Composition for preventing, improving or treating atopic dermatitis containing symphyocladia linearis as an active ingredient

Abstract

The present invention relates to a composition for the prevention, improvement, or treatment of atopic dermatitis comprising *Symphyocladia linearis* as an active ingredient. The extract of Symphyocladia linearis of the present invention has excellent antioxidant efficacy, exhibits low cytotoxicity against skin keratinocytes, and can alleviate pruritus found in atopic and psoriatic skin by improving skin barrier function. Therefore, the composition containing the extract of Symphyocladia linearis of the present invention can be utilized as a material for pharmaceuticals, cosmetics, food, etc., capable of preventing, improving, or treating atopic dermatitis based on its biological activity. The present invention is part of the Busan Technopark 2023 Industry-Academic Cooperation Corporate R&D Support Project.

Description

Technology Field

[0001] The present invention relates to a composition for the prevention, improvement, or treatment of atopic dermatitis comprising Symphyocladia linearis as an active ingredient and a method for preparing the same. More specifically, the invention relates to a composition comprising a Symphyocladia linearis extract for the prevention, improvement, or treatment of atopic dermatitis that can alleviate pruritus found in skin with atopic dermatitis and psoriasis by improving skin barrier function. Background Technology

[0003] Atopic dermatitis is a chronic inflammatory skin disease that begins in infancy or childhood and persists for a long time. It causes pruritus (itching) and dry skin, and a major characteristic is that when symptoms appear, the affected area is scratched or rubbed, which further exacerbates the skin symptoms. It also causes sleep disorders, emotional and physical stress, interferes with social life, lowers the quality of life, and results in economic losses. The prevalence is on the rise worldwide, with 20% to 25% of the population suffering from allergic diseases such as rhinitis, asthma, atopic dermatitis, and food allergies.

[0004] The causes of atopic dermatitis are known to include genetic predisposition due to a family history of atopic diseases such as atopic dermatitis, allergic rhinitis, and asthma; environmental factors due to increased exposure to allergens caused by air pollution and changes in the living environment; immunological abnormalities that cause abnormal hypersensitivity of immune cells involved in allergic reactions to allergens; and abnormalities in skin barrier function, but the cause has not been fully elucidated to date.

[0005] Currently, most medications used to treat atopic dermatitis are drugs that inhibit histamine secretion or treat and alleviate symptoms. In particular, steroids are widely used for their excellent therapeutic effects, but long-term use can lead to serious side effects. As atopic dermatitis predominantly affects children and adolescents, there is an urgent need to develop safe treatments free from toxicity and side effects. Although much research is currently being conducted, the reality is that developing a treatment remains challenging.

[0007] Accordingly, the inventors have completed the present invention, which can be utilized as a material for the prevention, improvement, and treatment of atopic dermatitis that is effective in alleviating pruritus found in atopic and psoriatic skin, by using a natural extract of fine purple agar, a seaweed inhabiting the rocky subtidal zone of the East Sea in Gangwon-do that has excellent anti-inflammatory and antioxidant properties. Prior art literature

[0009] Republic of Korea Registered Patent Publication No. 10-0745023 Republic of Korea Registered Patent Publication No. 10-1756020 The problem to be solved

[0010] The inventors aim to provide a composition for the prevention, improvement, or treatment of atopic dermatitis comprising fine purple agar (Symphyocladia linearis) as an active ingredient in order to overcome the above-mentioned problems.

[0011] The present invention aims to provide a pharmaceutical composition for the prevention, improvement, or treatment of atopic dermatitis comprising Symphyocladia linearis as an active ingredient.

[0012] The technical problems that the present invention aims to solve are not limited to those mentioned above, and other unmentioned technical problems can be clearly understood by those skilled in the art from the description of the present invention. means of solving the problem

[0014] To achieve the above objectives, the present invention provides a composition for preventing, improving, or treating atopic dermatitis comprising an extract of Symphyocladia linearis as an active ingredient.

[0015] The above composition is characterized by increasing the expression of one or more genes selected from the group consisting of keratin 1 (KRT1), keratin 2 (KRT2), keratin 10 (KRT10), involucrine (IVL), loricrine (LOR), claudin 1 (CLDN1) and filaggrin (FLG), which are differentiation genes of skin keratinocytes.

