Skin Improvement Complex

Abstract

The present invention relates to a cosmetic composition containing a GAGs (Glycosaminoglycans)-amino acid complex (GAGs-AA complex) as an active ingredient. In addition, the cosmetic composition according to the present invention contains a GAGs (Glycosaminoglycans)-amino acid complex (GAGs-AA complex) formed by physicochemical interaction as an active ingredient, and can therefore exhibit significant skin whitening, wrinkle reduction, elasticity improvement, moisturizing, skin barrier improvement, beneficial skin bacteria promotion, or skin soothing effects.

Description

[Technical Field]

[0001] The present invention relates to a polyion complex between GAGs (glycosaminoglycans), particularly chondroitin sulfate (CS) and amino acid (AA) (GAGs-AA complex or CS-AA complex). [Background technology]

[0002] One of the biggest concerns in the beauty field is age-related skin wrinkles, skin pigmentation, loss of skin elasticity, dry skin, hair loss and lack of hair shine.

[0003] In particular, skin undergoes various changes with aging. Before aging begins, skin cells are physiologically active, and fibroblasts in the dermis actively synthesize and metabolize collagen, which is important for skin elasticity and preventing wrinkle formation. GAGs (glucosaminoglycans), a type of glycoprotein containing hyaluronic acid essential for maintaining skin moisture, are also actively synthesized, providing skin with elasticity, softness, flexibility, and moisture retention. Furthermore, proliferation of basal layer cells in the epidermis is active, and the balanced production of adhesion proteins such as laminin, integrin, and desmosomes, which strengthen the bonds between skin cells, strengthens skin cell tissue and maintains healthy skin. However, modern people's skin is exposed to various harmful external environments and stresses, resulting in various damages. As we age, the physiological activity of the skin declines and the recovery rate of damaged skin slows, resulting in various skin aging phenomena such as loss of elasticity, dryness, wrinkle formation, pigmentation, age spots, dullness, etc. Therefore, various cosmetics have been developed to improve these aging symptoms.

[0004] Chondroitin sulfate is one of the various components that make up the human body, particularly joints. It has been studied as a biotransplant material and is also used as an ingestible arthritis treatment. However, due to its large molecular weight, chondroitin sulfate exhibits low skin absorption and permeability. Therefore, when used in topical skin preparations, the skin-improving effects experienced by the skin are limited, limiting its use in the cosmetics field. In this regard, biopharmaceuticals, drug delivery vehicles, and tissue engineering scaffolds, including chitosan-polymer complexes, have been shown to exhibit temperature- and pH-sensitive sol-gel transition properties in aqueous media (Korean Patent Registration No. 10-1335624). However, this approach suffers from the problem of the inherent effects of each component of the complex being lost during the complex formation process.

[0005] In addition, a cosmetic composition containing chondroitin sulfate is known that promotes keratinocyte differentiation, strengthens skin barrier function, or moisturizes the skin (Korean Patent Registration No. 10-1445644). However, when chondroitin sulfate is used alone or as a simple mixture as an active ingredient, the conventional limitations remain.

[0006] Therefore, the present invention aims to form a complex by electrostatically binding chondroitin sulfate with an amino acid monomer having a cationic functional group, thereby increasing the skin absorption of chondroitin sulfate and maximizing the skin-improving efficacy of chondroitin.

[0007] Therefore, when a chondroitin sulfate-amino acid complex (CS-AA complex) was produced, the zeta potential measured by DLS confirmed that chondroitin sulfate, which had previously been a wall-shaped compound due to linear agglomeration, had a particulate structure, leading to the completion of the present invention. [Prior art documents] [Patent documents]

[0008] [Patent Document 1] Korean Patent No. 10-0365242 [Patent Document 2] Korean Patent No. 10-1445644 Summary of the Invention [Problem to be solved by the invention]

[0009] Under the above circumstances, the present inventors have confirmed that GAGs (Glycosaminoglycans)-amino acid complexes formed by physicochemical interactions exhibit significant skin whitening, wrinkle reduction, elasticity improvement, moisturizing, skin barrier improvement, promotion of beneficial skin bacteria, or skin soothing effects, and have completed the present invention.

[0010] Therefore, an object of the present invention is to provide a cosmetic composition containing a GAGs (Glycosaminoglycans)-amino acid complex as an active ingredient. [Means for solving the problem]

[0011] As a means for solving the above problems, the present invention provides a GAGs (Glycosaminoglycans)-amino acid complex as a complex for improving skin.

[0012] To achieve the above object, there is provided an excellent method for improving skin, such as skin whitening, wrinkle reduction, elasticity improvement, moisturizing, improving the skin barrier, promoting beneficial skin bacteria, or skin soothing, which comprises the step of applying a GAGs (Glycosaminoglycans)-amino acid complex to the skin.

[0013] To achieve the above object, there is provided an excellent method for improving skin, such as skin whitening, wrinkle reduction, elasticity improvement, moisturizing, improving the skin barrier, promoting beneficial skin bacteria, or skin soothing, which comprises the step of administering a GAGs (Glycosaminoglycans)-amino acid complex to a subject.

[0014] To achieve the above object, the present invention provides a use of GAGs (Glycosaminoglycans)-amino acid complexes for the manufacture of cosmetic compositions for excellent skin improvement, such as skin whitening, wrinkle reduction, elasticity improvement, moisturizing, skin barrier improvement, promotion of beneficial skin bacteria, or skin soothing.

[0015] In the present invention, "skin improvement" means, but is not limited to, skin whitening, skin moisturizing, improvement of skin elasticity, improvement of skin wrinkles, improvement of skin barrier, promotion of beneficial skin bacteria, or skin soothing.

[0016] "Skin whitening" in the present invention means not only lightening the skin color but also alleviating or improving melasma, freckles, and hyperpigmentation caused by ultraviolet rays, hormones, or genetics.

