WO2023146123A1 - Composition for preventing or treating inflammatory diseases comprising composite extract of ocimum basilicum, cirsium nipponicum (maxim.) makino, and aruncus dioicus as active ingredient - Google Patents

Composition for preventing or treating inflammatory diseases comprising composite extract of ocimum basilicum, cirsium nipponicum (maxim.) makino, and aruncus dioicus as active ingredient Download PDF

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WO2023146123A1
WO2023146123A1 PCT/KR2022/019994 KR2022019994W WO2023146123A1 WO 2023146123 A1 WO2023146123 A1 WO 2023146123A1 KR 2022019994 W KR2022019994 W KR 2022019994W WO 2023146123 A1 WO2023146123 A1 WO 2023146123A1
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preventing
inflammatory diseases
water
composition
pharmaceutical composition
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PCT/KR2022/019994
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French (fr)
Korean (ko)
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송석진
차미란
손주아
오별님
방상우
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(주)유스케어팜
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9789—Magnoliopsida [dicotyledons]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/324—Foods, ingredients or supplements having a functional effect on health having an effect on the immune system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2300/00—Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
    • A61K2800/59—Mixtures
    • A61K2800/591—Mixtures of compounds not provided for by any of the codes A61K2800/592 - A61K2800/596

Definitions

  • the present invention Thai basil ( Ocimum basilicum ), water thistle ( Cirsium nipponicum (Maxim.) Makino), and snow horse riding ( Aruncus dioicus ) It relates to a composition for preventing or treating inflammatory diseases comprising a composite extract as an active ingredient.
  • Inflammation is a defense response of the body against infection from external infectious agents such as external physical and chemical stimuli, bacteria, fungi, viruses, and various allergens.
  • the inflammatory response is a part of the innate immune response, and during the inflammatory reaction, blood plasma accumulates at the inflammatory site to dilute the toxicity secreted by bacteria, blood flow increases, and symptoms such as erythema, pain, edema, and fever are accompanied.
  • Various biochemical phenomena are involved in this inflammatory response.
  • NOS nitric oxide synthase
  • COX cyclooxygenase
  • NOS there are three isomers of NOS: eNOS (endothelial NOS), nNOS (nerve NOS), and bacterial endotoxins such as LPS (lipopolysaccharide), IL-1 ⁇ , TNF- ⁇ , IL-6, IL-8, and IL-6.
  • eNOS endothelial NOS
  • nNOS nerve NOS
  • LPS lipopolysaccharide
  • IL-1 ⁇ IL-1 ⁇
  • TNF- ⁇ IL-6
  • IL-8 IL-6
  • iNOS inducible NOS
  • NO produced by eNOS or nNOS plays an important role in maintaining homeostasis in the human body by performing various physiological reactions related to blood pressure regulation, neurotransmission, learning, and memory. It is known to be involved in various inflammatory diseases such as transplant rejection, autoimmune diseases, and neuronal death.
  • COX enzyme has hydroperoxidase (HOX) activity along with COX function and synthesizes intermediates PGG 2 and PGH 2 from arachidonic acid. and thromboxane A 2 (thromboxane A2, TxA2).
  • HOX hydroperoxidase
  • thromboxane A 2 thromboxane A2, TxA2
  • COX-1 is constitutively expressed in most tissues
  • COX-2 is rapidly induced by inflammatory cytokines and plays an important role in the inflammatory response.
  • Bacterial endotoxins such as LPS (lipopolysaccharide) activate NF- ⁇ B, a transcription factor, by binding to TLR4 (toll-like receptor 4), and induce the expression of iNOS and COX-2 to induce NO, inflammatory cytokines, and PGE2. It secretes inflammatory mediators. Inflammatory cytokines such as NO, TNF- ⁇ , and IL-6, and PGE2 have been reported as important factors inducing an inflammatory response in arthritis, fibromyalgia, Sjogren's syndrome, and the like.
  • Non-steroidal anti-inflammatory drugs which are currently widely used as anti-inflammatory drugs, are known to cause serious side effects such as gastrointestinal disorders, liver disorders, and renal disorders. Therefore, there is a need to develop new drugs that have anti-inflammatory activity and have low side effects and are effective continuously.
  • An object of the present invention is Vietnamese basil ( Ocimum basilicum ), water thistle ( Cirsium nipponicum (Maxim.) Makino), and snow riding horse ( Aruncus dioicus )
  • a composition for preventing or treating inflammatory diseases comprising two or more extracts selected from the group consisting of as an active ingredient.
  • the present invention Thai basil ( Ocimum basilicum ), water thistle ( Cirsium nipponicum (Maxim.) Makino), and snow riding horse ( Aruncus diioicus ) It provides a pharmaceutical composition for preventing or treating inflammatory diseases comprising two or more extracts selected from the group consisting of as an active ingredient.
  • the present invention Thai basil ( Ocimum basilicum ), water thistle ( Cirsium nipponicum (Maxim.) Makino), and snow riding horse ( Aruncus dioicus ) It provides a health functional food composition for preventing or improving inflammatory diseases containing two or more extracts selected from the group consisting of as an active ingredient.
  • the present invention Thai basil ( Ocimum basilicum ), water thistle ( Cirsium nipponicum (Maxim.) Makino) and snow riding horse ( Aruncus dioicus ) It provides a cosmetic composition for preventing or improving inflammatory diseases comprising two or more extracts selected from the group consisting of as an active ingredient.
  • Figure 1 is a result of measuring the change in gastric mucosal PEG2 (Prostaglandin E2) concentration through treatment with a composite extract of Thai basil, water thistle, and snow-capped horsetail.
  • gastric mucosal PEG2 Prostaglandin E2
  • Figure 2 is the result of measuring the degree of TNF- ⁇ production in the gastric mucosal tissue through the treatment of the composite extract of Thai basil, water thistle and snow-capped horsetail.
  • Figure 3 is the result of measuring the degree of IL-6 production in the gastric mucosal tissue through the treatment of Thai basil, water thistle, and the composite extract of snow sage.
  • Figure 4 is a result of measuring the IL-6 inhibitory ability in skin fibroblasts of the composite extract of Thai basil, water thistle and snow horse riding.
  • Figure 5 is the result of measuring the MMP-1 inhibitory ability in skin fibroblasts of the composite extract of Thai basil, water thistle and snow horse riding.
  • the present invention Thai basil ( Ocimum basilicum ), water thistle ( Cirsium nipponicum (Maxim.) Makino), and snow riding horse ( Aruncus diioicus ) It provides a pharmaceutical composition for preventing or treating inflammatory diseases comprising two or more extracts selected from the group consisting of as an active ingredient.
  • the extract is a
  • the extract is characterized in that it is extracted with water, (C1 ⁇ C4) alcohol or a mixed solvent thereof, preferably extracted with water or 30% ethanol, but is not limited thereto.
  • the inflammatory diseases include acute and chronic gastritis, peptic ulcer, duodenitis, gastric ulcer, enteritis, inflammatory bowel disease, ulcerative colitis, Crohn's disease, hepatitis, bronchitis, allergic rhinitis, asthma, chronic obstructive pulmonary disease, contact dermatitis, allergy Dermatitis atopic, anaphylaxis, seborrheic dermatitis, systemic lupus erythematosus, scleroderma, stomatitis, osteoarthritis, rheumatoid arthritis, neuritis, diabetic retinopathy, retinitis, macular degeneration, uveitis, conjunctivitis, ankylosing spondylitis, osteoarthritis, nephritis , nephritis, Sjogren's syndrome, autoimmune pancreatitis, periodontal disease, chronic pelvic inflammatory disease, endometritis,
  • the extract is characterized in that it has an anti-inflammatory effect through inhibition of IL-6 (Interleukin 6) and TNF- ⁇ (tumor necrosis factor- ⁇ ) activity.
