Composition comprising hyaluronic acid

Abstract

The present invention relates to a hyaluronic acid composition having enhanced transdermal permeability and, more specifically, to a hyaluronic acid composition and a preparation method therefor, the composition comprising hyaluronic acid, a cationic oligopeptide and an exfoliating ingredient. The hyaluronic acid composition of the present invention has an effect of increasing the transdermal absorption rate of hyaluronic acid by 1.6 times or greater when applied to the skin.

Description

Composition containing hyaluronic acid

[0001] The present invention relates to a hyaluronic acid composition with enhanced percutaneous permeability, and more specifically, to a hyaluronic acid composition comprising hyaluronic acid, a cationic oligopeptide, and a keratin exfoliating component, and a method for producing the same.

[0002]

[0003] Hyaluronic acid is a major component of connective tissue, nerve tissue, cartilage tissue, and the dermis of the skin. It is a polymer of disaccharides composed of D-glucuronic acid and N-acetyl-D-glucosamine, and is an anionic non-sulfated linear glucosaminoglycan with alternating bonds. In the human body, hyaluronic acid is synthesized by hyaluronic acid synthetases such as HAS1, HAS2, and HAS3, and is mass-produced industrially through a fermentation process using Bacillus subtilis.

[0004] Due to the chemical structure of D-glucuronic acid and N-acetyl-D-glucosamine, which contain many hydrogen bonding donors and acceptors, they can interact with a large amount of water compared to other polymers, making them hygroscopic polymers that can bind water up to 1,000 times their own volume. As such, they are one of the most notable ingredients in the cosmetics industry for their effect of maintaining moisture and intercellular spaces within the skin.

[0005] Another property of hyaluronic acid is that it activates the skin repair process, depending on its molecular weight. Damaged skin undergoes four major stages of recovery: hemostasis, inflammation, proliferation, and remodeling. Hyaluronic acid participates in each stage, depending on its molecular weight, and aids in accelerated skin regeneration. Hyaluronic acid with a molecular weight of 400 kDa or greater is known to participate in the hemostasis stage, exerting coagulation and antibacterial effects. Studies have shown that hyaluronic acid with a molecular weight of less than 120 kDa participates in the inflammation, proliferation, and remodeling stages, promoting tissue regeneration and promoting healing after injury. Furthermore, due to its excellent biocompatibility and physiological properties, it is used in the medical field as a filler for the treatment of osteoarthritis, improving dry eyes, and alleviating sagging skin.

[0006] Although hyaluronic acid, which has such diverse properties, can be directly delivered to the necessary area through the skin to achieve localized improvement, it is well known that it has difficulty penetrating the stratum corneum, which has hydrophobic properties, except for low-molecular-weight hyaluronic acid, due to its high hydrophilicity, large charge, and large molecular weight. In a study evaluating the skin permeability of hyaluronic acid with different molecular weights using pig skin, it was confirmed that hyaluronic acid with different molecular weights could not penetrate the skin except for 5 kDa hyaluronic acid, and that high-molecular-weight hyaluronic acid could penetrate only when the stratum corneum was removed (Mol Pharm. 2015; 12(5): 1391-401.). In addition, it has been reported that only low-molecular-weight hyaluronic acid can penetrate the stratum corneum in human skin, while high-molecular-weight hyaluronic acid can penetrate when the stratum corneum is removed (Skin research and Technology. 2016; 22: 55-62.).

[0007] Low-molecular-weight hyaluronic acid (HA), which has excellent skin penetration, is known to be an inflammatory agent. Specifically, hyaluronic acid less than 20 kDa is known to bind to TLR-2 or TLR-4, inducing the expression of cytokines and chemokines such as TNF-α. Applying large amounts to the skin increases the likelihood of inducing inflammation.

[0008] Accordingly, various studies are being conducted to increase the skin permeability of hyaluronic acid with a higher molecular weight than low-molecular-weight hyaluronic acid. For example, methods for enhancing permeation through the stratum corneum using a chemical permeation enhancer or a carrier system (such as liposome) containing hyaluronic acid are known, but there are problems such as weakening of the skin barrier, irritation, low loading rate, and instability in cosmetic formulations.

[0009] Against this backdrop, the inventors of the present invention have made extensive efforts to enhance the percutaneous absorption of hyaluronic acid, and have confirmed that the percutaneous absorption of hyaluronic acid is enhanced when a hyaluronic acid composition is prepared using a cationic oligopeptide and a keratin exfoliating component together with hyaluronic acid, thereby completing the present invention.

[0010]

[0011] The present invention provides a hyaluronic acid composition comprising hyaluronic acid, cationic oligopeptide, and a keratin exfoliating component, and a method for producing the same.