[0017] In the present invention, "skin moisturizing" means supplying moisture to skin that has become dry and rough due to dry air and irritation, causing keratin formation, blocking evaporation of moisture, and maintaining the flexibility of the skin and maintaining a smooth surface.

[0018] The skin moisturizing may be for one or more selected from the group consisting of, for example, atopic dermatitis, psoriasis, dry skin, eczema, and xeroderma pigmentosum, but is not limited thereto.

[0019] In the present invention, "improving skin wrinkles" means preventing, suppressing or inhibiting the occurrence of wrinkles on the skin, or alleviating wrinkles that have already occurred.

[0020] In the present invention, "improving the skin barrier" means strengthening the barrier of the stratum corneum, which is located in the outer shell of the skin, and means improving skin gloss, keratin, skin protection from external factors, and the ability to maintain skin homeostasis.

[0021] In the present invention, "beneficial skin bacteria" refers to strains of normal skin bacteria that are useful for alleviating inflammation, regulating psoriasis-related inflammation, suppressing the growth of harmful bacteria, and inhibiting tumor growth. The beneficial skin bacteria may be, for example, but is not limited to, Staphylococcus epidermidis. The "harmful skin bacteria" refers to bacteria that cause inflammatory reactions or acne on the skin. The harmful skin bacteria may be, for example, but is not limited to, Propionibacterium acnes, Corynebacterium minutissimum, Staphlococcus aureus, etc.

[0022] In the present invention, "enhancement of beneficial skin bacteria" means promoting or improving the growth of the aforementioned beneficial skin bacteria, and improving the growth of beneficial bacteria also includes preventing the growth of beneficial bacteria from being inhibited. For the purposes of the present invention, the enhancement of beneficial skin bacteria may be, but is not limited to, the enhancement of Staphylococcus epidermidis.

[0023] In the present invention, "skin soothing" means suppressing or inhibiting skin inflammation, erythema, or skin itching caused by external stimuli, or soothing already occurring skin irritation symptoms.

[0024] The configuration of the present invention will be described in detail below.

[0025] In the present invention, the GAGs (Glycosaminoglycans) may be at least one selected from the group consisting of chondroitin sulfate, carrageenan, dextran sulfate, dermatan sulfate, heparan sulfate, keratan sulfate, heparin sulfate, and hyaluronic acid. In the present invention, at least one selected from the group consisting of sGAGs (sulfated glycosaminoglycans), including chondroitin sulfate, dextran sulfate, dermatan sulfate, heparan sulfate, keratan sulfate, and heparin sulfate, may be used, and preferably chondroitin sulfate and / or dextran sulfate may be used. Dextran sulfate is an isomer of chondroitin sulfate.

[0026] In the present invention, the amino acid may be at least one selected from the group consisting of serine, proline, threonine, cysteine, glycine, alanine, valine, leucine, isoleucine, aspartic acid, glutamic acid, glutamine, arginine, lysine, tyrosine, methionine, phenylalanine, histidine, tryptophan, and carnitine. Preferred amino acids include those that have a protonated -NH3 + At least one amino acid selected from the group consisting of glycine, serine, threonine, alanine, valine, leucine, cysteine, glutamine, arginine, and carnitine, which have a functional group, i.e., an amino group, and exhibit amphoteric properties, may be used. More preferably, at least one amino acid selected from the group consisting of arginine, glutamine, lysine, and carnitine, which are structurally small with a benzene ring structure or few branched chains and have a structure that is likely to interact with GAGs, may be used. Most preferably, arginine and / or glutamine may be used.

[0027] Commonly used GAGs have large molecular weights and low bioabsorption rates, making them difficult to apply at high concentrations to cosmetics, and limiting the formulation forms they can be applied in.

[0028] In the present invention, GAGs are not used alone, but are prepared and used as a complex that interacts physicochemically with amino acids. By including a GAGs-amino acid complex in a cosmetic composition according to the present invention, the bioabsorption rate of GAGs can be improved, thereby achieving even greater efficacy than the inherent efficacy of GAGs. Specifically, the GAGs-amino acid complex according to the present invention can improve skin-improving effects by at least 1.2-fold, 1.5-fold, 2-fold, 5-fold, 10-fold, or 15-fold compared to when the same amount of GAGs is used as the complex. Furthermore, the complex according to the present invention can improve skin-improving effects by at least 1.2-fold, 1.3-fold, 1.5-fold, 1.6-fold, 2-fold, 3-fold, or 5-fold compared to when the same amount of GAGs and amino acids are simply mixed together.

[0029] Furthermore, the cosmetic composition according to the present invention can improve the stability of the composition by containing a GAGs-amino acid complex.

[0030] In addition, in one embodiment of the present invention, a GAGs-amino acid complex was prepared and its skin improvement effects were evaluated in vitro and in vivo. As a result, it was confirmed that the complex has excellent effects on increasing collagen synthesis, increasing cell activity, increasing hyaluronic acid synthesis, inhibiting the production of inflammatory mediators, moisturizing, whitening, improving skin texture, improving pores, and promoting beneficial skin bacteria among the normal skin flora, and it was confirmed that the complex can be used as a skin improvement material.

[0031] The GAGs-amino acid complex is a polyion complex formed by physicochemical interactions between two components. That is, the physicochemical interaction of the GAGs-amino acid complex refers to an intermolecular interaction based on hydrophobic interaction, charge-charge interaction, or hydrogen bonding, while maintaining the individual molecular structures.

[0032] The ratio of substances required for the GAGs-amino acid complex is determined by the charge of each molecule, the type and number of functional groups on the molecule, and the polarity of the molecule or functional groups, as well as by the size and similarity of the molecules.

[0033] In the present invention, the GAGs-amino acid complex can be mixed at a molar ratio of GAGs:amino acid = 1:0.001 to 10, for example, 1:0.001 to 10, 1:0.01 to 1:8, 1:0.1 to 1:5, or 1:1 to 1:3. Experimental Example 1 confirmed that the GAGs-amino acid complex is stable within this molar ratio range.

