KR20230157782A - Compositions for preventing or improving skin damage comprising Zanthoxylum schinifolium oil as an active ingredient - Google Patents

Abstract

The present invention relates to a composition for preventing or improving skin damage using Sancho tree oil as an active ingredient. The composition according to one aspect inhibits the expression of MMP-1 and increases the expression of procollagen type I and skin barrier-related factors to prevent skin damage. It can be effectively used to prevent or improve.

Description

Compositions for preventing or improving skin damage comprising Zanthoxylum schinifolium oil as an active ingredient}

It relates to a composition for preventing or improving skin damage containing Sancho tree oil as an active ingredient.

As the elderly population increases, skin care, which can be a measure of aging, and antioxidants, which can prevent aging, are emerging as major areas of interest. The formation of wrinkles on the skin and the decrease in antioxidant activity in the body due to aging can be used as a measure of health. Depending on the factors, skin aging can be divided into intrinsic aging and extrinsic aging. Intrinsic aging is caused by changes in the physiological functions of the skin's epidermis and dermis with age, while extrinsic aging is known to be caused by changes in the physiological functions of the skin that occur from environments such as air pollution, exposure to ultraviolet rays, and stress. Among these aging mechanisms, oxidative stress induced by ultraviolet rays increases free radicals in the body, and MMP-1 (Matrix Metalloproteinase-1), which decomposes collagen, and hyaluronic acid ( Damage to the skin epidermis and dermis can be explained by increased activity of hyaluronidase, which decomposes hyaluronic acid.

The stratum corneum of the skin exists in the outermost layer of the skin and is in direct contact with the external environment, so it plays an important barrier function in protecting our body from external physical and chemical stresses. This barrier function is maintained by epidermal homeostasis. Epidermal homeostasis maintains continuous skin barrier function by forming a skin barrier called the stratum corneum through terminal differentiation through growth division and cell migration of keratinocytes in the basal layer (Korean J .Food.Sci.Technol.43:458-463, 2011).

As keratinocytes differentiate, they produce two factors that affect moisturization. First, during keratinocyte differentiation, the cell membrane is replaced by a structure called the cornified envelope. The keratinocyte membrane is a membrane structure in which various structural proteins, including filaggrin, are cross-linked. It provides skin protection against the external environment and at the same time suppresses moisture evaporation within the keratinocytes (Nat. Rev. Mol. Cell Biol. 6(4):328-340, 2005).

Additionally, during the differentiation process of keratinocytes, keratinocytes produce Natural Moisturizing Factor (NMF) and maintain the function of a skin barrier. The protein that is an important source for the production of natural moisturizing factors is filaggrin, which is decomposed into hydrophilic amino acids by CASP 14 (caspase 14) to form natural moisturizing factors. Natural moisturizing factors function to maintain moisture in the skin by providing water holding capacity and moisture absorption from the atmosphere (J. Cell Sci. 122:1285-1294, 2009). Therefore, maintaining an appropriate level of natural moisturizing factors in the skin is a very important factor for skin health through skin barrier function.

Therefore, research on mechanisms for preventing or improving UV skin damage has been actively conducted, and research is being conducted on cosmetics and health functional foods that demonstrate biochemical efficacy and effectiveness. With the development of research level and increase in technology, materials with practical efficacy and effectiveness are the main research targets.

Plant-derived materials have been used for a long time because they are excellent in terms of safety. In particular, in Korea, the development of functional materials mainly made of plants and herbal ingredients used in the private sector or in oriental medicine is actively being developed.

However, the use of Japanese pepper tree oil for preventing or improving skin damage is unknown, and no research on its mechanism has been conducted. Therefore, the present inventors completed the present invention by conducting direct efficacy studies on the oil's ability to prevent or improve skin damage.

One aspect is to provide a health functional food composition for preventing or improving skin damage containing Zanthoxylum schinifolium oil as an active ingredient.

Another aspect is to provide a cosmetic composition for preventing or improving skin damage containing Zanthoxylum schinifolium oil as an active ingredient.

Another aspect is to provide a pharmaceutical composition for preventing or treating skin damage comprising Zanthoxylum schinifolium oil.