[0012] The present invention also seeks to provide a cosmetic composition comprising the hyaluronic acid composition.

[0013]

[0014] This is explained in detail as follows. Meanwhile, each description and embodiment disclosed in the present invention can also be applied to each other description and embodiment. In other words, all combinations of the various elements disclosed in the present invention fall within the scope of the present invention. Furthermore, the scope of the present invention should not be considered limited by the specific descriptions described below.

[0015]

[0016] A first aspect of the present invention for achieving the aforementioned purpose is to provide a hyaluronic acid composition comprising hyaluronic acid, cationic oligopeptide, and a keratin exfoliating component.

[0017]

[0018] Hereinafter, the present invention will be described in more detail.

[0019]

[0020] The term "hyaluronic acid" of the present invention is a component that is one of the main components of the dermal layer of the skin, in addition to connective tissue, nerve tissue, and cartilage tissue, and is a polymer of disaccharides composed of D-glucuronic acid and N-acetyl-D-glucosamine, and is an anionic non-sulfated linear glucosaminoglycan that is alternately bonded. In the human body, hyaluronic acid is synthesized through hyaluronic acid synthetase such as HAS1, HAS2, and HAS3, and industrially, it can be mass-produced through a fermentation process using Bacillus subtilis. Specifically, it is a disaccharide polymer composed of D-glucuronic acid and N-acetyl-D-glucosamine, which are rich in hydrogen bonding donors and hydrogen bonding acceptors, and due to its chemical structural characteristics, it can interact with a large amount of water compared to other polymers, making it a hygroscopic polymer component that can bind water up to 1,000 times its own volume.

[0021] In the hyaluronic acid composition according to the present invention, the molecular weight of the hyaluronic acid may be 20 kDa to 10,000 kDa, preferably 20 kDa to 7,500 kDa, more preferably 20 kDa to 5,000 kDa or less, but is not limited thereto.

[0022] If the molecular weight of hyaluronic acid is less than 20 kDa, it may cause skin inflammation, and if it exceeds 10,000 kDa, it may be difficult to manufacture particles due to low solubility.

[0023] The hyaluronic acid may be included in an amount of 1 to 80 wt%, 1 to 70 wt%, 1 to 60 wt%, 3 to 80 wt%, 3 to 70 wt%, 3 to 60 wt%, 5 to 80 wt%, 5 to 70 wt%, 5 to 60 wt%, 10 to 80 wt%, 10 to 70 wt%, 10 to 60 wt%, 20 to 80 wt%, 20 to 70 wt%, 20 to 60 wt%, 40 to 80 wt%, 40 to 70 wt%, 40 to 60 wt%, 50 to 80 wt%, 50 to 70 wt%, or 50 to 60 wt% based on the solid content of the total composition.

[0024] If the weight % of the above hyaluronic acid is less than 1% or more than 80%, particle formation may be difficult due to difficulty in interaction with the cationic oligopeptide.

[0025]

[0026] As described above, hyaluronic acid can achieve local improvement effects when delivered directly into the body through the skin, but has the problem of having difficulty penetrating the stratum corneum, which has hydrophobic properties due to its high hydrophilicity, large charge, and large molecular weight.

[0027] Accordingly, the present invention sought to solve the above problem by using a cationic oligopeptide and a keratin exfoliating component together to improve the transdermal permeability of hyaluronic acid.

[0028] The term "cationic oligopeptide" of the present invention refers to a polymer composed of amino acids that have a positive charge in an aqueous solution of pH 6 to 8 and are capable of ionic interaction with anionic molecules.

[0029] The above cationic oligopeptide can increase the percutaneous absorption of hyaluronic acid through ionic interaction with hyaluronic acid and reduce the hydrophilicity and negative charge of hyaluronic acid.

[0030] Additionally, cationic oligopeptides can act as linkers that connect the interior of hyaluronic acid polymers by interacting with the carboxyl group (COOH), which is an anionic functional group in hyaluronic acid, thereby forming polyion particles or assisting in particle formation.

[0031] The cationic oligopeptide may comprise a neutral amino acid monomer, a cationic amino acid monomer, and an anionic amino acid monomer. The neutral amino acid monomer, the cationic amino acid monomer, and the anionic amino acid monomer may be an amino acid monomer that is neutral, positively charged, or negatively charged at around pH 7.

[0032] The neutral amino acid monomers may include, but are not limited to, Serine, Threonine, Asparagine, Glutamine, Cysteine, Selenocysteine, Glycine, Proline, Alanine, Valine, Isoleucine, Leucine, Methionine, Phenylalanine, Tyrosine, Tryptophan, and Tranexamic acid.

