Hyaluronic acid production promoter and collagen production promoter

Abstract

The purpose of the present disclosure is to provide a novel hyaluronic acid production promotor, collagen production promotor, composition for promoting the production of hyaluronic acid and / or collagen, agent for improving wrinkles, the firmness, the texture, and the moisture content of the skin, and cosmetic or non-therapeutic method for promoting the production of hyaluronic acid and / or collagen. More specifically, by using N-benzoyl glycylglycine as an active ingredient, a hyaluronic acid production promotor, collagen production promotor, composition for promoting the production of hyaluronic acid and / or collagen, and agent for improving wrinkles, the firmness, the texture, and the moisture content of the skin can be prepared and used in a cosmetic or non-therapeutic method for promoting the production of hyaluronic acid and / or collagen.

Description

Technical Field

[0001] The present invention relates to the technical field of improving skin conditions, and specifically relates to cosmetics, beauty or medical technologies.

Background Art

[0002] The skin is composed of the stratum corneum, epidermis, basement membrane, and dermis from the outside. The dermis is the largest part among them and is not as densely packed with cells as the epidermis. Rather, there are many extracellular spaces, which are filled by a network structure of macromolecules called the "extracellular matrix". This extracellular matrix is produced by fibroblasts in the dermis and is composed of polysaccharides called acidic mucopolysaccharides such as hyaluronic acid and dermatan sulfate, and fibrous proteins such as collagen and elastin. The extracellular matrix is directly involved in the elasticity, firmness, moisture, metabolism, etc. of the skin. When the production of the extracellular matrix in fibroblasts, etc. slows down, the elasticity and moisture of the skin are lost, skin roughness is likely to occur, which contributes to skin aging.

[0003] The above fibrous protein mainly consists of collagen, and among them, type I collagen accounts for 80% of the whole. In addition to type I collagen, the presence of types III, V, XII, and XIV collagen is also known. One of the causes of sagging seen in aging skin is the thinning of the skin tissue with aging. In aging skin, the decrease in collagen fibers, which are the main matrix components of the dermis, is remarkable, and this is likely to be the main cause of the thinning of the skin thickness. Therefore, it is considered effective to promote the production of collagen and maintain the amount of collagen for the prevention and improvement of sagging. In addition to the above, promoting the production of collagen is also effective for improving skin wound healing, treating osteoporosis, arthritis, tenosynovitis, etc., and preventing hypertension.

[0004] As components having such an effect of promoting collagen production, various components have been conventionally known. For example, chickpea and its solvent extract (Patent Document 1) and the like are known. Further, Patent Document 2 shows that N-benzoyl-glycylglycine has an effect of promoting the production of type I collagen.

[0005] On the other hand, hyaluronic acid in the above polysaccharide has many functions such as retention of moisture in the intercellular space, retention of cells based on formation of a jelly-like matrix in the tissue, retention of tissue lubricity and flexibility, resistance to external forces such as mechanical damage, and prevention of bacterial infection. Furthermore, recently, many studies have been made on the relationship between hyaluronic acid and cancer metastasis and the like.

[0006] The amount of hyaluronic acid in the skin is considered to decrease with aging, and accordingly, skin aging such as fine wrinkles and roughness is considered to appear. Therefore, it is considered effective in preventing skin aging to promote the production of hyaluronic acid and maintain the amount of hyaluronic acid.

[0007] Thus, the decrease in the amount and production ability of collagen and hyaluronic acid due to aging is considered to be one of the main causes of skin aging. Examination of components having an effect of promoting the production of collagen and hyaluronic acid has also been carried out. For example, a treated product of an organic solvent extract of konotegashiwa seeds has been reported (Patent Document 3). However, development of further novel components that can be widely applied to improvement, prevention, and treatment of symptoms related to the decrease in the amount and production ability of collagen and hyaluronic acid, including skin aging, has still been demanded.

Prior Art Documents

Patent Documents

[0008]

Patent Document 1

Patent Document 2

Patent Document 3

Summary of the Invention

Problems to be Solved by the Invention

[0009] The present invention aims to provide a novel collagen production promoter, a hyaluronic acid production promoter, a composition for promoting the production of collagen and / or hyaluronic acid, a wrinkle, firmness, texture, or skin moisture content improver, and a cosmetic or non-therapeutic method for promoting the production of collagen and / or hyaluronic acid.