[0034] If the molar ratio is outside the above range, the complex cannot be formed, and the remaining molecules may exceed their solubility in water and precipitate at low temperatures.

[0035] The present invention also relates to a cosmetic composition containing a GAGs (Glycosaminoglycans)-amino acid complex as an active ingredient.

[0036] In the present invention, the cosmetic composition may be in the form of a general emulsion preparation or solubilization preparation, for example, a lotion such as a softening lotion or a nutritious lotion, an emulsion such as a facial lotion or a body lotion, a cream such as a nutritious cream, a moisture cream, or an eye cream, an essence, an ointment, a balm, a spray, a gel, a pack, a sunscreen, a makeup base, a liquid, solid, or spray type foundation, a powder, a makeup remover such as a cleansing cream, a cleansing lotion, or a cleansing oil, or a cleanser such as a cleansing foam, a soap, or a body wash.

[0037] Furthermore, the cosmetic product may contain, in addition to the composition of the present invention, adjuvants commonly used in the cosmetic field, such as fatty substances, organic solvents, solubilizers, thickeners and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic or nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, humectants, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, liposomes, or any other ingredient commonly used in cosmetics.

[0038] The cosmetic preparation may contain a relatively high concentration of the composition of the present invention in the case of wash-off type cosmetics such as makeup removers and cleansers in which the active ingredients remain on the skin for a short period of time, whereas the cosmetic preparation may contain a lower concentration of the composition of the present invention in the case of leave-on type cosmetics such as lotions, emulsions, creams, and essences in which the active ingredients remain on the skin for a long period of time. Although not limited thereto, in one embodiment of the present invention, the composition of the present invention may contain 0.0001 to 30 parts by weight of a GAGs-amino acid complex, for example, 0.0001 to 0.001 parts by weight, 0.001 to 0.01 parts by weight, 0.01 to 0.05 parts by weight, 0.05 to 0.1 parts by weight, 0.1 to 0.5 parts by weight, 0.5 to 1 part by weight, 1 to 1.5 parts by weight, 1.5 to 3 parts by weight, 3 to 5 parts by weight, 5 to 8 parts by weight, 8 to 10 parts by weight, 10 to 15 parts by weight, 15 to 20 parts by weight, 20 to 25 parts by weight, or 25 to 30 parts by weight, per 100 parts by weight of the total composition.

[0039] If the GAGs-amino acid complex of the present invention contains less than 0.0001 parts by weight of the total composition, sufficient effects on improving skin whitening, wrinkles, elasticity, moisturizing, skin barrier, or skin soothing cannot be expected, and if it contains more than 30 parts by weight, undesirable reactions such as allergies may occur or there may be problems with skin safety, so this is intended to prevent these.

[0040] The above-mentioned components contained in the cosmetic composition according to the present invention can be contained in the cosmetic composition according to the present invention within the range not exceeding the maximum usage amount specified in the cosmetic safety standards of each country.

[0041] The present invention also provides a method for producing a cosmetic composition containing a GAGs (Glycosaminoglycans)-amino acid complex.

[0042] The manufacturing method of the present invention will be described in detail below.

[0043] The manufacturing method of the present invention can include the steps of: mixing an amino acid and a solvent to prepare a first mixture; first heating and stirring the first mixture; after the first heating and stirring is completed, adding GAGs and performing a second heating and stirring; after the second heating and stirring is completed, homogenizing to prepare a GAGs-amino acid complex; and adding the GAGs-amino acid complex to prepare a cosmetic composition.

[0044] In the step of preparing the first mixture, the amino acid may be at least one selected from the group consisting of serine, proline, threonine, cysteine, glycine, alanine, valine, leucine, isoleucine, aspartic acid, glutamic acid, glutamine, arginine, lysine, tyrosine, methionine, phenylalanine, histidine, tryptophan, and carnitine. Preferred amino acids include those that are protonated under physiological conditions with -NH3 +At least one amino acid selected from the group consisting of glycine, serine, threonine, alanine, valine, leucine, cysteine, glutamine, arginine, and carnitine, which have a functional group, i.e., an amino group, and exhibit amphoteric properties, may be used. More preferably, at least one amino acid selected from the group consisting of arginine, glutamine, lysine, and carnitine, which are structurally small with a benzene ring structure or few branched chains and have a structure that is likely to interact with GAGs, may be used. Most preferably, arginine and / or glutamine may be used.

[0045] The primary heating and stirring step includes a step of stirring at 30 to 80° C. and 500 to 1500 rpm for 3 to 30 minutes.

[0046] In the primary heating and stirring step, if the temperature is lower than 30°C, the complex will not be sufficiently formed, and if the temperature exceeds 70°C, the complex will be decomposed, resulting in a decrease in the skin improving effect.

[0047] In the step of producing the GAGs-amino acid complex, the GAGs (glycosaminoglycans) may include at least one selected from the group consisting of chondroitin sulfate, carrageenan, dextran sulfate, dermatan sulfate, heparan sulfate, keratan sulfate, heparin sulfate, and hyaluronic acid.

[0048] In the step of producing the GAGs-amino acid complex, the GAGs can be mixed at a molar ratio of GAGs:amino acid = 1:0.001 to 10, for example, 1:0.001 to 10, 1:0.01 to 1:8, 1:0.1 to 1:5, or 1:1 to 1:3.

[0049] The step of producing the GAGs-amino acid complex includes a step of stirring at 30 to 80° C. and 500 to 1500 rpm for 5 to 30 minutes.

[0050] The step of producing the GAGs-amino acid complex may further include a subsequent step of removing the solvent. The solvent removal step may be performed using a method for drying the cosmetic composition, such as freeze-drying. The solvent removal step allows the GAGs-amino acid complex to be obtained as a solid.