[0016] The above composition is characterized by restoring the expression of filaggrin (FLG) that is inhibited by cytokines IL-4 and IL-13, which are increased in atopic dermatitis.

[0017] In addition, the above composition is characterized by having an antioxidant effect.

[0018] The present invention provides a pharmaceutical composition for the prevention, improvement, or treatment of atopic dermatitis comprising an extract of *Symphyocladia linearis* as an active ingredient. Effects of the invention

[0020] The composition for preventing, improving, or treating atopic dermatitis containing Symphyocladia linearis as an active ingredient of the present invention has the effect of alleviating pruritus by increasing the gene expression of keratin 1 (KRT1), keratin 2 (KRT2), keratin 10 (KRT10), involucrine (IVL), loricrine (LOR), claudin 1 (CLDN1), and filaggrin (FLG), which are differentiation genes of skin keratinocytes, and by improving the skin barrier by restoring the expression of filaggrin (FLG), which is suppressed by cytokines IL-4 and IL-13 that are increased due to atopic dermatitis, and can be applied to the manufacture of pharmaceuticals, cosmetics, and food related to atopic dermatitis.

[0022] The effects of the present invention are not limited to those mentioned above, and other unmentioned effects will be clearly understood by those skilled in the art from the description in the claims. Brief explanation of the drawing

[0024] Figure 1 shows the antioxidant efficacy of Symphyocladia linearis extract according to preferred embodiment 1 and experimental example 1 of the present invention. Figure 2 shows the cytotoxicity of skin keratinocytes to the extract of Symphyocladia linearis according to preferred embodiments 1 and 2 and experimental example 2 of the present invention. FIG. 3 shows the expression of genes for maintaining skin barrier function by Symphyocladia linearis extract according to preferred embodiments 1 and 2 and experimental example 3 of the present invention. Figure 4 shows the protein expression of FLG by Symphyocladia linearis extract in an in vitro atopic dermatitis model according to preferred embodiments 1 and 2 and experimental example 3 of the present invention. Specific details for implementing the invention

[0025] Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings.

[0026] The terms used in this specification have been selected based on currently widely used general terms whenever possible, taking into account their functions in the present invention; however, these terms may vary depending on the intent of those skilled in the art, case law, the emergence of new technologies, etc. Additionally, in specific cases, terms have been arbitrarily selected by the applicant, and in such cases, their meanings will be described in detail in the relevant description of the invention. Therefore, the terms used in this invention should be defined not merely by their names, but based on their meanings and the overall content of the invention.

[0027] Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as generally understood by those skilled in the art to which the present invention pertains. Terms such as those defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the relevant technology, and should not be interpreted in an ideal or overly formal sense unless explicitly defined in this application.

[0028] When a part of a specification is described as "including" a certain component, this means that, unless specifically stated otherwise, it does not exclude other components but may include additional components.

[0029] Numerical ranges include the values ​​defined in the above ranges. All maximum numerical limits given throughout this specification include all lower numerical limits as clearly written. All minimum numerical limits given throughout this specification include all higher numerical limits as clearly written. All numerical limits given throughout this specification will include all better numerical ranges within a wider numerical range, as clearly written.

[0030] The embodiments of the present invention are described in detail below, but it is obvious that the present invention is not limited by the following embodiments.

[0032] The present invention provides a composition for preventing, improving, or treating atopic dermatitis comprising an extract of Symphyocladia linearis as an active ingredient.

[0033] In the present invention, “prevention” means any act of suppressing or delaying the onset of a target disease, “improvement” means any act of reducing parameters related to the target disease, such as the degree of symptoms, by administering a composition according to the present invention, and “treatment” means any act of improving or beneficially changing the target disease and the associated metabolic abnormalities by administering a pharmaceutical composition according to the present invention.

[0034] The above composition may be characterized by increasing the expression of one or more genes selected from the group consisting of keratin 1 (KRT1), keratin 2 (KRT2), keratin 10 (KRT10), involucrine (IVL), loricrine (LOR), claudin 1 (CLDN1) and filaggrin (FLG), which are differentiation genes of skin keratinocytes, but is not limited thereto.

[0035] The above composition may restore the expression of filaggrin (FLG) that is inhibited by cytokines IL-4 and IL-13, which are increased in atopic dermatitis.