Means for Solving the Problems

[0010] The present inventors conducted intensive studies to develop a novel component having an effect of promoting the production of collagen and hyaluronic acid, and as a result, they first found that N-benzoylglycylglycine can be used as an active ingredient of a collagen production promoter and a hyaluronic acid production promoter. Then, the inventors invented a composition for promoting the production of collagen and / or hyaluronic acid, a wrinkle, firmness, texture, or skin moisture content improver, and a cosmetic or non-therapeutic method for promoting the production of collagen and / or hyaluronic acid, which are characterized by containing N-benzoylglycylglycine.

[0011] Therefore, the present invention relates to the following inventions: [1] A hyaluronic acid production promoter containing N-benzoylglycylglycine as an active ingredient. [2] A collagen production promoter containing N-benzoylglycylglycine as an active ingredient. [3] The hyaluronic acid production promoter according to aspect 1, which is used for promoting the production of hyaluronic acid in epidermal cells, epidermal keratinocytes, dermal cells, and / or fibroblasts. [4] The collagen production promoter according to aspect 2, which is used for promoting the production of type I collagen and / or type III collagen. [5] A composition containing N-benzoylglycylglycine as an active ingredient for promoting the production of hyaluronic acid and / or collagen. [6] The composition according to aspect 5, which is a therapeutic agent for treating a condition or disease caused by a decrease in the ability to produce hyaluronic acid or collagen. [7] The composition according to aspect 5 or 6, which is a topical skin preparation. [8] The composition according to any one of aspects 5 to 7, which is a wrinkle-improving agent, firmness-improving agent, texture-improving agent, or skin moisture content-improving agent. [9] A cosmetic method or non-therapeutic method comprising applying a hyaluronic acid production promoter or collagen production promoter containing N-benzoylglycylglycine as an active ingredient.

[10] Use of N-benzoylglycylglycine for promoting the production of hyaluronic acid and / or collagen. [Effect of the Invention]

[0012] Collagen production and hyaluronic acid production can be promoted using N-benzoylglycylglycine. Furthermore, wrinkles, firmness, texture, or skin moisture content can be improved using N-benzoylglycylglycine. [Brief Description of the Drawings]

[0013]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Mode for Carrying Out the Invention

[0014] In one aspect, the present invention relates to a collagen production promoter or a hyaluronic acid production promoter, which is characterized by containing N-benzoylglycylglycine. Also, one aspect of the present invention relates to a composition containing N-benzoylglycylglycine as an active ingredient for promoting the production of hyaluronic acid and / or collagen. The composition according to this aspect can be the above-mentioned hyaluronic acid production promoter or collagen production promoter.

[0015] In this specification, the collagen production promoter, hyaluronic acid production promoter, and other compositions according to the present invention may sometimes be simply described as "the composition of the present invention" or "the composition according to the present invention".

[0016] N-benzoylglycylglycine is a compound represented by the following formula:

[0017]

Chemical Formula

[0018] N-Benzoylglycylglycine may be a commercially available product (for example, those sold by Wako Pure Chemical Industries, Ltd., etc.) or may be produced based on known methods.

[0019] In the present invention, the promotion of the production of collagen and hyaluronic acid means the promotion of the production of collagen and hyaluronic acid or the improvement of the production ability in cells, for example, cells constituting skin tissue. Therefore, the composition of the present invention can be used, although not limited thereto, for example, to promote the production of hyaluronic acid and / or collagen in epidermal cells, epidermal keratinocytes, dermal cells, fibroblasts, etc. In one embodiment, the composition of the present invention can promote the production of hyaluronic acid in both epidermal keratinocytes and fibroblasts.

[0020] In addition, in this specification, the method for evaluating the production ability of collagen and hyaluronic acid is not particularly limited. For example, it can be evaluated by visualizing the abundance of collagen (type I collagen and type III collagen) or hyaluronic acid in skin tissue to which the composition of the present invention is applied (coated) by immunohistochemical staining. Alternatively, it may be evaluated by measuring the abundance of collagen and / or hyaluronic acid in the cell culture medium (culture supernatant) after culturing in a culture solution containing the composition of the present invention by ELISA or the like.