Another aspect is to provide a quasi-drug composition for preventing or improving skin damage containing Zanthoxylum schinifolium oil as an active ingredient.

One aspect provides a health functional food composition for preventing or improving skin damage containing Zanthoxylum schinifolium oil as an active ingredient.

The Sancho tree is a deciduous broadleaf tree of the Rutaceae family, and may be a plant whose leaves, fruits, and seeds are specified as edible parts in the food raw materials notified by the Ministry of Food and Drug Safety.

In one embodiment, the Sancho tree oil may be obtained from the fruit of the Sancho tree.

In one embodiment, the Sancho tree fruit may include a pericarp or not.

In one embodiment, the fruit that does not contain the pericarp may refer to Sancho tree seeds.

In one embodiment, the Sancho tree oil may be an oil prepared by the following production method or a known commercially available Sancho tree oil.

The Sancho tree oil can be extracted by various methods commonly used in the field, such as extraction and concentration using organic solvents such as alcohol and hexane, or extraction by compression, for example, n-hexane or alcohol. It can be extracted by extraction/concentration or compression using a solvent, ultra-high pressure or supercritical conditions.

In the case of extraction by pressing, for example, dried Sancho tree fruits or seeds can be pulverized with a grinder to a size of about 50 to 1000 mesh, then placed in a mechanical press and milked at about 500 to 1000 kg/㎠, but it is limited to this. It doesn't work. Also, about 1 to 300°C, for example, 1 to 100°C, 1 to 50°C, 1 to 30°C, 5 to 200°C, 5 to 100°C, 5 to 50°C, 5 to 30°C or 5 to 20°C. It may be extracted by pressing under the conditions.

In one embodiment, the Sancho tree oil may increase the expression or activity of one or more selected from the group consisting of Filaggrin and Caspase14.

In one embodiment, the Sancho tree oil may inhibit the expression or activity of MMP-1.

In one embodiment, the Sancho tree oil may increase the expression or activity of Procollagen type I.

The present inventors confirmed that the Sancho tree oil is effective in moisturizing the skin by increasing the expression or activity of Filaggrin and/or Caspase14 and can further prevent, improve, or treat skin damage. In addition, it was confirmed that skin damage can be prevented, improved, or treated by increasing the expression or activity of procollagen type I and suppressing the expression or activity of MMP-1.

As used herein, the term "skin damage" includes death of human skin cells caused by external stimuli, such as ultraviolet rays, skin cell DNA damage, increased reactive oxygen species, increased lipid peroxidation, etc., and symptoms include erythema, This can include sunburn, pigmentation, photoaging, and skin cancer.

In one embodiment, the skin damage may be caused by ultraviolet rays.

As used herein, the term “prevention” refers to partially or completely delaying or preventing the onset or recurrence of a disease, disorder, or its secondary symptoms, preventing the acquisition or re-acquisition of a disease or disorder, or preventing the development or recurrence of a disease or disorder. A general term for reducing the risk of acquisition. The prevention refers to any action that suppresses or delays the occurrence of skin damage diseases, disorders, or symptoms by administering the composition according to the present invention.

As used herein, the term “improvement” refers to any action that results in at least a reduction in the severity of a parameter, such as a symptom, associated with the condition being treated.

In one embodiment, the Chinese pepper tree oil is 0.0001% to 99.0% by weight, for example, 0.01% to 60% by weight, 0.01% to 40% by weight, 0.01% to 30% by weight, based on the total weight of the composition. , 0.01% to 20% by weight, 0.01% to 10% by weight, 0.01% to 5% by weight, 0.05% to 60% by weight, 0.05% to 40% by weight, 0.05% to 30% by weight, 0.05% by weight. Weight % to 20 weight %, 0.05 weight % to 10 weight %, 0.05 weight % to 5 weight %, 0.1 weight % to 60 weight %, 0.1 weight % to 40 weight %, 0.1 weight % to 30 weight %, 0.1 weight % to 20% by weight, 0.1% to 10% by weight, 0.1% to 5% by weight, 1% to 50% by weight, 1% to 40% by weight, 2% to 40% by weight, 3% to 30% by weight. Weight % 4% to 30%, 5% to 30%, 5% to 20%, 5% to 18%, 6% to 15%, 8% to 15%, or It may be included in 7% by weight to 10% by weight. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount may be below the above range, and since there is no problem in terms of safety, the active ingredient may be used in amounts above the above range.

