KR20210116234A - topical composition comprising the extract of combined herbs comprising Longanae Arillus for TSLP inhibition and for the treatment or alleviation of skin inflammatory disease - Google Patents

Abstract

The present invention relates to a composition for inhibiting TLSP or for preventing and/or alleviating and/or treating inflammatory skin diseases and a skin itch, including a combined herbal medicine extract containing Longanae arillus, Angelica tenuissima Nakai. and Polygala tenuifolia Willd., as an active ingredient. Particularly, after carrying out various tests, such as a test for determining the effect of treating atopic dermatitis in vivo using BALB/C 6 week-aged female mice (test example 1), a test for determining the effect of inhibiting expression of various cytokines, including GM-CSF, IL-4, IL-10, IL-13, IL-31 and IL-33 (test example 2), and a test for determining the effect of inhibiting expression of TSLP cytokines (test example 3), for the combined herbal medicine extract, the samples are shown to realize a strong skin anti-inflammatory activity, or the like. Therefore, the composition can be used for an anti-inflammatory external skin application pharmaceutical composition or cosmetic composition.

Description

A topical composition comprising the extract of combined herbs comprising Longanae Arillus for TSLP inhibition and for the treatment or alleviation of skin inflammatory disease, containing as an active ingredient a combination herbal extract containing longan meat

The present invention relates to an external composition for inhibiting TSLP and for treating and improving skin inflammatory diseases, which contains a combination herbal extract containing longan meat as an active ingredient.

[Document 1] Mack et al., 2018 Trends Immunol. 2018 Dec; 39(12): 980-991. doi:10.1016/j.it. 2018.10.001. E pub 2018 Nov21.

[Document 2] Willoughby DA. (1975) Human arthritis applied to animal models. Towards a better therapy. Annals of the Rheumatic Disease. 34, 471-478

[Document 3] Yun HJ, Heo SK, Yi HS, Kim CH, Kim BW and Park SD. (2008) Anti-inflammatory effect of injinho-tang in Raw264.7 Cells. Kor. J. Herbology. 23(2), 169-178.

[Document 4] Bo Bo-seop, Shin Min-kyo, Dohaehyangyaksa Dictionary, Youngrimsa, pp371-373, 1998

[Document 5] One Information Recruiter, Younglim Publishing House, p428-429, 1998

[Document 6] One Information Recruiter, Younglim Publishing House, p798-799, 1998

[Document 7] Li et al., 2016 Drug Des Devel Ther. 2016 Feb 19;10:781-91. doi: 10.2147/DDDT.S94056. eCollection 2016.

[Document 8] Jean et al., 2019, J Invest Dermatol. 2019 May;139(5):1098-1109. doi: 10.1016/j.jid.2018.11.012. Epub 2018 Nov 29

Chronic recurrent skin inflammatory diseases such as atopic dermatitis and psoriasis are very difficult to treat because their causes are very diverse.

In particular, in the case of some patients with atopic dermatitis, mutations in the filaggrin gene cause damage to the skin barrier, and serious inflammatory reactions are induced by bacterial infection or infection with foreign substances derived from ticks. In addition to these genetic factors, a decrease in water retention due to a decrease in ceramide in the stratum corneum, abnormal loss of keratinocytes due to an increase in the pH of the skin surface, etc. can be factors that determine the severity of skin diseases along with damage to the skin barrier function. In addition, most of the onset of these skin diseases are concentrated in infancy and adolescence, resulting in emotional, educational and social problems. In particular, there is an increase in scratching due to unbearable itching (pruritus of the skin) accompanying these skin diseases, which further exacerbates the symptoms. Recently, the expression of various cytokines such as interleukin and thymic stromal lymphopoietin (TSLP) in the keratinocytes of the skin is increased, and it stimulates the sensory nerves present in the subcutaneous tissue, causing itching as well as dermatitis. (Mack et al., 2018 Trends Immunol. 2018 Dec; 39(12): 980-991. doi:10.1016/j.it. 2018.10.001. E pub 2018 Nov21.)

The inflammatory response is a mechanism to repair and regenerate the damaged area when an invasion that causes any organic change such as physical action, chemical substances, or bacterial infection is applied to a living body or tissue. Once a stimulus is applied, vasoactive substances such as inflammatory components are locally released and vascular permeability is increased, which causes inflammation. [Willoughby DA. (1975) Human arthritis applied to animal models. Towards a better therapy. Annals of the Rheumatic Disease. 34, 471-478.].

Tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), which are proinflammatory cytokines as inflammatory factors [Yun HJ, Heo SK, Yi HS, Kim CH, Kim BW and Park SD. (2008) Anti-inflammatory effect of injinho-tang in Raw264.7 Cells. Kor. J. Herbology. 23(2), 169-178.].

In general, antihistamines, steroids (cortisol, prednisolone, methylprednisolone, dexamethasone injections and ointments, etc.), moisturizers, etc. are used as therapeutic agents for atopic dermatitis, but they are not effective. In particular, in the case of steroids, capillaries are enlarged and the skin layer becomes thinner, resulting in more severe hypersensitivity reactions.

Therefore, there is a need for the development of an effective therapeutic agent for inflammatory skin diseases such as atopic dermatitis and few side effects.

Longan meat (Dimocarpus longan Loureiro) is the seed coat of Euphoria longan and plants belonging to the Sapindaceae family. Yes (Bobo-Sup, Shin Min-Kyo, Dohae-Hyang Pharmacy Dictionary, Youngrimsa, pp371-373, 1998).

Gobon (Ligusticum tenuissimum KITAG.) is a perennial herb belonging to the Umbelliferae family, and its rhizome and roots are called Gobon. It contains, and is known to have antifungal effects, etc. (Information Recruitment 1 person, Younglim Publishing House, p428-429, 1998).

Polygala tenuifolia WILLD. is a perennial herb belonging to the family Polygalaceae, and its root is called Polygalae Radix. It is known that there is (1 person in contact with information, Younglim Publishing House, p798-799, 1998).

However, none of the above documents discloses or teaches a therapeutic effect on TSLP inhibition or skin inflammatory diseases containing a combination extract composed of longan meat, gobon and raw paper as an active ingredient.

Accordingly, the present inventors conducted a cytokine expression inhibition effect test using human skin epithelial cells (HaCaT) for the combined herbal extract (Experimental Example 1), atopic dermatitis in vivo effect test using BALB/C 6-week-old female mice (Experimental Example) 2), GM-CSF, IL-4, IL-10, IL-13, IL-31, and various cytokine expression inhibitory effect tests such as IL-33 (Experimental Example 3), TSLP cytokine expression inhibitory effect test ( Experimental Example 4) and the like were conducted, and the present invention was completed by confirming that the samples showed strong skin anti-inflammatory activity, etc.

In order to achieve the above object, the present invention provides a pharmaceutical composition for external use on the skin for the treatment and prevention of TSLP inhibition or skin inflammatory disease, which contains a combination herbal extract composed of longan meat, gobon and raw paper as an active ingredient.

In addition, the present invention is a TLSP inhibitor or Provided is a cosmetic composition for preventing and improving skin inflammatory diseases.