[0051] In the step of producing the cosmetic composition, the GAGs-amino acid complex may be added in an amount of 0.0001 to 30 parts by weight, for example, 0.0001 to 0.001 parts by weight, 0.001 to 0.01 parts by weight, 0.01 to 0.05 parts by weight, 0.05 to 0.1 parts by weight, 0.1 to 0.5 parts by weight, 0.5 to 1 part by weight, 1 to 1.5 parts by weight, 1.5 to 3 parts by weight, 3 to 5 parts by weight, 5 to 8 parts by weight, 8 to 10 parts by weight, 10 to 15 parts by weight, 15 to 20 parts by weight, 20 to 25 parts by weight, or 25 to 30 parts by weight, relative to 100 parts by weight of the total composition.

[0052] If the step of preparing the GAGs-amino acid complex involves a subsequent step of removing the solvent, the step of preparing the cosmetic composition may include the GAGs-amino acid complex in an amount of 50 to 100 parts by weight, for example, 90 to 100 parts by weight, 95 to 100 parts by weight, or 99 to 100 parts by weight, per 100 parts by weight of the total composition. [Effects of the Invention]

[0053] The cosmetic composition according to the present invention contains a GAGs (Glycosaminoglycans)-amino acid complex formed by physicochemical interaction as an active ingredient, and can therefore exhibit significant skin whitening, wrinkle reduction, elasticity improvement, moisturizing, skin barrier improvement, skin beneficial bacteria promotion, or skin soothing effects. [Brief explanation of the drawings]

[0054] [Figure 1] 1 shows TEM analysis results of a) chondroitin sulfate alone, b) a simple mixture of chondroitin sulfate and glutamine, c) a chondroitin sulfate-glutamine complex, and d) a chondroitin sulfate-arginine complex according to one embodiment of the present invention. [Figure 2] 1 is a graph showing the results of FT-IR analysis of chondroitin sulfate, amino acids, a simple mixture of chondroitin sulfate and amino acids, and a chondroitin sulfate-amino acid complex according to one embodiment of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0055] The present invention provides a cosmetic composition for improving skin, which contains a GAGs (glycosaminoglycans)-amino acid complex as an active ingredient.

[0056] The present invention will be described in more detail below with reference to the following examples. However, the following examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following examples.

[0057] An example of the production of a GAGs (Glycosaminoglycans)-amino acid complex (GAGs-AA complex) according to the present invention is shown below.

[0058] Production Example 1. Production of GAGs (Glycosaminoglycans)-amino acid complex (GAGs-AA complex) Example 1. Preparation of chondroitin sulfate-arginine complex (CS-R complex) 1.75 g of arginine was added to 93.25 g of purified water and stirred at 750 rpm for 5 minutes at 50-60°C until dissolved. Once the arginine was completely dissolved and the solution was clear, 5 g of sodium chondroitin sulfate was added while maintaining the temperature at 45-50°C, and the mixture was stirred at 1000 rpm for 10 minutes. Once the chondroitin sulfate was completely dissolved and the solution turned pale yellow, the mixture was mixed in a homomixer at 5500 rpm for 5-15 minutes to produce the chondroitin sulfate-arginine complex (CS-R complex).

[0059] Example 2. Preparation of chondroitin sulfate-glutamine complex (CS-Q complex) 1.5 g of glutamine was added to 93.5 g of purified water and dissolved by stirring at 750 rpm for 5 minutes at 50-60°C. Once all of the glutamine had dissolved and the solution was clear, 5 g of sodium chondroitin sulfate was added while maintaining the temperature at 45-50°C, and the mixture was stirred at 1000 rpm for 10 minutes. Once all of the chondroitin sulfate had dissolved and the solution turned pale yellow, the mixture was homogenized using a homomixer at 5500 rpm for 5-15 minutes to produce the chondroitin sulfate-glutamine complex (CS-Q complex).

[0060] Comparative Example 1. Preparation of a simple chondroitin sulfate-arginine mixture (CS / R mixture) 5 g of sodium chondroitin sulfate was added to 40 g of purified water and dissolved by stirring at 750 rpm for 5 minutes at room temperature. In a separate container, 1.75 g of arginine was added to 53.25 g of purified water and dissolved by stirring at 750 rpm for 5 minutes at 50-60°C, and then allowed to cool to room temperature. Next, the sodium chondroitin sulfate aqueous solution was added to the arginine solution and stirred at 1000 rpm for 10 minutes to produce a chondroitin sulfate-arginine simple mixture (CS / R mixture).

[0061] Comparative Example 2. Preparation of a simple chondroitin sulfate-glutamine mixture (CS / Q mixture) 5 g of sodium chondroitin sulfate was added to 40 g of purified water and dissolved by stirring at 750 rpm for 5 minutes at room temperature. In a separate container, 1.5 g of glutamine was added to 53.5 g of purified water and dissolved by stirring at 750 rpm for 5 minutes at 50-60°C, and then allowed to cool to room temperature. Next, the sodium chondroitin sulfate aqueous solution was added to the glutamine solution and stirred at 1000 rpm for 10 minutes to produce a chondroitin sulfate-glutamine simple mixture (CS / Q mixture).

[0062] Experimental Example 1: Physicochemical characterization of GAGs (Glycosaminoglycans)-amino acid complexes <Experimental Example 1-1> Morphological analysis of GAGs (Glycosaminoglycans)-amino acid complexes To observe whether GAGs-amino acid complexes form specific structures, Examples 1 and 2 (chondroitin sulfate-amino acid complex (CS-AA complex)), Comparative Example 2 (a simple mixture of chondroitin sulfate and glutamine (CS / Q mixture)), and chondroitin sulfate (CS) were prepared. The structures of each of these were analyzed using a transmission electron microscope (TEM) (JEM-F200, JEOL Ltd., Japan). The results are shown in Figure 1 below.