In this specification, the term "health functional food" refers to a food manufactured or processed using specific ingredients as raw materials or specific ingredients contained in food ingredients by methods such as extraction, concentration, purification, mixing, etc., for the purpose of health supplementation. It refers to food designed and processed to fully exert bioregulatory functions on the living body, such as biodefense, regulation of biorhythm, and prevention and recovery of disease. The health functional food composition can perform functions related to preventing and improving skin damage caused by fine dust.

There are no special restrictions on the types of foods above. Examples of foods to which the oil can be added include formulations selected from the group consisting of powders, granules, tablets, capsules, pills, gels, jellies, suspensions, emulsions, syrups, tea bags, leached teas, gums, candies, and health drinks. and includes all health foods in the conventional sense.

The health drink composition may include various sweeteners, flavoring agents, or natural carbohydrates as additional ingredients, like ordinary drinks. The sweetener may be a natural sweetener or a synthetic sweetener. The natural sweetener may be thaumatin or stevia extract, and the synthetic sweetener may be saccharin or aspartame.

The natural carbohydrate may be monosaccharide, disaccharide, polysaccharide, xylitol, sorbitol, or erythritol. The monosaccharide may be glucose or fructose, and the disaccharide may be maltose or sucrose. The polysaccharide may be a dextrin or cyclodextrin. The proportion of the natural carbohydrate may generally be about 0.01 to 10 g, for example, about 0.01 to 0.1 g per 100 ml of the composition of the present invention.

The health functional food may include food supplements that are foodologically acceptable, and may include an appropriate carrier commonly used in the manufacture of health functional foods.

In addition to the above, the composition of the present invention contains various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, It may include carbonating agents used in carbonated drinks. Additionally, the composition of the present invention may include pulp for the production of natural fruit juice, fruit juice beverages, and vegetable beverages. These ingredients can be used independently or in combination. The ratio of these additives is not very important, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

Another aspect provides a cosmetic composition for preventing or improving skin damage containing Sancho tree oil as an active ingredient.

The cosmetic composition includes lotion (skin lotion), skin, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutritional lotion, massage cream, nutritional cream, moisture cream, hand cream, hand sanitizer, foundation, essence, It can be manufactured into formulations including nutritional essence, pack, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, suspension, gel, powder, paste, mask pack, and sheet. Compositions of this dosage form can be prepared according to methods conventional in the art. The mixing amount of additional ingredients such as the above-mentioned moisturizer can be easily selected by a person skilled in the art within a range that does not impair the purpose and effect of the present invention.

In addition to the active ingredients disclosed herein, the cosmetic composition may further include functional additives and ingredients included in general cosmetic compositions, including commonly used purified water, thickeners, preservatives, stabilizers, solubilizers, surfactants, carriers, It may further include fragrance or a combination thereof. The functional additive may include ingredients selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, sphingolipids, and seaweed extract. Examples of the carrier include alcohol, oil, surfactant, fatty acid, silicone oil, wetting agent, humectant, viscosity modifier, emulsion, stabilizer, ultraviolet scattering agent, ultraviolet absorber, coloring agent, fragrance, etc. Compounds/compositions that can be used as alcohols, oils, surfactants, fatty acids, silicone oils, wetting agents, moisturizers, viscosity modifiers, emulsions, stabilizers, UV scattering agents, UV absorbers, coloring agents, and fragrances are already known in the art. Therefore, a person skilled in the art can select and use the appropriate material/composition. In addition, the cosmetic composition may contain sunscreen, antioxidants (butylhydroxyanisole, propyl gallate, elisorbic acid, tocopheryl acetate, butylated hydroxytoluene, etc.), and preservatives (methylparaben, butylparaben) as necessary. , propylparaben, phenoxyethanol, imidazolidinyl urea, chlorphenesin, etc.), colorants, pH adjusters (triethanolamine, citric acid, citric acid, sodium citrate, malic acid, sodium malate, fmal acid, sodium fmalate, succinic acid) , sodium succinate, sodium hydroxide, sodium monohydrogen phosphate, etc.), moisturizers (glycerin, sorbitol, propylene glycol, butylene glycol, hexylene glycol, diglycerin, betaine, glycereth-26, methyl glue) Additional ingredients such as Seth-20, etc.) and lubricants can be added.