The combination herbal extract as defined herein preferably has a weight mixing ratio (w/w) of longan meat, gobon, and raw paper of 0.01 - 100 : 0.01 - 100 : 0.01 - 100 parts by weight (w/w), more preferably 0.5 -50: 0.5-50: 0.5-50 parts by weight (w/w), even more preferably 0.1-10: 0.1-10: 0.1-10 parts by weight (w/w), even more preferably 1-5 : 1-5: 1-5 parts by weight (w/w), even more preferably 1-3: 1-3: 1-3 parts by weight (w/w) .

In addition, the present invention provides a thymic stromal lymphopoietin (TSLP) cytokine expression inhibitor comprising a combination herbal extract composed of longan meat, gobon and raw paper as an active ingredient.

Hereinafter, the present invention will be described in detail. A combination herbal extract can be obtained through a conventional extraction process as follows.

Yonganyuk drying the yonganyuk, gobon and sheet herbal material obtained in the above step, the weight ratio of gobon and the base paper (w / w) of 0.01 - 100 0.01 - 100 0.01 - 100 parts by weight (w / w), more preferably is 0.5-50: 0.5-50: 0.5-50 parts by weight (w/w), even more preferably 0.1-10: 0.1-10: 0.1-10 parts by weight (w/w), even more preferably 1 -5: 1-5: 1-5 parts by weight (w/w), even more preferably 1-3: 1-3: 1-3 parts by weight (w/w) step; The formulation is washed and 1 to 20 times (w/v) weight, preferably 1 to 10 times (v/w) volume of the sample of the formulation is water, ethanol, methanol, propanol, butanol, acetone, ethyl acetate, hexane , butylene glycol, propylene glycol, hydrous butylene glycol, hydrous propylene glycol, at least one solvent selected from the group consisting of hydrous glycerin, preferably water or a mixed solvent of water and ethanol, most preferably 10% to 80% ethanol An agent for obtaining an extract by performing hot water extraction, cold extraction or ultrasonic extraction, preferably hot water extraction, at 50 to 120° C., preferably about 80-100° C. for 1 hour to 24 hours, preferably 2 hours to 12 hours Step 2; a third step of recovering the extract obtained by repeating the extraction process of the second step 2 to 10 times, preferably 3 to 5 times, and filtering it with a filter paper to obtain a filtrate; The combined herbal extract of the present invention can be prepared through the fourth step process of freeze-drying the filtrate, drying at room temperature or drying with hot air, preferably freeze-drying to obtain a combined herbal extract in a dry state.

The present invention provides a pharmaceutical composition for external use on the skin and a cosmetic composition having an anti-inflammatory effect, comprising the preparation method and the combined herbal extract obtained by the preparation method as an active ingredient.

The term "extract" as defined herein includes extracts of the root, stem, and leaf parts of the longan meat, gobon, and base branches, specifically, water, ethanol, methanol, propanol, butanol, acetone of the extract material. , ethyl acetate, hexane, butylene glycol, propylene glycol, hydrous butylene glycol, hydrous propylene glycol, at least one solvent selected from the group consisting of hydrous glycerin, preferably water or a mixed solvent of water and ethanol, most preferably 10% to 80% ethanol soluble extract.

The combined extract of longan meat, gobon, and raw paper is

(1) an extract of a combination of longan meat, gobon, and raw paper, or

(2) a mixture of individual longan meat extracts, gobon extracts, and raw paper extracts, or

(3) including combinations of these.

In one embodiment, the mixed extract of longanyuk, gobon, and raw paper may be an extract of a mixture of longanyuk, gobon, and raw paper.

In the present specification, "extract" means not only extracts obtained by adding an extraction solvent to herbal medicines, but also concentrates and dilutions of the extracts, and dried products of the extracts, concentrates, and dilutions. Unless otherwise specified, "extract" means at least one selected from the group consisting of an extract obtained by adding an extraction solvent to a crude drug, a dried product of the extract, a concentrate or dilution of the extract, and a dried product of the concentrate or dilution. can be interpreted as meaning

Skin inflammatory diseases as defined herein include chronic recurrent dermatitis, contact dermatitis, seborrheic dermatitis, neurodermatitis, dry skin, erythema, inflammatory dermatitis, psoriasis, wounds, such as pruritus due to aging or atopic dermatitis, atopic dermatitis or psoriasis. , atopic diseases, preferably chronic recurrent dermatitis such as pruritus, atopic dermatitis or psoriasis, contact dermatitis, seborrheic dermatitis, neurodermatitis , dry skin, Psoriasis, wounds, atopic disease, or inflammatory dermatitis, more preferably, chronic recurrent dermatitis such as atopic dermatitis or psoriasis, contact dermatitis, pruritus due to aging or atopic dermatitis due to increased expression of intracellular TLSP cytokine , seborrheic dermatitis, neurodermatitis, xerodermatitis, psoriasis, wounds, atopic disease, or inflammatory dermatitis.

As used herein, 'anti-inflammatory' refers to any mechanism that suppresses inflammation. Inflammation means that pathogens invade damaged skin or mucous membranes and exhibit symptoms such as fever, swelling, and pain, and the non-specific immune action showing these symptoms is called an inflammatory response. When macrophages in damaged tissue secrete cytokines, T lymphocytes are activated, and when mast cells, a type of white blood cell, secrete histamine, they initiate internal defense, and inflammation is induced in infected tissues. Therefore, the level of intracellular cytokine expression can be an indicator of inflammatory response activation (anti-inflammatory activity in another aspect). For example, the anti-inflammatory activity described herein may be an inhibitory activity against inflammation occurring in the skin.

Cytokine (cytokines) is a general term for immune substances secreted during the immune system is activated when pathogens penetrate the living body. Cytokines are secreted when pathogens such as viruses penetrate and stimulate other immune cells to block pathogens. Cytokines are usually secreted only in the early stages of infection, but may continue to be secreted if the immune system is overactive. When high concentrations of cytokines are secreted for a long period of time (eg, more than a week) (cytokine storm), the immune cells are overcrowded to the site of infection, causing inflammation and loosening of blood vessels, which can endanger the life of the infected person. have. The cytokine expression inhibitory activity provided herein may be interpreted as meaning including prevention, treatment and/or improvement of cytokine storm.

In one example, the cytokine may be one or more selected from cytokines related to dermatitis, for example, atopic dermatitis. For example, the cytokine is TSLP (thymic stromal lymphopoietin), CSF (colony stimulating factor; for example, GM (Granulocyte-macrophage)-CSF, M (Macrophage)-CSF, G (Granulocyte)-CSF, etc.), interleukin ( Interleukins; for example, IL-4, IL-10, IL-13, IL-31, IL-33, etc.) may be one or more selected from the group consisting of, but is not limited thereto.

The extract is characterized in that it contains 0.1 to 50% by weight based on the total weight of the pharmaceutical composition for external use.

The pharmaceutical composition for external use is a cream, gel, patch, spray, emulsion, ointment, warning agent, lotion, liniment agent, pasta agent, solvent, suspension, pack, patch or cataplasma. cataplasma formulations.

In addition, the cosmetic composition is an edible cosmetic for oral administration such as lotions, skins, lotions, nutritional lotions, nutritional creams, massage creams, essences, packs, soaps, liquid detergents, gels, patches, etc., tablets, capsules, powders, etc. including formulations.