  • IL-6 Interleukin 6
  • TNF- ⁇ tumor necrosis factor- ⁇
  • the extract is characterized in that it has an antioxidant effect through inhibition of catalase and SOD (superoxide dismutase) activity.
  • the extract is characterized in that it has a skin protection effect through inhibition of IL-6 (Interleukin 6) and MMP-1 (matrix metalloproteinase-1) activity.
  • IL-6 Interleukin 6
  • MMP-1 matrix metalloproteinase-1
  • the pharmaceutical composition of the present invention is prepared in a unit dose form or in a multi-dose container by formulating using a pharmaceutically acceptable carrier according to a method that can be easily performed by those skilled in the art. It can be prepared by incorporating into
  • the pharmaceutically acceptable carrier is one commonly used in formulation, and includes lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methyl hydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like.
  • the pharmaceutical composition of the present invention may further include lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, and the like, in addition to the above components.
  • the content of the additives included in the pharmaceutical composition is not particularly limited and may be appropriately adjusted within the range of content used in conventional formulations.
  • the pharmaceutical composition is an aqueous solution, suspension, emulsion, and the like for injection, pills, capsules, granules, tablets, creams, gels, patches, sprays, ointments, warning agents, lotions, liniment agents, pasta agents, and cataplasma agents. It may be formulated in the form of one or more external agents selected from the group consisting of
  • the pharmaceutical composition of the present invention may additionally contain pharmaceutically acceptable carriers and diluents for formulation.
  • pharmaceutically acceptable carriers and diluents include excipients such as starch, sugar, and mannitol, fillers and extenders such as calcium phosphate, cellulose derivatives such as carboxymethylcellulose and hydroxypropylcellulose, gelatin, alginates, and polyvinyl fibres.
  • binders such as rolidone, etc., lubricants such as talc, calcium stearate, hydrogenated castor oil and polyethylene glycol, disintegrants such as povidone, crospovidone, surfactants such as polysorbates, cetyl alcohol, and glycerol; don't
  • the pharmaceutically acceptable carrier and diluent may be biologically and physiologically compatible with the subject.
  • diluents include, but are not limited to, saline, aqueous buffers, solvents and/or dispersion media.
  • the pharmaceutical composition of the present invention may be administered orally or parenterally (eg, intravenous, subcutaneous, intraperitoneal or topical application) depending on the desired method.
  • parenterally eg, intravenous, subcutaneous, intraperitoneal or topical application
  • it may be formulated into tablets, troches, lozenges, aqueous suspensions, oily suspensions, powders, granules, emulsions, hard capsules, soft capsules, syrups or elixirs.
  • parenteral administration it may be formulated as an injection solution, suppository, powder for respiratory inhalation, aerosol for spray, ointment, powder for application, oil, cream, etc.
  • the dosage of the pharmaceutical composition of the present invention depends on the condition and weight of the patient, age, sex, health condition, dietary constitution specificity, the nature of the preparation, the severity of the disease, the administration time of the composition, the administration method, the administration period or interval, the excretion rate, And the range may vary depending on the type of drug, and may be appropriately selected by a person skilled in the art. For example, it may range from about 0.1 to 10,000 mg/kg, but is not limited thereto, and may be divided and administered once or several times a day.
  • the pharmaceutical composition may be administered orally or parenterally (eg, intravenous, subcutaneous, intraperitoneal or topical application) depending on the desired method.
  • the pharmaceutically effective amount and effective dose of the pharmaceutical composition of the present invention may vary depending on the formulation method, administration method, administration time and/or administration route of the pharmaceutical composition, and those skilled in the art may be able to use the desired treatment An effective dosage can be readily determined and prescribed.
  • Administration of the pharmaceutical composition of the present invention may be administered once a day, or may be divided and administered several times.
  • the “anti-inflammatory” means an effect of improving inflammation.
  • the “inflammation” is a mechanism for restoring and regenerating the damaged area when an invasion that causes any organic change such as physical action, chemical substance, bacterial infection, etc. is applied to a living body or tissue. Once stimulation is applied, histamine, Inflammation may be induced as vascular active substances such as serotonin, bradykinin, prostaglandin, HETE (hydroxyeicosatetraenoic acid), and leukotrienes are liberated to increase vascular permeability.
  • the “antioxidation” refers to an action of inhibiting oxidation.
  • oxidation promoters and antioxidants are in balance, but due to various factors, this balance is lost and tilted in the direction of promoting oxidation. , oxidative stress is induced in vivo, which can cause cell damage and pathological diseases.
  • the present invention Thai basil ( Ocimum basilicum ), water thistle ( Cirsium nipponicum (Maxim.) Makino), and snow riding horse ( Aruncus dioicus ) It provides a health functional food composition for preventing or improving inflammatory diseases containing two or more extracts selected from the group consisting of as an active ingredient.
  • the health functional food composition is characterized in that it is an inner beauty food.
  • the present invention can be generally used as a commonly used food.
  • the food composition of the present invention can be used as a health functional food.
  • health functional food refers to food manufactured and processed using raw materials or ingredients having useful functionalities for the human body in accordance with the Health Functional Food Act, and "functional” refers to food that is not related to the structure and function of the human body. It refers to intake for the purpose of obtaining useful effects for health purposes such as regulating nutrients or physiological functions.
  • the health functional food composition may include conventional food additives, and the suitability as the "food additive" is determined in accordance with the General Rules and General Test Methods of Food Additives approved by the Ministry of Food and Drug Safety, unless otherwise specified. It is judged according to the specifications and standards for the item.
  • Items listed in the "Food Additive Code” include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid, natural additives such as dark pigment, licorice extract, crystalline cellulose, goyang pigment, guar gum, and mixed preparations such as sodium L-glutamate preparations, noodle-added alkali preparations, preservative preparations, and tar color preparations.
  • chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid
  • natural additives such as dark pigment, licorice extract, crystalline cellulose, goyang pigment, guar gum
  • mixed preparations such as sodium L-glutamate preparations, noodle-added alkali preparations, preservative preparations, and tar color preparations.
  • the food composition of the present invention can be prepared and processed in the form of tablets, capsules, powders, granules, liquids, pills and the like.
  • hard capsules can be prepared by mixing and filling a composition according to the present invention with additives such as excipients in a conventional hard capsule, and soft capsules contain the composition according to the present invention. It can be prepared by mixing with additives such as excipients and filling in a capsule base such as gelatin.
  • the soft capsule may contain a plasticizer such as glycerin or sorbitol, a coloring agent, a preservative, and the like, if necessary.
  • prevention refers to all activities that inhibit or delay a disease by administering the composition according to the present invention.
  • treatment refers to all activities that improve or beneficially change the symptoms of a disease by administering the composition according to the present invention.
  • improvement means any action that improves the bad condition of a disease by administering or ingesting the composition of the present invention to a subject.