Additionally, in each type of cosmetic composition, appropriate ingredients can be selected and mixed depending on the formulation or purpose of use of the cosmetic. Since the mixing ingredients and methods may be in accordance with conventional techniques, detailed descriptions thereof are omitted in this specification.

Another aspect provides a pharmaceutical composition for preventing or treating skin damage containing Sancho tree oil as an active ingredient.

As used herein, the term “pharmaceutical composition” may refer to a molecule or compound that imparts some beneficial effect upon administration to a subject. Beneficial effects include enabling diagnostic decisions; Improvement of a disease, symptom, disorder or condition; Reducing or preventing the onset of a disease, symptom, disorder or condition; and generally responding to a disease, symptom, disorder or condition.

As used herein, the term “treatment” refers to any action that improves or beneficially changes a disease, disorder, or its accompanying symptoms.

The pharmaceutical composition may be formulated with a preparation selected from the group consisting of tablets, soft or hard capsules, pills, powders, suspending agents, syrups, injections, and granules.

The pharmaceutical composition may be for oral or parenteral administration.

The pharmaceutical composition includes conventional fillers, extenders, binders, disintegrants, anti-aggregants, lubricants, wetting agents, pH adjusters, nutrients, vitamins, electrolytes, alginic acid and its salts, pectic acid and its salts, protective chloride, glycerin, It may contain fragrances, emulsifiers, or preservatives.

The pharmaceutical composition may include a pharmaceutically acceptable carrier, examples of such carriers include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin. , calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, propylhydroxybenzoate, It may be one or more selected from the group consisting of talc, magnesium stearate, mineral oil, dextrin, calcium carbonate, propylene glycol, liquid paraffin, and physiological saline.

The formulation of the pharmaceutical composition may vary depending on the method of use, and may be formulated using methods well known in the art to provide rapid, sustained, or delayed release of the active ingredient after administration to a mammal. It can be.

Preparations for oral administration include tablets, soft or hard capsules, pills, powders, suspensions, syrups, injections and granules, and these preparations may contain one or more excipients such as starch, calcium carbonate, sucrose or lactose. It can be manufactured by mixing gelatin, etc. Additionally, in addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Preparations for parenteral administration may be creams, lotions, ointments, warning agents, solutions, aerosols, fluid extracts, elixirs, needles, sachets, patches, or injections.

The method is applicable to any animal, and the animals may include humans and primates, as well as domestic animals such as cattle, pigs, sheep, horses, dogs, and cats.

The dosage of the composition for treatment, prevention, or improvement can be determined considering the method of administration, the age and gender of the recipient, the severity and condition of the patient, the absorption of the active ingredient in the body, the inactivation rate, and the concomitant drug, Based on the daily active ingredient, 0.1 mg/kg (body weight) to 500 mg/kg (body weight), 0.1 mg/kg (body weight) to 400 mg/kg (body weight), or 1 mg/kg (body weight) to 300. It can be administered in mg/kg (body weight) and can be administered once or in several divided doses, but is not limited to this.

Another aspect provides a quasi-drug composition for preventing or improving skin damage containing Sancho tree oil as an active ingredient.

The term "quasi-drug" refers to fibers, rubber products or similar products used for the purpose of treating, alleviating, treating or preventing diseases in humans or animals, have a weak effect on the human body or do not directly act on the human body, and are not instruments or machines. Products that fall under one of the following: preparations used for sterilization, insecticide, and similar purposes to prevent infection, and are used for the purpose of diagnosing, treating, alleviating, treating, or preventing the condition or disease of humans or animals. It refers to articles that are not instruments, machines, or devices, and articles used for the purpose of having a pharmacological effect on the structure and function of humans or animals, excluding those that are not instruments, machines, or devices, including external skin preparations and personal hygiene products. It can be included.

