Anti-stress and skin improvement cosmetic composition comprising spinach extract as an active ingredient

Abstract

The present invention relates to a cosmetic composition for anti-stress and skin improvement comprising a *Seomcho* extract as an active ingredient. The composition comprising the *Seomcho* extract of the present invention as an active ingredient can enhance skin immunity and defense against various internal and external stresses, increase skin energy and vitality, and regulate skin biorhythms to activate slowed skin turnover cycles, thereby improving the function of damaged skin and maintaining healthy skin. It can be usefully utilized as a cosmetic material for neurocosmetics.

Description

Technology Field

[0001] The present invention relates to a cosmetic composition for anti-stress and skin improvement comprising an extract of *Seomcho* as an active ingredient. Background Technology

[0003] Neurocosmetics, one of the beauty trends recently gaining attention in various countries, refers to cosmetics that help improve the skin by eliciting reactions with skin cells through neurotransmitters that connect the brain. Neurocosmetics originated from "emotional cosmetics," which utilize the brain's nervous system to allow consumers to directly experience a variety of senses. The core of neurocosmetics lies in capturing consumers' emotions by acting on the nervous system—the mediator between the brain and the skin—through visual stimulation via intense colors, olfactory stimulation using fragrances, and tactile stimulation felt through textures. Subsequently, as research on neurotransmitters and their effects on the skin became more concrete, studies began in earnest that went beyond emotional cosmetics to demonstrate how cosmetics can significantly influence the signal transmission process between the brain and the nervous system.

[0004] In fact, skin condition is closely linked to emotional energy. Negative mental energy is immediately transmitted to the brain, which in turn causes pain or skin changes. When under stress, neurotransmitters (neuropeptides) such as Substance P, CGRP, and POMC are released. These neurotransmitters affect keratinocytes, mast cells, and vascular endothelial cells, causing them to release inflammatory cytokines, dilate blood vessels, and induce itching and rashes. Furthermore, emotions such as anger or hatred trigger the release of hormones like norepinephrine and cortisol, which constrict capillaries, obstruct blood flow, and cause facial flushing or skin problems. Neurocosmetics play a crucial role in regulating skin balance and improving signs of aging by activating or inhibiting these messengers between the nervous system and the brain.

[0005] Furthermore, neurocosmetics delivers messages to skin cells by incorporating neurotransmission processes that possess characteristics similar to those of nerve cells. For instance, neurocosmetics deactivates the sensitivity of specific nerve (sensory) receptors in the skin, thereby minimizing sensitive reactions to stimuli, maintaining skin health, preventing aging, and preserving beauty. Specifically, neurocosmetics inhibits the release of dopamine and substances that act as precursors to inflammatory responses, while stimulating the release of beta-endorphin, which revitalizes skin cell regeneration. This improves skin cell function and contributes to the homeostasis of the immune system and energy balance. Through these mechanisms, neurocosmetics allows the skin to feel comfortable and strengthens its defense capabilities, enabling it to protect itself from external stimuli while remaining soft and stable.

[0006] Therefore, we aim to develop cosmetic materials as neurocosmetics that prevent skin aging and maintain healthy skin by enhancing skin immunity and defense against various stresses, increasing skin energy and vitality, and helping to maintain homeostasis by regulating the skin's biorhythm. Prior art literature

[0008] Republic of Korea Published Patent No. 10-2020-0086395 (Published July 17, 2020) The problem to be solved

[0009] The inventors have made diligent research efforts to develop cosmetic materials as neurocosmetics that prevent skin aging and maintain healthy skin. As a result, it was confirmed that Seomcho extract enhances skin immunity and skin defense against various stresses. Furthermore, by identifying that it can increase skin energy and vitality, the present invention was completed. means of solving the problem

[0011] The inventors have made diligent research efforts to develop cosmetic materials as neurocosmetics that prevent skin aging and maintain healthy skin. As a result, it was confirmed that Seomcho extract enhances skin immunity and skin defense against various stresses. Furthermore, by identifying that it can increase skin energy and vitality, the present invention was completed.

[0012] According to one aspect of the present invention, the present invention provides a cosmetic composition for stress relief comprising an extract of *Seomcho* as an active ingredient.

[0013] In the present invention, "spinach, Spinacia oleracea "..." refers to spinach that has been bred in the Sinan region of South Jeolla Province. It is an annual or biennial vegetable native to Southwest Asia, and its roots are fleshy, thick, and long. The main stem is hollow and grows to a height of about 50 cm. The leaves are alternate and have petioles; they are characterized by being deeply lobed at the base and smooth at the top.

[0014] Seomcho is known as a treasure trove of vitamins as it contains a balanced mix of vitamins A, B, and C. In particular, its content of beta-carotene—vitamin A found in green and yellow vegetables—is significantly higher than that of other vegetables. Beta-carotene helps inhibit the onset of cancer by preventing the formation or proliferation of cancer cells.

[0015] The *Seomcho* extract used in the present invention can be obtained by purchasing or by direct extraction. When the *Seomcho* extract used in the composition of the present invention is obtained by treating with an extraction solvent, various extraction solvents, such as polar solvents or non-polar solvents, may be used.

[0016] Suitable polar solvents may include, but are not limited to, (i) water, (ii) alcohol (preferably methanol, ethanol, propanol, butanol, normal-propanol, iso-propanol, normal-butanol, 1-pentanol, 2-butoxyethanol or ethylene glycol), (iii) acetic acid, (iv) DMFO (dimethyl-formamide), and (v) DMSO (dimethylsulfoxide).