[0033] The cationic amino acid monomer may include, but is not limited to, arginine, histidine, and lysine.

[0034] The above anionic amino acid monomer may include, but is not limited to, aspartic acid and glutamic acid.

[0035]

[0036] In the present invention, the cationic oligopeptide has a ratio of (number of cationic amino acid monomers) / (number of anionic amino acid monomers) greater than 1.

[0037] In the above calculations, neutral amino acid monomers in cationic oligopeptides are not included because they do not affect the net charge.

[0038] In the present invention, the polymerization degree of the cationic oligopeptide may be 2 to 100, preferably 2 to 80, and more preferably 2 to 60.

[0039] If the polymerization degree of the above oligopeptide is less than 2, it is difficult to form particles due to interaction with hyaluronic acid, and if it exceeds 100, the molecular weight of the oligopeptide is reduced compared to hyaluronic acid, so unreacted hyaluronic acid may exist or non-uniform particles may be formed.

[0040]

[0041] The cationic oligopeptide may be included in an amount of 1 to 60 wt%, 1 to 45 wt%, 1 to 30 wt%, 3 to 60 wt%, 3 to 45 wt%, 3 to 30 wt%, 5 to 60 wt%, 5 to 45 wt%, 5 to 30 wt%, 10 to 60 wt%, 10 to 45 wt%, 10 to 30 wt%, 20 to 60 wt%, 20 to 45 wt%, or 20 to 30 wt% based on the solid content of the entire composition.

[0042] If the weight % of the above cationic oligopeptide is less than 1% based on the solid content in the entire composition, particle formation may be difficult, and if it exceeds 60%, the absolute value of the surface charge may decrease, resulting in reduced dispersibility or formation of non-uniform particles.

[0043] The hyaluronic acid composition according to the present invention comprises a cationic oligopeptide, for example, an arginine / lysine polypeptide, a polyarginine, a polylysine, a dipeptide-12, a tripeptide-1, a tripeptide-4, a tripeptide-8, a tripeptide-28, a tripeptide-30, a tetrapeptide-3, a tetrapeptide-8, a tetrapeptide-12, a tetrapeptide-17, a tetrapeptide-20, a tetrapeptide-22, It may include Pentapeptide-7, Pentapeptide-8, Pentapeptide-10, Pentapeptide-12, Pentapeptide-13, Pentapeptide-24, Pentapeptide-25, Hexapeptide-1, Hexapeptide-2, Hexapeptide-4, Hexapeptide-6, Hexapeptide-13, Hexapeptide-23, Hexapeptide-26, and preferably arginine / lysine. Polypeptide (Arginine / Lysine polypeptide), Tripeptide-1, Hexapeptide-1, Polyarginine,and may include at least one selected from the group consisting of polylysine, but is not limited thereto.

[0044]

[0045] The term "keratin exfoliation component" of the present invention may have the effect of weakening the bonding between keratinocytes by chelating calcium in desmosomes that connect keratinocytes, directly penetrating into the stratum corneum to increase flexibility and widen the space between cells, and promoting the keratin exfoliation process in the body, thereby improving the permeability of hyaluronic acid.

[0046] The exfoliating component may be included in an amount of 1 to 40 wt%, 1 to 25 wt%, 1 to 15 wt%, 2 to 40 wt%, 2 to 25 wt%, 2 to 15 wt%, 3 to 40 wt%, 3 to 25 wt%, 3 to 15 wt%, 5 to 40 wt%, 5 to 25 wt%, 5 to 15 wt%, 10 to 40 wt%, 10 to 25 wt%, or 10 to 15 wt% based on the total weight of the composition.

[0047] If the weight % of the above exfoliating component is less than 1%, skin permeability may decrease, and if it exceeds 40%, the absolute value of surface charge may decrease, resulting in decreased dispersibility or formation of non-uniform particles.

[0048] The above exfoliating ingredients are Glycolic acid, Lactic acid, Citric acid, Malic acid, Mandelic acid, Glucuronic acid, Gluconolactone, Succinic acid, Pyruvic acid, Tartaric acid, Propionic acid, α-hydroxypropionic acid, β-hydroxypropionic acid, β-hydroxybutyric acid, β-hydroxy β-methylbutyric acid, Carnitine, Salicylic acid, Papain enzyme, Dioic acid, It may contain urea, and preferably, it may contain at least one selected from the group consisting of succinic acid, glycolic acid, lactic acid, malic acid, gluconolactone, or salicylic acid, but is not limited thereto.

[0049]

[0050] The pH of the above hyaluronic acid composition may be 3 to 10.

[0051] When the pH of the above hyaluronic acid composition is less than 3 or more than 10, hydrolysis of hyaluronic acid may be accelerated.