When adding the oil to a quasi-drug composition, the oil can be added as is or used together with other quasi-drug ingredients, and can be used appropriately according to conventional methods. The mixing amount of the active ingredient can be appropriately determined depending on the purpose of use (prevention, health, or therapeutic treatment).

The quasi-drug composition is not particularly limited thereto, but may be a personal hygiene product, external skin preparation, humidifier filler, mask, ointment, or filter filler. The personal hygiene products may be soap, wet tissues, toilet paper, shampoo, toothpaste, hair care products, air freshener gel, or cleaning gel.

A composition containing Sancho tree oil according to one aspect can be effectively used to prevent or improve skin damage by inhibiting MMP-1 expression and increasing expression of procollagen type I and skin barrier-related factors.

Figure 1 is a chromatogram showing the results of analyzing Sancho tree oil by high-performance liquid chromatography.
Figure 2 is a graph showing the cell survival rate according to Sancho tree oil treatment in dermal cells damaged by UV-B.
Figure 3 is a graph showing the cell survival rate according to Sancho tree oil treatment in epidermal cells damaged by UV-B.
Figure 4 is a graph showing the amount of MMP-1 secretion according to Sancho tree oil treatment in dermal cells damaged by UV-B.
Figure 5 is a graph showing the secretion amount of Procollagen type I according to Chinese pepper oil treatment in dermal cells damaged by UV-B.
Figure 6 is a graph showing the expression level of Filaggrin and Caspase14 in epidermal cells damaged by UV-B according to treatment with Sancho tree oil:
Figure 6a is a graph showing the expression level of Filaggrin; and Figure 6b is a graph showing the expression level of Caspase14.

This will be described in more detail through examples below. However, these examples are for illustrative purposes only and the scope of the present invention is not limited to these examples.

Example 1. Sancho tree (Z anthoxylum schinifolium ) Buy oil

Sancho tree (Z anthoxylum schinifolium ) oil was purchased from Hadong Sancho Farm (product name: Hadong Sancho oil) and used.

Experimental Example 1. Sancho tree (Z anthoxylum schinifolium ) High-performance liquid chromatography analysis of oil

The Sancho tree oil obtained in Example 1 was analyzed using high-performance liquid chromatography (HPLC) and an ultraviolet light intensity detector (UV/Vis Detector). The HPLC equipment used was the Waters e2695 Series system and Waters 2489 UV/Vis detector (Waters, Milford, MA, USA), and all solvents used in the analysis were HPLC grade solvents purchased from Burdick & Jackson (Muskegon, MI, USA). used.

The analysis of Japanese pepper oil obtained in Example 1 used a Luna (5 μm, 250 Х 4.6 mm, Phenomenex) column, the column temperature was set to 30°C, the injection volume was 10 μL, and the measurement wavelength was set to 210 nm. Acetonitrile (ACN) and tertiary distilled water (D.W.) were used as mobile phases, and the analysis conditions were A: ACN, B: DW, 0-50 min 20% A, 51-61 min, 100% A, 62-72 Min 20% A was analyzed at a rate of 1 mL/min. Analysis sample: 1 mL of Sancho tree oil was taken, 5 mL of hexane was added, ultrasonic extraction was performed for 30 minutes at room temperature, and vortexing was performed for 30 seconds. Afterwards, post-use analysis was performed using a 0.2 μm membrane filter.

As shown in Figure 1, it was confirmed that a number of components were detected in Sancho tree oil.

Experimental Example 2. Sancho tree (Z anthoxylum schinifolium ) Evaluation of cytotoxicity of oil in dermal cells

The cytotoxicity of the Japanese pepper oil of Example 1 was confirmed through MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay.