The present inventors conducted a cytokine expression inhibition effect test using human skin epithelial cells (HaCaT) for the combined herbal extract of the present invention (Experimental Example 1), atopic dermatitis in vivo effect test using BALB/C 6-week-old female mice (Experimental Example) 2), GM-CSF, IL-4, IL-10, IL-13, IL-31, and various cytokine expression inhibitory effect tests such as IL-33 (Experimental Example 3), TSLP cytokine expression inhibitory effect test ( Experimental Example 4) and the like were conducted, and it was confirmed that the samples exhibited strong anti-inflammatory activity on the skin, and the composition was useful as an external pharmaceutical composition or cosmetic composition for anti-inflammatory use.

In addition, the above herbal medicines have been used for a long time as herbal medicines and food, and the extracts of the present invention extracted from them also do not have problems such as toxicity and side effects, and since it has been proven that they are non-irritating samples in a skin patch test, they can be safely used even for long-term use. can

The pharmaceutical composition for external use on the skin containing the extract of the present invention can be prepared and used as a pharmaceutical composition for external use on the skin of a cream, gel, patch, spray, ointment, warning agent, lotion, liniment, pasta, or cataplasma. However, the present invention is not limited thereto.

The preferred dosage of the extract of the present invention varies depending on the condition and weight of the patient, the degree of disease, the drug form, the route and duration of administration, but may be appropriately selected by those skilled in the art. However, for the desired effect, the extract of the present invention is preferably administered at 0.0001 to 100 mg/kg per day, preferably 0.001 to 10 mg/kg. Administration may be administered once a day, or may be administered in several divided doses. The above dosage does not limit the scope of the present invention in any way.

The extract of the present invention can be used in various ways, such as cosmetics and face washes having an anti-inflammatory effect.

Products to which the present composition can be added include, for example, cosmetics such as lotions, skin lotions, nutritional lotions, nourishing creams, massage creams, essences, packs, and cleansing, face wash, soap, treatment, and cosmetic products. have.

The cosmetic of the present invention includes a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, high-molecular peptides, high-molecular polysaccharides, sphingolipids and seaweed extract.

The water-soluble vitamin may be any as long as it can be formulated in cosmetics, but preferably vitamin B1, vitamin B2, vitamin B6, pyridoxine, pyridoxine hydrochloride, vitamin B12, pantothenic acid, nicotinic acid, nicotinic acid amide, folic acid, vitamin C, vitamin H, etc. and their salts (thiamine hydrochloride, sodium ascorbate salt, etc.) and derivatives (ascorbic acid-2-phosphate sodium salt, ascorbic acid-2-phosphate magnesium salt, etc.) are also included in the water-soluble vitamins that can be used in the present invention. Included. Water-soluble vitamins can be obtained by conventional methods such as a microbial transformation method, a purification method from a microbial culture, an enzymatic method or a chemical synthesis method.

The oil-soluble vitamins may be any as long as they can be formulated in cosmetics, but preferably include vitamin A, carotene, vitamin D2, vitamin D3, vitamin E (d1-alpha tocopherol, d-alpha tocopherol, d-alpha tocopherol). , their derivatives (ascorbine palmitate, ascorbine stearate, ascorbine dipalmitate, dl-alpha tocopherol acetate, dl-alpha tocopherol nicotinic acid, vitamin E, dl-pantothenyl alcohol, D-pantothenyl alcohol, pantothenyl ethyl ethers, etc.) are also included in the oil-soluble vitamins used in the present invention. Oil-soluble vitamins can be obtained by conventional methods such as a microbial transformation method, a purification method from a microbial culture, an enzyme or a chemical synthesis method, and the like.

The macromolecular peptide may be any compound as long as it can be formulated in cosmetics, and preferred examples include collagen, hydrolyzed collagen, gelatin, elastin, hydrolyzed elastin, and keratin. Polymeric peptides can be purified and obtained by conventional methods such as purification from a microbial culture solution, enzyme method or chemical synthesis method, or can be purified and used from natural products such as dermis of pigs or cattle, silk fibers of silkworms, etc.

The high molecular weight polysaccharide may be any compound as long as it can be formulated in cosmetics, but preferably hydroxyethyl cellulose, xanthan gum, sodium hyaluronate, chondroitin sulfate, or a salt thereof (such as sodium salt). For example, chondroitin sulfate or a salt thereof can be purified and used from mammals or fish.

The sphingolipids may be any as long as they can be formulated in cosmetics, and ceramides, phytosphingosine, sphingoglycolipids and the like are preferred. The sphingolipid can be purified by a conventional method from mammals, fish, shellfish, yeast or plants, or can be obtained by chemical synthesis.

The seaweed extract may be any type as long as it can be formulated in cosmetics, but preferably brown algae extract, red algae extract, green algae extract, etc., and calrageenan purified from these seaweed extracts, arginic acid, sodium alginate, Potassium alginate and the like are also included in the seaweed extract used in the present invention. The seaweed extract can be obtained by purification from seaweed by a conventional method.

In the cosmetic of the present invention, in addition to the above essential ingredients, if necessary, other ingredients usually blended in cosmetics may be blended.

Other ingredients that may be added include oils and fats, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, disinfectants, antioxidants, plant extracts, pH adjusters, alcohols, colorants, fragrances, A blood circulation promoter, a cooling agent, a restrictive agent, purified water, etc. are mentioned.

Examples of the fat and oil component include ester fats and oils, hydrocarbon oils and fats, silicone oils and fats, fluorine oils and fats, animal oils and fats, and vegetable oils and fats.

Examples of ester-based oils and fats include glyceryl tri-ethylhexanoate, cetyl 2-ethylhexanoate, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isocetyl isostearate, stearic acid. Butyl, ethyl linoleate, isopropyl linoleate, ethyl oleate, isocetyl myristate, isostearyl myristate, isostearyl palmitate, octyldodecyl myristate, isocetyl isostearate, diethyl sebacate, adipine Diisopropyl acid, isoalkyl neopentanoate, tri (caprylic acid, capric acid) glyceryl, trimethylol propane tri-2-ethylhexanoate, trimethylol propane triisostearate, pentaelysitol tetra2-ethylhexanoate , cetyl caprylate, decyl laurate, hexyl laurate, decyl myristate, myristyl myristate, cetyl myristate, stearyl stearate, decyl oleate, cetyl ricinoleate, isostea laurate Reel, isotridecyl myristate, isocetyl palmitate, octyl stearate, isocetyl stearate, isodecyl oleate, octyldodecyl oleate, octyldodecyl linoleate, isostearate isopropyl, 2-ethylhexanoate cetoster Aryl, 2-ethyl stearyl hexanoate, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate, propylene glycol dicapric acid, di(caprylic acid, capric acid) propylene glycol, dicaprylic acid propylene glycol, dicaprylate Neopentyl glycol phosphate, neopentyl glycol dioctanoate, glyceryl tricaprylate, glyceryl triundecylate, glyceryl triisopalmitate, glyceryl triisostearate, octyldodecyl neopentanoate, isostearyl octanoate , octyl isononanoate, hexyldecyl neodecanoate, octyldodecyl neodecanoate, isocetyl isostearate, isostearyl isostearate, octyldecyl isostearate, polyglycerol oleate, polyglycerol isostearate, Triisocetyl citrate, triisoalkyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactate, cetyl lactate, octyldecyl lactate, triethyl citrate, acetyl triethyl citrate, acetyl tributyl citrate, trioctyl citrate, dimalate Isostearyl, 2-ethylhexyl hydroxystearate, di2-ethylhexyl succinate, diisobutyl adipate, diisopropyl sebacate, dioctyl sebacate, Cholesteryl stearate, cholesteryl isostearate, cholesteryl hydroxystearate, cholesteryl oleate, dihydrocholesteryl oleate, phytsteryl isostearate, phytsteryl oleate, 12-stealoyl Ester types, such as isocetyl hydroxy stearate, 12-stealoyl hydroxy stearyl stearate, and 12-stealoyl hydroxy stearate isostearyl, etc. are mentioned.

