KR102256823B1 - Complex composition comprising DHA derivatives for anti-wrinkle, improving atopic dermatitis and enhancing skin barrier - Google Patents

Abstract

The present invention relates to an SPMs complex composition comprising 17-hydroxydocosahexaenoic acid, Resolvin D5 and Protectin DX as active ingredients, wherein the SPMs complex composition promotes collagen production, inhibits NO production and expression of TNF-α and IL-6, and increases expression of Filaggrin and Loricrin, thereby preventing or alleviating skin aging or wrinkles, strengthening the skin barrier, or being usefully used for preventing, alleviating or treating skin diseases including atopic dermatitis.

Description

Complex composition comprising DHA derivatives for anti-wrinkle, improving atopic dermatitis and enhancing skin barrier}

The present invention relates to the use of a combination composition comprising 17-hydroxydocosahexaenoic acid (17-HDHA), resolvin D5 and protectin DX to improve skin wrinkles, improve skin inflammation, or strengthen skin barriers.

The skin is an important organ that protects the human body from external harmful environments and is an immune organ with the largest area. The stratum corneum, which makes up the outermost part of the skin, has resistance to physical and chemical damage and prevents microbes and harmful chemicals from penetrating into the body. In addition, keratinocytes, Langerhans cells, fibroblasts, mast cells, dendritic cells and capillary endothelial cells form a complex immune system to protect the human body through an immune response when antigen invades from the outside.

The skin is constantly receiving external stimulation (ultraviolet rays, chemicals, heat, physical friction), and recently, new sources of irritation such as fine dust and blue light are increasing. In addition, internal factors such as mental stress, sleep disorders, nutritional imbalance, and medication are known to cause stress on the skin.

According to recent studies, inflammatory reactions occur due to skin irritation, and research results have been published that such inflammatory reactions cause skin aging. Franceschi first used the term inflammaging in 2000 to explain the relationship between inflammation and aging, and research results show that chronic inflammation is the cause of aging or aging-related diseases such as diabetes, arteriosclerosis, and dementia. Has become.

Pro-inflammatory cytokines secreted during the initiation of inflammation promote the expression of matrix metalloproteinases (MMPs). It is known that the increase in MMPs is directly related to skin aging and promotes aging such as wrinkle formation and elasticity reduction by decomposing collagen, which accounts for 70% of the skin's constituents.

In general, substances used to inhibit skin aging include enzymes such as Superoxide dismutase for removing active oxygen, antioxidants such as vitamin C and tocopherol, botulinum-related peptides, and vitamin A derivatives such as retinol. Among them, vitamin A derivatives are excellent in synthesizing collagen and have the most excellent wrinkle removal effect, but have side effects such as instable to light or heat and cause skin irritation, and the content in the formulation is unstable.

Atopic dermatitis is a chronic inflammatory skin disease that causes dryness, pruritus, and erythematous eczema. Atopic dermatitis is a disease caused by abnormal genetic, environmental, and immunological reactions and disruption of the skin barrier. Recently, as the prevalence rate is increasing worldwide due to environmental changes, dietary habits, and environmental pollution, interest in solutions is also increasing. However, since the cause is not identified, the underlying treatment is difficult, and most of them rely on symptomatic treatment.

On the other hand, the hydroxyl derivatives of docosahexaenoic acid (DHA, C22: 6n-3) and eicosapentaenoic acid (EPA, C20: 5n-3) act on the termination mechanism of the inflammatory response. It was confirmed that they can act as signaling substances, and those that act as signaling substances in the mechanism of terminating the inflammatory response were named SPM (specialized proresolving mediators; resolvins, protectins, maresins).

Recently, using the SPM, various chronic inflammatory diseases (e.g., blood vessels, myocardial infarction, stroke, dementia, osteoarthritis, lung diseases including asthma, gastroenteritis, periodontitis, dry eye syndrome, fatty liver, etc.), infection sepsis, burns, wound recovery, Although research has been actively conducted as a treatment for pain, cancer, and diabetes, studies and experiments on the effects on skin aging improvement, skin barrier strengthening, and atopic dermatitis are insignificant.

Non-Patent Document 1: Sima et al., Crit Rev Immunol. 38 (5); 343-365 (2018)

One object of the present invention is to provide a cosmetic composition, or a food composition for improving the skin condition by preventing or improving skin wrinkles, preventing or improving skin aging, preventing or improving skin inflammation, and strengthening the skin barrier.

Another object of the present invention is one selected from the group consisting of skin wounds, skin scars, dermatitis, atopic dermatitis, pruritus, eczematous skin disease, dry eczema, athlete's foot, erythema, urticaria, psoriasis, weak rash, acne, and hair loss It is to provide a pharmaceutical composition for preventing or treating the above skin diseases.

In order to achieve the above object, an aspect of the present invention provides a cosmetic composition for improving skin condition comprising 17-hydroxydocosahexaenoic acid (17-HDHA), resolvin D5, and protectin DX as active ingredients. do.

In addition, another aspect of the present invention provides a food composition for improving skin conditions comprising 17-hydroxy docosahexaenoic acid (17-HDHA), resolvin D5, and protectin DX as active ingredients.

In addition, another aspect of the present invention provides a pharmaceutical composition for the prevention or treatment of skin diseases comprising 17-hydroxy docosahexaenoic acid (17-HDHA), resolvin D5, and protectin DX as active ingredients.

The SPMs composite composition of the present invention increases collagen synthesis, restores damaged skin barrier, and reduces transdermal moisture loss, and has an effect of inhibiting skin aging or improving skin wrinkles.

In addition, the SPMs composite composition of the present invention remarkably improves the effect of inhibiting the expression of TNF-α, IL-6, increasing the expression of Filaggrin, Loricrin, and the antioxidant effect, thereby strengthening the skin barrier, and strengthening the skin barrier. It is effective in preventing or treating various skin diseases including dermatitis.

However, the effects of the present invention are not limited to the above-mentioned effects, and other effects not mentioned will be clearly understood by those skilled in the art from the following description.

Figure 1 shows the results of confirming the IL-6 expression inhibitory activity of each SPM alone substance or SPMs complex composition for human keratinocytes: Figure 1a is 17S-HDHA, protectin DX, and each concentration of resolvin D5 IL-6 expression inhibitory activity is shown, Figure 1b shows the IL-6 expression inhibitory activity by concentration of the SPMs composite composition 1 to 3 of the present invention, Figure 1c is the synergistic of the SPMs composite composition of the present invention compared to the SPM alone component It shows the effect of inhibiting IL-6 expression [Untreated control: group not irradiated with UVB, Con(-): group that did not treat anything after irradiation with UVB, Con(+) HC: treated with hydrocortisone after UVB irradiation. Positive control].
Figure 2 shows the results of confirming the procollagen synthesis effect of each of the SPM alone substance or the SPMs complex composition for human dermal fibroblasts: Figure 2a is a procollagen by concentration of 17S-HDHA, protectin DX, and resolvin D5, respectively. 2B shows the synergistic procollagen synthesis effect of the SPMs complex composition of the present invention compared to the SPM alone component, and FIG.2B shows the synergistic procollagen synthesis effect of the SPMS complex composition 1 to 3 of the present invention by concentration. [Untreated control: no treatment group, Con(+) RP: positive control group treated with retinyl palmitate].
Figure 3 shows the results of confirming the effect of improving the ability to inhibit the expression of collagen degrading enzyme (MMP-1) of the SPMs composite composition of the present invention with respect to human dermal fibroblasts [Untreated control: group not irradiated with UVB, Control: UVB After irradiation, no treatment group, Retinoic acid: positive control group treated with retinoic acid after UVB irradiation].
Figure 4 shows the results confirming the effect of improving the TNF-α expression inhibitory ability of the SPMs composite composition of the present invention with respect to human keratinocytes [Untreated control: group not irradiated with UVB, Control: no treatment after UVB irradiation Group, Hydrocortisone: positive control group treated with hydrocortisone after UVB irradiation].
5 shows the results of confirming the effect of reducing NO generation of the SPMs composite composition of the present invention [Untreated control: group not irradiated with UVB, Control: group not treated after irradiation with UVB, Hydrocortisone: Hydrocortisone after UVB irradiation Cortisone-treated positive control, Vit.C: UVB irradiated and then vitamin C-treated positive control].
Figure 6 shows the result of confirming the effect of reducing lipid peroxide production of the SPMs composite composition of the present invention with respect to the stimulation induced by fine dust [Untreated control: group not treated with fine dust, Control: after stimulation induced with fine dust No treatment group, Hydrocortisone: positive control group treated with hydrocortisone after induction of stimulation with fine dust, Vit.C: positive control group treated with vitamin C after induction of stimulation with fine dust].
7 shows the result of confirming the skin barrier improvement effect by the SPMs complex composition of the present invention through the expression level of the core protein constituting the skin barrier: FIG. 7A is the result of confirming the increase in expression of filaggrin, 7B is a result of confirming the increase in the expression of loricrin [Untreated control: group not treated with anything, Hyaluronic acid: positive control group treated with hyaluronic acid].
Figure 8 shows the results of confirming the skin barrier improvement effect by the SMPs composite composition of the present invention through the human efficacy evaluation: Figure 8a shows the skin barrier index change rate (%), Figure 8b is the skin barrier recovery rate (% ).
Figure 9 shows the results of confirming the atopic dermatitis improvement effect by the SMPs composite composition of the present invention through the human efficacy evaluation: Figure 9a shows the skin moisture change rate (%) at the lesion site, Figure 9b is at the lesion site It shows the rate of change in the amount of transdermal moisture loss (%), and FIG. 9C shows the rate of change in the skin itching index (%) at the lesion site.
Figure 10 shows the results of confirming the effect of improving wrinkles and skin hydration by the SPMs composite composition of the present invention through human efficacy evaluation: Figure 10a shows the rate of change of wrinkles around the eye (%), Figure 10b is the amount of moisture in the skin around the eye And Figure 10c shows the moisture improvement rate (%) around the eye area.