[0017] Suitable as non-polar solvents may include, but are not limited to, acetone, acetonitrile, ethyl acetate, methyl acetate, fluoroalkanes, pentane, hexane, 2,2,4-trimethylpentane, decane, cyclohexane, cyclopentane, diisobutylene, 1-pentene, 1-chlorobutane, 1-chloropentane, o-xylene, diisopropyl ether, 2-chloropropane, toluene, 1-chloropropane, chlorobenzene, benzene, diethyl ether, diethyl sulfide, chloroform, dichloromethane, 1,2-dichloroethane, aniline, diethylamine, ether, carbon tetrachloride, and THF (Tetrahydrofuran).

[0018] In one embodiment of the present invention, the Seomcho extract is extracted with any one extraction solvent selected from the group consisting of water, organic solvents and mixed solvents thereof, wherein the organic solvent may be selected from the group consisting of lower alcohols having 1 to 4 carbon atoms, petroleum ether, hexane, benzene, chloroform, methylene chloride, ether, ethyl acetate, and acetone, but is not limited thereto.

[0019] In one embodiment of the present invention, the alcohol is selected from the group consisting of methanol, ethanol, propanol, butanol, normal-propanol, iso-propanol, normal-butanol, and mixed solvents thereof.

[0020] In the present invention, the term "extract" has the meaning commonly used in the art as a crude extract as described above, but in a broader sense, it also includes fractions obtained by further fractionating the extract. That is, the extract of *Seomcho* includes not only that obtained using the extraction solvent described above, but also that obtained by additionally applying a purification process thereto. For example, fractions obtained through various additional purification methods, such as a fraction obtained by passing the extract through an ultrafiltration membrane having a certain molecular weight cut-off value, or separation by various chromatographs (designed for separation based on size, charge, hydrophobicity, or affinity), may also be included in the *Seomcho* extract of the present invention.

[0021] Preferably, ethanol may be selected as the extraction solvent, but is not limited thereto.

[0022] In the present invention, when extracting Seomcho, hot water extraction, pressurized hot water extraction, cold maceration extraction, reflux extraction, room temperature extraction, or ultrasonic extraction may be used, but is not limited thereto.

[0023] To prepare an extract according to one embodiment of the present invention, the amount of solvent used for extraction can be appropriately selected according to the amount of Seomcho used. Specifically, the volume may be 1 to 40 times the weight of Seomcho used for preparing the extract, more specifically 1 to 30 times, 1 to 25 times, 1 to 20 times, and even more specifically 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 times the weight of Seomcho, but is not limited thereto.

[0024] In one embodiment of the present invention, the ethanol is an ethanol solution of 60 to 100 volume%. In a more specific embodiment of the present invention, the ethanol may be 60 to 90 volume%, 60 to 80 volume%, 65 to 100 volume%, 65 to 90 volume%, 65 to 80 volume%, 70 volume%, 80 volume%, 90 volume%, 95 volume%, or 100 volume%, but is not limited thereto.

[0025] The extraction temperature of the ethanol extract according to one embodiment of the present invention is not particularly limited and, for example, may be 10°C to 110°C, and specifically, may be 20°C to 100°C, 30°C to 100°C, 40°C to 100°C, 50°C to 100°C, 60°C to 100°C, 20°C to 90°C, 30°C to 90°C, 40°C to 90°C, 50°C to 90°C, 60°C to 90°C, 20°C to 80°C, 30°C to 80°C, 40°C to 80°C, 50°C to 80°C, 60°C to 80°C, 20°C to 70°C, 30°C to 70°C, 40°C to 70°C, 50°C to 70°C, 60°C to 70°C, but therein It is not limited.

[0026] The extraction time of the ethanol extract according to one embodiment of the present invention is not particularly limited and, for example, may be 1 hour to 10 hours, and specifically, may be 1 hour to 9 hours, 1 hour to 8 hours, 1 hour to 7 hours, 1 hour to 6 hours, 1 hour to 5 hours, 2 hours to 9 hours, 2 hours to 8 hours, 2 hours to 7 hours, 2 hours to 6 hours, 2 hours to 5 hours, 3 hours to 9 hours, 3 hours to 8 hours, 3 hours to 7 hours, 3 hours to 6 hours, 3 hours to 5 hours, 4 hours to 9 hours, 4 hours to 8 hours, 4 hours to 7 hours, 4 hours to 6 hours, 4 hours to 5 hours, 5 hours to 9 hours, 5 hours to 8 hours, 5 hours to 7 hours, 5 hours to 6 hours, but is not limited thereto.

[0027] The sea mustard extract used in the present invention can be prepared in a powder state by additional processes such as vacuum distillation, freeze-drying, or spray-drying.

[0028] In the present invention, "including as an active ingredient" means including an amount sufficient to achieve the efficacy or activity of the said Seomcho extract.

[0029] In the present invention, "stress relief" means that when skin cells are damaged or stressed by internal (psychological stress, etc.) or external stimuli (ultraviolet rays, harmful environment, excessive use of cosmetics, etc.), or when skin cells are damaged or stressed by internal or external stimuli, such as skin that is naturally red or sensitive or skin that is sensitive to irritation, the skin is restored through anti-inflammatory, antioxidant, elasticity improvement, etc.

[0030] In one embodiment of the present invention, the composition reduces the expression of the 11β-HSD1 (11 beta-hydroxysteroid dehydrogenase type 1) gene.

[0031] In the present invention, "11β-HSD1 (11 beta-hydroxysteroid dehydrogenase type 1)" refers to an enzyme that converts inactive cortisone into active cortisol, which is known to be increased in aged skin and activated by inflammatory cytokines.

[0032] In one embodiment of the present invention, when a group of fibroblasts treated with TNF-α was treated with Seomcho extract, it was confirmed that 11β-HSD1 mRNA expression was reduced compared to the TNF-α treated group.