Examples of hydrocarbon-based oils and fats include hydrocarbon-based oils and fats such as squalene, liquid paraffin, alpha-olefin oligomer, isoparaffin, ceresin, paraffin, liquid isoparaffin, polybudene, microcrystalline wax, petrolatum.

Examples of silicone oils and fats include polymethyl silicone, methylphenyl silicone, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, dimethylsiloxane/methylcetyloxysiloxane copolymer, dimethylsiloxane/methylstealloxysiloxane copolymer, and alkyl Modified silicone oil, amino modified silicone oil, etc. are mentioned.

Perfluoropolyether etc. are mentioned as fluorine-type fats and oils.

Animal or vegetable oils include avocado oil, almond oil, olive oil, sesame oil, rice bran oil, saffron oil, soybean oil, corn oil, rapeseed oil, passerine oil, palm kernel oil, palm oil, castor oil, sunflower oil, grape seed oil , cottonseed oil, palm oil, kukui nut oil, wheat germ oil, rice germ oil, shea butter, cranberry colostrum, marcadeumia nut oil, meadow home oil, egg yolk oil, tallow oil, horse oil, mink oil, orange rapie oil, jojoba oil , animal or plant oils and fats, such as candelabra wax, carnauba wax, liquid lanolin, and hydrogenated castor oil.

Examples of the moisturizing agent include a water-soluble low-molecular moisturizer, a fat-soluble molecular moisturizer, a water-soluble polymer, and a fat-soluble polymer.

As water-soluble low molecular weight moisturizers, serine, glutamine, sorbitol, mannitol, pyrrolidone-sodium carboxylate, glycerin, propylene glycol, 1,3-butylene glycol, ethylene glycol, polyethylene glycol B (polymerization degree n = 2 or more), polypropylene Glycol (polymerization degree n=2 or more), polyglycerol B (polymerization degree n=2 or more), lactic acid, lactic acid salt, etc. are mentioned.

Cholesterol, cholesterol ester, etc. are mentioned as a fat-soluble low molecular weight humectant.

Examples of the water-soluble polymer include carboxyvinyl polymer, polyaspartate, tragacanth, xanthan gum, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, carboxymethylcellulose, water-soluble chitin, chitosan, and dextrin. can

Examples of the fat-soluble polymer include polyvinylpyrrolidone/eicocene copolymer, polyvinylpyrrolidone/hexadecene copolymer, nitrocellulose, dextrin fatty acid ester, and high molecular weight silicone.

Examples of the emollient include long-chain acylglutamic acid cholesteryl ester, hydroxystearic acid cholesteryl, 12-hydroxystearic acid, stearic acid, rosin acid, and lanolin fatty acid cholesteryl ester.

Nonionic surfactants, anionic surfactants, cationic surfactants, amphoteric surfactants, etc. are mentioned as surfactant.

Nonionic surfactants include self-emulsifying glycerin monostearate, propylene glycol fatty acid ester, glycerin fatty acid ester, polyglycerol fatty acid ester, sorbitan fatty acid ester, POE (polyoxyethylene) sorbitan fatty acid ester, POE sorbitan fatty acid ester, POE Glycerin fatty acid ester, POE alkyl ether, POE fatty acid ester, POE hydrogenated castor oil, POE castor oil, POE/POP (polyoxyethylene/polyoxypropylene) copolymer, POE/POP alkyl ether, polyether-modified silicone, lauric acid alkanolamides, alkylamine oxides, hydrogenated soybean phospholipids, and the like.

Examples of the anionic surfactant include fatty acid soap, alpha-acyl sulfonate, alkyl sulfonate, alkyl allyl sulfonate, alkyl naphthalene sulfonate, alkyl sulfate, POE alkyl ether sulfate, alkyl amide sulfate, alkyl phosphate, POE alkyl phosphate, alkyl amide. Phosphate, alkyloylalkyl taurine salt, N-acylamino acid salt, POE alkyl ether carboxylate, alkylsulfosuccinate, alkylsulfoacetate sodium, acylated hydrolyzed collagen peptide salt, perfluoroalkyl phosphate ester, etc. have.

Examples of the cationic surfactant include alkyltrimethylammonium chloride, stearyltrimethylammonium chloride, stearyltrimethylammonium bromide, cetostearyltrimethylammonium chloride, distearyldimethylammonium chloride, stearyldimethylbenzylammonium chloride, behenyltrimethylammonium bromide, chloride Benzalkonium, diethylaminoethylamide stearate, dimethylaminopropylamide stearate, quaternary ammonium salt of lanolin derivative, etc. are mentioned.

Examples of the amphoteric surfactant include carboxybetaine type, amidebetaine type, sulfobetaine type, hydroxysulfobetaine type, amidesulfobetaine type, phosphobetaine type, aminocarboxylate type, imidazoline derivative type, amidamine type, etc. Amphoteric surfactant etc. are mentioned.

Examples of organic and inorganic pigments include silicic acid, silicic anhydride, magnesium silicate, talc, sericite, mica, kaolin, bengala, clay, bentonite, titanium-coated mica, bismuth oxychloride, zirconium oxide, magnesium oxide, zinc oxide, titanium oxide, aluminum oxide. inorganic pigments such as calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, iron oxide, ultramarine blue, chromium oxide, chromium hydroxide, calamine, and complexes thereof; Polyamide, polyester, polypropylene, polystyrene, polyurethane, vinyl resin, urea resin, phenol resin, fluororesin, silicon resin, acrylic resin, melamine resin, epoxy resin, polycarbonate resin, divinylbenzene/styrene copolymer, and organic pigments such as silk powder, cellulose, CI pigment yellow and CI pigment orange, and composite pigments of these inorganic pigments and organic pigments.

Examples of the organic powder include metal soaps such as calcium stearate; alkyl phosphate metal salts such as sodium cetylate, zinc laurylate, and calcium laurylate; acylamino acid polyvalent metal salts such as calcium N-lauroyl-beta-alanine, zinc N-lauroyl-beta-alanine, and calcium N-lauroyl glycine; amidesulfonic acid polyvalent metal salts such as calcium N-lauroyl-taurine and calcium N-palmitoyl-taurine; N such as N-epsilon-lauroyl-L-lysine, N-epsilon-palmitoylizine, N-alpha-paritoylolnithine, N-alpha-lauroylarginine, N-alpha-hydrogenated beef fatty acid acylarginine, etc. -Acyl basic amino acid; N-acyl polypeptides, such as N-lauroyl glycyl glycine; alpha-amino fatty acids such as alpha-aminocaprylic acid and alpha-aminolauric acid; Polyethylene, polypropylene, nylon, polymethyl methacrylate, polystyrene, a divinylbenzene-styrene copolymer, ethylene tetrafluoride, etc. are mentioned.