Hereinafter, the present invention will be described in detail.

The present invention provides a composition for improving skin conditions comprising 17-hydroxydocosahexaenoic acid (17-HDHA), resolvin D5, and protectin DX as active ingredients.

The 17-hydroxydocosahexaenoic acid (17-HDHA), resolvin D5, and protectin DX are lipoxygenase derived from omega-3 fatty acids, that is, docosahexaenoic acid (DHA). ; LOX) is a metabolite, and is a specialized pro-resolving mediator (SPM).

The 17-hydroxydocosahexaenoic acid (17-HDHA) is (±) 17-hydroxy (dihydroxy)-4Z, 7Z, 10Z, 13Z, 15E, 19Z-docosahexaenoic acid, It is represented by the structure of the following formula (1).

[Formula 1]

The Resolvin D5 (RvD5) is 7S, 17S-dihydroxy-4Z, 8E, 10Z, 13Z, 15E, 19Z- docosahexaenoic acid, which has the structure of Formula 2 below. It is expressed as

[Formula 2]

The Protectin DX (PDX) is 10S,17S-dihydroxy-4Z,7Z,11E,13Z,15E,19Z-docosahexaenoic acid, which is an isomer of protectin D1. , Represented by the structure of the following formula (3).

[Formula 3]

The 17-HDHA, resolvin D5 and protectin DX can be chemically synthesized, biosynthesized, or commercially available.

In an embodiment of the present invention, as a result of treatment of the SPMs complex composition containing 17-hydroxydocosahexaenoic acid (17-HDHA), resolvin D5 and protectin DX on human dermal fibroblasts, by UV irradiation It was confirmed that suppressed the expression of increased inflammatory factors and increased procollagen synthesis. In particular, it was confirmed that the composite composition of the present invention synergistically induces the effect of synthesizing collagen and inhibiting inflammation compared to the single compound. Further, it was confirmed in human application experiments that the composite composition of the present invention restores damaged skin barrier and relieves skin dryness and itching caused by atopy by reducing transdermal moisture loss.

Therefore, the composition of the present invention may be for improving the skin condition.

In the present invention, the "skin condition" may be skin aging due to ultraviolet rays, melasma, freckles, skin wounds, dermatitis, atopic dermatitis, pruritus, eczema skin disease, dry eczema, erythema, urticaria, psoriasis, weak rash or acne.

The aging of the skin includes symptoms such as decreased skin elasticity, reduced gloss, wrinkles, weakened regenerative power, or severe dryness, and includes both endogenous aging and exogenous aging caused by the passage of time or external environment. In particular, the skin aging may be photoaging, and the photoaging refers to skin damage that occurs when the skin is repeatedly exposed to ultraviolet rays or for a long period of time.

In addition, the wrinkle refers to wrinkles occurring in all body parts including the face, and includes both static rhytides and dynamic rhytides.

In the present invention, the term "improving skin condition" means preventing or improving skin wrinkles, preventing or improving skin aging, preventing or improving skin inflammation, shrinking or reducing pores, or improving skin barrier function, alleviating skin irritation, skin cell proliferation, and It is a concept that includes all of the regeneration ability, antioxidant ability, and collagen synthesis enhancing ability.

In the present invention, the meaning of “including as an active ingredient” means an effective amount capable of showing skin improvement effects, for example, wrinkle improvement, inflammation improvement, atopic dermatitis improvement, skin regeneration or skin barrier strengthening, improvement It means to contain.

According to one embodiment of the present invention, the composition may promote the synthesis of collagen in cells, or inhibit collagen degradation by inhibiting the expression or activity of a matrix metalloproteinase (MMP). The composition may be, for example, inhibiting the expression or activity of MMP-1 that degrades type 1 collagen to inhibit the decomposition of extracellular matrix.

According to one embodiment of the present invention, the composition may have antioxidant activity. The composition may be one that inhibits the formation of lipid peroxide. According to another embodiment of the present invention, the composition may inhibit inflammatory factors. The composition may be one that inhibits NO, TNF-α, and IL-6, which are inflammation initiating factors.

The TNF-α (tumor necrosis factor-a) and IL-6 (Interleukin-6) are representative inflammation-inducing cytokines, and TNF-α is an endotoxin in the cell membrane of Gram-negative bacteria, Lipopolysaccharide: LPS) is a pro-inflammatory cytokine produced by activated lymphocytes, and IL-6 is secreted by macrophages and T cells to stimulate immune responses to promote inflammation.

The composite composition comprising 17-HDHA, resolbin D5 and protectin DX of the present invention can implement excellent anti-inflammatory action by inhibiting the expression of inflammatory cytokines TNF-α and IL-6, and oxidation occurring in the skin It has an excellent antioxidant effect that suppresses the generation of lipid peroxides caused by anxiety stress, and can improve skin resistance to external stimuli.

According to another embodiment of the present invention, the composition may be to strengthen the skin barrier.

The skin barrier is the stratum corneum, which is the outermost layer of the epidermis, mainly composed of nucleated flat corneocytes. A multi-lamella lipid layer formed of intercellular lipids such as ceramide, cholesterol, and fatty acids synthesized by keratinocytes of the skin barrier that is maintained through the process of division and differentiation of normal epidermal cells is a barrier to prevent moisture in the skin from evaporating. Plays a role. On the other hand, among these intercellular lipids, omega hydroxy ceramide is linked to involucrin, a protein in the outer layer of corneocytes, through a chemical covalent bond to form a corneocyte lipid envelope (CLE), thereby forming a multilayer lipid membrane. It plays a role in physically stabilizing the intercellular lipids in the form, thereby reinforcing the barrier function.

The composition of the present invention is delivered to the stratum corneum through skin application to promote the differentiation of keratinocytes, and has the effect of improving the thickness of the epidermal layer, as well as having an excellent effect of restoring damage to the skin barrier. It can be usefully used for the treatment and prevention of induced skin diseases. Skin diseases caused by the skin barrier damage include, but are not limited to, atopic dermatitis, xeroderma, psoriasis, ichthyosis, and acne.

In addition, the effect of strengthening the skin barrier was confirmed by increasing the expression of filaggrin and loliclean. Filaggrin is one of several structural proteins expressed by keratinocytes in the differentiation stage. It is involved in differentiation from the basal layer of the epidermis to the stratum corneum. It is the main body of the natural moisture factor (NMF), which is essential for the maintenance of moisture in the skin tissue. It is also used as a major indicator of skin moisture retention and skin membrane function.The composite composition comprising 17-HDHA, resolvin D5 and protectin DX according to the present invention has a superior gene expression of filaggrin or loliclean. As it increases, it has excellent skin barrier protection, strengthening, and improvement functions.