[0052]

[0053] The above hyaluronic acid composition may be in the form of a lyophilized product.

[0054]

[0055] Another aspect of the present invention for achieving the aforementioned object is to provide a method for producing a hyaluronic acid composition, comprising the step of forming hyaluronic acid particles by mixing hyaluronic acid, a cationic oligopeptide, and a keratin exfoliating component.

[0056] The method for manufacturing the above hyaluronic acid composition may be to first mix one or two of hyaluronic acid, cationic oligopeptide, and exfoliating ingredient, as needed, and then further mix the remainder.

[0057]

[0058] The hyaluronic acid particles provided by the method for manufacturing the hyaluronic acid composition may be formed with an average particle size of 5 to 80%, 5 to 70%, 5 to 60%, 5 to 55%, 5 to 50%, 5 to 40%, 5 to 30%, 5 to 25%, 10 to 80%, 10 to 70%, 10 to 60%, 10 to 55%, 10 to 50%, 10 to 40%, 10 to 30%, 10 to 25%, 15 to 80%, 15 to 70%, 15 to 60%, 15 to 55%, 15 to 50%, 15 to 40%, 15 to 30%, or 15 to 25% of the hyaluronic acid. there is.

[0059]

[0060] The average particle size of the above hyaluronic acid composition may be 100 nm to 800 nm.

[0061] If the average particle size of the hyaluronic acid composition is less than 100 nm or greater than 800 nm, it may be formed into non-uniform particles.

[0062] Specifically, the average particle size range of the hyaluronic acid composition is 100 nm to 800 nm, 100 nm to 700 nm, 100 nm to 650 nm, 100 nm to 500 nm, 100 nm to 450 nm, 100 nm to 350 nm, 100 nm to 300 nm, 100 nm to 250 nm, 150 nm to 800 nm, 150 nm to 700 nm, 150 nm to 650 nm, 150 nm to 500 nm, 150 nm to 450 nm, 150 nm to 350 nm, 150 nm to 300 nm, 150 nm to 250 nm, 200 nm to 800 nm, 200 nm to 700 nm, 200 nm to 650 nm, It may be 200 nm to 500 nm, 200 nm to 450 nm, 200 nm to 350 nm, 200 nm to 300 nm or 200 nm to 250 nm.

[0063]

[0064] The above manufacturing method may include raw material input, dissolution and mixing, defoaming, and filtering processes, and may include heating and cooling during dissolution to improve manufacturing efficiency and shorten the time of the continuous process.

[0065] It may further include a step of freeze-drying after the step of forming the hyaluronic acid particles.

[0066] The term "lyophilization" of the present invention not only obtains a composition with a high concentration of hyaluronic acid, but also further reduces the size of hyaluronic acid particles, thereby increasing the rate of percutaneous absorption. The freeze-drying process may be performed through conventional freeze-drying processes such as cooling, depressurization, and drying, and a grinding process may be added to facilitate storage and transport of the raw material after drying.

[0067]

[0068] A cosmetic product comprising the hyaluronic acid composition of the present invention can also be provided.

[0069]

[0070] The cosmetic composition (cosmetics) of the present invention comprises all kinds of ingredients usable in cosmetics, such as moisturizers such as glycerin, butylene glycol, propylene glycol, hexanediol, methyl gluceth-20, diglycerin, and ethylhexylglycerin; sunscreens such as ethylhexyl methoxycinnamate, ethylhexyl salicylate, ethylhexyl triazone, octocrylene, and bis-ethylhexyloxyphenol methoxyphenyl triazine; pH adjusters such as triethanolamine; thickeners such as carbomer, xanthan gum, acrylates / C10-30 alkyl acrylate crosspolymer, and hyaluronic acid; preservatives such as phenoxyethanol, methylparaben, and propylparaben; antioxidants such as BHT, ethyl ascorbyl ether, and ascorbic acid; skin conditioning agents such as beta-glucan; Surfactants such as cetearyl glucoside and sorbitan stearate; may additionally include fragrances or colors, but there is no limitation on the ingredients.

[0071] Each of the above-mentioned ingredients included in the cosmetic according to the present invention may be included in the cosmetic composition of the present invention within a range that preferably does not exceed the maximum usage amount stipulated in the “Cosmetics Safety and Technology Standards” established by each government.

[0072] The cosmetic according to the present invention can be manufactured in any formulation commonly manufactured in the art. For example, the cosmetic can have the formulation of, but is not limited to, a toner such as a flexible toner or a nourishing toner, a spray-type toner, an emulsion such as a facial lotion or a body lotion, a cream such as a nourishing cream, a moisturizing cream, an eye cream, a stick, an essence, a cosmetic ointment, a spray, a gel, a pack, a sunscreen, a makeup base, a foundation such as a liquid type or a spray type, a makeup remover such as a powder, a cleansing lotion, a cleansing oil, a cleanser such as a cleansing foam, a soap, a body wash, etc.