[0021] The composition of the present invention containing N-benzoylglycylglycine has, as disclosed in the examples described later, not only the effect of promoting the production of type I collagen but also the effects of promoting the production of type III collagen and further promoting the production of hyaluronic acid. Therefore, in one aspect, the composition of the present invention is used to promote the production of type I collagen and / or type III collagen. Also, in one aspect, the composition of the present invention is used to promote the production of hyaluronic acid. The fact that N-benzoylglycylglycine has not only the effect of promoting the production of type I collagen but also the effects of promoting the production of type III collagen and hyaluronic acid is a finding first discovered by the present inventors.

[0022] There are various types of collagen that make up the skin, mainly type I collagen and type III collagen. Type I collagen mainly maintains the structure of the skin, and type III collagen mainly functions to impart flexibility to the skin. Therefore, not only type I collagen but also type III collagen plays an important role in maintaining and restoring the elasticity of the skin. Type III collagen is also called fetal collagen and is known to rapidly decrease with aging. Therefore, if the amount of type III collagen in the skin can be increased using the composition of the present invention, it is possible to prevent skin aging due to aging and bring about excellent effects in improving wrinkles, firmness, texture, and skin moisture content. In addition, the effect of promoting the production of type III collagen by the composition of the present invention can also bring about effects such as promoting wound healing, improving skin atrophy, and improving conditions and diseases related to type III collagen synthesis deficiency.

[0023] One aspect of the present invention relates to a therapeutic agent for preventing, improving, or treating a condition or disease caused by abnormal degradation of hyaluronic acid or collagen, or a decrease in the production ability of hyaluronic acid or collagen. Examples of conditions or diseases caused by abnormal degradation of hyaluronic acid or collagen, or a decrease in the production ability of hyaluronic acid or collagen include skin aging, an increase in wrinkles, a decrease in firmness, a decrease in texture, or a decrease in skin moisture content, etc.; wounds caused by external and internal factors, such as incisions, lacerations, puncture wounds, bite wounds, abrasions, gunshot wounds, contusions, burns, pressure ulcers, diabetic ulcers, chemical injuries, etc.; various diseases caused by the deficiency of the above-mentioned type III collagen synthesis, such as clinical symptom type IV of Ehlers-Danlos syndrome and arterial rupture and intestinal perforation caused thereby; furthermore, hepatitis, gingivitis, rheumatism, osteoarthritis, tenosynovitis, osteoporosis, malignant tumors, liver cirrhosis, transplant rejection, scleroderma, liver cirrhosis, arteriosclerosis, hypertension, fibrosis, xeroderma, dry skin, rough skin, nephritis, keloid, overrepair, sepsis, etc. are included, but not limited thereto.

[0024] The composition of the present invention is particularly useful as a wrinkle-improving agent, firming agent, texture-improving agent, or skin moisture content-improving agent.

[0025] The content of N-benzoylglycylglycine in the composition according to the present invention can be any content within the range of 0.0001 to 10% by weight with respect to the total amount of the final products such as lotion and essence described later. For example, the composition according to the present invention may contain at least 0.0001% by weight, 0.001% by weight, 0.01% by weight, or 0.1% by weight of N-benzoylglycylglycine. Further, for example, the composition according to the present invention may contain 10% by weight or less, 5% by weight or less, 1% by weight or less, or 0.5% by weight or less of N-benzoylglycylglycine. Depending on the characteristics of the final product, any combination of these upper and lower limits can be defined as the range of the content of N-benzoylglycylglycine.

[0026] The composition of the present invention can be formulated into functional foods, cosmetics, quasi-drugs, and pharmaceuticals for the purpose of promoting the production of collagen and / or hyaluronic acid. In other words, the composition containing N-benzoylglycylglycine according to the present invention can be prepared as a functional food, a cosmetic, a quasi-drug, or a pharmaceutical.

[0027] The composition of the present invention can be used by any administration route including oral administration and intravenous injection, but transdermal administration is preferred from the viewpoint of directly acting on the skin. By transdermal administration, even a drug that causes side effects in systemic administration may be acceptable from the viewpoint of improving the skin condition.