[0036] The expression of the above filaggrin (FLG) may be gene expression or protein expression.

[0037] In addition, the above composition may have an antioxidant effect.

[0039] The present invention may provide a pharmaceutical composition for the prevention, improvement, or treatment of atopic dermatitis comprising an extract of Symphyocladia linearis as an active ingredient.

[0040] The pharmaceutical composition according to the present invention may be formulated and used in the form of external preparations such as powders, granules, sustained-release granules, enteric granules, liquids, eye drops, oxylic agents, emulsions, suspensions, ethanol tablets, troches, fragrances, limonene adzes, tablets, sustained-release tablets, enteric tablets, sublingual tablets, hard capsules, soft capsules, sustained-release capsules, enteric capsules, pills, tinctures, soft extracts, dry extracts, fluid extracts, injections, capsules, irrigation solutions, warning agents, lotions, pastes, sprays, inhalants, patches, sterile injectable solutions, or aerosols, according to conventional methods, and the external preparations may have formulations such as creams, gels, patches, sprays, ointments, warning agents, lotions, liniments, pastes, or cataplasms.

[0042] The present invention may provide a cosmetic composition for preventing, improving, or treating atopic dermatitis comprising an extract of Symphyocladia linearis as an active ingredient.

[0043] The cosmetic composition according to the present invention may be formulated and used in the form of, for example, lotion, ointment, patch, foam, spray, powder, soap, bath additive, spray, nourishing lotion, nourishing essence, massage cream, beauty bath additive, body lotion, body milk, bath oil, baby oil, baby powder, shower gel, shower cream, sunscreen lotion, sunscreen cream, tanning cream, sunscreen cosmetic, cleansing milk, skin whitening cream, hand lotion, hair lotion, cosmetic cream, bath soap, liquid soap, beauty soap, shampoo, hand sanitizer, cream soap, facial wash, hair rinse, cosmetic soap, teeth whitening gel or toothpaste, etc., according to conventional methods, but is not limited thereto.

[0044] The above cosmetic composition may further include a suitable carrier, excipient, or diluent commonly used in the manufacture of cosmetic compositions. The carrier, excipient, or diluent may include, but is not limited to, purified water, oil, wax, fatty acid, fatty acid alcohol, fatty acid ester, surfactant, hygroscopic agent, thickener, antioxidant, viscosity stabilizer, chelating agent, buffer, lower alcohol, etc. Additionally, it may include, as necessary, a whitening agent, moisturizer, vitamin, sunscreen, perfume, dye, antibiotic, antibacterial agent, or antifungal agent.

[0046] The present invention can provide a food composition for preventing, improving, or treating atopic dermatitis comprising an extract of Symphyocladia linearis as an active ingredient.

[0047] The food composition according to the present invention may be formulated and used in the form of powder, granules, pills, tablets, capsules, candy, syrup, or beverage, but is not limited thereto.

[0049] Hereinafter, the present invention will be described in detail with reference to examples to aid in understanding. However, the following examples are merely illustrative of the content of the present invention and the scope of the present invention is not limited to the following examples. The examples of the present invention are provided to more completely explain the present invention to those with average knowledge in the art.

[0051] <Example 1> Preparation of Symphyocladia linearis extract

[0052] The extract of *Symphyocladia linearis* (MABIK NP30210009) was obtained through the Marine Extract Bank of the National Institute of Marine Biological Resources. The above extract was obtained by washing *Symphyocladia linearis* collected in Gangneung-si, Gangwon-do in 2018 three times and freeze-drying it, then placing 30g of the dried powder into 400mL of 70% ethanol and extracting it three times repeatedly at room temperature using an ultrasonic extractor (40KHz, 60 min), followed by filtration and concentrating. The sample thus extracted was obtained from the Marine Extract Bank, 1 ml of dimethylsulfoxide (DMSO) was added and dissolved, and only the supernatant was used for the experiment using a centrifuge.