[0063] As shown in Figure 1, chondroitin sulfate (CS) (Comparative Example 3) did not exhibit a particulate form, but instead exhibited a broad, large, wall-like aggregated structure (Figure 1a). This is because chondroitin sulfate (CS) is a polymer and exists as an aggregate of molecules. A simple mixture of chondroitin sulfate and glutamine (CS / Q mixture) (Comparative Example 2) also exhibited large aggregates of approximately 1 μm in size, confirming a similar appearance to that of Comparative Example 3 (Figure 1b). On the other hand, the chondroitin sulfate-glutamine complex (CS-Q complex) (Example 2) was confirmed to contain almost no aggregates and to exist in the form of very small particles (Figure 1c). Based on these findings, a chondroitin sulfate-arginine complex (CS-R complex) (Example 1) was produced and analyzed, confirming a complex exhibiting the same appearance as that of Example 2 (Figure 1d). In other words, it was confirmed that when the chondroitin sulfate-amino acid complex (CS-AA complex) was formed, it existed as small particles, unlike a simple mixture of chondroitin sulfate and amino acids.

[0064] <Experimental Example 1-2> Zeta potential analysis of GAGs (Glycosaminoglycans)-amino acid complexes Particle size and zeta potential analyses were performed to analyze the binding characteristics within the complexes of Examples 1 and 2. Chondroitin sulfate (CS), arginine (R), and glutamine (Q), as well as simple mixtures of chondroitin sulfate and arginine or glutamine (CS / R or CS / Q mixtures), and complexes of chondroitin sulfate and arginine or glutamine (CS-R or CS-Q complexes), were analyzed by dynamic light scattering using a Zetasizer Nano-ZS (Malvern Instruments, Worcester, UK) to measure the particle size (effective dynamic diameter), particle size distribution, and zeta potential of the samples dispersed on the colloid. Samples were prepared by dispersion in distilled water, and a thermostated cell with a scattering angle of 90° was used, along with the manufacturer's software. After three measurements for each sample, the zeta potential was calculated, and the results are shown in Table 1 below.

[0065] [Table 1]

[0066] As shown in Table 1, chondroitin sulfate (CS) had a zeta potential of approximately -20 mV due to the sulfate functional group, confirming that the particle size was large and aggregated. On the other hand, the particle sizes of arginine (R) and glutamine (Q) were measured to be smaller than that of chondroitin sulfate (CS). In the case of zeta potential, both arginine and glutamine have both -NH2 and -COOH functional groups, and the zeta potential was close to "0" or showed both (+) and (-) values.

[0067] In the case of the simple mixtures of Comparative Examples 1 and 2, two types of zeta potential values ​​were measured: the (-) zeta potential value of chondroitin sulfate (CS) and the (+) zeta potential value of arginine (R) or glutamine (Q). This indicates that chondroitin sulfate (CS) and amino acids (AA) partially interact with each other, but maintain their respective properties.

[0068] On the other hand, in the case of the complex, the particle size was measured to be the smallest, and the zeta potential was not measured, indicating that a complex with properties different from those of chondroitin sulfate (CS) was formed.

[0069] That is, similar to the transmission electron microscope image in Experimental Example 1-1, the dynamic light scattering method also confirmed that the chondroitin sulfate-amino acid complex (CS-AA complex) existed as small particles, and the zeta potential measurement results confirmed that this complex had properties different from those of either chondroitin sulfate (CS) or amino acids.

[0070] <Experimental Example 1-3> FT-IR analysis of GAGs (Glycosaminoglycans)-amino acid complexes The individual components, Comparative Examples 1 and 2 (simple mixtures), and Examples 1 and 2 (complexes) were analyzed using an infrared spectrophotometer (FT-IR, Spectrum Two, Perkin Elmer) to confirm whether interactions occurred during the reaction period. The FT-IR spectrum of each sample is shown in Figure 2.

[0071] Figure 2a compares the FT-IR spectra of chondroitin sulfate (CS), arginine (R), a simple mixture of chondroitin sulfate and arginine (CS / R mixture), and a chondroitin sulfate-arginine complex (CS-R complex). -1It can be seen that a peak shift occurs around 1000 cm. According to the FT-IR peak table, this position corresponds to the C=O functional group adjacent to the NH amide functional group, and it is considered that the peak shift occurs due to a change in the energy of the functional group caused by the interaction between the C=O functional group of chondroitin sulfate (CS) and the -NH2 or =NH functional group of arginine (R). In addition, the complex has a peak shift of 1000 cm. -1 Below 3000cm -1 It can be seen that the spectrum differs from other comparison groups around 1000cm. -1 The following is a portion containing -OH and -NH functional groups, which is considered to show the effect of the interaction between the -OH functional group of chondroitin sulfate (CS) and the -NH functional group in arginine (R). -1 The vicinity is related to the -NH functional group bonded to the ring structure, and it is expected that the energy of the -NH functional group bonded to the ring structure of the complex has changed.

[0072] Figure 2b compares the FT-IR spectra of chondroitin sulfate (CS), glutamine (Q), a simple mixture of chondroitin sulfate and glutamine (CS / Q mixture), and a chondroitin sulfate-glutamine complex (CS-Q complex). -1 The NH stretching (amine salt) peak near 1250 cm -1 A peak shift can be seen in the S=O stretching (sulfate) peak near 0.15, which is thought to be the result of the interaction between the functional groups of chondroitin sulfate (CS) and glutamine (Q), as in Figure 2a.

[0073] In other words, the FT-IR spectrum confirmed the existence of interactions between the functional groups of chondroitin sulfate (CS) and amino acids.

[0074] Taking the results of Experimental Examples 1-1 to 1-3 together, it was concluded that the chondroitin sulfate-amino acid complex (CS-AA complex) is a small particle formed by the interaction of chondroitin sulfate (CS) with amino acids through hydrogen bonding or charge-charge interaction, and that this complex has properties different from those of each of its constituent components.