[0033] In one embodiment of the present invention, the composition reduces the mRNA expression level of TRPV1.

[0034] In the present invention, "TRPV1 (Transient Receptor Potential Vanilloid 1; capsaicin receptor)" is a cutaneous neuropain sensory receptor found in all nervous systems, including the skin, spinal cord, and brain. It is activated by various internal and external factors (heat, ultraviolet rays, etc.) and chemical stimuli (capsaicin). When TRPV1 is activated, it detects heat, triggers inflammation, and transmits pain. This causes the secretion of cytokines, which are signaling molecules that control and stimulate the body's defense system, resulting in skin rashes, burning sensations, and itching.

[0035] In one embodiment of the present invention, when a group of capsaicin-treated keratinocytes was treated with a sea mustard extract, it was confirmed that TRPV1 mRNA expression was reduced compared to the capsaicin-treated group.

[0036] The inhibitory effect of Seomcho extract on TRPV1 expression means that it can inhibit the induction of inflammation, itching, and pain, and soothe sensitive skin.

[0037] According to another aspect of the present invention, the present invention provides a cosmetic composition for strengthening skin defense, comprising a sea mustard extract as an active ingredient.

[0038] In the present invention, "strengthening skin defense" refers to the ability to protect the skin from external stimuli and toxins, and the stimuli refer to stimuli harmful to the skin, such as bacteria, viruses, ultraviolet rays, dryness, and wounds. When bacteria invade the skin, the skin fights against them on its own by secreting antimicrobial peptides. Therefore, a substance that promotes the antimicrobial peptide activity of the skin can enhance the skin's antimicrobial and anti-inflammatory activities by enhancing the skin's own defense without irritating the skin.

[0039] In one embodiment of the present invention, the composition increases the expression of the HBD-2 (Human beta-defensin-2) gene.

[0040] In the present invention, "HBD-2 (Human beta-defensin-2)" is an antimicrobial peptide that performs a primary defense function by rapidly increasing in response to infection or inflammatory reactions, and acts as a chemical barrier that enhances immunity by activating immune cells. A decrease in the expression of HBD-2 is a cause of high levels of bacterial infection and inflammation.

[0041] In one embodiment of the present invention, when a group of keratinocytes treated with TNF-α, IFN-γ, and IL-4 was treated with Seomcho extract, it was confirmed that HBD-2 mRNA expression, which was reduced by TNF-α+IFN-γ+IL-4, was increased.

[0042] According to another aspect of the present invention, the present invention provides a cosmetic composition for improving itching comprising an extract of *Seomcho* as an active ingredient.

[0043] In the present invention, "pruritus" is also referred to as skin itching and is not particularly limited, but examples include itching caused by dry skin; itching caused by urticaria; itching caused by insect bites; itching caused by prickly heat, acne, chafing, frostbite, contact dermatitis, seborrheic dermatitis, or psoriasis; itching caused by various skin diseases; scalp itching, etc. However, it goes without saying that the present invention does not exclude skin itching of various causes other than those listed above. The present invention includes all types of itching known in the art, such as dermatological itching, systemic itching, neuropathic itching, and psychogenic itching.

[0044] In one embodiment of the present invention, the composition reduces the expression of the NGF (Nerve growth factor) gene.

[0045] In this invention, "NGF (Nerve growth factor)" refers to a nerve growth factor that induces itching by penetrating nerve fibers present in the dermis into the epidermis.

[0046] Continuous secretion of NGF accelerates epidermal hypertrophy caused by the proliferation of keratinocytes, induces the activation of mast cells in atopic dermatitis lesions, and further intensifies skin pruritus.

[0047] In the present invention, "TNF-α (Tumor necrosis factor-α)" and "IL-1β (Interleukin-1β)" are cytokines involved in inflammation and immune responses, and are known to have increased expression in inflammatory diseases.

[0048] NGF (Nerve growth factor), an itching factor, and TNF-α (Tumor necrosis factor-α), an inflammatory factor, are increased by Substance P, a neurotransmitter secreted when under stress.

[0049] In one embodiment of the present invention, when a group of keratinocytes treated with Substance P was treated with Seomcho extract, it was confirmed that the expression of NGF, TNF-α, and IL-1β mRNA was reduced compared to the group treated with Substance P.

[0050] According to another aspect of the present invention, the present invention provides a cosmetic composition for improving skin condition by activating skin energy metabolism, comprising a sea mustard extract as an active ingredient.

[0051] In one embodiment of the present invention, the composition increases the expression of the PGC-1α (Peroxisome proliferator-activated receptor-gamma coactivator 1 alpha) gene.

[0052] In the present invention, "PGC-1α (Peroxisome proliferator-activated receptor-gamma coactivator 1 alpha)" is an important factor that induces metabolic processes such as mitochondrial biosynthesis, enhancement of antioxidant response, and enhancement of fatty acid oxidation function, and plays a role in promoting energy production and enhancing skin vitality.

[0053] In the present invention, "skin vitality" refers to the degree of activation of skin cells, and "enhancement of skin vitality" means enabling the skin to regain vitality by enhancing the activation of skin cells.

[0054] In one embodiment of the present invention, when a group of UVB-irradiated fibroblasts was treated with a sea mustard extract, it was confirmed that the PGC-1α mRNA expression, which was reduced by UVB irradiation, increased.

[0055] In the present invention, the Seomcho extract may increase the mRNA expression levels of COL-1 (Collagen Type I) and Elastin.