[0073] In one specific example, the formulation of the cosmetic may be a balm, a water-in-oil (W / O) formulation, an oil-in-water (O / W) formulation, a solubilized formulation, or an oil formulation.

[0074] The cosmetic of the present invention can be used according to a conventional method of use, and the number of times it is used can vary depending on the user's skin condition or preference.

[0075]

[0076] In a specific embodiment, it was confirmed that the hyaluronic acid composition of the present invention provides high percutaneous absorption of hyaluronic acid, thereby providing damaged skin regeneration, anti-inflammation, improved moisturizing, wrinkle and elasticity improvement, and soothing effects.

[0077]

[0078] The advantages and features of the present invention, and the methods for achieving them, will become clearer with reference to the experimental examples and manufacturing examples described in detail below. However, the present invention is not limited to the experimental examples and manufacturing examples disclosed below, but may be implemented in various different forms. These examples are provided solely to ensure the complete disclosure of the present invention and to fully inform those skilled in the art of the invention of the scope of the invention.

[0079]

[0080] The hyaluronic acid composition of the present invention, when applied to the skin, increases the percutaneous absorption rate of hyaluronic acid by at least 1.6 times. This hyaluronic acid composition with enhanced percutaneous permeability provides excellent skin regeneration, anti-inflammation, improved moisturizing, wrinkle and elasticity improvement, and soothing effects.

[0081]

[0082] Figure 1 is a graph showing the particle size measurement results of Examples 1 and 2 of the present invention.

[0083] Figure 2 is a microscopic photograph showing a comparative evaluation of keratin exfoliation of Example 2 and Comparative Example 1 of the present invention.

[0084] Figure 3 is a microscopic photograph showing the skin regeneration efficacy evaluation of Example 2 and Comparative Examples 5 to 6 of the present invention.

[0085] Figure 4 is a Raman peak graph showing the skin penetration rate of Example 2 and Comparative Examples 4 to 6 of the present invention.

[0086]

[0087] Hereinafter, the present invention will be described in more detail through examples. These examples are intended to explain the present invention more specifically, and the scope of the present invention is not limited by these examples.

[0088]

[0089] Example 1

[0090] Hyaluronic acid particles of Example 1 were manufactured based on the composition shown in Table 1 below.

[0091] Specifically, hyaluronic acid, succinic acid, and arginine / lysine polypeptide were added to purified water at room temperature, and then mixed for 30 minutes to completely dissolve, thereby preparing Example 1.

[0092] Raw material name Example 1 Comparative example 1 Hyaluronic acid 5 5 Arginine / Lysine polypeptide 2.5 Succinic acid 1 Purified water 91.5 95 Total 100 100

[0093] Experimental Example 1: Evaluation of physical property changes

[0094] The hyaluronic acid used in the manufacture of Example 1 was used as Comparative Example 1 as shown in Table 1 above. The particle size and shape were compared with those of Example 1.

[0095] Specifically, Example 1 and Comparative Example 1 were dissolved in purified water to a concentration of 1% each, diluted again by the same ratio to a measurable concentration, and then the particle size was measured using a Zetasizer from Malvern.

[0096] As a result, the particle size of Example 1 was measured to be 636 nm. On the other hand, the particle size of Comparative Example 1 was measured to be 1220 nm, and accordingly, it was found that the hyaluronic acid particles of the present invention were formed to have a smaller size.

[0097]

[0098] Example 2

[0099] Example 2 was performed by removing moisture from Example 1 using a freeze-drying device and pulverizing the same to obtain hyaluronic acid particles in powder form.

[0100]

[0101] Experimental Example 2: Changes in particle size due to freeze-drying application

[0102] The particle sizes were compared according to the application of the freeze-drying process after particle formation.

[0103] As shown in Fig. 1, the particle size of Example 1 was measured to be 636 nm, and the particle size of Example 2 was measured to be 286 nm. Therefore, it was confirmed that the particle size could be made smaller by adding a freeze-drying process.

[0104]

[0105] Experimental Example 3: Comparative Evaluation of Manufacturing Availability by Type of Oligopeptide

[0106] The particle formation was compared depending on the type of oligopeptide. Specifically, arginine / lysine polypeptide (Example 2), tripeptide-1 (Example 3), hexapeptide-1 (Example 4), polyarginine (Example 5), and polylysine (Example 6) were used as cationic oligopeptides, and polyglutamic acid (Comparative Example 2) was used as a comparison target as an anionic oligopeptide. In addition, succinic acid was used as a keratin exfoliating ingredient under all conditions. The particle size of the manufactured particles was measured using a Zetasizer from Malvern, and the shape and uniformity of the formed particles were confirmed through a microscope (Transmission Electron Microscope).