UV absorbers include para-aminobenzoic acid, ethyl para-aminobenzoate, amyl para-aminobenzoate, octyl para-aminobenzoate, ethylene glycol salicylate, phenyl salicylate, octyl salicylate, benzyl salicylate, butylphenyl salicylate, homomentyl salicylate, benzyl cinnamate. , para-methoxycinnamic acid-2-ethoxyethyl, para-methoxycinnamic acid octyl, dipara-methoxycinnamic acid mono-2-ethylhexane glyceryl, para-methoxycinnamic acid isopropyl, diisopropyl diisopropyl cinnamic acid ester mixture, uro Cannic acid, ethyl urokanate, hydroxymethoxybenzophenone, hydroxymethoxybenzophenonesulfonic acid and salts thereof, dihydroxymethoxybenzophenone, sodium dihydroxymethoxybenzophenonedisulfonate, dihydroxybenzophenone , tetrahydroxybenzophenone, 4- tert -butyl-4'-methoxydibenzoylmethane, 2,4,6-trianilino- p- (carbo-2'-ethylhexyl-1'-oxy)-1 and 3,5-triazine and 2-(2-hydroxy-5-methylphenyl)benzotriazole.

Examples of disinfectants include hinokitiol, triclosan, trichlorohydroxydiphenyl ether, chlorhexidine gluconate, phenoxyethanol, resorcin, isopropylmethylphenol, azulene, salicylic acid, zinc phyllithione, benzalkonium chloride, photosensitizer. So, No. 301, mononitroguaial sodium, undecyrenic acid, etc. are mentioned.

Examples of the antioxidant include butylhydroxyanisole, propyl gallic acid, and ellisorbic acid.

Examples of the pH adjuster include citric acid, sodium citrate, malic acid, sodium malate, fmalic acid, sodium fumarate, succinic acid, sodium succinate, sodium hydroxide, sodium monohydrogen phosphate, and the like.

Examples of the alcohol include higher alcohols such as cetyl alcohol.

In addition, blending components that may be added are not limited thereto, and any of the above components can be blended within a range that does not impair the object and effect of the present invention, but preferably 0.01 to 5% by weight relative to the total weight, more Preferably it is blended in 0.01 - 3% by weight.

The cosmetic of the present invention may take the form of a solution, an emulsion, a viscous mixture, or the like.

Components included in the cosmetic composition of the present invention may include components commonly used in cosmetic compositions in addition to the extract as an active ingredient, for example, conventional adjuvants such as stabilizers, solubilizers, vitamins, pigments and fragrances and carriers.

The cosmetic composition of the present invention may be prepared in any formulation conventionally prepared in the art, for example, emulsions, creams, lotions, packs, foundations, lotions, cosmetics, hair cosmetics, and the like.

Specifically, the cosmetic composition of the present invention is a skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisture cream, hand cream, foundation, essence, nourishing essence, It includes formulations of packs, soaps, cleansing foams, cleansing lotions, cleansing creams, body lotions and body cleansers.

When the formulation of the present invention is a paste, cream or gel, animal fiber, vegetable fiber, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component. can

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

When the formulation of the present invention is a solution or emulsion, a solvent, solvating agent or 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 fatty esters, fatty acid esters of polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, as a carrier component, water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, microcrystalline Cellulose, aluminum metahydroxide, bentonite, agar or tracanth may be used.

When the formulation of the present invention is a surfactant-containing cleansing agent, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide as carrier components Ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, linolin derivative or ethoxylated glycerol fatty acid ester and the like can be used.

Cytokine expression inhibition effect test using human skin epithelial cells (HaCaT) for the combined herbal extract of the present invention (Experimental Example 1), atopic dermatitis in vivo effect test using BALB/C 6-week-old female mice (Experimental Example 2), GM-CSF, IL-4, IL-10, IL-13, IL-31, and various cytokine expression inhibitory effect tests such as IL-33 (Experimental Example 3), TSLP cytokine expression inhibitory effect test (Experimental Example 4) ) and the like, it was confirmed that the samples exhibit strong skin anti-inflammatory activity, and the like, so that the composition can be used as an anti-inflammatory external pharmaceutical composition or cosmetic composition.

1 is a photograph showing the dermatitis-inducing site of a test animal in which dermatitis was induced and treated with herbal mixture extract (WIN), dexamethasone (DEX), or deionized water (DIW) according to an embodiment, respectively;
2 is hematoxylin & eosin (H&E) and It is a photograph showing the result of toluidine blue (TB) staining.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, these are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.

Example 1. Preparation of Combination Herbal Extracts

After washing and cutting 20 g of dried longan meat (Buyeong Pharm), 20 g of raw paper (Buyeong Pharm), and 20 g of Gobon (Buyeong Pharm), 20% (v/v) ethanol/ Add water solvent and extract under reflux at 90±5°C for 3 hours in Onggi yaktanggi, then filter (10 μm or less) and add 4 times the volume (v/w) of 20% (v/v) ethanol/water solvent to the remaining solid content After secondary extraction under the same conditions as above, it was filtered (10 μm or less).

The three extracts were mixed, concentrated under reduced pressure (16-21 brix), and the concentrate was sterilized (80-90°C) and then cooled. After freeze-drying, the mixed ethanol extract powder of longan meat, raw paper and gobon was crushed (50 mesh or less) and used as an experimental sample (20.5 g, yield 33.4%, hereinafter, named "WIN-1001X").

Example 2-6. Preparation of Combination Herbal Extracts

Based on the dry weight of longan meat (Buyeong Pharmaceutical Company), raw paper (Buyeong Pharmaceutical Company), and old book in the dry state, Example Samples 2-5 were prepared respectively with the composition shown in Table 1 below, except that the mixing ratio and extraction solvent were different. Experimental Example was used as a sample.

Combination herbal extract sample Sample weight (g) Example longan meat paper Gobon Extraction solvent* person extract weight final yield Example 2 10 5 50 10% EtOH WIN-1002X 16.6g 25.6% Example 3 20 50 5 Distilled water WIN-1003X 24.7g 32.9% Example 4 10 80 20 70% BuOH WIN-1004X 32.3g 29.4% Example 5 5 50 20 50% EtOH WIN-1005X 21.5g 28.7% Example 6 30 10 2 hexane WIN-1006X 12.6g 30.1%

Experimental Example 1. Cytokine expression inhibitory effect test of herbal mixture extract (in vitro)

In order to confirm the inhibitory effect on the cytokine expression of the sample of the above example, an experiment was performed as follows by applying the method described in the existing literature. (Jeong et al., 2019, J Invest Dermatol. 2019 May;139(5):1098-1109. doi: 10.1016/j.jid.2018.11.012. Epub 2018 Nov 29.)

Human skin epithelial cells (HaCaT, 300493, CLS) were placed in DMEM medium (D6429, Sigma-Aldrich) containing 10% fetal bovine serum (FBS), 100 units/ml penicillin, and 100 μg/ml streptomycin at an appropriate humidity (85-95). %) and 5% CO ₂ were maintained in a 37° C. incubator (HERA cell 150i, Thermo Fisher Scientific).

For gene expression investigation, the cells were passaged into 12 wells and treated with TNFα (RC214-12, Biobasic) 50ng/ml for 1 hour to induce an inflammatory response.

One hour after induction of inflammation, the herbal mixture extract was treated at a concentration of 1 μg/ml each in the same medium and cultured for an additional hour.