[0056] In the present invention, "COL-1 (Collagen Type I)" is the most representative form of collagen that makes up most of the dermis and plays a role in making the skin firm and tight. In the case of COL-1, when subjected to internal or external stress, its expression decreases, causing skin elasticity to decline and sagging.

[0057] In this invention, "Elastin" is one of the components that make up the dermis and makes the skin flexible, but skin elasticity decreases when under stress.

[0058] In one embodiment of the present invention, when a group of cortisol-treated fibroblasts was treated with Seomcho extract, it was confirmed that COL-1 and Elastin mRNA expression increased compared to the cortisol-treated group.

[0059] In the present invention, the above-mentioned sea mustard extract can exhibit antioxidant efficacy.

[0060] In the present invention, "antioxidant efficacy" means inhibiting the production of reactive oxygen species that increase due to oxidative stress.

[0061] In one embodiment of the present invention, when a group of H2O2-treated keratinocytes was treated with a sea mustard extract, it was confirmed that the generated ROS was reduced compared to the H2O2-treated group.

[0062] The cosmetic composition of the present invention may be prepared in any formulation conventionally manufactured in the art, for example, as a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, and spray, but is not limited thereto. More specifically, it may be prepared in the form of a softening lotion, a nourishing lotion, a lotion, a nourishing cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray, or a powder.

[0063] The cosmetic composition of the present invention may include a carrier acceptable in a cosmetic formulation in addition to the active ingredient. The term "carrier acceptable in a cosmetic formulation" refers to an additional ingredient that is known and used to be included in a cosmetic formulation and can improve the application of the active ingredient to the skin, user convenience, and preference without significantly attenuating the main efficacy of the active ingredient or causing adverse effects on the human body.

[0064] The above carrier may be included in an amount of about 1% to about 99.99% by weight, preferably about 50% to about 99% by weight, based on the total weight of the composition of the present invention. However, the content of the above carrier may be appropriately adjusted according to the formulation of the cosmetic, the specific application site, and the preferred amount of application, and is not particularly limited.

[0065] In the case where the formulation of the present invention is a paste, cream, lotion, or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanth gum, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide may be used as a carrier component.

[0066] In the case where the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder may be used as a carrier component, and in particular, in the case of a spray, it may additionally include a propellant such as chlorofluorohydrocarbon, propane / butane, or dimethyl ether.

[0067] When the formulation of the present invention is a solution or an emulsion, a solvent, a solubilizing agent, or an emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, or fatty acid ester of sorbitan.

[0068] In the case where the formulation of the present invention is a suspension, liquid diluents such as water, ethanol, or propylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tragacanth gum, etc. may be used as carrier components.

[0069] In the case where the formulation of the present invention is a surfactant-containing cleansing agent, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester, etc. may be used as a carrier component.

[0070] In addition to the active ingredient and the carrier ingredient, the ingredients included in the cosmetic composition of the present invention include ingredients commonly used in cosmetic compositions, and may include, for example, conventional auxiliary agents such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances.

[0071] In one embodiment of the present invention, the composition of the present invention may include not only the active ingredient, the above-mentioned sea mustard extract, but also ingredients commonly used in cosmetic compositions, such as conventional auxiliary agents like antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances, and a carrier.

[0072] As the above carrier, purified water, monohydric alcohols (ethanol or propyl alcohol), polyhydric alcohols (glycerol, 1,3-butylene glycol or propylene glycol), higher fatty acids (palmityl acid or linolenic acid), oils and fats (wheat germ oil, camellia oil, jojoba oil, olive oil, squalene, sunflower oil, macadamia peanut oil, Avogad oil, soybean water-added lecithin or fatty acid glycerides) may be used, but are not limited thereto. In addition, surfactants, disinfectants, antioxidants, UV absorbers, anti-inflammatory agents, and cooling agents may be added as needed.

[0073] The above surfactants may include polyoxyethylene, hydrogenated castor oil, polyoxyethylene, oleyl ether, polyoxyethylene monooleic acid, polyoxyethylene, glyceryl monostearate, sorbitan monostearate, polyoxyethylene monooleic acid, sorbitan, sucrose fatty acid ester, hexaglycerin monolauric acid, polyoxyethylene reduced lanolin, POE, glyceryl pyroglutamic acid, isostearic acid, diester, N-acetylglutamine, and isostearyl ester.

[0074] The above disinfectants may include hinoxyol, triclosan, chlorhexidine gluconate, phenoxyethanol, resorcin, isopropylmethylphenol, azulene, salicylic acid, and zinc pyriteon.

[0075] Any of butylhydroxyanisole, gallic acid, propyl gallic acid, and erythorbic acid may be used as the above antioxidant.

[0076] The above-mentioned ultraviolet absorbers may include benzophenones such as dihydroxybenzophenone, melanin, ethyl para-aminobenzoate, 2-ethylhexyl ester of para-dimethylaminobenzoate, cinoxidase, 2-ethylhexyl ester of paramethoxycinnamic acid, 2-(2-hydroxy-5-methylphenyl)benzotriazole, urocanic acid, and metal oxide microparticles.

[0077] The above anti-inflammatory agent may be dipotassium glycyrrhizinate or allantoin, and the cooling agent may be capsicum tincture or 1-menthol.

[0078] In another embodiment of the present invention, the cosmetic composition of the present invention may be prepared in a formulation selected from the group consisting of external ointments, gels, creams, foams, nourishing lotions, softening lotions, packs, softening waters, emulsions, makeup bases, essences, skin ampoules, hair ampoules, scalp treatments, hair tonics, hair conditioners, hair treatments, hair lotions, hair shampoos, hair rinses, shampoos that serve as rinses, hair nourishing lotions, hair gels, hair waxes, hair sprays, hair dyes, soaps, liquid cleansers, bath additives, sunscreen creams, sun oils, suspensions, emulsions, pastes, skin lotions, powders, skin cleansing products, powder foundations, emulsion foundations, wax foundations, patches, and sprays, but is not limited thereto. Effects of the invention

[0080] The features and advantages of the present invention are summarized as follows:

[0081] (a) The present invention provides a cosmetic composition for stress relief comprising an active ingredient of sea mustard extract.