Specifically, Hs68 fibroblasts, which are dermal cells, were cultured in DMEM medium containing 10% FBS and 1% antibiotic-antimycotic at 5% CO ₂ and 37°C. And Hs68 fibroblast cells were stabilized in a 24-well plate at a concentration of 8.0 Х 10 ⁴ cells/500㎕/well for 24 hours. To irradiate UV-B, the existing medium was removed, washed with PBS (Phosphate Buffer Solution), and then treated with UV-B at 22 mJ/cm ² . Then, they were treated with the Japanese pepper oil of Example 1 at concentrations of 25, 50, and 100 ppm, respectively, and cultured for 24 hours. Afterwards, each cell was treated with MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) solution to achieve a final concentration of 0.5 mg/ml and incubated for 4 hours. Afterwards, the solution was completely removed and the absorbance of the plate dissolved in DMSO (Dimethyl sulfoxide) was measured at 590 nm.

Figure 2 is a graph showing the cell survival rate according to Sancho tree oil treatment in dermal cells damaged by UV-B.

As shown in Figure 2, Sancho tree oil did not show cytotoxicity to dermal cells, and the treatment concentration in the subsequent experimental example was determined based on the cytotoxicity test.

Experimental Example 2. Sancho tree (Z anthoxylum schinifolium ) Evaluation of cytotoxicity of oil in epidermal cells

The cytotoxicity of the Japanese pepper oil of Example 1 was confirmed through MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay.

Specifically, Hacat, an epidermal cell, was cultured in DMEM medium containing 10% FBS and 1% antibiotic-antimycotic at 5% CO ₂ and 37°C. And Hacat cells were stabilized in a 96-well plate at a concentration of 1.0 Х 10 ⁴ cells/200㎕/well for 24 hours. To irradiate UV-B, the existing medium was removed, washed with PBS (Phosphate Buffer Solution), and treated with UV-B at 20 mJ/cm ² . Then, the Japanese pepper tree oil of Example 1 was treated at concentrations of 25, 50, and 100 ppm and cultured for 24 hours. Afterwards, each cell was treated with MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) solution to achieve a final concentration of 0.5 mg/ml and incubated for 4 hours. Afterwards, the solution was completely removed and the absorbance of the plate dissolved in DMSO (Dimethyl sulfoxide) was measured at 590 nm.

Figure 3 is a graph showing the cell survival rate according to treatment with Sancho tree oil in epidermal cells damaged by UV-B.

As shown in Figure 3, Sancho tree oil did not show cytotoxicity to epidermal cells, and the treatment concentration in the subsequent experimental example was determined based on the cytotoxicity test.

Experimental Example 3. Sancho tree (Z anthoxylum schinifolium ) Confirmation of inhibition of MMP-1 (Matrix Metalloproteinase-1) expression in oil

In order to confirm the efficacy of the Japanese pepper oil of Example 1 in suppressing secretion of MMP-1 on cells damaged by UV-B, the amount of MMP-1 secretion was measured using the Human MMP1 ELISA Kit (ab100603, abcam, US). It was measured.

Specifically, Hs68 fibroblasts, which are dermal cells, were cultured in DMEM medium containing 10% FBS and 1% antibiotic-antimycotic at 5% CO ₂ and 37°C. And Hs68 fibroblast cells were stabilized in a 24-well plate at a concentration of 8.0 Х 10 ⁴ cells/500㎕/well for 24 hours. To irradiate UV-B, the existing medium was removed, washed with PBS (Phosphate Buffer Solution), and then treated with UV-B at 22 mJ/cm ² . Next, the Sancho tree oil of Example 1 was treated at concentrations of 25, 50, and 100 ppm, respectively, and cultured for 24 hours. MMP-1 in the supernatant obtained after 24 hours was quantified using an ELISA kit, and the results are shown in Figure 4.

Figure 4 is a graph showing the amount of MMP-1 secretion according to Sancho tree oil treatment in dermal cells damaged by UV-B.

As shown in Figure 4, it was confirmed that all Sancho tree oils significantly inhibited MMP-1 secretion in proportion to their concentration.

The above results mean that Sancho tree oil according to one embodiment effectively inhibits MMP-1 secretion and is effective in preventing or improving skin damage caused by ultraviolet rays.