The composite composition comprising 17-HDHA, resolbin D5 and protectin DX of the present invention contains the same concentration of each of these compounds by the complex synergistic action of 17-HDHA, resolbin D5 and protectin DX, which are contained as active ingredients. Compared to that used as, it is possible to achieve a particularly remarkably improved IL-6 expression inhibitory effect and procollagen synthesis effect (FIGS. 1 and 2).

In one embodiment of the present invention, the composite is 3 to 85: 10 to 60: 5 to 50 weight ratio of the 17-HDHA, resolvin D5, and protectin DX, for example, 3:10:5 weight ratio, 5: It may be included in a weight ratio of 60:50 and a weight ratio of 85:10:5, but is not limited thereto. In an embodiment of the present invention, a composite composition comprising 17-HDHA, resolvin D5, and protectin DX in a weight ratio of 3:47:50, a weight ratio of 25:56:19, and a weight ratio of 84:10:6 is used. The skin improvement effect was confirmed.

On the other hand, the composition for improving the skin condition can be used as a cosmetic composition, a pharmaceutical composition, or a food composition.

Accordingly, in one aspect, the present invention provides a cosmetic composition for improving skin condition comprising 17-hydroxydocosahexaenoic acid (17-HDHA), resolvin D5, and protectin DX as active ingredients.

The composite composition containing the 17-hydroxydocosahexaenoic acid (17-HDHA), resolvin D5, and protectin DX, and the contents related to the improvement of the skin condition are as described above.

When the composite composition of the present invention is used as a cosmetic composition, in addition to the 17-HDHA, resolbin D5 and protectin DX, ingredients commonly used in cosmetic compositions are included, such as a sulfating agent, a stabilizer, a solubilizing agent, and a vitamin. , Conventional auxiliaries such as pigments and fragrances, and carriers.

The cosmetic composition of the present invention can be prepared in any formulation conventionally prepared in the art, for example, solutions, suspensions (anhydrous and aqueous systems), anhydrous products (oils and glycols), emulsions, pastes, gels. , Mask, pack, powder, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, spray, etc., but is not limited thereto. More specifically, it may be prepared in the form of a flexible lotion (skin), nutritional lotion (milk lotion), nutritional cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray, or powder. . In this case, the accessibility of the cosmetic composition may be further improved, and a moisturizing effect may be implemented to prevent chronic recurrence of atopic dermatitis.

When the formulation of the cosmetic composition is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanta, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide, etc. are used as carrier components. Can be used.

When the formulation of the cosmetic composition is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder may be used as a carrier component. In particular, in the case of a spray, additional chlorofluorohydrocarbon, Propellants such as propane/butane or dimethyl ether.

When the formulation of the cosmetic composition is a solution or 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-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan.

When the formulation of the cosmetic composition is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, an ethoxylated isostearyl alcohol, a suspending agent such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, micro Crystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacantha, and the like may be used.

When the formulation of the cosmetic composition is a surfactant containing cleansing, as a carrier component, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid Amide ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters may be used.

When the cosmetic composition is in the form of a soap, it may be prepared including a skin moisturizer, an emulsifier, a water softener, etc. as an additive, and as a base of the soap, vegetable oils such as palm oil, palm oil, soybean oil, olive oil, palm kernel oil, jooba oil, etc. Animal fats and oils such as beef tallow, pork fat, sunny fat, and fish oil can be used, and as a moisturizing agent, glycerin, detipritol, propylene glycol, butylene glycol, hexyl glycol, isopropyl myristate, aloe vera, sorbitol, etc. can be used, and emulsifiers As examples, natural oils, waxes, hydrocarbons, etc. may be used, and tetrasodium EDTA may be used as the water softener, but the present invention is not limited thereto.

The soap may additionally include an antibacterial agent, a foam inhibitor, a solvent, a corrosion inhibitor, a fragrance, a colorant, a metal ion sequestering agent, a preservative, and the like as an additive.

When the composition of the present invention is used as a cosmetic composition, the 17-HDHA, resolvin D5 and protectin DX in the composition may be included in a concentration of 30 μM or more, specifically 35 μM or more, and more specifically 40 μM or more, but this It is not limited thereto, since it varies depending on the form in which the composition is prepared and its specific application site (face or hand) or application dose.

In another aspect, the present invention provides a pharmaceutical composition for preventing or treating skin diseases comprising 17-hydroxydocosahexaenoic acid (17-HDHA), resolvin D5, and protectin DX as active ingredients.

The skin diseases may be skin wounds, skin scars, dermatitis, atopic dermatitis, pruritus, eczematous skin disease, dry eczema, athlete's foot, erythema, urticaria, psoriasis, weak rash, acne, or hair loss.

The composite composition containing the 17-hydroxydocosahexaenoic acid (17-HDHA), resolvin D5, and protectin DX, and the contents related to skin diseases are as described above.

When the composition is used as a pharmaceutical composition, in addition to the above, 17-HDHA, resolbin D5, and protectin DX may further include a pharmaceutically acceptable carrier or additive.

The meaning of'pharmaceutically acceptable' means that the application (prescription) does not have toxicity beyond adaptable without inhibiting the activity of the active ingredient. The'carrier' is defined as a compound that facilitates the addition of the compound into cells or tissues.

The 17-HDHA, resolvin D5 and protectin DX of the present invention may be administered alone or in admixture with any convenient carrier and the like, and such dosage forms may be single-dose or repeat-dose formulations. The pharmaceutical composition may be a solid formulation or a liquid formulation. Solid preparations include, but are not limited to, powders, granules, tablets, capsules, and suppositories. The solid preparation may include a carrier, a flavoring agent, a binder, a preservative, a disintegrant, a lubricant, a filler, etc., but is not limited thereto. Liquid formulations include, but are not limited to, solutions such as water and propylene glycol solutions, suspensions, and emulsions, and may be prepared by adding suitable coloring agents, flavoring agents, stabilizers, and viscosifying agents. For example, the powder can be prepared by simply mixing the active ingredients of the present invention 17-HDHA, resolvin D5, and protectin DX with a suitable pharmaceutically acceptable carrier such as lactose, starch, and microcrystalline cellulose. The granules are prepared by mixing the 17-HDHA of the present invention, resolvin D5 and protectin DX, a suitable pharmaceutically acceptable carrier, and a suitable pharmaceutically acceptable binder such as polyvinylpyrrolidone and hydroxypropyl cellulose, It can be prepared using a wet granulation method using a solvent such as water, ethanol, isopropanol, or a dry granulation method using a compressive force. In addition, tablets may be prepared by mixing the granules with a suitable pharmaceutically acceptable lubricant such as magnesium stearate, and then tableting them using a tablet press.

The 17-HDHA, resolvin D5, and protectin DX of the present invention are oral, injection (e.g., intramuscular injection, intraperitoneal injection, intravenous injection, infusion, subcutaneous injection) depending on the disease to be treated and the condition of the individual. Injection, implant), inhalation, nasal administration, vaginal, rectal, sublingual, transdermal, topical, and the like, but are not limited thereto. Depending on the route of administration, it may be formulated into a suitable dosage unit dosage form comprising a pharmaceutically acceptable vehicle, excipient, vehicle, which is conventionally used and non-toxic.

The pharmaceutical composition of the present invention may be administered at a daily dose of about 0.0001 mg/kg to about 10 g/kg, and about 0.001 mg/kg to about 1 g/kg. However, the dosage may vary depending on the degree of purification of the mixture, the patient's condition (age, sex, weight, etc.), the severity of the condition being treated, and the like. If necessary, for convenience, the total daily dose may be divided and administered several times during the day.

When the composition of the present invention is used as a pharmaceutical composition, the 17-HDHA, resolvin D5, and protectin DX in the composition may be included in a concentration of 30 μM or more, specifically 35 μM or more, and more specifically 40 μM or more.

In another aspect, the present invention provides a food composition for improving skin conditions, including 17-hydroxydocosahexaenoic acid (17-HDHA), resolvin D5, and protectin DX as active ingredients.

The composite composition containing the 17-hydroxydocosahexaenoic acid (17-HDHA), resolvin D5, and protectin DX, and the contents related to the improvement of the skin condition are as described above.