[0075] Experimental Example 2: Evaluation of the in vitro skin improvement efficacy of GAGs (Glycosaminoglycans)-amino acid complex For the in vitro evaluation of skin improvement efficacy, chondroitin sulfate (CS), glutamine (Q), arginine (R), Examples 1 and 2 (CS-R complex and CS-Q complex), and Comparative Examples 1 and 2 (CS / R mixture and CS / Q mixture) were used as mother solutions, and the final concentrations for each evaluation were as shown in Table 2 below.

[0076] [Table 2]

[0077] <Experimental Example 2-1> Confirmation of in vitro collagen synthesis effect To compare the wrinkle-improving effects of chondroitin sulfate (CS), arginine (R), and glutamine (Q), a simple mixture of chondroitin sulfate and amino acids (CS / AA mixture), and a chondroitin sulfate-amino acid complex (CS-AA complex), we investigated their effect of promoting type I collagen synthesis in skin fibroblasts. For evaluation, we quantified the C-terminal peptide of type I precursor collagen (P1CP), which is typically synthesized in the most amount by fibroblasts.

[0078] Specifically, human fibroblasts were cultured in a 48-well cell culture plate at 1 × 10 5The cells were dispensed at 100 cells / well and cultured for 24 hours in DMEM (Dulbecco's Modified Eagle's Medium, Gibco) containing 10% fetal bovine serum (FBS). The medium was then replaced with serum-free DMEM containing 2% by weight of each mother solution according to Table 2, and the cells were treated with the target compounds for 48 hours. Afterwards, 20 μl of the culture supernatant was sampled and the degree of collagen increase at each concentration was measured at 450 nm using a P1CP EIA kit (Procollagen Type-1-C-peptide Enzyme Immuno Assay kit). A negative control group was used as an experimental group containing no target compound. The collagen synthesis activity of the sample was calculated by (sample-treated group value - negative control group value) ÷ negative control group value × 100. The results are shown in Table 3 below.

[0079] [Table 3]

[0080] As shown in the above results, collagen synthesis was significantly increased in the complexes (Examples 1 and 2) compared to the single components or simple mixtures (Comparative Examples 1 and 2). Therefore, the composition of the present invention can exert effects of improving skin wrinkles, skin elasticity, and dermal density through collagen synthesis.

[0081] <Experimental Example 2-2> Confirmation of in vitro cell activation effect To compare the effects of chondroitin sulfate (CS), arginine (R), and glutamine (Q) on skin cell activity, a simple mixture of chondroitin sulfate and amino acids (CS / AA mixture), and a chondroitin sulfate-amino acid complex (CS-AA complex), human fibroblast cultures were used to determine whether cell activity increased.

[0082] Specifically, 5 × 10 human fibroblasts 3The cells were dispensed at 1000 cells / well and cultured in DMEM medium containing 10% FBS for 24 hours. The culture medium was then removed, and the cells were treated with each sample at the designated concentration according to Table 2 for 48 hours. The reagent provided in the CCK-8 kit (DOJINDO, Cell Counting Kit-8, catalog number CK04) was then added and the cells were cultured for 4 hours, after which the absorbance was measured at 450 nm using an ELISA reader. The negative control group was an experimental group that did not contain the compound to be tested for efficacy, and the cell activity value of the sample was calculated by (sample-treated group value - negative control group value) ÷ negative control group value × 100. The results are shown in Table 4 below.

[0083] [Table 4]

[0084] As shown in the above results, the cell activity of the complex (Examples 1 and 2) was significantly increased compared to the single component or simple mixture (Comparative Examples 1 and 2). Therefore, the composition of the present invention can exert wound healing and skin barrier recovery effects through cell activity.

[0085] <Experimental Example 2-3> Confirmation of in vitro hyaluronic acid synthesis effect To compare the skin moisturizing effects of chondroitin sulfate (CS), arginine (R), and glutamine (Q), a simple mixture of chondroitin sulfate and amino acids (CS / AA mixture), and a chondroitin sulfate-amino acid complex (CS-AA complex), we investigated whether hyaluronic acid production increased using human dermal fibroblasts.

[0086] Specifically, fibroblasts were cultured in DMEM medium containing 10% FBS until the cells occupied 80% of the culture dish. After 24 hours of treatment with each sample at the designated concentration (Table 2), the hyaluronic acid concentration was measured using a Hyaluronan ELISA kit (R&D Systems, Catalog No. DY3614-05). The negative control group was an experimental group that did not contain the component to be confirmed for efficacy. The hyaluronic acid synthesis capacity of the sample was calculated by (sample-treated group value - negative control group value) ÷ negative control group value × 100. The results are shown in Table 5 below.

[0087] [Table 5]

[0088] As shown in the above results, the synthesis of hyaluronic acid in the complex (Examples 1 and 2) was significantly increased compared to the single component. Therefore, the composition of the present invention can exert a skin moisturizing effect through the synthesis of hyaluronic acid.

[0089] <Experimental Example 2-4> Confirmation of the inhibitory effect on the production of inflammatory mediators in vitro To compare the skin soothing effects of chondroitin sulfate (CS), arginine (R), and glutamine (Q), a simple mixture of chondroitin sulfate and amino acids (CS / AA mixture), and a chondroitin sulfate-amino acid complex (CS-AA complex), we investigated whether NO production was suppressed using mouse macrophage cell line Raw 264.7 cells (ATCC, catalog number CRL-2788).