  • the present invention Thai basil ( Ocimum basilicum ), water thistle ( Cirsium nipponicum (Maxim.) Makino) and snow riding horse ( Aruncus dioicus ) It provides a cosmetic composition for preventing or improving inflammatory diseases comprising two or more extracts selected from the group consisting of as an active ingredient.
  • the cosmetic composition includes skin lotion, skin softener, skin toner, astringent, milk lotion, moisture lotion, nutrient lotion, massage cream, nutrient cream, moisture cream, hand cream, foundation, essence, nutrient essence, pack, soap, cleansing foam, It is characterized in that it has one or more formulations selected from the group consisting of cleansing lotion, cleansing cream, body lotion and body cleanser, but is not limited thereto.
  • the cosmetic composition may be prepared in general emulsified and solubilized formulations.
  • cosmetic lotion such as softening lotion or nourishing lotion
  • emulsions such as facial lotion and body lotion
  • creams such as nourishing creams, moisture creams, and eye creams; essence; cosmetic ointment; spray; gel; pack; sunscreen; makeup base; foundations such as liquid type, solid type or spray type; powder
  • makeup removers such as cleansing creams, cleansing lotions, and cleansing oils
  • it may be formulated as a cleanser such as a cleansing foam, soap, body wash, etc., but is not limited thereto.
  • the external skin preparation may be formulated as an ointment, patch, gel, cream or spray, but is not limited thereto.
  • the cosmetic composition may be appropriately mixed with other components within a range that does not impair the purpose of the present invention depending on the type of formulation or purpose of use.
  • the cosmetic composition may include a conventionally acceptable carrier, and for example, oil, water, a surfactant, a moisturizer, a lower alcohol, a thickener, a chelating agent, a colorant, a preservative, a flavoring agent, and the like may be appropriately blended, but are not limited thereto. no.
  • the acceptable carrier may vary depending on the formulation.
  • animal oil, vegetable oil, wax, paraffin, starch, tracanthate, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide or any of these as a carrier component when formulated into an ointment, paste, cream or gel extracts may be used.
  • lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder or extracts thereof may be used as a carrier component, and in the case of a spray, chlorofluorohydro It may further contain a propellant such as carbon, propane, butane or dimethyl ether.
  • a solvent, solubilizing agent, or emulsifying agent may be used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil may be used, and in particular, cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, fatty acid esters of glycerol, polyethylene glycol or sorbitan may be used.
  • a liquid diluent such as water, ethanol or propylene glycol, an ethoxylated isostearyl alcohol, a suspending agent such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tracanth and the like can be used.
  • the cosmetic composition is formulated as a soap, alkali metal salts of fatty acids, fatty acid hemiester salts, fatty acid protein hydrolyzates, isethionates, lanolin derivatives, fatty alcohols, vegetable oils, glycerol, sugars, etc. can be used
  • the cosmetic composition includes fatty substances, organic solvents, solubilizers, thickeners, gelling agents, softeners, antioxidants, suspending agents, stabilizers, foaming agents, and fragrances commonly used in the industry depending on the quality or function of the final product. , surfactants, water, ionic or nonionic emulsifiers, fillers, sequestering agents, chelating agents, preservatives, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives commonly used in cosmetics It may additionally contain adjuvants commonly used in cosmetology or dermatology, such as any other ingredient.
  • the adjuvant and its mixing ratio may be appropriately selected so as not to affect desirable properties of the cosmetic composition according to the present invention.
  • LPS lipopolysaccharide
  • MTT 3-(4,5-dime thylthiazol-2-yl)-2,5-diphenyltetrazolium
  • DMSO dimethyl sulfoxide
  • Tumor necrosis factor (TNF)- ⁇ , IL-1 ⁇ , and IL-6 ELISA kits were purchased from Pierce Endogen (Rockford, IL., USA).
  • TNF Tumor necrosis factor
  • IL-1 ⁇ IL-1 ⁇
  • IL-6 ELISA kits were purchased from Pierce Endogen (Rockford, IL., USA).
  • special grade reagents were used, and measurements were performed using a UV/vis spectrophotometer (Hitachi, Japan), centrifuge (Hitachi, Japan), ELISA reader (Bio Rad, Japan), and the like.
  • Raw 264.7 cells a murine macrophage cell line, were purchased from ATCC (American Type Culture Collection), and Dulbecco's modified Eagle's medium (DMEM) was mixed with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 ⁇ g/mL streptomycin. 37 °C, 5% CO 2 was cultured in an incubator. During the experiment, the conditions of 80% confluency and less than 20 passages were observed, and the cells were cultured in a medium without FBS for 12 hours before the experiment.
  • DMEM Dulbecco's modified Eagle's medium
  • Nitric Oxide (hereinafter referred to as NO) was measured by measuring the amount of NO in the supernatant of cells as nitrite and nitrate.
  • the safe form after oxidation from nitrite to nitrate was measured using a grease reagent (Sigma, USA).
  • 500 ⁇ M of lipopolysaccharide (LPS) was added to all wells except for the normal group to stimulate them.
  • LPS lipopolysaccharide
  • the single or combined extracts of Examples 1 to 34 of Examples 1 to 34 were treated with 200 ⁇ g/mL concentration of Thai basil, snow sage, and water thistle, and then incubated for 24 hours.
  • the amount of NO production was measured by absorbance at 540 nm after collecting the supernatant over time and reacting by blocking light with a grease reagent for 20 minutes.
  • the NO inhibition rate was calculated according to the following formula.
  • Example NO inhibitory ability ( % )
  • Example NO inhibitory ability ( % )
  • LPS lipopolysaccharide
  • Example 29 to 31 showed high TNF- ⁇ inhibitory ability, and in particular, Example 30 showed the highest TNF- ⁇ inhibitory ability at 75.8% (Table 3).
  • Example TNF - ⁇ inhibitory ability ( % )
  • Example TNF - ⁇ inhibitory ability ( % )
  • Examples 25 to 34 a comparative experiment was conducted for NO and TNF- ⁇ inhibition rate by changing the extraction solvent.
  • the extraction solvent conditions and the weight ratio of the three composite extracts were set as shown in Table 4 to prepare Comparative Examples 1 to 30.
  • the water extract (Examples 25 to 34) showed superior NO and TNF- ⁇ inhibitory ability than the ethanol extract for each condition, and among the ethanol extracts, the 30% ethanol extract (Comparative Example 1 ⁇ 10) showed better inhibitory activity than other ethanol extraction conditions, and Comparative Example 6 showed 83.1% and 70.0% inhibitory activity, respectively, showing the best inhibitory activity among ethanol extracts (Table 4).
  • Seven-week-old female rats (Sprage-Dawley rats) were purchased from Coretech. The rats were placed in a rat cage at a temperature of 23 ⁇ 3 ° C, relative humidity of 55 ⁇ 15%, ventilation frequency of 10 to 20 times / hr, lighting time of 12 hours (8:00 am to 8:00 pm off), and illumination of 150 to 300 Lux. They were reared in a set room and fed a normal laboratory diet.
  • the area of the damaged area and the total area of the stomach on the mucosal surface of the lesion-prone gastric mucosa photographed with a digital camera were analyzed using ImageJ software (NIH, Bethesda, MD), and the Ulcer index (%) and gastritis inhibition rate were calculated. Ulcer index (%) and gastritis inhibition rate were calculated according to the following formula.