[0054] <Example 2> Skin Keratinocyte Culture

[0055] Human normal primary epidermal keratinocytes (NHEK) were KGM SingleQuot containing bovine pituitary extract, hEGF, recombinant human insulin, hydrocortisone, gentamicin, and amphotericin-B in Keratinocyte Basal Medium (KBM). ™Supplements were mixed to form the culture medium, and the cells were cultured in a 37°C, 5% CO2 incubator (ARA P150, Hanil Science Co., Ltd., Korea) with the medium replaced every 2 days. When the cells grew to cover approximately 80% of the plate area, the existing culture medium was removed, the cells were washed once with HEPES Buffered Saline Solution, and Trypsin / EDTA (2 ml) was added and reacted in the incubator for 7 minutes to detach the attached cells. After inhibiting the action of trypsin with Trypsin Neutralizing Solution (TNS; 4 ml), the cells were centrifuged at 37°C and 3,000 rpm for 5 minutes and subcultured. In this invention, cells from passages 3 to 6 were used.

[0057] <Experimental Example 1> Confirmation of Antioxidant Efficacy of Symphyocladia linearis Extract

[0058] Experimental Example 1-1. DPPH assay

[0059] To investigate the antioxidant effects of Symphyocladia linearis extract, DPPH radical scavenging activity was verified using a DPPH assay. 200 μL of 0.2 mM DPPH solution was mixed with samples of Symphyocladia linearis extract according to Example 1 at different concentrations (0.5, 1, 5, 10, 50 μg / μl) and reacted at room temperature for 30 minutes. The absorbance was measured at 517 nm using a spectrophotometer (INNO-M, LTek, Korea). The absorbance of the experimental control group was measured after treatment with DMSO. The results of the absorbance measurements are shown as a graph in Figure 1. The DPPH radical scavenging activity was calculated using the following Equation 1 and expressed as IC50, which is the concentration that inhibits DPPH radicals by 50% for each sample. 50 obtained.

[0061] (Mathematical Formula 1)

[0062]

[0064] As shown in Figure 1, it was confirmed that the antioxidant capacity increased as the extract of Symphyocladia linearis of Example 1 was treated at different concentrations (N=5, p ≤ 0.05 (*), p ≤ 0.01 (**), p ≤ 0.001 (***)), and it was found that it exhibited an antioxidant capacity of over 70% at 50 μg / µl. This indicates that the extract of Symphyocladia linearis has the effect of alleviating atopy by removing reactive oxygen species that cause atopy.

[0066] Experimental Example 1-2. ABTS assay

[0067] To investigate the antioxidant effect of the Symphyocladia linearis extract, the extract prepared in Example 1 was tested using the ABTS assay. A 7 mM aqueous ABTS solution and a 2.45 mM aqueous potassium persulfate solution were mixed in a 1:1 ratio (v / v) and reacted for 12 to 16 hours to generate radical cations. ABTS +· The solution was diluted in 10% ethanol so that the absorbance value was 0.7 ± 0.05. Then, 200 μL of ABTS was added to the samples of the fine purple agar extract from Example 1 at different concentrations (0.5, 1, 5, 10, 50 μg / μl). +· After adding the solution, the mixture was reacted at room temperature for 5 minutes, and the absorbance was measured at 734 nm using a spectrophotometer. The absorbance of the control group was measured after treatment with DMSO. The ABTS radical scavenging activity of the samples at different concentrations of Symphyocladia linearis extract was calculated using the above mathematical formula (1), and the IC50, which is the concentration that inhibits ABTS radicals by 50% for each sample, was 50 obtained.

[0068] As shown in Figure 1, it was confirmed that the antioxidant capacity increased as the extract of Symphyocladia linearis of Example 1 was treated at different concentrations, and it showed an antioxidant capacity of up to 92% at 50 μg / µl (N = 5, p ≤ 0.05 (*), p ≤ 0.01 (**), p ≤ 0.001 (***)). This indicates that the extract of Symphyocladia linearis has the effect of alleviating atopy by removing reactive oxygen species that cause atopy.

[0070] <Experimental Example 2> Evaluation of Cytotoxicity of Symphyocladia linearis Extract

[0071] NHEK cells subcultured according to Example 2 were placed in a 96-well plate at a ratio of 1 × 10 4 Cells were dispensed into wells and cultured in a 37°C incubator for 24 hours. Afterward, the Symphyocladia linearis extract according to Example 1 was treated at different concentrations (0.5, 1, 5, 10, 50 μg / ml) and cultured in an incubator for 48 hours. The existing culture medium was removed, and 100 μL of a new culture medium and a CCK-8 solution to confirm cytotoxicity were mixed in a 10:1 ratio and dispensed. After reacting in an incubator for 3 hours, the absorbance was measured at 450 nm. Cells treated with DMSO were used as a control, and the cytotoxicity of each sample of the Symphyocladia linearis extract was calculated using the following mathematical formula (2) and is shown in Figure 2.