[0107] Examples 3 to 6 and Comparative Example 2 were manufactured using the compositions shown in Table 2 below. Specifically, the preparation was carried out using the same freeze-drying method as Example 2, and Examples 3 to 6 and Comparative Example 2 were manufactured using different oligopeptides.

[0108] Raw material nameExample 2Example 3Example 4Example 5Example 6Comparative example 2Hyaluronic acid555555Arginine / Lysine polypeptide2.5Tripeptide-12.5Hexapeptide-12.5Polyarginine2.5Polylysine2.5Polyglutamic acid2.5Succinic acid111111Purified water91.591.591.591.591.591.591.5Total100100100100100100Particle size(nm)2864354102622141105UniformityUniformUniformUniformUniformUniform

[0109] As a result, the particle sizes of Examples 2 to 6 were measured to be 286 nm, 435 nm, 410 nm, 262 nm, and 214 nm, respectively, and it was confirmed through TEM that the particles were formed uniformly. On the other hand, in the case of Comparative Example 2, the particle size was 1105 nm and non-uniform particles were confirmed.

[0110] Experimental Example 4: Comparative evaluation of manufacturing and exfoliation effects according to the type of exfoliating ingredient.

[0111] The formation of particles was compared depending on the type of exfoliating ingredients. Specifically, exfoliating ingredients such as succinic acid (Example 2), glycolic acid (Example 7), lactic acid (Example 8), malic acid (Example 9), gluconolactone (Example 10), and salicylic acid (Example 11) were used differently and manufactured by freeze-drying. In addition, arginine / lysine polypeptide was used as the oligopeptide under all conditions. The particle size of the manufactured particles was measured using a Zetasizer from Melvern, and the shape and uniformity of the formed particles were confirmed through a microscope after manufacturing.

[0112] The exfoliation effect of Examples 7 to 11 was compared and evaluated after manufacturing the exfoliating agent with the composition shown in Table 3 below. Specifically, the hyaluronic acid particles of each Example or Comparative Example were dissolved in purified water to an amount of 8.5%, and then applied to the skin of an experimental pig for 24 hours at 37°C and 50% relative humidity. Then, 10 μL of the supernatant was collected, and the number of exfoliated ...

[0113] Example 2 Example 7 Example 8 Example 9 Example 10 Example 11 Hyaluronic acid 5 5 5 5 5 Arginine / Lysine polypeptide 2.5 2.5 2.5 2.5 2.5 2.5 Succinic acid 1 Glycolic acid 1 Lactic acid 1 Malic acid 1 Gluconolactone 1 Salicylic acid 1 Purified water 1 8 1.5 8 1.5 8 1.5 8 1.5 8 1.5 8 Purified water 2 1 0 1 0 1 0 1 0 1 0 Total 1 0 1 0 1 0 1 0 1 0 1 0 1 0 Particle size 2 8 6 2 9 1 3 0 5 2 7 4 2 5 5 2 8 Uniformity Uniform Uniform Uniform Uniform Number of peeled keratin 2 0 9 2 8 2 4 0 1 4 2 3 4 5 7 1 9 7 5 8 1 4 7 5 9 1 8 4 3 7

[0114] As shown in Table 3, the particle sizes of Examples 2, 7 to 11 were measured to be 286 nm, 291 nm, 305 nm, 274 nm, 255 nm, and 282 nm, respectively, and it was confirmed that the particles were all uniformly formed. In addition, in the evaluation of exfoliation of comparative examples 1 and 3, the number of exfoliated keratin was confirmed to be 304 and 792, respectively, confirming that the exfoliation efficacy was insufficient.

[0115] As shown in Fig. 2, when the surface of the pig skin to which Example 2 and Comparative Example 1 were applied was observed under a microscope, it was difficult to confirm the shed keratin in the case of Comparative Example 1, whereas in the case of Example 2, it was clearly observed that the keratin had shed.