  • Gastritis inhibition rate ( % ) [ 1 - ( oral administration group ulcer index / Gastritis provoking group ulcer index)] X 100
  • a portion of the extracted stomach was cut to a certain weight, dissolved in RIPA buffer (Cell Signaling Technology, Danvers, MA, USA) to separate proteins, and then centrifuged for 10 minutes at 4°C and 13,000 ⁇ g in a centrifuge. Protein lysates were measured using a PGE2 ELISA kit (Cusabio Biotech, Wuhan, China).
  • Example 30 showed the highest value among all experimental groups including the positive control group at 140.9 pg/mg/protein (FIG. 1).
  • SOD activity was measured using the SOD assay kit (Biomax, Korea) according to the manufacturer's protocol, and in the other, the supernatant was centrifuged at 4°C and 10,000 rpm for 15 minutes, and then the CAT assay kit (Biomax, Korea) was used. ) was measured according to the manufacturer's protocol.
  • Example Thai basil Snow Riding water thistle SOD activity (U/g of protein)
  • Catalase activity (U/g of protein)
  • Vehicle 68.3 ⁇ 3.8 11.2 ⁇ 4.0 One One 80.6 ⁇ 5.5 19.5 ⁇ 2.2 2 One 82.1 ⁇ 5.1 20.5 ⁇ 3.4 3 One 83.7 ⁇ 4.0 16.8 ⁇ 2.6 25 One One One 97.5 ⁇ 7.1 21.5 ⁇ 3.5 29 One 2 One 99.1 ⁇ 3.7 22.4 ⁇ 4.0 30 One 4 One 116.9 ⁇ 5.7 30.4 ⁇ 3.5 31
  • One 6 One 104.5 ⁇ 4.6 25.5 ⁇ 1.5 Positive control group: Ayeop ethanol soft-textured extract 100.1 ⁇ 6.2 22.1 ⁇ 0.5
  • Examples 25, 29, 30 and 31 showed SDO and catalase activities similar to those of the positive control group, and in particular, Example 30 was a positive control group in both SOD and catalase activities. It was confirmed that it showed the highest activity among all experimental groups including (Table 6).
  • TNF- ⁇ and IL-6 in gastric mucosal tissue were measured. 100mM Tris-HCl (pH 7.4), 5mM Tris-HCl (pH 7.5), 2mM MgCl 2 , 15mM CaCl 2 , 1.5M sucrose and 0.1M DTT protease inhibitor cocktail were added to obtain cytoplasm from gastric tissue frozen in liquid nitrogen. After adding one buffer A and grinding with a tissue grinder (Bio Spec Product, USA), a 10% NP-40 solution was added. After standing on ice for 20 minutes, the supernatant containing cytoplasm was separated by centrifugation at 12,000 rpm for 2 minutes. TNF- ⁇ and IL-6 enzyme immunoassay (EIA) kit (RayBiotech, Norcross, GA, USA) were used to measure the supernatant. As a positive control group, ethanol soft extract was used.
  • EIA enzyme immunoassay
  • Example 25 showed lower TNF- ⁇ values than the positive control group, and among them, Example 30 showed the lowest value at 20.3 pg/mg/protein (Fig. 2).
  • Example 29 showed lower IL-6 values than the positive control group, and among them, Example 30 showed the lowest value at 107.7 pg/mg/protein (FIG. 3) .
  • CCD986sk a human skin fibroblast cell line
  • ATCC American Type Culture Collection
  • IMDM Iscove Modified Dulbecco Media
  • FBS fetal bovine serum
  • penicillin 100 U/mL penicillin
  • streptomycin 100 ⁇ g/mL streptomycin.
  • 37 °C, 5% CO 2 was cultured in an incubator.
  • the cells were cultured in a medium without FBS for 12 hours before the experiment in compliance with the conditions of 80% confluency and less than 10 passages.
  • IL-6 a pro-inflammatory cytokine
  • 1 ⁇ 10 6 cells were cultured for 24 hours and then stimulated by adding 20 ng/ml of TNF-a to all wells except for the normal group. After 1 hour, each well was treated with single or combined extracts of 200 ⁇ g/mL of Thai basil, snow sage, and water thistle, and then incubated for 24 hours.
  • the amount of IL-6 produced was measured by collecting the cell culture medium and using an IL-6 enzyme immunoassay (EIA) kit (R&D Systems Inc. Minneapolis, MN, USA) according to the manufacturer's protocol.
  • EIA enzyme immunoassay
  • Example 30 showed the lowest amount of IL-6 production among all experimental groups including the untreated group at 81.8% (FIG. 4).
  • MMP-1 matrix metalloproteinase-1
  • TNF-a matrix metalloproteinase-1
  • each well was treated with single or combined extracts of 200 ⁇ g/mL of Thai basil, snow sage, and water thistle, and then incubated for 24 hours.
  • the cell culture medium was collected and measured using the Human MMP1 ELISA Kit (Abcam, Cambridge, MA, USA) according to the manufacturer's protocol.
  • Example 30 showed the smallest amount of MMP-1 production at 101.3% (FIG. 5).

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Abstract

The present invention relates to a composition for preventing or treating inflammatory diseases comprising a composite extract of Ocimum basilicum, Cirsium nipponicum (Maxim.) Makino, and Aruncus dioicus as an active ingredient. It has been confirmed that the composite extract obtained by extracting the medicinal plants at a specific weight ratio and extraction conditions has excellent anti-inflammatory effect compared to single extracts of the medicinal plants, and as a result, the composition can be effectively used as a pharmaceutical composition, a functional health food, or a cosmetic composition for preventing or treating inflammatory diseases.

Description

타이바질, 물엉겅퀴 및 눈개승마의 복합추출물을 유효성분으로 포함하는 염증질환 예방 또는 치료용 조성물A composition for the prevention or treatment of inflammatory diseases containing complex extracts of Thai basil, water thistle and sagebrush as an active ingredient
본 발명은 타이바질(Ocimum basilicum), 물엉겅퀴(Cirsium nipponicum(Maxim.) Makino) 및 눈개승마(Aruncus dioicus)의 복합추출물을 유효성분으로 포함하는 염증질환 예방 또는 치료용 조성물에 관한 것이다.The present invention Thai basil ( Ocimum basilicum ), water thistle ( Cirsium nipponicum (Maxim.) Makino), and snow horse riding ( Aruncus dioicus ) It relates to a composition for preventing or treating inflammatory diseases comprising a composite extract as an active ingredient.