As a comparison group, dexamethasone (200 nM; positive control; denoted by DEX; D4902, Sigma-Aldrich) or DIW (deionized water; negative control) was used, respectively.

After treating the cells with the herbal mixture extract for 1 hour, RNA was extracted (FATRR-001, Favorgen), and cDNA was synthesized from the extracted RNA using a cDNA synthesis kit (RR036A, ,TAKARA).

Polymerization was performed using synthesized cDNA and Sybrgreen kit (RT500M, Enzynomics), and Real-time PCR using primers for cytokines related to skin inflammation (TSLP, GM-CSF and IL-1β) was performed. The primers used in the PCR are shown in Table 2.

PCR used primers human* direction sequence seq. no. RPLP0 forward 5'- AGC CCA GAA CAC TGG TCT C-3' One reverse 5’- ACT CAG GAT TTC AAT GGT GCC-3’ 2 TSLP forward 5'-TAT GAG TGG GAC CAA AAG TAC CG-3' 3 reverse 5'-GGG ATT GAA GGT TAG GCT CTG G-3' 4 GM-CSF forward 5'-TCC TGA ACC TGA GTA GAG ACA C-3' 5 reverse 5'-TGC TGC TTG TAG TGG CTG G-3' 6 IL-1β forward 5'-CTC CAG GGA CAG GAT ATG GA-3' 7 reverse 5'-TCT TTC AAC ACG CAG GAC AG-3' 8
*: abbreviation- RPLP0 (Ribosomal Protein Lateral Stalk Subunit P0); thymic stromal lymphopoietin (TSLP); GM (Granulocyte-macrophage)-CSF (colony stimulating factor); IL (interleukin)

Table 3 shows the results of quantification by real-time PCR using the primers. As shown in Table 3 , the test group treated with the extract of Example 1 significantly reduced the expression level of skin inflammation-related cytokines compared to the DIW-treated negative control group, and the degree of decrease was treated with dexamethasone (DEX) It was confirmed that it was equivalent to the positive control group. (Table 3)

Inhibitory effect on the expression of cytokines TSLP -
- TNFα
DIW TNFα
WIN-1001X TNFα
WIN-1002X TNFα
WIN-1003X TNFα
WIN-1005X TNFα
Dex One 132.4692 47.43735 60.85783 48.9323 55.34286 52.49334 0.462769 26.91228 9.645089 24.95619 19.85678 26.59252 11.2336 GM-CSF -
- TNFα
DIW TNFα
WIN-1001X TNFα
WIN-1002X TNFα
WIN-1003X TNFα
WIN-1005X TNFα
Dex One 4.473627 1.982161 2.069408 2.384771 1.917569 1.935997 0.111 0.817826 0.889233 0.326074 0.871501 0.599711 0.581338 IL-1β -
- TNFα
DIW TNFα
WIN-1001X TNFα
WIN-1002X TNFα
WIN-1003X TNFα
WIN-1005X TNFα
Dex One 4.152715 1.407169 1.437399 2.064964 1.662578 1.080503 0.483565 1.087056 0.394622 0.1926 0.620225 0.193175 0.413136

Statistical processing

Experimental results were expressed as mean ± SD after three repeated measurements. Statistical analysis was performed using SAS program (SAS Institute, Cary, NC, USA), and then Duncan's multiple test was performed, and correlation was analyzed.

Experimental Example 2. Atopic dermatitis effect test of herbal mixture extract (in vivo)

In order to confirm the effect of the sample of the above example on atopic dermatitis, an experiment was conducted by applying the method described in the literature using a mouse as follows. (Li et al., Drug Des Devel Ther. 2016 Feb 19;10:781-91. doi: 10.2147/DDDT.S94056. eCollection 2016.)

0.15% (w/v) DNFB (2,4-dinitrofluorobenzene, D1529, Sigma-Aldrich) 100uL (microliter) was applied to the abdominal cavity of BALB/C 6-week-old female mice (Daehan Biolink), and from the 7th to the 16th day Dermatitis was induced by removing the hair on the back and applying 100uL of 0.15% (w/v) DNFB every 3 days.

The drug was applied to the dermatitis-induced skin area every day from the 7th day to the 16th day after the first application of DNFB to the animal.

As the drug, WIN-1001X (10 mg/ml; expressed as WIN), dexamethasone (200 μM; positive control; expressed as DEX (D4902, Sigma-Aldrich), or DIW (deionized water; negative control) prepared in Example 1 above as the drug) were used respectively.

For comparison, instead of DNFB

Acetone-coated mice were treated with DIW and tested in the same manner.

1 shows a photograph of the skin area of the back induced by dermatitis on the 16th day from the day the test animal was first treated with the drug.

In addition, the level of atopic dermatitis was measured on the back region where dermatitis of the test animal was induced on the 16th day from the day the test animal was first treated with the drug. Atopic dermatitis scores were 0 (not at all) and 1 (slightly present) for 4 symptoms: (i) erythema/bleeding, (ii) edema, (iii) abrasions/burns, and (iv) wounds/dryness, respectively. ), 2 points (moderate), and 3 points (severe), and a total of 12 points was calculated. The obtained atopic dermatitis values (total 12 points) are shown in FIG. 1B .

As a result of the above experiment, as shown in FIGS. 1 and 4, the test group treated with the extract of Example 1 showed an excellent dermatitis improvement effect compared to the DIW-treated negative control group, and dexamethasone (DEX) treated positive It can be confirmed that the dermatitis improvement effect is equivalent to that of the control group. (See Table 2)

Atopic dermatitis effect (Fig. 1) Acetone
DIW DNFB
DIW DNFB
WIN-1001X DNFB
DEX 0 8.8 5 2.2 0 1.30384 0.707107 0.83666

In addition, histological analysis was performed on the skin tissues of the test animals. Specifically, on the 16th day from the day the test animal was first treated with the drug, the skin tissue of the test animal induced with dermatitis was treated with 4% (w/v) paraformaldehyde (P6148, Sigma-Aldrich) at 4° C. shaker (CR300, FINEPCR), and 12 hours later, the dehydrated and cleared tissue was impregnated with paraffin, and then sliced to a thickness of 5 μm to prepare tissue sections.

The prepared tissue sections were treated with hematoxylin (S3309, DAKO) & eosin (109844, Millipore) (H&E; skin epidermal layer staining) or toluidine blue (TB, 185426, Sigma-Aldrich; mast cell staining infiltrated into skin inflammatory sites). Stained and observed under a microscope (EVOS XL, Life Technologies).

The observation results are shown in FIG. 2 (micrograph) and Tables 5 to 6, respectively.

As shown in Figure 2 and Tables 5 to 6, the test group treated with the extract (WIN-1001X) of Example significantly reduced the thickness of the epidermal layer of the skin compared to the negative control group treated with DIW, and obesity that penetrated into the inflammatory site It can be seen that the number of cells (mast cells) is also significantly reduced, the thickness of the epidermal layer and the degree of reduction in the number of mast cells (mast cells) were similar to the positive control group treated with dexamethasone (DEX).