When the composition is used as a food composition, it may contain acceptable food additives, and may further include suitable carriers, excipients, and diluents commonly used in the manufacture of food.

In the present invention, food refers to a natural product or processed product containing one or more nutrients, and specifically refers to a state that can be eaten directly through a certain amount of processing process, and as a general meaning, various foods, Functional foods, beverages, food additives, and beverage additives are all included. Examples of the foods include various foods, beverages, gum, tea, vitamin complexes, and functional foods. In addition, the food of the present invention includes special nutritional foods (e.g., formula, infants, infant food, etc.), processed meat products, fish meat products, tofu, muk, noodles (e.g., ramen, noodles, etc.), health supplement food, seasoning food ( Examples, soy sauce, miso, red pepper paste, mixed sauce, etc.), sauces, confectionery (e.g. snacks), dairy products (e.g. fermented milk, cheese, etc.), other processed foods, kimchi, pickles (various kimchi, pickles, etc.), beverages ( Examples include, but are not limited to, fruit, vegetable beverages, soy milk, fermented beverages, ice cream, etc.), natural seasonings (eg, ramen soup, etc.), vitamin complexes, alcoholic beverages, alcoholic beverages, and other health supplements. The functional food, beverage, food additive or beverage additive may be prepared by a conventional manufacturing method.

The term'functional food' refers to a food group or food composition that has added value to act and express the function of the food for a specific purpose by using physical, biochemical, biotechnological techniques, etc. to control the biological defense rhythm of the food composition, and to prevent and recover diseases. It refers to a food that is designed and processed to sufficiently express the body's control function related to the body to the living body, and specifically, it may be a health functional food.

The term'health functional food' as used in the present invention refers to food manufactured and processed in the form of tablets, capsules, powders, granules, liquids and pills using raw materials or ingredients having useful functions for the human body. Here, the term'function' refers to obtaining useful effects for health purposes, such as controlling nutrients or physiological effects on the structure and function of the human body. The health functional food of the present invention can be prepared by a method commonly used in the art, and at the time of manufacture, it can be prepared by adding raw materials and ingredients commonly added in the art. In addition, the formulation of the health functional food may be prepared without limitation as long as it is a formulation recognized as a health functional food. The food composition of the present invention can be prepared in various forms of formulation, and unlike general drugs, it has the advantage of not having side effects that may occur when taking drugs for a long time using food as a raw material, and is excellent in portability, and the present invention The health functional food of can be consumed as a supplement to enhance the skin improvement effect.

In addition, the functional food may include food additives acceptable for food, and may further include suitable carriers, excipients, and diluents commonly used in the manufacture of functional foods.

In addition, in the food composition, the 17-HDHA, resolbin D5, and protectin DX in the composition may be included in a concentration of 30 μM or more, specifically 35 μM or more, and more specifically 40 μM or more.

In addition to the active ingredients, the food composition of the present invention may contain sweetening agents, flavoring agents, physiologically active ingredients, minerals, and the like. The sweetener may be used in an amount to give the food a suitable sweet taste, and may be natural or synthetic. Specifically, in the case of using a natural sweetener, examples of the natural sweetener include corn syrup solids, honey, sucrose, fructose, lactose, and sugar sweeteners such as maltose. Flavoring agents can be used to enhance taste or aroma, and both natural and synthetic can be used. Specifically, it is the case of using a natural one. In the case of using natural ones, the purpose of nutrient enhancement can be combined in addition to flavor. As a natural flavoring agent, it may be obtained from apples, lemons, tangerines, grapes, strawberries, peaches, or the like, or from green tea leaves, roundtails, bamboo leaves, cinnamon, chrysanthemum leaves, jasmine, and the like. In addition, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, and ginkgo may be used. The natural flavoring agent may be a liquid concentrate or a solid extract. In some cases, synthetic flavoring agents may be used, and synthetic flavoring agents may include esters, alcohols, aldehydes, terpenes, and the like. As the physiologically active substance, catechins such as catechin, epicatechin, gallocatechin, and epigallocatechin, and vitamins such as retinol, ascorbic acid, tocopherol, calciferol, thiamine, riboflavin, and the like can be used. As the mineral, calcium, magnesium, chromium, cobalt, copper, fluoride, germanium, iodine, iron, lithium, magnesium, manganese, molybdenum, phosphorus, potassium, selenium, silicon, sodium, sulfur, vanadium, zinc, and the like can be used.

In addition, the food composition of the present invention may contain preservatives, emulsifiers, acidulants, thickeners, and the like, if necessary, in addition to the sweetener. These preservatives, emulsifiers, and the like are preferably added and used in a very small amount as long as the application to which it is added can be achieved. When expressed numerically, the trace amount means a range of about 0.0005% by weight to about 0.5% by weight based on the total weight of the food composition. Preservatives that can be used include sodium calcium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), and the like. Examples of emulsifiers that can be used include gum acacia, carboxymethylcellulose, xanthan gum, pectin, and the like. Examples of acidulants that can be used include arithmetic, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, and phosphoric acid. These acidulants may be added so that the food composition has an appropriate acidity for the purpose of suppressing the growth of microorganisms in addition to the purpose of enhancing taste. As a thickener that can be used, a suspending agent, a settling agent, a gel-forming agent, a swelling agent, and the like may be mentioned.

Hereinafter, the present invention will be described in detail by examples.

However, the following examples specifically illustrate the present invention, and the contents of the present invention are not limited by the following examples.

[Production Example 1]

Preparation of a composite composition of 17-HDHA, resolvin D5 and protectin DX (hereinafter referred to as'SPMs composite composition')

Lipoxygenase having the amino acid sequence of SEQ ID NO: 1 was added to docosahexaenoic acid (DHA) at various concentrations and reacted at pH 7, room temperature for 30 minutes, and the final concentration of 1 M was 50 mM. The reaction product was reduced by adding sodium borohydride, and then 5 µl/ml of acetic acid was added to terminate the reaction. After the reaction product was purified using a solid cartridge (SPE, 500 mg of C18), the types of compounds in the reaction product were analyzed using normal phase HPLC.

As a result, it was confirmed that 17S-HDHA, resolbin D5 (7S, 17S-diHDHA), and protectin DX (10S, 17S-diHDHA) were produced from DHA, from which they were mixed in the ratio as shown in Table 1 below. A composite composition of SPMs that existed in the state was prepared.

Composition 1 Composition 2 Composition 3 17S-HDHA 3 25 84 Resolvin D5 47 56 10 Protectin DX 50 19 6

[Comparative Examples 1 to 3]

Isolation and purification of 17-HDHA, resolvin D5 and protectin DX

17S-HDHA, Resolvin D5 (7S, 17S-diHDHA), Protectin DX (10S, 17S-diHDHA) separated each product from the SPMs composite composition of the above preparation through prep HPLC equipped with a C18 column or a Diol column. It was purified and used.

[Example 1]

Confirmation of synergistic IL-6 expression inhibition effect by SPMs complex composition

In order to confirm the synergistic inflammation alleviating effect of the SPMs complex composition, the complex composition of 17-HDHA, resolvin D5 and protectin DX (compositions 1 to 3) and each SPM alone substance were used as human keratinocytes, HaCaT. Cells were treated to confirm changes in IL-6 expression.

[1-1] Experiment method

Specifically, first, human keratinocytes were dispensed into a 48-well plate at ^{a concentration of 4x10 4} cells/well and incubated at 5% CO ₂ , 37°C for 24 hours, and then the medium was removed and the starvation state was eliminated with DMEM Serum Free media. It was held for 24 hours.

The SPMs composite composition was diluted with DMEM (FBS 2%) to make 0.5, 1, 2, and 5 μg/ml, and then cultured as described above and treated on human keratinocytes from which the medium was removed for 1 hour. Then, for IL-6 expression, the medium was exchanged with 250µl of DPBS (WelGENE), irradiated with 160mJ/cm2 of UVB, and then treated with SPMs again and cultured for 24 hours. And after collecting the medium, the amount of IL-6 was measured using Human IL-6 DuoSet ELISA (R&D system), and the cells attached to the bottom were washed with DPBS, and then dissolved with 1N NaOH for BCA analysis. The amount of protein was measured by measuring the amount of IL-6 synthesized per certain protein. At this time, the case where nothing was treated after UVB irradiation was Con(-), and the case of 10 μM hydrocortisone was treated as Con(+). The effect of inhibiting IL-6 expression was confirmed.