염증은 외부의 물리·화학적 자극, 박테리아, 곰팡이, 바이러스, 각종 알레르기 유발 물질 등 외부 감염원의 감염에 대한 생체의 방어 반응이다. 염증 반응은 선천성 면역 반응의 일부이며, 염증 반응 시 염증 부위에 혈장이 축적되어 세균이 분비한 독성을 희석시키며, 혈류가 증가하고, 홍반, 통증, 부종, 발열 등의 증상이 수반되게 된다. 이러한 염증 반응에는 다양한 생화학적 현상이 관여하는데, 특히 산화질소 합성효소(nitric oxide synthase, NOS)와 다양한 프로스타글란딘(prostaglandins)의 생합성과 관련되는 사이클로옥시제나제(cyclooxygenase, COX)가 염증 반응의 중요한 매개체로 알려져 있다.Inflammation is a defense response of the body against infection from external infectious agents such as external physical and chemical stimuli, bacteria, fungi, viruses, and various allergens. The inflammatory response is a part of the innate immune response, and during the inflammatory reaction, blood plasma accumulates at the inflammatory site to dilute the toxicity secreted by bacteria, blood flow increases, and symptoms such as erythema, pain, edema, and fever are accompanied. Various biochemical phenomena are involved in this inflammatory response. In particular, nitric oxide synthase (NOS) and cyclooxygenase (COX) related to the biosynthesis of various prostaglandins are important mediators of the inflammatory response. is known as
NOS는 세 가지 이성질체가 존재하는데, eNOS(내피성 NOS), nNOS(신경성 NOS) 및 LPS(lipopolysaccharide)와 같은 세균의 내독소나 IL-1β, TNF-α, IL-6, IL-8 및 IL-12와 같은 여러 염증성 사이토카인에 의해 유도되는 iNOS(유도성 NOS)가 있으며, L-아르기닌(L-arginine)으로부터 산화질소(NO)를 생성한다. eNOS나 nNOS에 의해 생성되는 NO는 혈압 조절 작용, 신경 전달 작용, 학습, 기억 등과 관련된 다양한 생리 반응을 수행함으로써 인체의 항상성 유지에 중요한 역할을 하지만, iNOS에 의해 생성되는 NO는 관절염, 패혈증, 조직이식거부반응, 자가면역질환, 신경세포의 사멸 등 다양한 염증성 질환에 관여하는 것으로 알려져 있다.There are three isomers of NOS: eNOS (endothelial NOS), nNOS (nerve NOS), and bacterial endotoxins such as LPS (lipopolysaccharide), IL-1β, TNF-α, IL-6, IL-8, and IL-6. There is iNOS (inducible NOS), which is induced by several inflammatory cytokines, such as -12, and produces nitric oxide (NO) from L-arginine. NO produced by eNOS or nNOS plays an important role in maintaining homeostasis in the human body by performing various physiological reactions related to blood pressure regulation, neurotransmission, learning, and memory. It is known to be involved in various inflammatory diseases such as transplant rejection, autoimmune diseases, and neuronal death.
COX 효소는 COX의 기능과 함께 하이드로퍼옥시다제(hydroperoxidase, HOX) 활성을 가지고 아라키돈산으로부터 중간체인 PGG2와 PGH2를 합성하며, 이들 화합물로 PGE2, PGF2, PGD2, 프로스타시클린 및 트롬복신A2(thromboxane A2, TxA2)를 만든다. COX의 기능 중 PGH 합성효소의 기능은 PGE2의 합성을 통해 통증과 염증 반응에 관여한다. COX에는 두 가지 아형이 있고 COX-1은 대부분의 조직에 항시 발현되는데 비해, COX-2는 염증성 사이토카인에 의해 신속히 발현이 유도되어 염증 반응에서 중요한 역할을 한다.COX enzyme has hydroperoxidase (HOX) activity along with COX function and synthesizes intermediates PGG 2 and PGH 2 from arachidonic acid. and thromboxane A 2 (thromboxane A2, TxA2). Among the functions of COX, the function of PGH synthetase is involved in pain and inflammatory responses through the synthesis of PGE 2 . There are two subtypes of COX, and COX-1 is constitutively expressed in most tissues, whereas COX-2 is rapidly induced by inflammatory cytokines and plays an important role in the inflammatory response.
LPS(lipopolysaccharide) 등의 세균 내독소는 TLR4(toll-like receptor 4)와의 결합함으로써 전사인자인 NF-κB를 활성화시키며, iNOS 및 COX-2의 발현을 유도하여 NO, 염증성 사이토카인, PGE2 등 여러 염증 조절 물질을 분비하게 한다. NO, TNF-α, IL-6 등의 염증성 사이토카인, PGE2 등은 관절염, 섬유근통, 쇼그렌 증후군 등에서 염증 반응의 유도하는 중요한 인자로 보고되어 있다. Bacterial endotoxins such as LPS (lipopolysaccharide) activate NF-κB, a transcription factor, by binding to TLR4 (toll-like receptor 4), and induce the expression of iNOS and COX-2 to induce NO, inflammatory cytokines, and PGE2. It secretes inflammatory mediators. Inflammatory cytokines such as NO, TNF-α, and IL-6, and PGE2 have been reported as important factors inducing an inflammatory response in arthritis, fibromyalgia, Sjogren's syndrome, and the like.
이러한 연구 결과는 NO 생성을 억제하거나 TNF-α, IL-6 등의 염증성 사이토카인의 생성을 억제하는 약물, iNOS나 COX-2의 발현을 억제하는 약물은 유효한 항염증제로서의 가능성을 가짐을 시사한다.These findings suggest that drugs that inhibit NO production or production of inflammatory cytokines such as TNF-α and IL-6, and drugs that inhibit the expression of iNOS or COX-2 have potential as effective anti-inflammatory agents.
현재 항염증제로서 널리 사용되고 있는 비스테로이드성 소염제(non-steroidal anti-inflammatory drugs, NSAIDS)는 위장관 장애, 간장애, 신장애 등의 심각한 부작용을 야기한다고 알려져 있다. 따라서 항염 활성을 가지면서 부작용이 적고 효과가 지속적인 새로운 약물의 개발이 필요한 실정이다.Non-steroidal anti-inflammatory drugs (NSAIDS), which are currently widely used as anti-inflammatory drugs, are known to cause serious side effects such as gastrointestinal disorders, liver disorders, and renal disorders. Therefore, there is a need to develop new drugs that have anti-inflammatory activity and have low side effects and are effective continuously.
본 발명의 목적은 타이바질(Ocimum basilicum), 물엉겅퀴(Cirsium nipponicum(Maxim.) Makino) 및 눈개승마(Aruncus dioicus)로 이루어진 군에서 선택된 둘 이상의 추출물을 유효성분으로 포함하는 염증질환 예방 또는 치료용 조성물을 제공하는 것이다.An object of the present invention is Thai basil ( Ocimum basilicum ), water thistle ( Cirsium nipponicum (Maxim.) Makino), and snow riding horse ( Aruncus dioicus ) To provide a composition for preventing or treating inflammatory diseases comprising two or more extracts selected from the group consisting of as an active ingredient.
상기 목적을 달성하기 위해, 본 발명은 타이바질(Ocimum basilicum), 물엉겅퀴(Cirsium nipponicum(Maxim.) Makino) 및 눈개승마(Aruncus dioicus)로 이루어진 군에서 선택된 둘 이상의 추출물을 유효성분으로 포함하는 염증질환 예방 또는 치료용 약학 조성물을 제공한다.In order to achieve the above object, the present invention Thai basil ( Ocimum basilicum ), water thistle ( Cirsium nipponicum (Maxim.) Makino), and snow riding horse ( Aruncus diioicus ) It provides a pharmaceutical composition for preventing or treating inflammatory diseases comprising two or more extracts selected from the group consisting of as an active ingredient.
또한, 본 발명은 타이바질(Ocimum basilicum), 물엉겅퀴(Cirsium nipponicum(Maxim.) Makino) 및 눈개승마(Aruncus dioicus)로 이루어진 군에서 선택된 둘 이상의 추출물을 유효성분으로 포함하는 염증질환 예방 또는 개선용 건강기능식품 조성물을 제공한다.In addition, the present invention Thai basil ( Ocimum basilicum ), water thistle ( Cirsium nipponicum (Maxim.) Makino), and snow riding horse ( Aruncus dioicus ) It provides a health functional food composition for preventing or improving inflammatory diseases containing two or more extracts selected from the group consisting of as an active ingredient.