[0082] (b) The present invention provides a cosmetic composition for strengthening skin defense, comprising an extract of *Seomcho* as an active ingredient.

[0083] (c) The present invention provides a cosmetic composition for improving itching that includes an extract of *Seomcho* as an active ingredient.

[0084] (d) The present invention provides a cosmetic composition for improving skin condition by activating skin energy metabolism, comprising a sea mustard extract as an active ingredient.

[0085] (e) The composition containing the Seomcho extract of the present invention as an active ingredient has excellent efficacy in enhancing skin immunity and defense, and can be usefully used as a cosmetic material for neurocosmetics to prevent skin aging and maintain healthy skin. Specific details for implementing the invention

[0087] The present invention will be described in more detail below through examples. These examples are intended solely to explain the invention more specifically, and it will be obvious to those skilled in the art that the scope of the invention is not limited by these examples according to the gist of the invention.

[0088] Throughout this specification, “%” used to indicate the concentration of a particular substance is (weight / weight) % for solid / solid, (weight / volume) % for solid / liquid, and (volume / volume) % for liquid / liquid, unless otherwise noted.

[0090] Examples

[0092] Example 1: Method for preparing Seomcho extract

[0093] Dried sea mustard was immersed in 70% ethanol at a 1:20 ratio and extracted at 60°C for 5 hours. The resulting powder was then filtered through filter paper (Whatman No. 2) and concentrated. The concentrated powder was dissolved in 50% butylene glycol to achieve a concentration of 1.0% (g / g), filtered through filter paper (Durapore 0.22㎛, PVDF), and used in the test.

[0095] Example 2: Evaluation of Cytotoxicity of Seomcho Extract

[0096] To evaluate the cytotoxicity of the *Seomcho* extract on human keratinocytes, the inventors measured cell viability according to extract concentration using an MTT assay.

[0097] Specifically, human keratinocytes or human fibroblasts were inoculated into a 24-well plate and cultured for 24 hours. Afterward, the culture medium was replaced with serum-free medium, and the extract was added at various concentrations (0, 0.1, 0.25, 0.5, 1, 2, 3, 5%) and cultured for 24 hours. After the culture was complete, the medium was replaced with a 10-fold diluted medium containing 2.5 mg / mL of MTT solution, and the reaction was carried out. The supernatant was then removed, and DMSO was added to dissolve the formed MTT-formazan crystals. Subsequently, the absorbance was measured at 570 nm using an ELISA reader. Cytotoxicity was expressed as a percentage of absorbance compared to the control group. The results of cytotoxicity measurements of the Seomcho extract on human keratinocytes are shown in Table 1, and the results of cytotoxicity measurements of the Seomcho extract on human fibroblasts are shown in Table 2.

[0099] Sample Conc. Average Stdev. p-value Control 0 100.0 4.37 1.000 Seomcho extract (%) 0.1 112.8 2.84 0.013 0.25 109.2 0.48 0.023 0.5 108.0 2.79 0.055 1 98.8 0.92 0.675 2 108.8 3.77 0.058 3 105.0 6.62 0.337 5 41.5 4.01 0.001

[0101] Sample Conc. Average Stdev. p-value Control 0 100.0 8.01 1.000 Seomcho extract (%) 0.1 103.0 6.80 0.645 5 107.7 4.48 0.219 1 108.0 1.50 0.164 2 107.2 3.64 0.228 3 104.0 3.61 0.475 4 103.1 2.65 0.560 5 103.7 3.81 0.511

[0103] As shown in Table 1, when human keratinocytes were treated with Seomcho extract at different concentrations, it was confirmed that no cytotoxicity occurred up to a concentration of 3%, and as shown in Table 2, it was confirmed that there was no toxicity in human fibroblasts up to a concentration of 5%. In subsequent experiments, the efficacy of Seomcho extract was evaluated at concentrations of 0.1, 0.5, 1, and 2%.

[0105] Example 3: Evaluation of the efficacy of Seomcho extract in inhibiting skin nerve pain receptor expression

[0106] To confirm the inhibitory effect of Seomcho extract on the expression of TRPV1 (Transient receptor potential family V 1: capsaicin receptor), a skin neuropathic pain sensory receptor, the inventors confirmed the mRNA expression level of TRPV1 following treatment with Seomcho extract using Real-time PCR.

[0107] Specifically, NHEK (Human normal keratinocyte cells) were seeded into a 6-well plate and cultured for 24 hours; subsequently, the cell medium was replaced with serum-free medium, and 200 μM of capsaicin was added. After 2 hours, the medium was replaced with fresh serum-free medium, and Seomcho extract was added at various concentrations (0.5, 1, 2%) and cultured for 24 hours. After 24 hours, the cultured cells were treated with QIAzol TM RNA was isolated using Lysis Reagent (Qiagen) according to the manufacturer's instructions. After quantifying the isolated RNA, cDNA was synthesized and Real-Time PCR was performed.

[0108] The results are shown in Table 3.