[1-2] Confirmation of IL-6 inhibitory effect of SPM alone component

As a result, as shown in Table 2 and FIG. 1A, the expression of IL-6 was increased by irradiating UVB on human keratinocytes, and the increased expression of IL-6 was 2 μg/ml of 17S-HDHA and protectin. When treated with DX and resolbin D5, respectively, compared to Con(-), it decreased by 10.8%, 11.8%, and 0.5%, respectively, and showed a tendency dependent on the treatment concentration of SPM.

From this, it can be seen that all of 17-HDHA, Protectin DX, and Resolvin D5 have inflammation-relieving effects.

Sample UVB density
(μg/ml) IL-6 expression
(% of control) Average Untreated control - - 23.64 Con(-) + - 100 Con(+) H.C. + 10 μM 31.23 17S-HDHA + 0.1 89.63 + One 83 + 2 89.17 + 5 81.66 PDX + 0.1 95.36 + One 102.69 + 2 88.13 + 5 86.72 RvD5 + 0.1 99.58 + One 109.86 + 2 99.49 + 5 83.38

[1-3] Confirmation of IL-6 inhibitory effect of SPMs composite composition

Next, in the case of the SPMs composite composition, as shown in Table 3 and FIG. 1b, the expression of IL-6 was increased by irradiation with UVB, and thus the increased expression of IL-6 was the SPMs composite composition of the present invention (composition 1, 2, 3) When 2 μg/ml was treated, it was reduced by 18%, 33.8%, and 29.5%, respectively, compared to Con(-), and showed a tendency dependent on the treatment concentration of SPM.

From this, it can be seen that all of the SPMs composite compositions of the present invention have excellent inflammation alleviating effects.

Sample UVB density
(μg/ml) IL-6 expression
(% of control) Average Untreated control - - 15.62 Con(-) + - 100 Con(+) H.C. + 10 μM 42.82 Composition 1
(17S-HDHA: RvD5: PDX
= 3: 47:50) + 0.1 74.66 + One 72.49 + 2 72.98 + 5 64.93 Composition 2
(17S-HDHA: RvD5: PDX
= 25:56:19) + 0.1 71.28 + One 64.98 + 2 66.18 + 5 62.03 Composition 3
(17S-HDHA: RvD5: PDX
= 84:10:6) + 0.1 80.61 + One 78.63 + 2 70.52 + 5 68.19

[1-4] Confirmation of the synergistic IL-6 inhibitory effect of the SPMs composite composition

As shown in Table 4 and FIG. 1C below, when treating the composite composition of 2 μg/ml, composition 1 is 18%, composition 2 is 33.8%, and composition 3 is 29.5% compared to Con(-). It was confirmed that the effect of reducing the expression of IL-6 was increased compared to when the SPM alone component was treated at the same concentration.

UVB 20mJ/cm ² Con (-) 17S-HDHA PDX RvD5 Composition 1 Composition 2 Composition 3 100 89.17 88.13 99.49 72.98 66.18 70.52

From this, it can be seen that the SPMs composite composition of the present invention has a remarkably increased inflammation alleviating effect compared to the single component. In addition, the above results mean that due to the synergistic effect of 17S-HDHA, PDX and RvD5, the amount of use can be reduced when used as a composite composition than when using them alone. It can be seen that phosphorus side effects can be prevented.

[Example 2]

Confirmation of synergistic procollagen synthesis effect by SPMs composite composition

Collagen, a major component of the extracellular matrix, is synthesized in the form of a precursor called procollagen. Since the procollagen is known to be cleaved and separated from the collagen molecule when the polymerization reaction occurs, the degree of collagen biosynthesis in the cell can be estimated by measuring the amount of procollagen.

[2-1] Experimental method

Human dermal fibroblasts were treated with the SPM alone substance and their composite composition (compositions 1 to 3) at concentrations of 0.1, 1, 2 and 5 μg/ml, respectively, and cultured for 24 hours, and then the cultured cell medium was Collected and measured the amount of procollagen with procollagen typeI c-peptide (PIP) EIA kit (TAKARA, MK101). At this time, the case where nothing was treated as Untreated control, and the case where 50 μM of retinyl palmitate was treated as Con(+), the SPM alone component and their composite composition (composition 1 to 3 ) Was confirmed.

[2-2] Confirmation of the effect of synthesizing procollagen of SPM alone

As shown in Table 5 below and FIG. 2A, a higher amount of procollagen was identified in all of 17-HDHA, Protectin DX, and Resolvin D5 compared to the Untreated control. In particular, a higher amount of procollagen was confirmed in Protectin DX and Resolvin D5 than in the positive control (50 μM of retinyl palmitate) at a concentration of 1 μg/ml or more.

From this, it can be seen that 17-HDHA, Protectin DX, and Resolbin D5 all improve the synthesis of procollagen.

Sample density
(μg/ml) Procollagen synthesis
(% of control) Average Untreated control - 100 Con(+) R.P. 50 μM 249.88 17S-HDHA 0.01 212.76 0.1 237.76 One 235.8 2 255.32 PDX 0.01 237.15 0.1 244.1 One 251.41 2 254.1 RvD5 0.01 245.68 0.1 248 One 249.34 2 255.32

[2-3] Confirmation of the effect of synthesizing procollagen of the SPMs composite composition

Next, in the case of the SPMs composite composition, as shown in Table 6 and FIG. 2b, both compositions 1 to 3 having different mixing ratios were found to have a greater amount of procollagen than the Untreated control by 2 to 3 times. In particular, the SPMs composite composition was found to have a greater amount of procollagen compared to Con(+) treated with 50 μM of retinyl palmitate at a concentration of 0.1 μg/ml or more for all of the compositions 1 to 3.

From this, it can be seen that the SPMs composite composition of the present invention remarkably improves the synthesis ability of procollagen.

Sample density
(μg/ml) Procollagen synthesis
(% of control) Average Untreated control - 100 Con(+) R.P. 50 μM 240.2 Composition 1
(17S-HDHA: RvD5: PDX
= 3: 47:50) 0.01 218.2 0.1 259.3 One 289.5 2 302.7 Composition 2
(17S-HDHA: RvD5: PDX
= 25:56:19) 0.01 225.6 0.1 265.2 One 302 2 307.1 Composition 3
(17S-HDHA: RvD5: PDX
= 84:10:6) 0.01 220.7 0.1 267.3 One 301.1 2 304.4

[2-4] Synergistic Procollagen Synthesis Effect Confirmation of SPMs Composite Composition

As shown in Table 7 and FIG. 2C below, the SPMs composite composition (compositions 1, 2, and 3) all showed a greater amount of procollagen compared to 17-HDHA, protectin Dx or resolvin D5 alone component. .

Untreated control 17S-HDHA PDX RvD5 Composition 1 Composition 2 Composition 3 100 255.32 254.1 255.32 302.7 307.1 304.4

From this, it can be seen that the SPMs composite composition of the present invention has significantly increased procollagen synthesis compared to the single component. In addition, the above results mean that due to the synergistic effect of 17S-HDHA, PDX and RvD5, the amount of use can be reduced when used as a composite composition than when using them alone. It can be seen that phosphorus side effects can be prevented.

[Example 3]

Confirmation of the effect of improving the ability to inhibit the expression of collagen-degrading enzyme (MMP-1) by the SPMs complex composition

MMPs (matrix metalloproteinases) are enzymes that have proteolytic activity, and degrade proteins constituting the extracellular matrix to have an important effect on the binding of proteins constituting the extracellular matrix to cells. There are 28 types of MMPs, of which MMP-1 is an enzyme that degrades collagen, and in this example, the degree of expression of MMP-1 in cells was confirmed.

Specifically, after culturing human dermal fibroblasts for 24 hours, the medium was exchanged with DPBS (WelGENE) at a concentration of 250 µl/well and irradiated with 100 mJ/cm 2 of UVB, and the composition 2 was 0.1, 1, 2, and DMEM (2% FBS) diluted to a concentration of 5 ppm was treated and incubated for 48 hours. After collecting the cultured cell medium, the amount of MMP-1 was measured using a human total MMP-1 ELISA kit (R&D systems, DY901). At this time, when nothing was treated as a negative control, and 20 μM of retinoic acid was treated as a positive control, the effect of Composition 2 was confirmed.