[0028] Therefore, the composition of the present invention is preferably formulated as a topical skin preparation that can be directly applied to the skin. In such a topical skin preparation, in addition to N-benzoylglycylglycine, optional components used in cosmetics, pharmaceuticals, etc. can be appropriately formulated as needed. Examples of such optional components include, for example, oils, surfactants, powders, colorants, water, alcohols, thickeners, chelating agents, silicones, antioxidants, ultraviolet absorbers, moisturizers, fragrances, various skin nutrients, pH adjusters, neutralizing agents, powder components, colorants, aqueous components, water, and the like.

[0029] Furthermore, metal ion sequestering agents such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, gluconic acid, etc., preservatives such as methyl paraben, ethyl paraben, butyl paraben, etc., caffeine, tannin, verapamil, tranexamic acid and its derivatives, anti-inflammatory components, licorice extract, glabridin, hot water extract of the fruit of Paliurus ramosissimus, various crude drugs, tocopheryl acetate, glycyrrhizic acid and its derivatives or salts thereof, various medicinal components such as salicylic acid and its derivatives or salts thereof, vitamin C, magnesium ascorbyl phosphate, ascorbyl glucoside, 3-O-ethylascorbic acid, disodium adenosine monophosphate OT, arbutin, kojic acid, 4-methoxysalicylic acid, and salts thereof, 4-n-butylresorcinol and resorcinol derivatives, linoleic acid, ellagic acid, chamomile extract, 5,5'-dipropyl-biphenyl-2,2'-diol, whitening agents such as dexpanthol, retinoic acid, retinal, retinol, retinol acetate, retinol propionate, retinol palmitate, and other retinol derivatives, nicotinamide and its derivatives, sodium isopropyl oxopropylaminocarbonyl pyrrolidine carbonylmethylpropylaminocarbonyl benzoylaminoacetate, pyridoxine and its salts, rice power extract, and furthermore, saccharides such as glucose, fructose, mannose, sucrose, trehalose, etc. can also be appropriately formulated.

[0030] The topical skin preparation can be widely applied to cosmetics, quasi-drugs, etc. applied to the skin, and particularly preferably to cosmetics. Any dosage form that can be applied to the skin may be used, including solution systems, solubilized systems, emulsified systems, powder dispersion systems, water-oil two-layer systems, water-oil-powder three-layer systems, ointments, lotions, gels, aerosols, etc. Examples of the topical skin preparation include solid external preparations, external powders, external liquid preparations, liniments, sprays, external aerosol preparations, pump sprays, ointment preparations, cream preparations, gel preparations, patches, tape preparations, poultices, etc.

[0031] The form of use is also arbitrary. For example, facial cosmetics such as lotions, essences, milks, creams, packs, etc., makeup cosmetics such as foundations, lipsticks, eyeshadows, etc., sunscreen, aftercare lotions, body oils, etc. for the whole body, fragrance cosmetics, bath preparations, etc. can be mentioned. However, it can be incorporated into any cosmetic as long as it is applicable to the skin.

[0032] Also, if it is a makeup cosmetic, it can be widely applied to forms such as foundations, etc., and as toiletry products, it can be widely applied to forms such as body soaps, soaps, etc. Furthermore, if it is a quasi-drug, it can be widely applied to forms such as various ointment preparations. And the form that the composition of the present invention can take is not limited to these dosage forms and forms.

[0033] Furthermore, one aspect of the present invention relates to a beauty method or non-therapeutic method including applying the composition of the present invention containing N-benzoylglycylglycine (for example, a hyaluronic acid production promoter or a collagen production promoter). The beauty method or non-therapeutic method of this aspect can be a method for preventing, improving, or treating a condition or disease caused by abnormal degradation of hyaluronic acid or collagen, or a decrease in the production ability of hyaluronic acid or collagen in a subject. In this aspect, the subject can be, but is not limited to, mammals, particularly humans, and non-human mammals such as dogs, cats, cows, horses, pigs, etc. The subject is preferably a human.

[0034] In the beauty or non-therapeutic method according to this aspect, the composition of the present invention, for example, a topical skin preparation, is applied to a site where a desired effect is to be obtained, such as the face, neck, hands, arms, legs, etc. The application can be performed in any manner, such as by applying with hands or an applicator. The number of applications, frequency, amount, etc. are appropriately set according to the desired effect, etc. In one aspect, the beauty or non-therapeutic method according to the present invention does not include so-called medical acts.