[0110] Sample Conc. Average Stdev. p-value Control 42.8 0.47 0.000 Capsaicin (μM) 200 100.0 0.00 1.000 Capsaicin + Capsazepine (μM) 5 61.5 1.65 0.000 Capsaicin + Seomcho Extract (%) 0.5 81.4 1.92 0.000 1 72.8 3.16 0.000 2 70.5 1.12 0.000

[0112] As shown in Table 3, when TRPV1 mRNA expression was confirmed following treatment with Seomcho extract, it was confirmed that TRPV1 mRNA expression was reduced compared to the capsaicin-treated group.

[0113] Therefore, these results indicate that Seomcho extract can be used as an ingredient to soothe sensitive skin by inhibiting the expression of the skin sensory receptor TRPV1, which causes inflammation, itching, and pain.

[0115] Example 4: Evaluation of the efficacy of Seomcho extract in inhibiting stress hormone-converting enzyme expression

[0116] 11β-HSD1 (11 beta-hydroxysteroid dehydrogenase type 1), a stress hormone conversion enzyme, is an enzyme that converts inactive cortisone into active cortisol, is increased in aged skin, and is activated by inflammatory cytokines.

[0117] To confirm the inhibitory effect of Seomcho extract on the expression of 11β-HSD1 (11 beta-hydroxysteroid dehydrogenase type 1), a stress hormone-converting enzyme, the inventors confirmed the mRNA expression level of 11β-HSD1 following treatment with Seomcho extract using Real-time PCR.

[0118] Specifically, fibroblasts were inoculated into 60 mm dishes and cultured for 24 hours; subsequently, the cell medium was replaced with serum-free medium, and TNF-α and Seomcho extract were treated at various concentrations and cultured for 6 hours. Afterward, the cultured cells were treated with QIAzol TM RNA was isolated using Lysis Reagent (Qiagen) according to the manufacturer's instructions. After quantifying the isolated RNA, cDNA was synthesized and Real-Time PCR was performed, and the results are shown in Table 4.

[0120] Sample Conc. Average Stdev. p-value Control 56.6 1.21 0.000 TNF-α (ng / ㎖) 10 100.0 0.00 1.000 TNF-α + EGCG (μg / ㎖) 5 68.5 1.19 0.000 TNF-α + Seomcho Extract (%) 0.1 81.1 0.98 0.000 0.5 73.5 2.49 0.000 1 73.6 2.04 0.000 2 72.2 2.11 0.000

[0122] As shown in Table 4, when the mRNA expression of 11β-HSD1 was examined following treatment with Seomcho extract, it was confirmed that 11β-HSD1 mRNA expression decreased compared to the TNF-α treated group.

[0123] Therefore, these results indicate that Seomcho extract can be used as an ingredient to prevent skin aging caused by psychological stress.

[0125] Example 5: Evaluation of the efficacy of Seomcho extract in improving elasticity reduced by stress hormones

[0126] COL-1 (Collagen Type I) is the most representative form of collagen, which makes up most of the dermis; it makes the skin firm and tight, but under stress, skin elasticity decreases and it sags. Elastin is one of the components that make up the dermis; it makes the skin flexible, but under stress, skin elasticity decreases.

[0127] To confirm the efficacy of Seomcho extract in improving COL-1 (Collagen Type I) and Elastin, the inventors used Real-time PCR to determine the mRNA expression levels of COL-1 and Elastin following treatment with Seomcho extract.

[0128] Specifically, fibroblasts were inoculated into a 60 mm dish and cultured for 24 hours; subsequently, the cell medium was replaced with serum-free medium, and the cells were treated with Cortisol and Seomcho extract at various concentrations and cultured for 6 hours. Afterward, the cultured cells were treated with QIAzol TM RNA was isolated using Lysis Reagent (Qiagen) according to the manufacturer's instructions. After quantifying the isolated RNA, cDNA was synthesized and Real-Time PCR was performed. The mRNA expression results for COL-1 are shown in Table 5, and the mRNA expression results for Elastin are shown in Table 6.

[0130] Sample Conc. Average Stdev. p-value Control 225.3 0.00 0.000 Cortisol (mM) 0.3 100.0 2.39 1.000 Cortisol + Seomcho Extract (%) 0.1 103.2 0.88 0.094 0.5 134.5 3.47 0.000 1 138.6 2.38 0.000 2 143.3 2.15 0.000

[0132] Sample Conc. Average Stdev. p-value Control 230.9 0.00 0.000 Cortisol (mM) 0.3 100.0 4.36 1.000 Cortisol + Seomcho Extract (%) 0.1 117.8 9.91 0.046 0.5 145.7 4.12 0.000 1 150.2 7.33 0.001 2 164.5 3.15 0.000

[0134] As shown in Tables 5 and 6, when the mRNA expression of COL-1 and Elastin was examined following treatment with Seomcho extract, it was confirmed that the expression of COL-1 mRNA and Elastin mRNA, which were lowered by Cortisol, increased.

[0135] Therefore, these results suggest that Seomcho extract can help maintain elastic skin by increasing the expression of COL-1 mRNA and Elastin mRNA, which are reduced by Cortisol.

[0137] Example 6: Evaluation of the skin defense-enhancing efficacy of Seomcho extract

[0138] HBD-2 (Human beta-defensin-2) is an antimicrobial peptide that performs a primary defense function by rapidly increasing in response to infection or inflammation. It performs a chemical barrier function that enhances immunity by activating immune cells, and a decrease in HBD-2 expression is a cause of high bacterial infection and inflammation.

[0139] To confirm the efficacy of Seomcho extract in increasing HBD-2 expression, the inventors used Real-time PCR to determine the mRNA expression levels of HBD-2 following treatment with Seomcho extract.