또한, 본 발명은 타이바질(Ocimum basilicum), 물엉겅퀴(Cirsium nipponicum(Maxim.) Makino) 및 눈개승마(Aruncus dioicus)로 이루어진 군에서 선택된 둘 이상의 추출물을 유효성분으로 포함하는 염증질환 예방 또는 개선용 화장료 조성물을 제공한다.In addition, the present invention Thai basil ( Ocimum basilicum ), water thistle ( Cirsium nipponicum (Maxim.) Makino) and snow riding horse ( Aruncus dioicus ) It provides a cosmetic composition for preventing or improving inflammatory diseases comprising two or more extracts selected from the group consisting of as an active ingredient.
본 발명에 따르면, 타이바질(Ocimum basilicum), 물엉겅퀴(Cirsium nipponicum(Maxim.) Makino) 및 눈개승마(Aruncus dioicus)을 특정 중량 비율 및 추출 조건으로 추출한 복합추출물이 상기 약용 식물들의 단독 추출물보다 항염 효과가 우수하다는 것을 확인함으로써, 염증질환 예방 또는 치료용 약학 조성물, 건강기능식품 또는 화장료 조성물로서 유용하게 활용될 수 있다.According to the present invention, Thai basil ( Ocimum basilicum ), water thistle ( Cirsium nipponicum (Maxim.) Makino), and snow riding horse ( Aruncus dioicus ) by confirming that the composite extracts extracted under specific weight ratios and extraction conditions have superior anti-inflammatory effects than single extracts of the medicinal plants, thereby reducing inflammation It can be usefully used as a pharmaceutical composition for preventing or treating a disease, a health functional food, or a cosmetic composition.
도 1은 타이바질, 물엉겅퀴 및 눈개승마의 복합추출물 처리를 통한 위 점막 PEG2(Prostaglandin E2) 농도 변화를 측정한 결과이다.Figure 1 is a result of measuring the change in gastric mucosal PEG2 (Prostaglandin E2) concentration through treatment with a composite extract of Thai basil, water thistle, and snow-capped horsetail.
도 2는 타이바질, 물엉겅퀴 및 눈개승마의 복합추출물 처리를 통한 위 점막 조직 내 TNF-α생성 정도를 측정한 결과이다.Figure 2 is the result of measuring the degree of TNF-α production in the gastric mucosal tissue through the treatment of the composite extract of Thai basil, water thistle and snow-capped horsetail.
도 3은 타이바질, 물엉겅퀴 및 눈개승마의 복합추출물 처리를 통한 위 점막 조직 내 IL-6 생성 정도를 측정한 결과이다.Figure 3 is the result of measuring the degree of IL-6 production in the gastric mucosal tissue through the treatment of Thai basil, water thistle, and the composite extract of snow sage.
도 4는 타이바질, 물엉겅퀴 및 눈개승마의 복합추출물의 피부섬유아세포에서의 IL-6 억제능을 측정한 결과이다.Figure 4 is a result of measuring the IL-6 inhibitory ability in skin fibroblasts of the composite extract of Thai basil, water thistle and snow horse riding.
도 5는 타이바질, 물엉겅퀴 및 눈개승마의 복합추출물의 피부섬유아세포에서의 MMP-1 억제능을 측정한 결과이다.Figure 5 is the result of measuring the MMP-1 inhibitory ability in skin fibroblasts of the composite extract of Thai basil, water thistle and snow horse riding.
이하, 본 발명을 보다 상세하게 설명한다. Hereinafter, the present invention will be described in more detail.
본 발명은 타이바질(Ocimum basilicum), 물엉겅퀴(Cirsium nipponicum(Maxim.) Makino) 및 눈개승마(Aruncus dioicus)로 이루어진 군에서 선택된 둘 이상의 추출물을 유효성분으로 포함하는 염증질환 예방 또는 치료용 약학 조성물을 제공한다.The present invention Thai basil ( Ocimum basilicum ), water thistle ( Cirsium nipponicum (Maxim.) Makino), and snow riding horse ( Aruncus diioicus ) It provides a pharmaceutical composition for preventing or treating inflammatory diseases comprising two or more extracts selected from the group consisting of as an active ingredient.
상기 추출물은 The extract is
(1) 타이바질 : 눈개승마 = 1 : (1~6); (1) Thai basil: Snow riding = 1: (1~6);
(2) 타이바질 : 물엉겅퀴 = 1 : (2~6);(2) Thai basil: water thistle = 1: (2~6);
(3) 물엉겅퀴 : 타이바질 = 1 : (2~6);(3) Water thistle : Thai basil = 1 : (2~6);
(4) 물엉겅퀴 : 눈개승마 = 1 : (2~6) 또는(4) Water Thistle: Snow Riding = 1: (2~6) or
(5) 타이바질 : 물엉겅퀴 : 눈개승마 = (1~6) : (1~6) : 1로 이루어진 군에서 선택된 하나의 중량 비율로 추출한 것을 특징으로 하고, 바람직하게는 타이바질 : 물엉겅퀴 : 눈개승마 = 1 : (2~6) :1의 중량 비율로 추출한 것을 특징으로 하나, 이에 한정되는 것은 아니다.(5) Thai basil: Water thistle: It is characterized in that it is extracted in a weight ratio selected from the group consisting of (1-6): (1-6): 1, preferably Thai basil: Water thistle: It is characterized in that it is extracted in a weight ratio of snow riding = 1: (2 to 6): 1, but is not limited thereto.
또한, 상기 추출물은 물, (C1~C4) 알콜 또는 이의 혼합용매로 추출한 것을 특징으로 하고, 바람직하게는 물 또는 30% 에탄올로 추출한 것을 특징으로 하나, 이에 한정되는 것은 아니다.In addition, the extract is characterized in that it is extracted with water, (C1 ~ C4) alcohol or a mixed solvent thereof, preferably extracted with water or 30% ethanol, but is not limited thereto.
상기 염증질환은 급성 및 만성 위염, 소화성궤양, 십이지장염, 위궤양, 장염, 염증성 장질환, 궤양성 대장염, 크론병, 간염, 기관지염, 알레르기성 비염, 천식, 만성폐쇄성폐질환, 접촉성 피부염, 알레르기성 피부염, 아토피성 피부염, 아나필락시스, 지루성 피부염, 전신성 홍반성 루푸스, 경피증, 구내염, 퇴행성 과절염, 류마티스성 관절염, 신경염, 당뇨병성 망막증, 망막염, 황반 변성, 포도막염, 결막염, 강직성 척추염, 골관절염, 신염, 신장염, 쇼그렌 증후군, 자가 면역 췌장염, 치주질환, 만성 골반 염증 질환, 자궁내막염, 비염, 편도염, 중이염, 인후염, 방광염 및 만성 전립선염으로 이루어진 군에서 선택된 하나 이상일 수 있고, 바람직하게는 위염, 위궤양 및 피부염으로 이루어진 군에서 선택된 하나 이상일 수 있으나, 이에 한정되는 것은 아니다.The inflammatory diseases include acute and chronic gastritis, peptic ulcer, duodenitis, gastric ulcer, enteritis, inflammatory bowel disease, ulcerative colitis, Crohn's disease, hepatitis, bronchitis, allergic rhinitis, asthma, chronic obstructive pulmonary disease, contact dermatitis, allergy Dermatitis atopic, anaphylaxis, seborrheic dermatitis, systemic lupus erythematosus, scleroderma, stomatitis, osteoarthritis, rheumatoid arthritis, neuritis, diabetic retinopathy, retinitis, macular degeneration, uveitis, conjunctivitis, ankylosing spondylitis, osteoarthritis, nephritis , nephritis, Sjogren's syndrome, autoimmune pancreatitis, periodontal disease, chronic pelvic inflammatory disease, endometritis, rhinitis, tonsillitis, otitis media, sore throat, cystitis, and chronic prostatitis, and preferably one or more selected from the group consisting of gastritis and gastric ulcer And it may be one or more selected from the group consisting of dermatitis, but is not limited thereto.