[0116]

[0117] Experimental Example 5: Evaluation of Collagen Synthesis Efficacy

[0118] Example 2 and Comparative Examples 4 to 8 were prepared and the collagen synthesis promotion effect was evaluated. Specifically, Comparative Examples 4 to 8 were prepared by adding individual raw materials to purified water at room temperature according to the composition in Table 4 below, mixing for 30 minutes to completely dissolve, removing moisture using a freeze dryer, and pulverizing. Next, Example 2 and Comparative Examples 4 to 6 were diluted to 100 ppm based on hyaluronic acid, Comparative Example 7 to 50 ppm based on arginine / lysine polypeptide, and Comparative Example 8 to 20 ppm based on succinic acid, respectively, to prepare samples. After applying each sample to human fibroblast culture for 48 hours, the amount of synthesized collagen was quantified using a procollagen type I C-peptide (PIP) EIA kit. The collagen synthesis efficacy of the sample was calculated and measured as a numerical value compared to the control group (untreated group).

[0119] Raw material nameExample 2Comparative example 4Comparative example 5Comparative example 6Comparative example 7Comparative example 8Hyaluronic acid5555Arginine / Lysine polypeptide2.52.52.5Succinic acid111Purified water91.592.5949597.599Total100100100100100100Collagen synthesis rate(%)41.8410.8810.8020.6915.210

[0120] As shown in Table 4, it was confirmed that the collagen synthesis efficacy of Example 2 was the best.

[0121] Experimental Example 6: Evaluation of Damaged Skin Regeneration Efficacy

[0122] In the same manner as in Experimental Example 5, Example 2 and Comparative Examples 4 to 8 were manufactured and diluted to prepare samples. Then, equal amounts of each sample were applied to artificial skin with a 2 × 2 mm square wound induced on the epidermis, and the degree of improvement in epidermal thickness was compared after 3 days of wound recovery. The thickness of the epidermis was examined using a microscope by cutting a cross-section after H&E staining, and the change was calculated as a percentage (%) compared to the thickness of the control group (untreated test group).

[0123] Example 2 Comparative Example 4 Comparative Example 5 Comparative Example 6 Comparative Example 7 Comparative Example 8 Epidermal layer improvement rate (%) 152.1133.997.70112.712.4

[0124] As shown in Table 5 and Figure 3, it can be seen that Example 2 has the best effect on damaged skin regeneration.

[0125] Experimental Example 7: Evaluation of Anti-Inflammatory Efficacy

[0126] In the same manner as in Experimental Example 5, Example 2 and Comparative Examples 4 to 8 were prepared and diluted to prepare samples.

[0127] Afterwards, the NO production inhibitory effect of the above samples was evaluated using Raw 264.7 cells. The above samples were treated with 1 μg / ml LPS and cultured for 24 hours. Then, the NO production inhibitory effect was evaluated using a NO quantification kit using the following formula.

[0128] NO production inhibition ability (%) = (1-(NO production amount when sample is added ÷ NO production amount when LPS is treated only)) × 100

[0129] Example 2 Comparative Example 4 Comparative Example 5 Comparative Example 6 Comparative Example 7 Comparative Example 8 NO production inhibition ability (%) 37.1 24.0 22.0 1.7 15.2 10.7

[0130] As shown in Table 6, it can be seen that the anti-inflammatory effect of Example 2 of the present invention is the best.

[0131] Experimental Example 8: Comparative Evaluation of Percutaneous Absorption

[0132] A comparative evaluation of percutaneous absorption was conducted for Example 2 and Comparative Examples 4 to 6. Specifically, Example 2 and Comparative Examples 4 to 6 were prepared in the same manner as Experimental Example 5, and then diluted to a level of 1% based on hyaluronic acid to prepare samples.

[0133] Thereafter, to compare the skin penetration enhancement effect, each sample was applied to human skin and left on for 30 minutes. Then, Raman spectroscopy was used to measure the Raman peak of hyaluronic acid according to skin depth. Based on the results, the skin penetration rates of HA of Example 2 and Comparative Examples 4 to 6 were compared.

[0134] Example 2 Comparative Example 4 Comparative Example 5 Comparative Example 6 Absorption rate (μm / h) 81.649.627.227.2

[0135] As shown in Table 7 and Figure 4, it can be seen that the skin penetration speed of Example 2 of the present invention is the fastest.

[0136] Experimental Example 9: Wrinkle and Elasticity Improvement Effect

[0137] Example 2 was applied directly to the skin to evaluate whether it improved skin wrinkles and elasticity. Specifically, Example 2 was diluted to 1% hyaluronic acid as an aqueous solution and applied to the faces of 6 subjects for 4 weeks, and the wrinkle and elasticity improvement effect was evaluated by comparing the before and after application. In order to determine the degree of skin wrinkle improvement, a 3D skin photographing device, PRIMOS CR Small Field (Canfield Imaging Systems, USA), was used, and the wrinkle area was measured and the wrinkle improvement rate of the stored image was measured using the PRIMOS software program. In order to determine the degree of skin elasticity improvement, an elasticity measuring device, Cutometer, was used, and the measured values ​​were analyzed to measure the elasticity improvement rate.