[0035] Furthermore, one aspect of the present invention relates to the use of N-benzoylglycylglycine for promoting the production of collagen and / or hyaluronic acid. From another perspective, the aspect of the present invention includes N-benzoylglycylglycine for use in improving a decrease or deficiency in the production of collagen and / or hyaluronic acid. The data disclosed herein demonstrate that N-benzoylglycylglycine is useful for improving a decrease or deficiency in the production of collagen and / or hyaluronic acid. Therefore, a composition of the present invention can be prepared using N-benzoylglycylglycine for use in improving abnormal degradation of collagen and / or hyaluronic acid, or a decrease in the production ability of collagen and / or hyaluronic acid. That is, one aspect of the present invention also relates to the use of N-benzoylglycylglycine in the production of a composition for promoting the production of collagen and / or hyaluronic acid. In these aspects, the improvement of abnormal degradation of collagen and / or hyaluronic acid, or a decrease in the production ability of collagen and / or hyaluronic acid can be an improvement in wrinkles, firmness, texture, or skin moisture content.

[0036] Formulation examples of the composition of the present invention containing N-benzoylglycylglycine are illustrated, but it goes without saying that the present invention is not limited to these formulation examples. The numerical values of each component are in mass%.

[0037] Formulation Example 1 Cream (Formulation) Stearic acid 5.0 Stearyl alcohol 4.0 Isopropyl myristate 18.0 Glycerin monostearate 3.0 Retinol 0.1 Dipropylene glycol 10.0 N-Benzoylglycylglycine (Collagen production promoter or hyaluronic acid production promoter) 0.1 Potassium hydroxide 0.35 Sodium bisulfite appropriate amount Preservative appropriate amount Fragrance appropriate amount Ion-exchanged water remainder (Production method) Add dipropylene glycol, potassium hydroxide, and N-benzoylglycylglycine to ion-exchanged water, heat and maintain at 70°C (aqueous phase). Mix the other components, heat and melt, and maintain at 70°C (oil phase). Gradually add the oil phase to the aqueous phase, keep at that temperature for a while after all is added to cause a reaction. Then, emulsify uniformly with a homomixer and cool to 30°C while stirring well.

[0038] Formulation Example 2 Cream (Formulation) Stearic acid 2.0 Stearyl alcohol 7.0 Macadamia nut fatty acid phytosterol 2.0 Squalane 5.0 Pentaerythrityl tetraethylhexanoate 6.0 Polyoxyethylene (25 moles) cetyl ether 3.0 Cetyl ether Glycerin monostearate 2.0 Retinol acetate 0.1 Dipropylene glycol 5.0 N-Benzoylglycylglycine (Collagen production promoter or hyaluronic acid production promoter) 0.001 Potassium hydroxide 0.14 Sodium bisulfite appropriate amount Ethylparaben appropriate amount Fragrance appropriate amount Ion-exchanged water remainder (Production method) Add dipropylene glycol, caustic potash, and N-benzoyl-glycylglycine to ion-exchanged water, and heat to maintain at 70°C (aqueous phase). Mix the other components, heat and melt them, and maintain at 70°C (oil phase). Add the oil phase to the aqueous phase for preliminary emulsification, emulsify uniformly with a homomixer, and then cool to 30°C while stirring well.

[0039] Formulation Example 3 Emulsion (Wrinkle improver, firmness improver, texture improver, or skin moisture content improver) (Formulation) Stearic acid 2.5 Cetyl alcohol 1.5 Petrolatum 5.0 Liquid paraffin 10.0 Stearic acid PEG-5 glyceryl 2.0 Polyethylene glycol 1500 3.0 Triethanolamine 1.3 Carboxyvinyl polymer 0.05 N-benzoyl-glycylglycine (Collagen production promoter or hyaluronic acid production promoter) 0.05 Tranexamic acid 1.0 Sodium bisulfite appropriate amount Ethylparaben appropriate amount Fragrance appropriate amount Ion-exchanged water remainder (Production method) Dissolve carboxyvinyl polymer in a small amount of ion-exchanged water (Phase A). Add polyethylene glycol 1500, triethanolamine, N-benzoyl-glycylglycine, and tranexamic acid to the remaining ion-exchanged water, heat and dissolve, and maintain at 70°C (aqueous phase). Mix the other components, heat and melt, and maintain at 70°C (oil phase). Add the oil phase to the aqueous phase for preliminary emulsification, add Phase A, emulsify uniformly with a homomixer, and cool to 30°C while stirring well after emulsification.