Relative area of epidermis (fold) Acetone
DIW DNFB
DIW DNFB
WIN-1001X DNFB
DEX One 7.06432606 3.55861484 1.638117 0.173221 2.07300369 0.42330352 0.875806

Relative number of mast cells (fold) Acetone
DIW DNFB
DIW DNFB
WIN-1001X DNFB
DEX One 3.182796 1.72043 0.854839 0.312754 1.003359 0.263386 0.270532

Experimental Example 3. Cytokine expression inhibition effect test of herbal mixture extract (Real-time PCR)

In order to confirm the inhibitory effect on the cytokine expression of the sample of the above example, an experiment was conducted by applying the method described in the literature using a mouse as follows. (Li et al., 2016 Drug Des Devel Ther. 2016 Feb 19;10:781-91. doi: 10.2147/DDDT.S94056. eCollection 2016.)

RNA was extracted from the skin tissue such as DNFB was applied to the test animal of Experimental Example 2 on the 16th day from the day the test animal was first treated with the drug (FATRR-001, Favorgen), and a cDNA synthesis kit (RR036A, ,TAKARA) cDNA was synthesized from the extracted RNA using

Polymerization was performed using the synthesized cDNA and Sybrgreen kit (RT500M, Enzynomics), and cytokines related to skin inflammation (GM-CSF, IL-4, IL-10, IL-13, IL-31, and IL- 33), real-time PCR was performed using a primer. The primers used in the PCR are shown in Table 7:

PCR used primers mouse* direction sequence seq. no. GAPDH forward 5'- AGG TCG GTG TGA ACG GAT TTG-3' 9 reverse 5'-TGT AGA CCA TGT AGT TGA GGT CA-3' 10 TSLP forward 5'-AGC TTG TCT CCT GAA AAT CGA G-3' 11 reverse 5'-AGG TTT GAT TCA GGC AGA TG TT-3' 12 CSF2 forward 5'-AGG GTC TAC GGG GCA ATT TC-3' 13 reverse 5'-TCA CAG TCC GTT TCC GGA GTT-3' 14 IL-4 forward 5'-GGT CTC AAC CCC CAG CTA GT-3' 15 reverse 5'-GCC GAT GAT CTC TCT CAA GTG AT-3' 16 IL-10 forward 5'-GCT CTT ACT GAC TGG CAT GAG-3' 17 reverse 5'-CGC AGC TCT AGG AGC ATG TG-3' 18 IL-13 forward 5'-CCT GGC TCT TGC TTG CCT T-3' 19 reverse 5'-GGT CTT GTG TGA TGT TGC TCA-3' 20 IL-22 forward 5’-ATG AGT TTT TCC CTT ATG GGG AC-3’ 21 reverse 5’-GCT GGA AGT TGG ACA CCT CAA-3’ 22 IL-31 forward 5'-TCA GCA GAC GAA TCA ATA CAG C-3' 23 reverse 5'-TCG CTC AAC ACT TTG ACT TTC T-3' 24 IL-33 forward 5'-GCT GCA GAA GGG AGA AAT CAC G-3' 25 reverse 5'-GGA GTT GGA ATA CTT CAT TCT AGG TCT CAT-3' 26
* : abbreviation- GAPDH (Glyceraldehyde-3-phosphate dehydrogenase); thymic stromal lymphopoietin (TSLP); GM (Granulocyte-macrophage)-CSF (colony stimulating factor); IL (interleukin)

Table 6 shows the results of quantification by real-time PCR using the primers. As shown in Table 6, the test group treated with the extract of Example 1 (WIN-1001X) significantly decreased the expression level of skin inflammation-related cytokines compared to the DIW-treated negative control group, and the degree of reduction was dexamethasone (DEX) was confirmed to be equal to or greater than that of the treated positive control group (Table 8).

Inhibitory effect on the expression of cytokines CSF2 Acetone
DIW DNFB*
DIW** DNFB
WIN-1001X DNFB
DEX*** One 72.14648 4.676082 7.817781 0.210396 12.96696 1.723294 0.98556 IL-4 Acetone
DIW DNFB
DIW DNFB
WIN-1001X DNFB
DEX One 11.36284 1.06316 4.798023 0.543902 3.453585 0.037212 2.312695 IL-10 Acetone
DIW DNFB
DIW DNFB
WIN-1001X DNFB
DEX One 19.29307 8.810186 20.44472 0.354329 1.973736 0.411741 4.157348 IL-13 Acetone
DIW DNFB
DIW DNFB
WIN-1001X DNFB
DEX One 48.62991 6.488924 12.56024 0.465986 10.37666 3.599525 8.866739 IL-22 Acetone
DIW DNFB
DIW DNFB
WIN-1001X DNFB
DEX One 29.18521 1.303359 13.08191 0.577444 2.801416 0.465745 1.945029 IL-31 Acetone
DIW DNFB
DIW DNFB
WIN-1001X DNFB
DEX One 3.933601 1.217765 1.000609 0.473134 0.44744 0.304217 0.13203 IL-33 Acetone
DIW DNFB
DIW DNFB
WIN-1001X DNFB
DEX One 9.471517 0.541047 0.736505 0.487178 2.797898 0.460806 0.486004 *DNFB: 2,4-dinitrofluorobenzene; **DIW: distilled water; ***DEX: Dexamethasone

Experimental Example 4. TSLP cytokine expression inhibitory effect test of herbal mixture extract (Real-time PCR)

In order to confirm the inhibitory effect on the TSLP cytokine expression of the sample of the above example, an experiment was conducted by applying the method described in the literature using a mouse as follows. (Li et al., 2016 Drug Des Devel Ther. 2016 Feb 19;10:781-91. doi: 10.2147/DDDT.S94056. eCollection 2016.)

RNA was extracted from the skin tissue such as DNFB was applied to the test animal of Experimental Example 2 on the 16th day from the day the test animal was first treated with the drug (FATRR-001, Favorgen), and a cDNA synthesis kit (RR036A, ,TAKARA) cDNA was synthesized from the extracted RNA using

Polymerization was performed using the synthesized cDNA and Sybrgreen kit (RT500M, Enzynomics), and real-time PCR was performed using a primer for TSLP, a cytokine related to skin inflammation. The primers used in the PCR are shown in Table 7:

Table 7 shows the results of quantification by real-time PCR using the primers. As shown in Table 7, the test group treated with the extract (WIN) of Example 1 significantly reduced the expression level of the skin inflammation-related cytokine TSLP compared to the negative control group treated with DIW, and the degree of reduction was dexamethasone ( DEX) was confirmed to be equal to or greater than that of the treated positive control group (Table 9).

Inhibitory effect on expression of TSLP TSLP Acetone
DIW DNFB
DIW DNFB
WIN-1001X DNFB
DEX One 7.514841 1.525431 1.310709 0.45284 1.954331 0.784039 0.32691

Statistical processing

Experimental results were expressed as mean ± SD after three repeated measurements. Statistical analysis was performed using SAS program (SAS Institute, Cary, NC, USA), and then Duncan's multiple test was performed, and correlation was analyzed.

Hereinafter, formulations of creams, massage creams, lotions, skin lotions, essences, packs, and cleansing foams are exemplified as formulation examples of the present invention, but the formulation including the cosmetic composition of the present invention is not limited thereto.

Formulation Example 1. Cream composition

The oil phase and the aqueous phase are heated and mixed at 75 °C, respectively, and then cooled to room temperature.

Formulation Example 2. Massage cream composition

The oil phase and the aqueous phase are each heated to 75 ° C., dissolved and mixed, and then cooled to room temperature.