[0090] Specifically, Raw264.7 cells were cultured in a 24-well tissue culture plate at 1–2 × 10 5Cells were aliquoted at 1000 cells / ml and cultured in DMEM containing 10% FBS for 24 hours in a 5% CO2 incubator. The medium was then replaced with serum-free medium, and cells were starved for 12 hours. Afterward, the cells were treated with each sample at the indicated concentration (Table 2) for 30 minutes. After sample treatment, the cells were treated with 500 ng / ml LPS (Lipopolysaccharides, SIGMA, Cat. No. L43901) and cultured for another 18 hours. The culture supernatant was then transferred to a 96-well tissue culture plate and incubated with GREISS reagent (SIGMA, Cat. No. G4410) at room temperature for 15 minutes. The absorbance was then measured at 540 nm using an ELISA reader. The NO was then quantified using the measured absorbance, and the NO production inhibitory activity was calculated as the reduction ratio of NO produced in the positive control group treated with 20 μg / ml of L-NMMA compared to the positive control group by (positive control group - sample treatment group value) ÷ negative control group value × 100. The results are shown in Table 6 below.

[0091] [Table 6]

[0092] As shown in the above results, the NO suppression ability of the complex (Examples 1 and 2) was significantly increased compared to the individual components or simple mixtures (Comparative Examples 1 and 2). Therefore, the composition of the present invention can exert anti-inflammatory, skin soothing, and skin trouble relieving effects through NO suppression.

[0093] <Experimental Example 2-5> Confirmation of efficacy of in vitro tight junction protein mRNA expression To compare the skin barrier recovery effects of chondroitin sulfate (CS), glutamine (Q), a simple mixture of chondroitin sulfate and glutamine (CS / Q mixture), and a chondroitin sulfate-glutamine complex (CS-Q complex), we investigated whether the expression of ZO-1 (Zonula Occludens-1), one of the proteins that make up the tight junctions at the intercellular junctions in the skin barrier, was increased.

[0094] Specifically, human keratinocyte cell line HaCaT cells were cultured in DMEM medium (Gibco, Waltham, MA, USA) supplemented with 10% FBS (fetal bovine serum, Gibco, Waltham, MA, USA), 100 mg / ml penicillin, and 100 mg / ml streptomycin at 37°C under a 5% CO environment. Then, 4.0 × 10 cells were cultured per well. 5 The cells were seeded into 6-well plates at 1000 μg per well and cultured for 24 hours. The medium was then replaced with FBS-free DMEM medium, and each sample was treated at the designated concentration according to Table 2. After further culturing for 24 hours, the cells were washed twice with PBS and RNA was extracted using the Qiagen RNeasy Mini kit. The extracted RNA concentration was quantified, and cDNA was synthesized by reacting 1 μg of RNA with a cDNA synthesis kit (ET21100, Philekorea Technology Inc.) at 42°C for 30 minutes and 72°C for 10 minutes. The synthesized cDNA was mixed with Taqman Universal Master Mix II with UNG (4440038, Applied Biosystems) and Taqman Probe (4331182, Applied Biosystems) and reacted using a StepOnePlus Real-Time PCR System (Applied Biosystems). Based on the results of the analysis using the software of the device, the relative expression level of the tight junction protein ZO-1 (Zonula Occludens-1) mRNA was calculated. Specifically, the negative control group was an experimental group that did not contain the component to be confirmed for efficacy, and the ZO-1 expression potential value of the sample was calculated by (sample-treated group value - negative control group value) ÷ negative control group value × 100. The results are shown in Table 7 below.

[0095] [Table 7]

[0096] As shown in the above results, the complex (Example 2) was found to have a superior ability to increase the mRNA expression level of the tight junction protein ZO-1 compared to the individual components or simple mixture (Comparative Example 1). Therefore, the composition of the present invention can exert the effect of strengthening, improving, or restoring the skin barrier by increasing the expression of the tight junction protein ZO-1.

[0097] <Experimental Example 2-6> In vitro evaluation of the efficacy of GAGs (Glycosaminoglycans)-amino acid complex (CS-AA complex) in increasing beneficial bacteria on the skin To evaluate the prebiotic potential of Example 2, the degree of increase in beneficial bacteria among normal skin flora was compared before and after treatment with the CS or Example 2. Staphylococcus epidermidis was selected as a representative beneficial bacterium, and the growth rate of beneficial bacteria was compared before and after treatment with the substance.

[0098] The growth rate of beneficial bacteria was evaluated as follows. A sterile liquid medium was prepared using commercially available TSB (Tryptic soy broth, DFICO) medium, and a glycerol stock solution of S. epidermidis was inoculated into the sterile liquid TSB medium and pre-cultured at 30-37°C for 6-18 hours. This pre-culture was performed once or twice. Then, chondroitin sulfate (CS) and Example 2 were added to the sterile liquid TSB medium at the specified concentrations according to Table 2, and the pre-culture solution was inoculated and cultured at 30-37°C for 6-18 hours.

[0099] After incubation, the optical density (OD) immediately after inoculation and the optical density during the stationary growth phase were measured, and the ratio of these values ​​(growth rate) was compared. The growth rates of the experimental groups treated with chondroitin sulfate (CS) and Example 2 were compared to those of the negative control group, which was an experimental group not containing the component to be confirmed for efficacy, to determine whether the sample had a significant effect on bacterial growth. Specifically, this was calculated as (sample-treated group value - negative control group value) ÷ negative control group value × 100. Evaluations were performed three times, and the mean and standard deviation of the growth rates were calculated, followed by a t-test to verify significance. The evaluation results for the growth rates of the beneficial bacteria are shown in Table 8 below.

[0100] [Table 8]

[0101] As shown in the above results, chondroitin sulfate (CS) itself has a certain degree of beneficial bacteria-promoting effect, but the complex (Example 2) showed a significantly superior beneficial bacteria-promoting effect, with a beneficial bacteria growth rate of 17.521% compared to the control group without addition.