[0040] Formulation Example 4 Jelly (Formulation) Alcohol 10.0 Dipropylene glycol 15.0 Polyoxyethylene (50 mol) oleyl ether 2.0 Carboxyvinyl polymer 0.3 Caustic potash 0.12 N-Benzoylglycylglycine (Collagen production promoter or hyaluronic acid production promoter) 0.1 Tranexamic acid methylamide hydrochloride 0.2 Ethylenediaminetetraacetate· 3 Sodium·2 water (EDTA-3Na) 0.05 Methylparaben appropriate amount Fragrance appropriate amount Ion-exchanged water remainder (Production method) Dissolve carboxyvinyl polymer uniformly in ion-exchanged water. On the other hand, dissolve polyoxyethylene (50 mol) oleyl ether in alcohol and add it to the aqueous phase. Then, add the other components and neutralize with caustic potash and L-arginine to thicken it.

[0041] Formulation example 5 Cream (Formulation) Stearic acid 5.0 Stearyl alcohol 4.0 Isopropyl myristate 18.0 Glycerin monostearate 3.0 Dipropylene glycol 10.0 N-Benzoylglycylglycine (Collagen production promoter or hyaluronic acid production promoter) 0.01 Potassium 4-methoxysalicylate 1.0 Caustic potash 0.35 Sodium bisulfite 0.01 Preservative appropriate amount Fragrance appropriate amount Ion-exchanged water remainder (Production method) Add dipropylene glycol, potassium hydroxide, and N-benzoyl-glycylglycine to ion-exchanged water, and heat while adding potassium 4-methoxysalicylate to maintain the temperature at 70 °C (aqueous phase). Mix the other components, heat and melt them, and maintain the temperature at 70 °C (oil phase). Gradually add the oil phase to the aqueous phase while stirring the oil phase, and after adding all of it, maintain the temperature for a while to cause a reaction. Then, emulsify it uniformly with a homomixer and cool it to 30 °C while stirring well.

[0042] Formulation Example 6 Emulsion (Wrinkle improver, firmness improver, skin texture improver, or skin moisture content improver) (Formulation) Microcrystalline wax 1.0 Beeswax 2.0 Liquid paraffin 1.0 Squalane 5.0 Polyoxy monooleate 4.0 Ethylene sorbitan (20 E.O.) Sorbitan monooleate 1.0 Dipropylene glycol 7.0 N-benzoyl-glycylglycine (Collagen production promoter or hyaluronic acid production promoter) 10.0 Hydrolyzed extract of Prunus mume 0.1 Potassium hydroxide 2.0 Saccharoglucan 1.0 Sodium bisulfite Appropriate amount Ethylparaben Appropriate amount Fragrance Appropriate amount Ion-exchanged water Remainder (Production method) Add dipropylene glycol, saccharoglucan, potassium hydroxide, and N-benzoyl-glycylglycine to ion-exchanged water, heat, and maintain the temperature at 70 °C (aqueous phase). Mix the other components, heat and melt them, and maintain the temperature at 70 °C (oil phase). Gradually add the aqueous phase to this while stirring the oil phase, and emulsify it uniformly with a homomixer. After emulsification, cool it to 30 °C while stirring well.

[0043] Formulation Example 7 Pack (Formulation) (Phase A) Dipropylene glycol 5.0 Polyoxyethylene (60 mol) hydrogenated castor oil 5.0 (Phase B) Olive oil 5.0 Tocopherol acetate 0.2 Ethylparaben appropriate amount Fragrance appropriate amount (Phase C) Sodium bisulfite appropriate amount Polyvinyl alcohol 13.0 (Saponification degree 90, polymerization degree 2,000) Ethanol 7.0 N-Benzoylglycylglycine (Collagen production promoter or hyaluronic acid production promoter) 5.0 Dimethylpyrazolyl dimethylpyrimidine hydrochloride 0.1 Caustic potash 1.0 Purified water remainder (Production method) Mix Phase A, Phase B, and Phase C respectively. Keep Phase C heated at 70 °C. Add Phase B to Phase A and mix. Then add this to Phase C, mix well, and then cool and fill.