As a result, as shown in FIG. 3, the expression of MMP-1 increased by about 2 times by irradiation with UVB, and the expression of MMP-1 thus increased was reduced by about 70% by treatment with 20 μM of retinoic acid. On the other hand, in the case of treatment with the SPMs composite composition, it was reduced by 11.4% at 0.01ppm, 26.1% at 0.1ppm, and 63.3% at 1ppm, and the effect of inhibiting the expression of MMP-1 is also dependent on the treatment concentration of the SPMs composite composition. Shown.

From this, it can be seen that the SPMs composite composition of the present invention has an excellent effect of improving skin wrinkles.

[Example 4]

Confirmation of the effect of improving the ability to inhibit TNF-α expression by the SPMs complex composition

TNF-α is a representative inflammatory cytokine and is known to intensify inflammatory skin diseases when the amount of expression increases. TNF-α, a tumor necrosis factor, is an important factor that plays a pivotal role in many inflammatory responses, and the regulation of TNF-α expression plays an important role in inflammatory diseases. Therefore, in order to confirm the anti-inflammatory effect of the SPMs composite composition of the present invention, the expression pattern of TNF-α was confirmed.

Specifically, human keratinocytes were dispensed in a 48-well plate at ^{a concentration of 4x10 4} cells/well and incubated at 5% CO ₂ , 37°C for 24 hours, and then the medium was removed and the starvation state was 24 with DMEM Serum Free media. Held for hours.

The composition 2 was diluted with DMEM (FBS 2%) to make 0.5, 1, 2 and 5 μg/ml, and then cultured as described above and treated with human keratinocytes from which the medium was removed for 1 hour. Then, for TNF-α expression, the medium was exchanged with 250µl of DPBS (WelGENE), irradiated with 160mJ/cm2 of UVB, and then treated with SPMs again and cultured for 24 hours. And after collecting the medium, the amount of TNF-α was measured using Human TNF-α DuoSet ELISA (R&D system), and the cells attached to the bottom were washed with DPBS, and then dissolved with 1N NaOH for BCA analysis. Through measuring the amount of protein was measured the amount of synthesis of TNF-α per certain protein. At this time, the effect of the SPMs composite composition was confirmed by using a case where nothing was treated as a negative control, and a case treated with 10 μM of hydrocortisone as a positive control.

As a result, in the case of TNF-α, as shown in FIG. 4, the expression of TNF-α was increased by about 5 times by irradiation with UVB, and thus the increased expression of TNF-α was treated with 10 μM of hydrocortisone. Accordingly, it was reduced by about 52%, while it was reduced by about 49% by 1 ppm of the SPMs complex composition, and the effect of inhibiting the expression of TNF-α showed a tendency dependent on the treatment concentration of the SPMs complex composition.

From this, it can be seen that the SPMs composite composition of the present invention has an excellent effect of terminating inflammation.

[Example 5]

Confirmation of the effect of improving the NO generation reduction ability by the SPMs composite composition

Nitric oxide (NO), which is known to play an important role in inducing inflammation as a kind of reactive oxygen species, is a highly reactive bio-generated molecule. NO, a key molecule involved in the inflammatory response, is an inflammatory mediator, accelerating the inflammatory response and further exacerbating the inflammatory response. Therefore, the amount of NO generation was confirmed for the stimulation induced by fine dust.

Specifically, the amount of NO generated from the cells was measured using the Griess Reagent System for the nitrite concentration present in the cell culture medium, and the NO concentration in the culture medium was determined using a standard curve for each concentration of sodium nitrite.

As a result, it was confirmed that the generation of NO in the cells treated with the SPMs composite composition 0.1 to 5 ppm was reduced to 39.2 to 70% as shown in FIG. 5.

From this, it can be seen that the SPMs composite composition of the present invention has an effect of alleviating inflammation.

[Example 6]

Checking the effect of improving the ability to reduce the amount of lipid peroxide produced by the SPMs complex composition

Lipid peroxiation of cells is one of the mechanisms of cellular damage that occurs in aging or disease states in animals and plants, and the degree of cellular damage can be determined by measuring total malondialdehyde (MDA), a product of lipid peroxidation. Therefore, the amount of MDA produced by the SPMs composite composition was confirmed for the stimulation induced by fine dust.

Specifically, the amount of total malondialdehyde (MDA), a product of Lipid peroxidation, was reacted with thiobarbituric Acid (TBA) to measure MDA-TBA adduct by the TBARS Method.

As a result, it was confirmed that, as shown in FIG. 6, the production of MDA in the cells treated with the SPMs complex composition at 0.1 to 5 ppm was reduced to 19.1 to 62.7%.

From this, it can be seen that the SPMs composite composition of the present invention has an effect of inhibiting skin aging.

[Example 7]

Checking the effect of improving skin barrier improvement by SPMs complex composition

Key proteins that make up the skin barrier include Filaggrin, Loricrin, and Involucrin. Among them, filaggrin is a protein produced by the decomposition of propyllagrin, which constitutes keratohyaline granule (KG) present in the granular cells of the epidermis, and plays an important role in the skin barrier by aggregation of keratin filaments in keratinocytes. Therefore, it was attempted to confirm the effect of strengthening the skin barrier by confirming the expression of filaggrin and loliclean by the sample.

Specifically, Human keratinocytes were cultured in DMEM medium under the same conditions as in Example 4. In order to confirm the expression of genes related to skin barrier improvement, the cells were cultured with the SPMs complex composition of the present invention at different concentrations, and then RNA was extracted and RT-PCR was performed. The amplified genes were analyzed using the Gel documentation system and Image J program.

As a result, as shown in FIGS. 7A and 7B, when 50 ppm of hyaluronic acid, which is a positive control, was treated, the expression of pilagrin was increased by 52.6% and loliclean by 97.7%. It was confirmed that Pilargreen increased 31.3~52.8%, and LoliClean increased 31.8~88.5%.

From this, it can be seen that the SPMs composite composition of the present invention has an effect of strengthening the skin barrier.

[Example 8]

Evaluation of human efficacy using SPMs cream

Using the SPMs composite composition, a cream (Cream S) was prepared in the composition shown in Table 8 below. For the comparative experiment, ceramide cream (Cream C) and base cream (Cream B) were also prepared in the following composition.

Cream S Cream C Cream B SPMs composite composition 2.00 - - Ceramide AC45 - 1.00 - glycerin 5.00 5.00 5.00 1,2-hexanediol 2.00 2.00 2.00 oil Appropriate amount Appropriate amount Appropriate amount antiseptic Appropriate amount Appropriate amount Appropriate amount Purified water Appropriate amount Appropriate amount Appropriate amount

(Unit: parts by weight)

[8-1] Checking the effect of improving skin barrier with SPMs cream

12 women (average age 45 years ± 5.54) were evaluated for the improvement of skin barrier human efficacy. The forearm was the test site, and the physical damage was divided into 3 parts of the test area and the control area (no application) and applied to 4 areas, and then Cream S (SPM cream), Cream C (Ceramide cream), and Cream B (Base cream) were applied. Each designated site was used for 1 week, twice a day, and instrument measurement was performed by comparing it with the control site (uncoated). Measurements and efficacy questionnaire evaluation were conducted before skin damage, immediately after skin damage, 1 day, 4 days, and 7 days after using the test product. The skin barrier improvement effect was confirmed by measuring TEWL (Transepidermal Water Loss) using Tewameter TM300.

As a result, as shown in Fig.8a, the ceramide cream, which is most widely used as a skin barrier improvement material, has no significant improvement compared to the base cream, whereas the SPM cream has statistically significant skin barrier improvement effect compared to the non-application group and the base cream. (Equal variance group two-sided test, t-test; p<0.034 after 4 days of use, p<0.027 after 7 days of use). In addition, as a result of comparing the skin barrier recovery rate (%), it was confirmed that the SPM cream has a comparative advantage compared to the base cream and ceramide cream as shown in FIG. 8B (Wilcoxon code ranking test; * p≤0.05, ** p≤ 0.01, ***p≤0.001).