[0102] Experimental Example 3: Evaluation of the in vivo skin improvement efficacy of GAGs (Glycosaminoglycans)-amino acid complex (GAGs-AA complex) <Experimental Example 3-1> Confirmation of in vivo moisturizing effect To examine the moisturizing effect of Example 2, the skin moisture content was measured using a corneometer (Corneometer Cm 825 Combi 3, Courage & Khazaka, Germany). A sample for in-vivo evaluation was prepared by diluting Example 2 with water at a ratio of 1 / 50. The test was conducted on both arms using the skin moisture meter. The evaluation was conducted on eight subjects. Before the evaluation, the subjects' arms were washed thoroughly and left to stand for 30 minutes under constant temperature and humidity conditions (25°C, humidity 40-50%), and then the moisture content was measured. The moisture content of the arms at week 0, where nothing was applied, and after applying the prepared samples to the same areas daily for 2 weeks and 4 weeks, was measured using the same method. The results are shown in Table 9.

[0103] [Table 9]

[0104] As a result, it was confirmed that the moisture content of the skin was improved by about 30% after 4 weeks from the application of Example 2 of the present invention. Therefore, it can be seen that the topical skin preparation containing the complex of the present invention has a skin moisturizing effect.

[0105] <Experimental Example 3-2> Confirmation of in vivo whitening, pigmentation improvement, skin texture improvement, and pore improvement effects To confirm the moisturizing effect of Example 2, as well as various skin improvement effects such as skin brightness, texture, color, and pore size, skin condition was measured using an ANTERA 3D (Miravex, Ireland). Measurement methods followed the instrument manual. Samples for in-vivo evaluation were prepared by diluting Example 2 with water at a ratio of 1 / 50. Evaluation was performed by comparing the results of week 0 (no application) with the results measured after 2 and 4 weeks of daily application of the prepared samples to the same areas. Three measurement points were set on the cheeks on both sides of the face, and each point was measured three times consecutively for analysis. Skin brightness was measured using the "Color - L*average" value, skin texture using the "Roughness, Rq" value, and skin pores using the "pore index" value. The measurement results are shown in Table 10.

[0106] [Table 10]

[0107] As shown in the results, after 4 weeks of application of Example 2 of the present invention, skin brightness improved by 3.68%, skin texture improved by 10.5%, and skin pores improved by 62.85%. Therefore, it can be seen that the topical skin preparation containing the complex of the present invention has a skin-improving effect.

Claims

1. A cosmetic composition for improving skin, characterized by containing a GAGs (Glycosaminoglycans)-amino acid complex as an active ingredient.

2. 2. The skin-improving cosmetic composition according to claim 1, wherein the GAGs (Glycosaminoglycans) are at least one selected from the group consisting of chondroitin sulfate, carrageenan, dextran sulfate, dermatan sulfate, heparan sulfate, keratan sulfate, heparin sulfate, and hyaluronic acid.

3. 2. The skin-improving cosmetic composition according to claim 1, wherein the amino acid is at least one selected from the group consisting of serine, proline, threonine, cysteine, glycine, alanine, valine, leucine, isoleucine, aspartic acid, glutamic acid, glutamine, arginine, lysine, tyrosine, methionine, phenylalanine, histidine, tryptophan, and carnitine.

4. 2. The skin-improving cosmetic composition according to claim 1, wherein the GAGs (Glycosaminoglycans)-amino acid complex is a polyion complex.

5. 5. The skin-improving cosmetic composition according to claim 4, wherein the polyion complex is formed by physicochemical interaction.

6. 6. The skin-improving cosmetic composition according to claim 5, wherein the physicochemical interaction is a hydrophobic interaction, a charge-charge interaction, or a hydrogen bond.

7. 2. The skin-improving cosmetic composition according to claim 1, wherein the GAGs (Glycosaminoglycans)-amino acid complex is contained in an amount of 0.0001 to 30 parts by weight per 100 parts by weight of the total composition.

8. The cosmetic composition for improving skin according to claim 1, characterized in that the GAGs (Glycosaminoglycans)-amino acid complex is a polyion complex formed at a molar ratio of GAGs:amino acid = 1:0.001 to 10.

9. The cosmetic composition for improving skin according to claim 1, wherein the skin improvement is skin whitening, wrinkle improvement, elasticity improvement, moisturizing, skin barrier improvement, promotion of beneficial bacteria on the skin, or skin soothing.

10. mixing an amino acid and a solvent to produce a first mixture; a step of first heating and stirring the first mixture; After the first heating and stirring is completed, GAGs (glycosaminoglycans) are added and the mixture is heated and stirred for a second time; After the second heating and stirring is completed, homogenization is performed to prepare a GAGs-amino acid complex; and adding the GAGs-amino acid complex to prepare a cosmetic composition.

11. 11. The method for producing a cosmetic composition according to claim 10, wherein in the step of preparing the first mixture, the amino acid is at least one selected from the group consisting of serine, proline, threonine, cysteine, glycine, alanine, valine, leucine, isoleucine, aspartic acid, glutamic acid, glutamine, arginine, lysine, tyrosine, methionine, phenylalanine, histidine, tryptophan, and carnitine.

12. 11. The method for producing a cosmetic composition according to claim 10, wherein the primary heating and stirring step comprises stirring at 30 to 80°C and 500 to 1500 rpm for 3 to 30 minutes.

13. 11. The method for producing a cosmetic composition according to claim 10, wherein in the second heating and stirring step, the GAGs (Glycosaminoglycans) are at least one selected from the group consisting of chondroitin sulfate, carrageenan, dextran sulfate, dermatan sulfate, heparan sulfate, keratan sulfate, heparin sulfate, and hyaluronic acid.

14. The method for producing a cosmetic composition according to claim 10, wherein the secondary heating and stirring step includes a step of adding GAGs (Glycosaminoglycans) at a molar ratio of GAGs:amino acids = 1:0.001 to 10.

15. 11. The method for producing a cosmetic composition according to claim 10, wherein the second heating and stirring step comprises stirring at 30 to 80°C and 500 to 1500 rpm for 5 to 30 minutes.

16. The method for producing a cosmetic composition according to claim 10, characterized in that the step of producing the cosmetic composition includes a step of adding the GAGs (Glycosaminoglycans)-amino acid complex in an amount of 0.0001 to 30 parts by weight per 100 parts by weight of the total composition.

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