[0044] Formulation Example 8 Cream (Formulation) Alcohol 5.0 Glycerin 3.0 1,3-Butylene glycol 7.0 Potassium hydroxide 0.26 Carboxyvinyl polymer 0.25 (Acrylates / alkyl acrylate (C10-30)) Cross polymer 0.1 Xanthan gum 0.05 PPG-13 decyltetradeceth-24 1.0 Pentaerythrityl tetraethylhexanoate 15.0 Hydrogenated polydecene 7.0 Squalane 3.0 N-Benzoylglycylglycine (Collagen production promoter or hyaluronic acid production promoter) 0.48 Phenoxyethanol Appropriate amount EDTA-3Na Appropriate amount Ion-exchanged water The remainder

[0045] All documents referred to in this specification are hereby incorporated by reference in their entirety into this specification.

[0046] The examples of the present invention described below are for illustrative purposes only and do not limit the technical scope of the present invention. The technical scope of the present invention is limited only by the description in the claims. Changes to the present invention, for example, addition, deletion, and substitution of the constituent elements of the present invention, can be made on the condition that the gist of the present invention is not deviated from.

Example

[0047] 1. Immunohistochemical staining test The test composition was applied to the skin tissue donated by the consenting subjects as follows and cultured for 10 days. The cultured skin tissue was evaluated by an immunohistochemical staining test. [Skin tissue] Site: Non-exposed part (abdomen) Skin tissue size: 1 cm × 1 cm Donor: Caucasian female (40 years old) [Test composition] Composition of the present invention: The composition of the present invention was prepared in the same manner as in Formulation Example 8 above. Comparative composition (placebo): A comparative composition was prepared in the same manner as the composition of the present invention above, except that it did not contain N-benzoylglycylglycine. Incidentally, the structure of N-benzoylglycylglycine (Wako Pure Chemical Industries, Ltd.) used in this test is shown below.

[0048]

Chemical formula

[0049] [Immunohistochemical staining] Reagents, etc. Primary antibody: Anti-human collagen type I (Rabbit) Anti-Collagen Type III (RABBIT) Antibody Hyaluronic Acid Binding Protein, Bovine Nasal Cartilage, Biotinylated Chromogenic substrate: DAB (3,3'-diaminobenzidine) (manufactured by DAKO) Counterstain: Hematoxylin solution (manufactured by Fujifilm Wako Pure Chemical Corporation) The skin tissue to which the test composition was applied was fixed with 4% PFA, and paraffin blocks were prepared. The tissue was sectioned to a thickness of 5 μm and subjected to immunohistochemical staining. Briefly, after deparaffinization, inactivation of endogenous peroxidase, and blocking treatment, the reaction of the primary antibody, the chromogenic reaction with the chromogenic substrate DAB, and counterstaining with hematoxylin were performed, and then it was mounted.

[0050] The results are shown in Figures 1 to 3. As shown in Figures 1 and 2, it can be seen that the application of the composition of the present invention containing N-benzoyl-glycyl-glycine promotes the production of both type I collagen and type III collagen. Also, as shown in Figure 3, it can be seen that the application of the composition of the present invention containing N-benzoyl-glycyl-glycine promotes the production of hyaluronic acid in both the epidermis and the dermis.

[0051] 2. In vitro test using epidermal keratinocytes Using a 24-well plate, human epidermal keratinocytes were seeded at 2×10 4 cells / well in HuMedia medium and left standing for 24 hours while culturing until the cells adhered sufficiently. One day later, it was replaced with HuMedia medium containing a sample at a predetermined concentration and cultured for 2 days. After culturing, the culture supernatant was removed, and the amount of hyaluronic acid produced was measured by ELISA (product name: Hyaluronic Acid, ELISA Kit, QnE (96well)). Note that N-benzoyl-glycyl-glycine (concentration in the medium: 0.00001% or 0.0001%) was used as the sample, DMSO ("Blank (DMSO)" in Fig. 4) as the negative control, and retinol ("RO" in Fig. 4, concentration in the medium: 0.01 μM) as the positive control.