Experimental Example 4. Sancho tree (Z anthoxylum schinifolium ) Confirmation of promotion of Procollagen type I expression in oil

In order to confirm the effectiveness of the Japanese pepper oil of Example 1 in increasing the secretion of Procollagen on cells damaged by UV-B, the amount of Procollagen secretion was measured using the Procollagen Type I C-peptide (PIP) ELISA Kit (Mk101, Takara, Japan) ) was measured using .

Specifically, Hs68 fibroblasts, which are dermal cells, were cultured in DMEM medium containing 10% FBS and 1% antibiotic-antimycotic at 5% CO ₂ and 37°C. And Hs68 fibroblast cells were stabilized in a 24-well plate at a concentration of 8.0 Х 10 ⁴ cells/500㎕/well for 24 hours. To irradiate UV-B, the existing medium was removed, washed with PBS (Phosphate Buffer Solution), and then treated with UV-B at 22 mJ/cm ² . Next, the Sancho tree oil of Example 1 was treated at concentrations of 25, 50, and 100 ppm, respectively, and cultured for 24 hours. Procollagen type I in the supernatant obtained after 24 hours was quantified using an ELISA kit, and the results are shown in Figure 5.

Figure 5 is a graph showing the secretion amount of Procollagen type I according to Chinese pepper oil treatment in dermal cells damaged by UV-B.

As shown in Figure 5, it was confirmed that all Sancho tree oils significantly increased the amount of Procollagen type I secretion in proportion to the concentration.

The above results mean that Sancho tree oil according to one embodiment effectively increases Procollagen type I secretion and is effective in preventing or improving skin damage caused by ultraviolet rays.

Experimental Example 5. Sancho tree ( Zanthoxylum schinifolium ) Evaluation of changes in skin barrier-related gene expression levels due to oil

In order to confirm the skin moisturizing effect of the Japanese pepper oil of Example 1 on cells damaged by UV-B, the skin barrier-related genes Filaggrin and Caspase14 were measured.

Specifically, Hacat, an epidermal cell, was cultured in DMEM medium containing 10% FBS and 1% antibiotic-antimycotic at 5% CO ₂ and 37°C. And Hacat cells were stabilized in a 6-well plate at a concentration of 8.0 Х 10 ⁵ cells/2 mL/well for 24 hours. To irradiate UV-B, the existing medium was removed, washed with PBS (Phosphate Buffer Solution), and treated with UV-B at 20 mJ/cm ² . Next, the Sancho tree oil of Example 1 was treated at concentrations of 25, 50, and 100 ppm, respectively, and cultured for 24 hours. After 24 hours, mRNA was extracted from the cells, synthesized into cDNA, and quantitative real-time-PCR was performed using a target template (primer) to finally evaluate the level of Filaggrin and Caspase14 gene expression, and the results are shown in Figure 6. . Primer sequences used to synthesize cDNA are shown in Table 1.

gene Primer direction Primer sequence (5'→3') sequence number Filaggrin Forward AGT GCA CTC AGG GGG CTC ACA One Reverse CCG GCT TGG CCG TAA TGT GT 2 Caspase14 Forward CAA ACA CAT GGG TCA CTT GC 3 Reverse CAG AAC TGC TGA GCC TAC CC 4 β-actin Forward CAT GTA CGT TGC TAT CCA GGC 5 Reverse CTC CTT AAT GTC ACG CAC GAT 6

Figure 6 is a graph showing the expression levels of Filaggrin and Caspase14 in epidermal cells damaged by UV-B according to treatment with Sancho tree oil.

The above results mean that Sancho tree oil according to one embodiment effectively increases the expression levels of Filaggrin and Caspase14, and is effective in preventing or improving skin damage caused by ultraviolet rays.

Preparation Example 1. Preparation of tablets

Tablets were prepared by mixing the Japanese pepper oil of Example 1 in the ingredient ratio shown in Table 2 below and compressing it into tablets according to a conventional tablet manufacturing method. The manufactured tablets were confirmed to be stable under all formulation test conditions.