[0140] Specifically, HaCaT was inoculated into a 60 mm dish and cultured for 24 hours; subsequently, the cell medium was replaced with serum-free medium, and TNF-α, IFN-γ, IL-4, and Seomcho extract were treated at various concentrations and cultured for 25 hours. Afterward, the cultured cells were treated with QIAzol TM RNA was isolated using Lysis Reagent (Qiagen) according to the manufacturer's instructions. After quantifying the isolated RNA, cDNA was synthesized and Real-Time PCR was performed, and the results are shown in Table 7.

[0142] Sample Conc. TNF-α + IFN-γ treatment IL-4 treatment Average Stdev. p-value Normal skin + - 360.6 21.80 0.000 Disrupted skin + + 100.0 0.00 1.000 Lactoferrin (μg / ml) 25 + + 162.0 15.06 0.002 Seomcho extract (%) 0.1 + + 99.4 9.47 0.917 0.5 + + 125.2 2.42 0.000 1 + + 160.3 2.02 0.000 2 + + 179.7 4.30 0.000

[0144] As shown in Table 7, when HBD-2 mRNA expression was confirmed following treatment with Seomcho extract, it was confirmed that HBD-2 mRNA expression, which was lowered by TNF-α+IFN-γ+IL-4, increased.

[0145] Therefore, these results indicate that Seomcho extract can be used as an ingredient to strengthen skin defense by increasing the expression of HBD-2.

[0147] Example 7: Evaluation of Cell Vitality Efficacy of Seomcho Extract

[0148] PGC-1α (Peroxisome proliferator-activated receptor-gamma coactivator 1alpha) is an important factor that induces mitochondrial biosynthesis and promotes energy production.

[0149] To confirm the efficacy of Seomcho extract in increasing PGC-1α expression, the inventors used Real-time PCR to determine the mRNA expression levels of PGC-1α following treatment with Seomcho extract.

[0150] Specifically, fibroblasts were seeded into 60 mm dishes and cultured for 24 hours, after which they were washed with HBSS and irradiated with UVB. Subsequently, the medium was replaced with serum-free medium, and the cells were treated with various concentrations of *Seomcho* extract and cultured for 20 hours. Afterward, the cultured cells were treated with QIAzol TM RNA was isolated using Lysis Reagent (Qiagen) according to the manufacturer's instructions. After quantifying the isolated RNA, cDNA was synthesized and Real-Time PCR was performed, and the results are shown in Table 8.

[0152] Sample Conc. Average Stdev. p-value Control 541.4 9.71 0.000 UVB 50 mJ / cm³ 100.0 0.00 1.000 UVB+ Island Grass Extract (%) 0.1 100.5 1.54 0.625 0.5 99.7 0.13 0.019 1 105.6 1.78 0.006 2 182.6 10.48 0.000

[0154] As shown in Table 8, when examining the mRNA expression of PGC-1α following treatment with Seomcho extract, it was confirmed that the PGC-1α mRNA expression, which was reduced by UVB irradiation, increased.

[0155] Therefore, these results indicate that Seomcho extract can be used as an ingredient to increase skin vitality by inducing mitochondrial biosynthesis.

[0157] Example 8: Evaluation of Neuro-inflammation Inhibitory Efficacy of Seomcho Extract

[0158] NGF is a nerve growth factor that induces itching by causing nerve fibers in the dermis to penetrate into the epidermis. Continuous secretion of NGF accelerates epidermal hypertrophy caused by the proliferation of keratinocytes, induces the activation of mast cells in atopic dermatitis lesions, and further exacerbates skin pruritus. TNF-α and IL-1β are cytokines involved in inflammation and immune responses, and their expression is known to increase in inflammatory diseases.

[0159] NGF (Nerve growth factor), an itching factor, and TNF-α (Tumor necrosis factor-α), an inflammatory factor, are increased by Substance P, a neurotransmitter secreted when under stress.

[0160] To confirm the inhibitory effect of Seomcho extract on the expression of NGF, TNF-α, and IL-1β, the inventors used Real-time PCR to determine the mRNA expression levels of NGF, TNF-α, and IL-1β following treatment with Seomcho extract.

[0161] Specifically, HaCaT was inoculated into a 6-well plate and cultured for 24 hours, after which the medium was replaced with fresh medium. Substance P and Seomcho extract were then treated at various concentrations (0.1, 0.5, 1, 2%) and cultured for 16 hours. The cultured cells were then treated with QIAzol TM RNA was isolated using Lysis Reagent (Qiagen) according to the manufacturer's instructions. After quantifying the isolated RNA, cDNA was synthesized and Real-Time PCR was performed.

[0162] The results of NGF mRNA expression are shown in Table 9, the results of TNF-α mRNA expression are shown in Table 10, and the results of IL-1β mRNA expression are shown in Table 11.

[0164] Sample Conc. Average Stdev. p-value Control 52.6 4.95 0.000 Substance P (nM) 35 100.0 0.00 1.000 SP + Dexamethasone (ng / ㎖) 40 65.3 6.06 0.002 SP + Seomcho Extract (%) 0.1 88.5 6.62 0.040 0.5 82.4 6.39 0.009 1 81.7 7.75 0.015 2 73.4 5.77 0.001

[0166] Sample Conc. Average Stdev. p-value Control 20.3 0.98 0.000 Substance P (nM) 35 100.0 0.00 1.000 SP + Dexamethasone (ng / ㎖) 40 59.4 3.68 0.000 SP + Seomcho Extract (%) 0.1 105.7 2.75 0.030 0.5 84.1 4.58 0.004 1 83.3 1.95 0.000 2 82.6 3.60 0.001

[0168] Sample Conc. Average Stdev. p-value Control 54.4 1.11 0.000 Substance P (nM) 35 100.0 0.00 1.000 SP + Dexamethasone (ng / ㎖) 40 39.4 1.63 0.000 SP + Seomcho Extract (%) 0.1 88.8 1.46 0.000 0.5 83.5 1.37 0.000 1 80.4 0.92 0.000 2 53.1 1.44 0.000

[0170] As shown in Tables 9, 10, and 11, when the mRNA expression of NGF, TNF-α, and IL-1β was examined following treatment with Seomcho extract, it was confirmed that the mRNA expression of NGF, TNF-α, and IL-1β was reduced compared to the group treated with the neurotransmitter Substance P.

