68 1.9 Example 5 Week 0 none 0 100 0.7 23.1 18.4 4.7 1 week none 0.21 99.79 0.7 2 weeks none 0.54 99.46 1.1 3 weeks none 0.90 99.10 1.3 4 Weeks none 1.57 98.43 1.6

From the results of Table 2, when the antioxidants easily discolored according to the present invention prepared with a composite with hyaluronic acid, it exhibits high stability by maintaining a high residual content of at least 98.43% in an aqueous medium as a whole, there is little change in appearance In addition, the difference in the amount of percutaneous moisture retention is up to 5.2 g / m2 / h or less, the excellent moisturizing ability, and it can be seen that there is little problem of deodorization.

Claims (10)

Retinol, coenzyme Q10, α-tocopherol, ascorbic acid, butylated hydroxy toluene, genistein, cucetin, propyl gallate, epigallocatechin gallate, gallocatechin gallate, silybin, diosmethine, camphorol, epicatechin And an antioxidant selected from the group consisting of galangin and hyaluronic acid in the presence of 3-dimethyl-aminopropyl-N-ethyl carbodiimide. The antioxidant-hyaluronic acid complex according to claim 1, wherein the antioxidant is retinol, coenzyme Q10, or α-tocopherol. The antioxidant-hyaluronic acid complex according to claim 1, wherein the reaction is carried out in the presence of an acid as a catalyst. The antioxidant-hyaluronic acid complex according to claim 3, wherein the acid is hydrochloric acid, acetic acid, sulfonic acid, sulfuric acid, or phosphoric acid. The method according to any one of claims 1 to 4, wherein after the reaction is carried out, the precipitate separated from the reaction mixture is dispersed in water, filtered through a semipermeable membrane having a molecular weight cut-off of 2,000 to 10,000, and then drying. Antioxidant-hyaluronic acid complex, characterized in that it further comprises. (a) reacting hyaluronic acid with adipic acid dihydrazide in the presence of 3-dimethyl-aminopropyl-N-ethyl carbodiimide; And (b) reacting α-lipoic acid or glutathione with the product obtained in step (a) in the presence of 3-dimethyl-aminopropyl-N-ethyl carbodiimide. The antioxidant-hyaluronic acid complex according to claim 6, wherein the reaction of step (b) is carried out in the presence of an acid. 8. The antioxidant-hyaluronic acid complex according to claim 7, wherein the acid is hydrochloric acid, acetic acid, sulfonic acid, sulfuric acid, or phosphoric acid. The process according to any one of claims 6 to 8, wherein the reaction product of step (a) is subjected to the reaction of step (a) and then the precipitate separated from the reaction mixture is dispersed in water and the molecular weight cut-off is 2,000 to 10,000. An antioxidant-hyaluronic acid complex, which is separated by filtration through a semipermeable membrane and then drying. The process according to any one of claims 6 to 8, wherein the reaction product of step (b) is subjected to the reaction of step (b) and then the precipitate separated from the reaction mixture is dispersed in water and the molecular weight cut-off is 2,000 to 10,000. An antioxidant-hyaluronic acid complex, which is separated by filtration through a semipermeable membrane and then drying.