[0138] As a result, the wrinkle improvement rate was confirmed to be 8.06% and the elasticity improvement rate was confirmed to be 3.13%. Through this, it was confirmed that Example 2 has a wrinkle and elasticity improvement effect.

[0139]

[0140] Experimental Example 10: Improved moisturizing effect

[0141] Example 2 was applied directly to the skin to evaluate whether skin hydration was improved. Specifically, Example 2 was diluted to a 1% hyaluronic acid level in an aqueous solution and applied to the faces of four subjects. The moisturizing effect was evaluated by comparing the results before application and after 30, 180, and 360 minutes. A Corneometer was used to determine the degree of improvement in skin hydration, and the measured values ​​were analyzed to determine the moisturizing improvement rate.

[0142] As a result, it was confirmed that the improvement was 14.51% at 30 minutes, 18.40% at 180 minutes, and 18.90% at 360 minutes after application compared to before application. Through this, it was confirmed that Example 2 has a skin moisturizing improvement effect.

[0143]

[0144] Experimental Example 11: Calming Effect

[0145] Example 2 was directly applied to the skin to evaluate whether it had a skin soothing effect. Specifically, 1 MED (Minimun Erythemal Dose) of UV rays was irradiated on the upper arm of the subject in a 1x1cm size, and erythema was measured 24 hours later, which was taken as the E.I0 value. After that, Example 2 was diluted to a 1% level based on hyaluronic acid, and an aqueous solution was applied to the evaluation area twice a day for t days, and when erythema was measured on day t, the value was taken as the EI t The erythema improvement effect was evaluated using the value. To determine the skin soothing effect, a Mexameter was used, and the measured value was analyzed and the erythema index reduction was measured using the formula below.

[0146] Erythema index reduction (%) = (E.I0- EI t ) ÷ E.I0× 100,

[0147] E.I0: Erythema index, EI 24 hours after UV exposure t : Erythema index after using the product for t days

[0148] As a result, it was confirmed that the erythema improvement effect increased by 109% (day 7) and 167% (day 14) compared to the no-application group that did not apply anything after erythema was created. Through this, it was confirmed that Example 2 had a skin soothing improvement effect.

[0149]

[0150] In summary of the examples and experimental examples of the present invention, it was confirmed that the hyaluronic acid composition of the present invention provides high percutaneous absorption capacity of hyaluronic acid, thereby providing damaged skin regeneration, anti-inflammation, moisturizing improvement, wrinkle and elasticity improvement, and soothing effects.

[0151]

[0152] From the above description, those skilled in the art will understand that the present invention can be implemented in other specific forms without altering its technical spirit or essential characteristics. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be interpreted as encompassing all changes or modifications derived from the meaning and scope of the following claims and their equivalent concepts, rather than the detailed description above.

Claims

1. A hyaluronic acid composition comprising hyaluronic acid, cationic oligopeptide, and a keratin exfoliating component.

2. In paragraph 1, A hyaluronic acid composition wherein the molecular weight of the hyaluronic acid is 20 kDa to 10,000 kDa.

3. In paragraph 1, A hyaluronic acid composition, wherein the hyaluronic acid is contained in an amount of 1 to 80 wt% based on the solid content of the entire composition.

4. In paragraph 1, A hyaluronic acid composition wherein the cationic oligopeptide comprises a neutral amino acid monomer, a cationic amino acid monomer, and an anionic amino acid monomer, and the cationic oligopeptide has a ratio of (number of cationic amino acid monomers) / (number of anionic amino acid monomers) of greater than 1.

5. In paragraph 1, A hyaluronic acid composition wherein the above-mentioned exfoliating component has the function of weakening the bond between keratinocytes, directly penetrating into the stratum corneum to increase flexibility, widening the space between cells in the stratum corneum, or promoting the exfoliation process in the body.

6. A method for producing a hyaluronic acid composition, comprising the step of forming hyaluronic acid particles by mixing hyaluronic acid, cationic oligopeptide, and a keratin exfoliating component.

7. In paragraph 6, A method for producing a hyaluronic acid composition, wherein the hyaluronic acid particles are formed with an average particle size reduced by 5 to 80% compared to the hyaluronic acid.

8. In paragraph 6, A method for producing a hyaluronic acid composition, wherein the average particle size of the hyaluronic acid composition is 100 nm to 800 nm.

9. In paragraph 6, A method for producing a hyaluronic acid composition, further comprising a step of freeze-drying after the step of forming the hyaluronic acid particles.

10. A cosmetic comprising a hyaluronic acid composition according to any one of claims 1 to 5.

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