[8-2] Checking the efficacy of improving atopic dermatitis with SPMs cream

A human efficacy evaluation was conducted on 22 subjects to relieve itching caused by dryness. After measuring the skin moisture, the average value between groups was adjusted, and the test products Cream S and Cream C were randomly assigned to 11 subjects to use each product. The forearm and dry lesions were used as test sites, and instrument measurement and efficacy questionnaire evaluation were conducted before product use, 2 weeks after product use, and 4 weeks after product use.

The effect of improving atopic dermatitis was confirmed by measuring skin moisture using Comeometer CM825, measuring skin moisture loss using Tewameter TM300, and evaluating the degree of improvement during itching (VAS).

Checking the skin moisturizing effect

As shown in Figure 9a, the most widely used ceramide cream for relieving dry symptoms of atopic dermatitis compared to before use of the product, the increase in skin moisture level is visible, but there is no statistical significance, whereas SPM cream is statistically significant at the lesion site. It was found to show the skin hydration improvement effect of (2 weeks, 4 weeks of use; <0.014, p<0.0115).

Confirmation of the effect of improving the skin barrier of the lesion area

As shown in Figure 9b, in terms of the rate of change in the amount of transdermal moisture loss, ceramide cream compared to before use of the product, the decrease in the amount of transdermal moisture loss was seen, but there was no statistical significance, whereas the SPM cream statistically significantly improved the transdermal moisture loss at the lesion site Showed efficacy (4 weeks of use; P<0.01451).

Checking the effect of itching improvement

As shown in Fig. 9c, in terms of the itching improvement effect, it was confirmed that the itching index was statistically significantly improved after 2 weeks and 4 weeks of using both SPM cream and ceramide cream.

From this, the SPM cream of the present invention moisturizes the skin, improves skin barrier, and itching at an equal level or higher than that of ceramide, which is commonly used as a functional material for improving skin barrier and improving skin pruritus, moisture, and itching caused by dry skin such as atopy. It can be seen that the improvement effect is shown.

[8-3] Checking the effect of improving eye wrinkles and skin moisture by SPMs cream

The human efficacy of Cream A and Cream S was evaluated for the improvement of wrinkles and skin moisture in the eyes of 20 selected subjects. The eye area was set as the test site, and 10 subjects used Cream A and the remaining 10 subjects used Cream S twice a day for 8 weeks. Before using the test product, after 4 weeks of use, and after 8 weeks of use, the effect of improving wrinkles and skin moisture was confirmed by measuring the test subjects.

Specifically, the eye wrinkles were measured using Antera 3D CS, and the skin moisture was measured using a Comeometer.

Checking the effect of improving wrinkles around the eyes

As shown in FIG. 10A, the skin wrinkle improvement rate (%) for each subject of Cream A containing retinol, a functional anti-wrinkle ingredient, was not statistically significant compared to before using the product, but SPM cream was used at 4 weeks and 8 weeks of use. It was confirmed that the skin wrinkle improvement rate (%) was significantly compared to the previous one (t-test (equal variance group, two-sided test); *, p≤0.05); **, p≤0.01; ***, p≤0.001).

Checking the moisturizing effect around the eyes

As shown in FIGS. 10B and 10C, the moisture of the skin around the eyes increased statistically significantly at 8 weeks of using SPM cream and retinol cream, and there was no statistically significant difference between SPM cream and retinol cream.

From this, it can be seen that the SPM cream of the present invention exhibits an effect of improving the amount of skin moisture around the eyes and skin wrinkles around the eyes at a level equal to or higher than that of retinol, a raw material for wrinkle improvement notice.

In the above, preferred embodiments of the present invention have been exemplarily described, but the scope of the present invention is not limited to the specific embodiments as described above, and those of ordinary skill in the relevant field have the scope described in the claims of the present invention. It will be possible to make appropriate changes within it.