Formulation Example 3. Lotion composition

After emulsifying the oil phase and the water phase by heating at 75 °C, respectively, it is cooled to room temperature.

Formulation Example 4. Skin lotion composition

The aqueous phase and the ethanol phase are prepared and mixed, respectively, and then filtered.

Formulation Example 5. Essence composition

The aqueous phase and the ethanol phase are prepared and mixed, respectively, and then filtered.

Formulation Example 6. Pack composition

The aqueous phase and the ethanol phase are each dispersed and dissolved, mixed, and then cooled to room temperature.

Formulation Example 7. Cleansing Foam Composition

After dispersing and dissolving the aqueous phase and the oil phase, mixing and saponification, the mixture is cooled to room temperature.

<110> PARK, Ok Nam MEDI HELP LINE Co., LTD. <120> topical composition comprising the extract of combined herbs comprising Longanae Arillus for TSLP inhibition and for the treatment or alleviation of skin inflammatory disease <130> DIF/2021-02-002/EK <150> KR 10-2020-0031482 <151> 2020-03-13 <160> 26 <170> KoPatentIn 3.0 <210> 1 <211> 19 <212> DNA <213> Homo sapiens <400> 1 agcccagaac actggtctc 19 <210> 2 <211> 21 <212> DNA <213> Homo sapiens <400> 2 actcaggatt tcaatggtgc c 21 <210> 3 <211> 23 <212> DNA <213> Homo sapiens <400> 3 tatgagtggg accaaaagta ccg 23 <210> 4 <211> 22 <212> DNA <213> Homo sapiens <400> 4 gggattgaag gttaggctct gg 22 <210> 5 <211> 22 <212> DNA <213> Homo sapiens <400> 5 tcctgaacct gagtagagac ac 22 <210> 6 <211> 19 <212> DNA <213> Homo sapiens <400> 6 tgctgcttgt agtggctgg 19 <210> 7 <211> 20 <212> DNA <213> Homo sapiens <400> 7 ctccagggac aggatatgga 20 <210> 8 <211> 20 <212> DNA <213> Homo sapiens <400> 8 tctttcaaca cgcaggacag 20 <210> 9 <211> 21 <212> DNA <213> Mus musculus <400> 9 aggtcggtgt gaacggattt g 21 <210> 10 <211> 23 <212> DNA <213> Mus musculus <400> 10 tgtagaccat gtagttgagg tca 23 <210> 11 <211> 22 <212> DNA <213> Mus musculus <400> 11 agcttgtctc ctgaaaatcg ag 22 <210> 12 <211> 22 <212> DNA <213> Mus musculus <400> 12 aggtttgatt caggcagatg tt 22 <210> 13 <211> 20 <212> DNA <213> Mus musculus <400> 13 agggtctacg gggcaatttc 20 <210> 14 <211> 21 <212> DNA <213> Mus musculus <400> 14 tcacagtccg tttccggagt t 21 <210> 15 <211> 20 <212> DNA <213> Mus musculus <400> 15 ggtctcaacc cccagctagt 20 <210> 16 <211> 23 <212> DNA <213> Mus musculus <400> 16 gccgatgatc tctctcaagt gat 23 <210> 17 <211> 21 <212> DNA <213> Mus musculus <400> 17 gctcttactg actggcatga g 21 <210> 18 <211> 20 <212> DNA <213> Mus musculus <400> 18 cgcagctcta ggagcatgtg 20 <210> 19 <211> 19 <212> DNA <213> Mus musculus <400> 19 cctggctctt gcttgcctt 19 <210> 20 <211> 21 <212> DNA <213> Mus musculus <400> 20 ggtcttgtgt gatgttgctc a 21 <210> 21 <211> 23 <212> DNA <213> Mus musculus <400> 21 atgagttttt cccttatggg gac 23 <210> 22 <211> 21 <212> DNA <213> Mus musculus <400> 22 gctggaagtt ggacacctca a 21 <210> 23 <211> 22 <212> DNA <213> Mus musculus <400> 23 tcagcagacg aatcaataca gc 22 <210> 24 <211> 22 <212> DNA <213> Mus musculus <400> 24 tcgctcaaca cttggacttt ct 22 <210> 25 <211> 22 <212> DNA <213> Mus musculus <400> 25 gctgcagaag ggagaaatca cg 22 <210> 26 <211> 30 <212> DNA <213> Mus musculus <400> 26 ggagttggaa tacttcattc taggtctcat 30

Claims (11)

A pharmaceutical composition for external use on the skin for the treatment and prevention of TLSP inhibition or skin inflammatory diseases, containing a combination herbal extract composed of longan meat, gobon and raw paper as an active ingredient. The pharmaceutical composition for external use on the skin according to claim 1, wherein the weight mixing ratio (w/w) of the longan meat, the gobon, and the base paper is 0.01-100: 0.01-100: 0.01-100 parts by weight (w/w). According to claim 1, wherein the combined herbal extract is water, ethanol, methanol, propanol, butanol, acetone, ethyl acetate, hexane, butylene glycol, propylene glycol, hydrous butylene glycol, hydrous propylene glycol, hydrous glycerin from the group consisting of A pharmaceutical composition for external use on the skin extracted with one or more selected solvents. The method of claim 1, wherein the skin inflammatory disease is pruritus, atopic dermatitis, chronic recurrent dermatitis, contact dermatitis, seborrheic dermatitis, dermatitis nervosa, dry skin, erythema, inflammatory dermatitis, psoriasis, wounds, or atopic disease. A pharmaceutical composition for external use. According to claim 1, wherein the pharmaceutical composition is a cream, gel, patch, spray, emulsion, ointment, warning agent, lotion, liniment agent, pasta agent, solvent, suspension, pack (pack), patch (patch) ) or cataplasmase (cataplasma) pharmaceutical composition for external use, characterized in that the formulation. A cosmetic composition for inhibiting TLSP or improving and preventing skin inflammatory diseases, comprising a combination herbal extract composed of longan meat, gobon and raw paper as an active ingredient. The cosmetic composition according to claim 6, wherein the weight mixing ratio (w/w) of the longan meat, the gobon, and the base paper is 0.01-100: 0.01-100: 0.01-100 parts by weight (w/w). The method of claim 6, wherein the combined herbal extract is water, ethanol, methanol, propanol, butanol, acetone, ethyl acetate, hexane, butylene glycol, propylene glycol, hydrous butylene glycol, hydrous propylene glycol, hydrous glycerin from the group consisting of A cosmetic composition extracted with one or more selected solvents. The method of claim 6, wherein the skin inflammatory disease is chronic recurrent dermatitis such as pruritus, atopic dermatitis or psoriasis due to aging or atopic dermatitis, contact dermatitis, seborrheic dermatitis, neurodermatitis, dry skin, erythema, inflammatory dermatitis, psoriasis A cosmetic composition that is a disease, wound, or atopic disease. The cosmetic composition according to claim 6, wherein the cosmetic composition is a lotion, skin, lotion, nutritional lotion, nourishing cream, massage cream, essence, pack, soap, liquid detergent, gel, patch formulation, or an edible cosmetic preparation for oral administration. cosmetic composition. A thymic stromal lymphopoietin (TSLP) cytokine expression inhibitor comprising a combination herbal extract composed of longan meat, gobon, and raw paper as an active ingredient.

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