[0073] (Mathematical Formula 2)

[0074]

[0076] As shown in Figure 2, when Symphyocladia linearis extract was treated at different concentrations (0.5, 1, 5, 10, 50 μg / ml), approximately 30% cytotoxicity was observed at 50 μg / ml, but no cytotoxicity was observed at the other concentrations of 0.5 to 10 μg / ml (N=3, p ≤ 0.05 (*)).

[0078] <Experimental Example 3> Changes in Differentiation Gene Expression of Skin Keratinocytes by Extract of Symphyocladia linearis

[0079] NHEK cells subcultured according to Example 2 were placed in a 12-well plate at a ratio of 6 × 10 4Cells were seeded into wells, and after 24 hours, 10 μg / mL of Symphyocladia linearis extract according to Example 1 was added and cultured for 72 hours to confirm the appearance of differentiated cells. Subsequently, the existing culture medium was removed, and after washing once with Phosphate Buffered Saline (PBS, GenDepot, TX, USA), RNA was isolated using the Quick-RNA Micro Prep Kit (Zymo Research, CA, USA). The concentration and purity of the isolated RNA were measured using a nano-spectrophotometer (Genova Nano Micro-Spectrophotometer, Jenway, London, UK), and a total of 1 to 2 μg of RNA was synthesized into cDNA following the instructions of the High-capacity cDNA Kit (ThermoFisher Scientific, MA, USA). The synthesized cDNA was subjected to quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) on a QuantStudio-3 instrument (Applied Biosystems, Carlsbad, CA) according to the manufacturer's protocol using TaqMan™ Fast Advanced Master Mix (ThermoFisher Scientific) and Taqman Probe (ThermoFisher Scientific).The genes used in the present invention were KRT1 (Hs00196158_m1), KRT2 (Hs00166294_m1), KRT10 (Hs00166289_m1), IVL (Hs00846307_s1), LOR (Hs01894962_s1), FLG (Hs00856927_g1), and CLDN1 (Hs00221623_m1), and were normalized using ribosomal protein lateral stalk subunit P0 (RPLP0, Hs00420895) as a loading control, and qRT-PCR was performed a total of 40 times with 5 minutes at 50 ℃, 20 seconds at 95 ℃, 3 seconds at 95 ℃, and 20 seconds at 60 ℃ set as one cycle. In addition, for quantitative analysis, target genes were quantified using the ddCt method and are shown in Figure 3.

[0080] As a result of confirming the gene expression of keratinocyte differentiation factors (keratin (KRT)1, KRT2, KRT10, filaggrin (FLG), involucrin (IVL), loricrin (LOR), cloudin1 (CLDN1)) in Fig. 3, it was found that their gene expression was sparse purple agar (Symphyocladia linearis) extract (10 μ It was confirmed that levels increased from approximately 1.8 to a maximum of 5 times (N = 4, p ≤ 0.05 (*), p ≤ 0.01 (**), p ≤ 0.001 (***)). In particular, FLG, LOR, and IVL, which are important for the skin barrier, increased by 4, 2.9, and 3.3 times, respectively, confirming that the extract of *Symphyocladia linearis* increases gene expression essential for maintaining skin barrier function.

[0082] <Experimental Example 4> Protein expression of FLG induced by Symphyocladia linearis extract in an in vitro atopic dermatitis model

[0083] In order to investigate the recovery of FLG protein expression caused by the overexpression of cytokines IL-4 and IL-13, which are involved in the pathophysiological characteristics of atopic dermatitis, using the extract of Symphyocladia linearis from Example 1 in an atopic dermatitis environment, the following was performed using the Western blotting method.