<110> Korea Research Institute of Bioscience & Biotechnology KOLON LIFE SCIENCE INC. <120> Complex composition comprising DHA derivatives for anti-wrinkle, improving atopic dermatitis and enhancing skin barrier <130> 2019DPA3549 <160> 2 <170> KoPatentIn 3.0 <210> 1 <211> 571 <212> PRT <213> Artificial Sequence <220> <223> enzyme (K2) <400> 1 Met Val Asp Asn Met Lys Pro Ser Leu Pro Gln Asp Asp Pro Asn Gln 1 5 10 15 Glu Gln Arg Lys Asp Ser Leu Asn Arg Gln Gln Gln Ala Tyr Gln Phe 20 25 30 Asp Tyr Glu Ser Leu Ser Pro Leu Ala Leu Leu Lys Asn Val Pro Ala 35 40 45 Val Glu Asn Phe Ser Ser Lys Tyr Ile Gly Glu Arg Ile Leu Ala Thr 50 55 60 Ser Glu Leu Pro Ala Asn Met Leu Ala Ala Asp Ser Arg Thr Phe Leu 65 70 75 80 Asp Pro Leu Asp Glu Leu Gln Asp Tyr Glu Asp Phe Phe Thr Leu Leu 85 90 95 Pro Leu Pro Ala Val Ala Lys Ile Tyr Gln Thr Asp Arg Ser Phe Ala 100 105 110 Glu Gln Arg Leu Ser Gly Ala Asn Pro Met Val Leu Arg Leu Leu Asp 115 120 125 Ala Gly Asp Pro Arg Ala Gln Thr Leu Ala Gln Ile Ser Ser Phe His 130 135 140 Pro Leu Phe Asp Leu Gly Gln Glu Leu Gln Gln Lys Asn Ile Tyr Val 145 150 155 160 Ala Asp Tyr Thr Gly Thr Asp Glu His Tyr Arg Ala Pro Ser Lys Ile 165 170 175 Gly Gly Gly Ser Tyr Glu Lys Gly Arg Lys Phe Leu Pro Lys Pro Arg 180 185 190 Ala Phe Phe Ala Trp Arg Trp Thr Gly Ile Arg Asp Arg Gly Glu Met 195 200 205 Thr Pro Ile Ala Ile Gln Leu Asp Pro Thr Pro Asp Ser His Val Tyr 210 215 220 Thr Pro Phe Asp Pro Pro Val Asp Trp Leu Phe Ala Lys Leu Cys Val 225 230 235 240 Gln Val Ala Asp Ala Asn His His Glu Met Ser Ser His Leu Gly Arg 245 250 255 Thr His Leu Val Met Glu Pro Ile Ala Ile Val Thr Ala Arg Gln Leu 260 265 270 Ala Gln Asn His Pro Leu Ser Leu Leu Leu Lys Pro His Phe Arg Phe 275 280 285 Met Leu Thr Asn Asn Glu Leu Ala Arg Ser Tyr Leu Ile Ala Pro Gly 290 295 300 Gly Pro Val Asp Glu Leu Leu Gly Gly Thr Leu Pro Glu Thr Met Glu 305 310 315 320 Ile Ala Arg Glu Ala Cys Ser Thr Trp Ser Leu Asp Glu Phe Ala Leu 325 330 335 Pro Ala Glu Leu Lys Asn Arg Gly Met Asp Asp Thr Asn Gln Leu Pro 340 345 350 His Tyr Pro Tyr Arg Asp Asp Gly Leu Leu Leu Trp Asp Ala Ile Glu 355 360 365 Thr Phe Val Ser Gly Tyr Leu Lys Phe Phe Tyr Pro Thr Glu Ile Ala 370 375 380 Ile Val Gln Asp Val Glu Leu Gln Thr Trp Ala Gln Glu Leu Ala Ser 385 390 395 400 Asp Arg Gly Gly Lys Val Lys Gly Met Pro Pro Arg Ile Asn Thr Val 405 410 415 Glu Gln Leu Ile Lys Ile Val Thr Thr Ile Ile Phe Thr Cys Gly Pro 420 425 430 Gln His Ser Ala Val Asn Phe Pro Gln Tyr Glu Tyr Met Ser Phe Ala 435 440 445 Ala Asn Met Pro Leu Ala Ala Tyr Arg Asp Ile Pro Lys Ile Thr Ala 450 455 460 Ser Gly Asn Leu Glu Val Ile Thr Glu Lys Asp Ile Leu Arg Leu Leu 465 470 475 480 Pro Pro Tyr Lys Arg Ala Ala Asp Gln Leu Lys Ile Leu Phe Thr Leu 485 490 495 Ser Ala Tyr Arg Tyr Asp Arg Leu Gly Tyr Tyr Asp Lys Ser Phe Arg 500 505 510 Glu Leu Tyr Arg Met Ser Phe Asp Glu Val Phe Ala Gly Thr Pro Ile 515 520 525 Gln Leu Leu Ala Arg Gln Phe Gln Gln Asn Leu Asn Met Ala Glu Gln 530 535 540 Lys Ile Asp Ala Asn Asn Gln Lys Arg Val Ile Pro Tyr Ile Ala Leu 545 550 555 560 Lys Pro Ser Leu Val Ile Asn Ser Ile Ser Met 565 570 <210> 2 <211> 2595 <212> DNA <213> Artificial Sequence <220> <223> enzyme (K2) <400> 2 atgtttggca tcttcgacaa ggggcagaag ataaagggga ccgtggtgtt gatgccgaag 60 aacgtgttgg acttcaacgc cataacctcc atcggtaaag gtggtgttat tgacacagcc 120 accggcatct taggccaggg cgttagctta gttggtggag taattgatac cgccacttcc 180 tttttaggcc gtaatatctc catgcaattg atcagtgcta ctcagaccga tggctctgga 240 aatgggaaag taggaaaaga agtctatttg gaaaaacatc ttccgaccct accaacgttg 300 ggagcacgtc aggatgcgtt tagtattttc tttgagtggg acgcttcgtt tggaattccg 360 ggagcatttt atatcaaaaa ctttatgacg gatgagttct tcctcgtcag tgttaaactc 420 gaagacattc cgaaccatgg aaccattgaa tttgtttgca actcatgggt ttacaacttc 480 cgatcttaca aaaagaatcg gattttcttt gtcaatgaca catatcttcc aagtgccaca 540 ccagctccgc tgcttaagta ccgcaaagaa gaattggagg ttttaagagg agatggtaca 600 ggaaagcgca aggactttga cagaatctat gattatgatg tctataatga tttgggcaat 660 ccagatggtg gtgatcctcg cccaattctt ggtgggtcta gcatctatcc ctaccctcgc 720 cgggttcgta cagggagaga aaggacccgt accgatccca actcagagaa accgggcgag 780 gtttatgttc cacgcgacga aaattttggt cacctgaagt catcggattt ccttacatac 840 ggaatcaaat ctctgtccca cgatgtgata cctttgttca aaagcgcgat ttttcagtta 900 cgggtcacat cgagtgagtt tgaaagcttc gaagatgtgc gttcacttta cgagggtgga 960 attaaattgc caacagatat actgagccaa attagcccct taccagccct caaagaaatc 1020 tttcgcactg acggtgaaaa tgttcttcag tttccacctc cacatgtagc caaagtttcc 1080 aagtctgggt ggatgactga tgaggaattt gctcgtgagg tgattgccgg cgtaaaccca 1140 aatgtcatcc gtcggcttca ggaattcccg ccaaagagca cacttgatcc cacactctat 1200 ggtgatcaga ccagcaccat aacaaaagaa cagttggaaa ttaacatggg tggggtcaca 1260 gtagaagagg cgcttagtac ccagcggtta ttcatactgg actatcaaga tgcatttatc 1320 ccatatttga cgcggataaa cagcctacct actgcaaaag cgtatgccac aaggacaatc 1380 ctgttcttga aagacgatgg cactttgaag ccattagcta tcgaattaag caaaccacat 1440 ccagatggag ataatttggg tcctgagtcg atagtggtgc tgcctgcaac ggaaggtgtt 1500 gatagtacca tctggctatt ggccaaggct catgttattg tgaatgactc tggttatcac 1560 cagctggtaa gccattggtt aaacactcat gcagtgatgg aaccatttgc cattgcaacc 1620 aataggcacc tcagtgtgct ccaccctatt tataaacttt tgtatcctca ctaccgcgac 1680 accataaata ttaatggctt ggctcgccag tccctgatta atgcggatgg cattatcgaa 1740 aaatcatttt tgcccggaaa gtactccatt gagatgtctt catcagtcta caaaaattgg 1800 gtttttactg accaggcatt accagccgat ctggtcaagc gaggattggc aattgaagat 1860 ccctctgcac cccatggcct tcgccttgtg atagaggact acccttatgc tgttgatgga 1920 cttgaaatat gggatgctat taagacatgg gttcatgaat atgtctcctt gtattaccca 1980 acagatgcag ccgttcaaca ggacactgaa ctacaagcat ggtggaagga agcggtggag 2040 aagggtcacg gtgacctgaa agaaaagcct tggtggccta aaatgcagac aacggaagat 2100 ctgattcaat catgttctat tatcgtttgg acagcttcgg ctctccatgc tgcggttaat 2160 tttggacaat atccttacgg gggcttaatc ctgaaccgtc caactctagc ccgccgtttt 2220 atcccagcgg agggaacccc agagtatgat gaaatggtga agaatcctca aaaagcttat 2280 ctgcgaacaa tcacacccaa gtttgagacc ctgattgatt tatcggtaat tgaaatattg 2340 tcacgacatg cttctgatga gatatacctg ggagagcgcg aaaccccaaa ttggacgact 2400 gataagaagg cattagaagc attcaaacgt tttggcagca aactgactgg tattgaagga 2460 aaaatcaatg cgaggaacag tgatccgagt cttagaaacc ggacggggcc agttcagctg 2520 ccatatactt tgctccatcg ttcatcggag gaagggttga ctttcaaggg gatcccgaac 2580 agtatctcta tctaa 2595

Claims (12)

As a complex composition for improving skin condition comprising 17-hydroxydocosahexaenoic acid, resolvin D5 and protectin DX as active ingredients,
The 17-hydroxydocosahexaenoic acid, resolbin D5 and protectin DX are included in a weight ratio of 3 to 85: 10 to 60: 6 to 50,
The total concentration of 17-hydroxydocosahexaenoic acid, resolvin D5, and protectin DX of the complex composition for improving skin condition is 0.1 μg/ml or more, a complex composition for improving skin condition. The method according to claim 1,
The improvement of the skin condition is one or more selected from the group consisting of preventing or improving skin wrinkles, preventing or improving skin aging, preventing or improving skin inflammation, regenerating skin, and strengthening the skin barrier. The method according to claim 1,
The skin condition is any one or more selected from the group consisting of photoaging due to ultraviolet rays, melasma, freckles, skin wounds, dermatitis, atopic dermatitis, pruritus, eczema skin disease, dry eczema, erythema, urticaria, psoriasis, weak rash, and acne. That, a composite composition for improving skin conditions. The method according to claim 1,
The composition i) promotes the synthesis of collagen in cells, ii) inhibits collagen degradation, iii) inhibits the expression or activity of metalloproteinase-1 (MMP-1). , Complex composition for improving skin condition. The method according to claim 1,
The composition will have antioxidant activity, a composite composition for improving skin conditions. The method according to claim 1,
The composition is to suppress the inflammatory factor, a composite composition for improving skin conditions. The method according to claim 1,
The composition is to increase the expression of filaggrin or loricrin, a composite composition for improving skin conditions. delete The method according to any one of claims 1 to 7,
The composition is a cosmetic composition, a composite composition for improving skin condition. The method according to any one of claims 1 to 7,
The composition is a food composition, a composite composition for improving skin conditions. As a pharmaceutical composition for preventing or treating skin diseases comprising 17-hydroxydocosahexaenoic acid, resolvin D5 and protectin DX as active ingredients,
The 17-hydroxydocosahexaenoic acid, resolbin D5 and protectin DX are included in a weight ratio of 3 to 85: 10 to 60: 6 to 50,
The total concentration of 17-hydroxydocosahexaenoic acid, resolvin D5 and protectin DX of the pharmaceutical composition for preventing or treating skin diseases is 2ppm or more, a pharmaceutical composition for preventing or treating skin diseases. The method of claim 11,
The skin disease is one or more selected from the group consisting of skin wounds, skin scars, dermatitis, atopic dermatitis, pruritus, eczematous skin disease, dry eczema, athlete's foot, erythema, urticaria, psoriasis, weak rash, acne, and hair loss, pharmaceuticals Ever composition.

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