또한, 상기 추출물은 IL-6(Interleukin 6) 및 TNF-α(tumor necrosis factor-α) 활성 억제를 통한 항염 효과가 있는 것을 특징으로 한다.In addition, the extract is characterized in that it has an anti-inflammatory effect through inhibition of IL-6 (Interleukin 6) and TNF-α (tumor necrosis factor-α) activity.
또한, 상기 추출물은 카탈라아제(catalase) 및 SOD(superoxide dismutase) 활성 억제를 통한 항산화 효과가 있는 것을 특징으로 한다.In addition, the extract is characterized in that it has an antioxidant effect through inhibition of catalase and SOD (superoxide dismutase) activity.
또한, 상기 추출물은 IL-6(Interleukin 6) 및 MMP-1(matrix metalloproteinase-1) 활성 억제를 통한 피부 보호 효과가 있는 것을 특징으로 한다.In addition, the extract is characterized in that it has a skin protection effect through inhibition of IL-6 (Interleukin 6) and MMP-1 (matrix metalloproteinase-1) activity.
본 발명의 약학 조성물은 당해 발명이 속하는 기술 분야에서 통상의 지식을 가진 자가 용이하게 실시할 수 있는 방법에 따라, 약제학적으로 허용되는 담체를 이용하여 제제화함으로써 단위 용량 형태로 제조되거나 또는 다용량 용기 내에 내입시켜 제조될 수 있다.The pharmaceutical composition of the present invention is prepared in a unit dose form or in a multi-dose container by formulating using a pharmaceutically acceptable carrier according to a method that can be easily performed by those skilled in the art. It can be prepared by incorporating into
상기 약제학적으로 허용되는 담체는 제제시에 통상적으로 이용되는 것으로서, 락토스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산 칼슘, 알기네이트, 젤라틴, 규산 칼슘, 미세결정성 셀룰로스, 폴리비닐피롤리돈, 셀룰로스, 물, 시럽, 메틸 셀룰로스, 메틸 히드록시벤조에이트, 프로필히드록시벤조에이트, 활석, 스테아르산 마그네슘 및 미네랄 오일 등을 포함하나, 이에 한정되는 것은 아니다. 본 발명의 약학 조성물은 상기 성분들 이외에 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제, 보존제 등을 추가로 포함할 수 있다.The pharmaceutically acceptable carrier is one commonly used in formulation, and includes lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methyl hydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like. The pharmaceutical composition of the present invention may further include lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, and the like, in addition to the above components.
본 발명에 있어서, 상기 약학 조성물에 포함되는 첨가제의 함량은 특별히 한정되는 것은 아니며 통상의 제제화에 사용되는 함량 범위 내에서 적절하게 조절될 수 있다.In the present invention, the content of the additives included in the pharmaceutical composition is not particularly limited and may be appropriately adjusted within the range of content used in conventional formulations.
상기 약학 조성물은 수용액, 현탁액, 유탁액 등과 같은 주사용 제형, 환약, 캡슐, 과립, 정제, 크림, 젤, 패취, 분무제, 연고제, 경고제, 로션제, 리니멘트제, 파스타제 및 카타플라스마제로 이루어진 군으로부터 선택되는 하나 이상의 외용제 형태로 제형화될 수 있다.The pharmaceutical composition is an aqueous solution, suspension, emulsion, and the like for injection, pills, capsules, granules, tablets, creams, gels, patches, sprays, ointments, warning agents, lotions, liniment agents, pasta agents, and cataplasma agents. It may be formulated in the form of one or more external agents selected from the group consisting of
본 발명의 약학 조성물은 제형화를 위해 추가로 있는 약학적으로 허용 가능한 담체 및 희석제를 포함할 수 있다. 상기 약학적으로 허용 가능한 담체 및 희석제는 전분, 당, 및 만니톨과 같은 부형제, 칼슘 포스페이트 등과 같은 충전제 및 증량제, 카르복시메틸셀룰로오스, 히드록시프로필셀룰로오스 등과 같은 셀룰로오스 유도체, 젤라틴, 알긴산염, 및 폴리비닐 피롤리돈 등과 같은 결합제, 활석, 스테아린산 칼슘, 수소화 피마자유 및 폴리에틸렌 글리콜과 같은 윤활제, 포비돈, 크로스포비돈과 같은 붕해제, 폴리소르베이트, 세틸알코올, 및 글리세롤 등과 같은 계면활성제를 포함하나, 이에 한정되지 않는다. 상기 약학적으로 허용 가능한 담체 및 희석제는 대상체에게 생물학적 및 생리학적으로 친화적인 것일 수 있다. 희석제의 예로는 염수, 수용성 완충액, 용매 및/또는 분산제(dispersion media)를 들 수 있으나, 이에 한정되는 것은 아니다.The pharmaceutical composition of the present invention may additionally contain pharmaceutically acceptable carriers and diluents for formulation. The pharmaceutically acceptable carriers and diluents include excipients such as starch, sugar, and mannitol, fillers and extenders such as calcium phosphate, cellulose derivatives such as carboxymethylcellulose and hydroxypropylcellulose, gelatin, alginates, and polyvinyl fibres. binders such as rolidone, etc., lubricants such as talc, calcium stearate, hydrogenated castor oil and polyethylene glycol, disintegrants such as povidone, crospovidone, surfactants such as polysorbates, cetyl alcohol, and glycerol; don't The pharmaceutically acceptable carrier and diluent may be biologically and physiologically compatible with the subject. Examples of diluents include, but are not limited to, saline, aqueous buffers, solvents and/or dispersion media.
본 발명의 약학 조성물은 목적하는 방법에 따라 경구 투여하거나 비경구 투여(예를 들어, 정맥 내, 피하, 복강 내 또는 국소에 적용)할 수 있다. 경구 투여일 경우, 정제, 트로키제(troches), 로젠지(lozenge), 수용성 현탁액, 유성 현탁액, 조제 분말, 과립, 에멀젼, 하드 캡슐, 소프트 캡슐, 시럽 또는 엘릭시르제 등으로 제형화될 수 있다. 비경구 투여일 경우, 주사액, 좌제, 호흡기 흡입용 분말, 스프레이용 에어로졸제, 연고, 도포용 파우더, 오일, 크림 등으로 제형화 될 수 있다.The pharmaceutical composition of the present invention may be administered orally or parenterally (eg, intravenous, subcutaneous, intraperitoneal or topical application) depending on the desired method. For oral administration, it may be formulated into tablets, troches, lozenges, aqueous suspensions, oily suspensions, powders, granules, emulsions, hard capsules, soft capsules, syrups or elixirs. In the case of parenteral administration, it may be formulated as an injection solution, suppository, powder for respiratory inhalation, aerosol for spray, ointment, powder for application, oil, cream, etc.