[0052] The results are shown in Fig. 4. The results were calculated as relative values to the hyaluronic acid production amount of Blank without N-benzoyl-glycyl-glycine being 100. Also, the significant difference between groups was verified by a t-test (threshold of p-value 0.05). As shown in Fig. 4, it can be seen that the addition of N-benzoyl-glycyl-glycine significantly (p < 0.01 or p < 0.001) promotes hyaluronic acid production in epidermal keratinocytes. Also, it can be seen that N-benzoyl-glycyl-glycine has an equivalent or better effect of promoting hyaluronic acid production compared to retinol (p < 0.01).

[0053] 3. In vitro test using fibroblasts Using a 24-well plate, human fibroblasts were seeded at 6 × 10 4 cells / well in DMEM medium containing 10% FBS and left standing for 6 hours while culturing until the cells adhered sufficiently. The serum concentration was reduced and cultured with DMEM medium containing 0.5% FBS. One day later, it was replaced with DMEM medium containing 0.5% FBS containing the sample at a predetermined concentration and cultured for 3 days. The culture supernatant was extracted, and the hyaluronic acid production amount was measured by ELISA (product name: Hyaluronic Acid, ELISA Kit, QnE (96well)). Note that N-benzoyl-glycyl-glycine (concentration in the medium: 0.0001% or 0.001%) was used as the sample, DMSO ("Blank (DMSO)" in Fig. 5) as the negative control, and 5% FBS as the positive control.

[0054] The results are shown in Fig. 5. The results were calculated as relative values to the hyaluronic acid production amount of Blank not containing N-benzoyl-glycylglycine, which was set to 100. Also, the significant difference between groups was verified by a t-test (threshold of p-value: 0.05). As shown in Fig. 5, it can be seen that N-benzoyl-glycylglycine has an effect of promoting hyaluronic acid production with a significant difference equivalent to that of 5% FBS (both p < 0.001).

[0055] From the above results, since the composition of the present invention containing N-benzoyl-glycylglycine has an effect of promoting the production of type I collagen, type III collagen, and hyaluronic acid, it is found to be useful for improving wrinkles, firmness, texture, and skin moisture content. These are findings discovered for the first time in the present invention.

Industrial Applicability

[0056] The hyaluronic acid production promoter, collagen production promoter, and other compositions of the present invention have excellent effects of promoting collagen production and hyaluronic acid production, and are useful for improving wrinkles, firmness, texture, and skin moisture content. Therefore, they can be used in various fields related to cosmetics, beauty, pharmaceuticals, etc.

Claims

1. A hyaluronic acid production promoter containing N-benzoyl-glycylglycine as an active ingredient.

2. The hyaluronic acid production promoter according to claim 1, which can further promote the production of collagen.

3. The hyaluronic acid production promoter according to claim 1 or 2, which is used to promote the production of hyaluronic acid in epidermal cells, epidermal keratinocytes, dermal cells, and / or fibroblasts.

4. The hyaluronic acid production promoter according to claim 2, which is used to promote the production of type I collagen and / or type III collagen.

5. A composition containing N-benzoyl-glycylglycine as an active ingredient for promoting the production of hyaluronic acid.

6. The composition according to claim 5, which is further for promoting the production of collagen.

7. The composition according to claim 5 or 6, which is a therapeutic agent for treating a condition or disease caused by a decrease in the ability to produce hyaluronic acid.

8. The composition according to claim 6, which is a therapeutic agent for treating a condition or disease caused by a decrease in the ability to produce collagen.

9. The composition according to any one of claims 5 to 8, which is a topical skin preparation.

10. The composition according to claim 5 or 6, which is a wrinkle-improving agent, firmness-improving agent, skin texture-improving agent, or skin moisture content-improving agent.

11. The composition according to claim 7 or 8, which is a wrinkle-improving agent, firmness-improving agent, skin texture-improving agent, or skin moisture content-improving agent.

12. A cosmetic method or non-therapeutic method including applying a hyaluronic acid production promoter containing N-benzoyl-glycylglycine as an active ingredient.

13. Use of N-benzoyl-glycylglycine for promoting the production of hyaluronic acid.

14. The use according to claim 13, which is further for promoting the production of collagen.

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