Raw material name Unit weight (mg) Sancho tree oil 50 corn starch 100 lactose 100 stearic acid 2

Preparation Example 2. Preparation of capsules

The Sancho tree oil of Example 1 was mixed in the ingredient ratio shown in Table 3 below and filled into a gelatin capsule to prepare a soft capsule. The manufactured capsule was confirmed to be stable under all formulation test conditions.

Raw material name Unit weight (mg) Sancho tree oil 50 corn starch 100 lactose 100 stearic acid 2

Preparation Example 3. Preparation of liquid

A liquid was prepared by mixing the Japanese pepper oil of Example 1 in the ingredient ratio shown in Table 4 below and filling it into a fruit bottle or pouch according to a beverage preparation method suited to taste. The prepared liquid was confirmed to be stable under all formulation test conditions.

Raw material name Unit weight (g) Sancho tree oil 2.5050 xanthan gum 0.0075 Fructooligosaccharide solution 0.7500 Coconut flower essence powder 1.0500 Ssanghwa Concentrate 1.5000 Red ginseng scent 0.0450 Purified water 9.1425

Preparation Example 4. Preparation of jelly

Jelly was prepared by mixing the Japanese pepper oil of Example 1 in the ingredient ratio shown in Table 5 below and filling it into a three-sided cloth according to a jelly manufacturing method suitable to taste. The prepared jelly was confirmed to be stable under all formulation test conditions.

Raw material name Unit weight (g) Sancho tree oil 2.0000 food gel 0.3600 Carrageenan 0.0600 calcium lactate 0.1000 sodium citrate 0.0600 Complex golden extract 0.0200 Enzyme-processed stevia 0.0440 Fructooligosaccharide solution 5.0000 red grape concentrate 2.4000 Purified water 13.9560

The composition ratios of Preparation Examples 1 to 4 above were generally prepared by mixing appropriate ingredients as formulation examples, but the mixing ratio and raw materials may be arbitrarily changed as needed.

The Sancho tree oil of the present invention was stable under all formulation test conditions, so there was no problem with the stability of the formulation.

<110> COSMAX BIO CO., LTD. <120> Compositions for preventing or improving skin damage comprising Zanthoxylum schinifolium oil as an active ingredient <130> PN220391KR <160> 6 <170> KoPatentIn 3.0 <210> 1 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> forward primer for Filaggrin <400> 1 agtgcactca gggggctcac a 21 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> reverse primer for Filaggrin <400> 2 ccggcttggc cgtaatgtgt 20 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> forward primer for Caspase14 <400> 3 caaacacatg ggtcacttgc 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> reverse primer for Caspase14 <400> 4 cagaactgct gagcctaccc 20 <210> 5 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> forward primer for beta-actin <400> 5 catgtacgtt gctatccagg c 21 <210> 6 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> reverse primer for beta-actin <400> 6 ctccttaatg tcacgcacga t 21

Claims (10)

A health functional food composition for preventing or improving skin damage containing Zanthoxylum schinifolium oil as an active ingredient. In claim 1,
Sancho tree oil is a health functional food composition that increases the expression or activity of one or more selected from the group consisting of Filaggrin and Caspase14. In claim 1,
Sancho tree oil is a health functional food composition that inhibits the expression or activity of MMP-1. In claim 1,
Sancho tree oil is a health functional food composition that increases the expression or activity of procollagen type I. In claim 1,
A health functional food composition wherein the skin damage is caused by ultraviolet rays. In claim 1,
A health functional food composition wherein Sancho tree oil is included in an amount of 0.0001% to 99.0% by weight based on the total weight of the composition. In claim 1,
The health functional food is a health functional food having a dosage form selected from the group consisting of powders, granules, tablets, capsules, pills, gels, jellies, suspensions, emulsions, syrups, tea bags, leached teas, gums, candies and health drinks. Composition. A cosmetic composition for preventing or improving skin damage containing Sancho tree oil as an active ingredient. A pharmaceutical composition for preventing or treating skin damage containing Sancho tree oil as an active ingredient. A quasi-drug composition for preventing or improving skin damage containing Sancho tree oil as an active ingredient.