[0171] Therefore, these results indicate that Seomcho extract can be used as an ingredient for improving skin itching by reducing the expression of NGF, an itching-related factor increased by neurotransmitters (neuropeptides) secreted under stress, and inhibiting the expression of inflammatory cytokines.

[0173] Example 9: Evaluation of the Antioxidant Efficacy of Seomcho Extract

[0174] The inventors performed an ROS assay to confirm the inhibitory effect of the Seomcho extract on the production of reactive oxygen species that increase due to oxidative stress.

[0175] Specifically, HaCaT was inoculated into a 24-well plate and cultured for 24 hours, after which the medium was replaced with serum-free medium and treated with Seomcho extract at various concentrations and cultured for 24 hours. Subsequently, H2DcFDA was added and incubated for 30 minutes, followed by treatment with H2O2. After 30 minutes, the plates were washed with HBSS, and HBSS was added to measure the fluorescence value; the results are shown in Table 12.

[0177] Sample conc. H2O2 Avg. Stdev. p-value Control - 24.5 2.57 0.001 H2O2(mM) 1 + 100.00 3.82 1.000 Vitamin E (μM) 100 + 55.9 7.79 0.011 Seomcho extract (%) 0.5 + 125.6 16.21 0.196 1 + 90.4 7.88 0.393 2 + 66.4 9.50 0.031

[0179] As shown in Table 12, the ROS inhibitory efficacy of the Seomcho extract was confirmed, and it was confirmed that ROS was reduced compared to the H2O2-treated group.

[0180] Therefore, from these results, it was found that Seomcho extract exhibits excellent antioxidant efficacy.

[0182] Example 10: Evaluation of the skin biorhythm activation efficacy of Seomcho extract

[0183] The skin's recovery mechanism (light-dark cycle) occurs through core clock genes that are expressed differently during daytime and nighttime on a 24-hour cycle. This is called the skin's circadian rhythm, and through this process, not only is skin homeostasis maintained, but overall skin functions such as recovery, hydration, proliferation, and barrier function are also restored.

[0184] To investigate the skin biorhythm activating efficacy of Seomcho extract, the inventors confirmed the mRNA expression level of Bmal-1, which is related to skin biorhythm, using Real-time PCR.

[0185] Specifically, HaCaT was inoculated into a 60 mm dish and cultured for 24 hours, after which it was washed with HBSS and irradiated with UVB. Subsequently, the medium was replaced with serum-free medium, and the cells were treated with various concentrations of Seomcho extract and cultured for 8 hours. The cultured cells were collected using QIAzol® Lysis Reagent (Qiagen), RNA was isolated according to the manufacturer's instructions, and after quantifying the isolated RNA, cDNA was synthesized and Real-Time PCR was performed.

[0186] The results of confirming the mRNA expression level of Bmal-1 are shown in Table 13.

[0188] Sample Conc. Average Stdev. p-value UVB irradiated group 100.00 0.00 1.000 UVB + Retinoic acid (μM) 0.5 185.9 12.86 0.000 UVB + Island Grass Extract (%) 0.1 118.6 15.62 0.108 0.5 142.7 17.12 0.012 1 188.1 11.60 0.000

[0190] As shown in Table 13, when Bmal-1 mRNA expression, a skin biorhythm-related factor, was examined, it was confirmed that Bmal-1 mRNA expression increased compared to the UVB-treated group when treated with Seomcho extract.

[0191] Therefore, these results suggest that Seomcho extract can be used as an ingredient that positively contributes to damaged skin repair by activating the skin's biorhythm in skin with an imbalanced skin.

Claims

Claim 1 A cosmetic composition for improving skin damaged by stress, comprising Seomcho extract as an active ingredient. Claim 2 A cosmetic composition for improving skin damaged by stress, wherein, in claim 1, the composition reduces the expression of the 11B-HSD1 (11 beta-hydroxysteroid dehydrogenase type 1) gene. Claim 3 A cosmetic composition for improving skin damaged by stress, wherein, in claim 1, the composition reduces the expression of the TRPV1 (transient receptor potential vanilloid 1) gene. Claim 4 A cosmetic composition for strengthening skin defense, comprising Seomcho extract as an active ingredient. Claim 5 In claim 4, the above composition is a cosmetic composition for enhancing skin defense, which increases the expression of the HBD-2 (Human beta-defensin-2) gene. Claim 6 A cosmetic composition for improving neurogenic pruritus containing Seomcho extract as an active ingredient. Claim 7 In claim 6, the above composition is a cosmetic composition for improving neurogenic pruritus, wherein the composition reduces the expression of the NGF (Nerve growth factor) gene. Claim 8 A cosmetic composition for improving skin condition by activating skin energy metabolism, comprising Seomcho extract as an active ingredient. Claim 9 A cosmetic composition for improving skin condition according to claim 8, wherein the composition increases the expression of the PGC-1α (Peroxisome proliferator-activated receptor-gamma coactivator 1 alpha) gene. Claim 10 A cosmetic composition according to any one of claims 1 to 9, wherein the extract is extracted with any one extraction solvent selected from the group consisting of water, organic solvents and mixed solvents thereof.