[0084] NHEK cells subcultured according to Example 2 were placed in a 6-well plate at a volume of 1.5 × 10 5Cells were seeded into wells and cultured for 24 hours. Then, 10 μg / mL of the Symphyocladia linearis extract from Example 1 and 10 ng / mL of IL-4 and IL-13 were added simultaneously, and the cells were cultured for 96 hours. The existing culture medium was removed, and after washing once with PBS, the cells were harvested using a scraper. RIPA lysis buffer (ThermoFisher Scientific) containing a protease inhibitor (GenDepot) and a phosphatase inhibitor (GenDepot) was added, and the proteins were lysed using an ultrasonic extractor (Q125, Qsonica, CT, USA). The separated lysate was centrifuged at 15,000 rpm at 4°C for 10 minutes, and only the supernatant was separated. Protein was quantified using the BCA Protein Assay Kit (ThermoFisher Scientific), and 20 μg of protein was mixed with Sample Reducing Agent, LDS Sample Buffer, and distilled water to denature the protein at 95°C for 10 minutes. Electrophoresis was performed on an 8% SDS-polyacrylamide gel at 120 V, 200 mA, and 40 W. The separated protein was transferred to a polyvinylidene difluoride (PVDF) membrane (Cytiva, Korea) at 30 V, 300 mA, and 40 W for 1 hour. Non-Fat Dried Milk (NFDM, Cellconic, USA) was dissolved in TBS-T, prepared by adding 0.1% Tween 20 (SAMCHUN, Korea) to Tris Buffered Saline (TBS, GenDepot), to make 5% NFDM, which was then used. After the transfer was complete, the membrane was blocked in 5% NFDM for 1 hour and washed three times with TBS-T for 5 minutes each.Subsequently, the samples were reacted with primary antibodies (anti-β-actin, 1:2000, Santa Cruz, TX, USA; anti-Filaggrin, 1:2000, Santa Cruz) overnight at 4°C. Unattached primary antibodies were removed by washing three times with TBST for 10 minutes, and the samples were reacted with secondary antibodies (goat anti-mouse IgG (H+L), 1:5000, Invitrogen, CA, USA) for 1 hour. After washing three times with TBST for 10 minutes, protein expression was confirmed using Femto ECL chemiluminescent substrate (Biomax, Korea) and an iBright CL750 instrument (Invitrogen). Protein expression levels were quantified using iBright Analysis Software and then analyzed for β-actin, as shown in Figure 4.

[0085] As shown in Figure 4, in an in vitro atopic dermatitis model, FLG expression was increased in the experimental group treated with Symphyocladia linearis extract compared to the experimental control KGM (undifferentiated keratinocytes) and KBM (differentiated keratinocytes). It was also confirmed that FLG expression was inhibited by IL4 and IL13 but restored by the Symphyocladia linearis extract. This indicates that FLG expression is induced for skin barrier restoration (p ≤ 0.05 (*), p ≤ 0.01 (**), p ≤ 0.001 (***)).

[0086] According to the results of Figures 1 to 4, it can be seen that the extract of *Symphyocladia linearis* exhibits excellent antioxidant efficacy, has low cytotoxicity against skin keratinocytes, has excellent skin keratinocyte differentiation ability, and is effective in restoring the skin barrier. Based on these biological activities, it is expected that it can be utilized as a material for treating atopic dermatitis.

[0088] As specific parts of the present invention have been described in detail above, it is evident to those skilled in the art that such specific descriptions are merely preferred embodiments and do not limit the scope of the invention. Accordingly, the actual scope of the invention shall be defined by the appended claims and their equivalents. The scope of the invention is defined by the claims set forth below, and all modifications or variations derived from the meaning and scope of the claims and their equivalents shall be interpreted as being included within the scope of the invention.

Claims

Claim 1 A composition for preventing, improving, or treating atopic dermatitis comprising an extract of *Symphyocladia linearis* as an active ingredient, wherein the composition increases the expression of one or more genes selected from the group consisting of keratin 1 (KRT1), keratin 2 (KRT2), keratin 10 (KRT10), involucrin (IVL), loricrin (LOR), claudin 1 (CLDN1), and filaggrin (FLG), which are differentiation genes of skin keratinocytes, wherein the composition restores the expression of filaggrin (FLG) that is suppressed by cytokines IL-4 and IL-13, which are increased due to atopic dermatitis, and wherein the composition has an antioxidant effect. Claim 2 delete Claim 3 delete Claim 4 delete Claim 5 A pharmaceutical composition for the prevention, improvement, or treatment of atopic dermatitis comprising an extract of *Symphyocladia linearis* as an active ingredient.

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