본 발명의 약학 조성물의 투여량은 환자의 상태 및 체중, 연령, 성별, 건강상태, 식이 체질 특이성, 제제의 성질, 질병의 정도, 조성물의 투여시간, 투여방법, 투여기간 또는 간격, 배설율, 및 약물 형태에 따라 그 범위가 다양할 수 있으며, 이 분야 통상의 기술자에 의해 적절하게 선택될 수 있다. 예컨대, 약 0.1 내지 10,000mg/kg의 범위일 수 있으나 이제 제한되지 않으며, 하루 일회 내지 수회에 나누어 투여될 수 있다.The dosage of the pharmaceutical composition of the present invention depends on the condition and weight of the patient, age, sex, health condition, dietary constitution specificity, the nature of the preparation, the severity of the disease, the administration time of the composition, the administration method, the administration period or interval, the excretion rate, And the range may vary depending on the type of drug, and may be appropriately selected by a person skilled in the art. For example, it may range from about 0.1 to 10,000 mg/kg, but is not limited thereto, and may be divided and administered once or several times a day.
상기 약학 조성물은 목적하는 방법에 따라 경구 투여되거나 비경구 투여(예를 들면, 정맥 내, 피하 내, 복강 내 또는 국소에 적용)될 수 있다. 본 발명의 약학 조성물의 약학 유효량, 유효 투여량은 약학 조성물의 제제화 방법, 투여 방식, 투여 시간 및/또는 투여 경로 등에 의해 다양해질 수 있으며, 당해 기술 분야에서 통상의 지식을 가진 자는 목적하는 치료에 효과적인 투여량을 용이하게 결정하고 처방할 수 있다. 본 발명의 약학 조성물의 투여는 하루에 1회 투여될 수 있고, 수회에 나누어 투여될 수도 있다.The pharmaceutical composition may be administered orally or parenterally (eg, intravenous, subcutaneous, intraperitoneal or topical application) depending on the desired method. The pharmaceutically effective amount and effective dose of the pharmaceutical composition of the present invention may vary depending on the formulation method, administration method, administration time and/or administration route of the pharmaceutical composition, and those skilled in the art may be able to use the desired treatment An effective dosage can be readily determined and prescribed. Administration of the pharmaceutical composition of the present invention may be administered once a day, or may be divided and administered several times.
상기 “항염”은 염증 개선 효과를 의미한다. 상기 “염증”은 생체나 조직에 물리적 작용이나 화학적 물질, 세균 감염 등의 어떠한 기질적 변화를 가져오는 침습이 가해질 때 그 손상부위를 수복 재생하려는 기전으로, 일단 자극이 가해지면 국소적으로 히스타민, 세로토닌, 브라디키닌, 프로스타글란딘, HETE(hydroxyeicosatetraenoic acid) 및 류코트리엔과 같은 혈관 활성 물질이 유리되어 혈관 투과성이 증대되면서 염증이 유발될 수 있다.The “anti-inflammatory” means an effect of improving inflammation. The "inflammation" is a mechanism for restoring and regenerating the damaged area when an invasion that causes any organic change such as physical action, chemical substance, bacterial infection, etc. is applied to a living body or tissue. Once stimulation is applied, histamine, Inflammation may be induced as vascular active substances such as serotonin, bradykinin, prostaglandin, HETE (hydroxyeicosatetraenoic acid), and leukotrienes are liberated to increase vascular permeability.
상기 “항산화”는 산화를 억제하는 작용을 의미하는 것으로, 인체는 산화촉 진물질과 산화억제물질이 균형을 이루고 있으나, 여러 가지 요인들로 인하여 이러한 균형 상태를 잃고 산화를 촉진하는 방향으로 기울게 되면, 생체 내에 산화적 스트레스(oxidative stress)가 유발되어 세포손상 및 병리적 질환을 유발할 수 있다.The “antioxidation” refers to an action of inhibiting oxidation. In the human body, oxidation promoters and antioxidants are in balance, but due to various factors, this balance is lost and tilted in the direction of promoting oxidation. , oxidative stress is induced in vivo, which can cause cell damage and pathological diseases.
또한, 본 발명은 타이바질(Ocimum basilicum), 물엉겅퀴(Cirsium nipponicum(Maxim.) Makino) 및 눈개승마(Aruncus dioicus)로 이루어진 군에서 선택된 둘 이상의 추출물을 유효성분으로 포함하는 염증질환 예방 또는 개선용 건강기능식품 조성물을 제공한다. In addition, the present invention Thai basil ( Ocimum basilicum ), water thistle ( Cirsium nipponicum (Maxim.) Makino), and snow riding horse ( Aruncus dioicus ) It provides a health functional food composition for preventing or improving inflammatory diseases containing two or more extracts selected from the group consisting of as an active ingredient.
상기 건강기능식품 조성물은 이너뷰티 푸드(inner beauty food)인 것을 특징으로 한다.The health functional food composition is characterized in that it is an inner beauty food.
본 발명은 통상적으로 이용되는 식품으로써 일반적으로 사용될 수 있다.The present invention can be generally used as a commonly used food.
본 발명의 식품 조성물은 건강기능식품으로서 사용될 수 있다. 상기 "건강기능 식품"이라 함은 건강기능 식품에 관한 법률에 따른 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 제조 및 가공한 식품을 의미하며, "기능성"이라 함은 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건용도에 유용한 효과를 얻을 목적으로 섭취하는 것을 의미한다.The food composition of the present invention can be used as a health functional food. The term "health functional food" refers to food manufactured and processed using raw materials or ingredients having useful functionalities for the human body in accordance with the Health Functional Food Act, and "functional" refers to food that is not related to the structure and function of the human body. It refers to intake for the purpose of obtaining useful effects for health purposes such as regulating nutrients or physiological functions.
상기 건강기능식품 조성물은 통상의 식품 첨가물을 포함할 수 있으며, 상기 "식품 첨가물"로서의 적합 여부는 다른 규정이 없는 한, 식품의약품안전처에 승인된 식품 첨가물 공전의 총칙 및 일반시험법 등에 따라 해당 품목에 관한 규격 및 기준에 의하여 판정한다.The health functional food composition may include conventional food additives, and the suitability as the "food additive" is determined in accordance with the General Rules and General Test Methods of Food Additives approved by the Ministry of Food and Drug Safety, unless otherwise specified. It is judged according to the specifications and standards for the item.
상기 "식품 첨가물 공전"에 수재된 품목으로는 예를 들어, 케톤류, 글리신, 구연산칼륨, 니코틴산, 계피산 등의 화학적 합성물, 감색소, 감초추출물, 결정셀룰로오스, 고량색소, 구아검 등의 천연첨가물, L-글루타민산나트륨 제제, 면류첨가알칼리제, 보존료제제, 타르색소제제 등의 혼합제제류들을 들 수 있다.Items listed in the "Food Additive Code" include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid, natural additives such as dark pigment, licorice extract, crystalline cellulose, goyang pigment, guar gum, and mixed preparations such as sodium L-glutamate preparations, noodle-added alkali preparations, preservative preparations, and tar color preparations.
본 발명의 식품 조성물은 정제, 캡슐, 분말, 과립, 액상, 환 등의 형태로 제조 및 가공할 수 있다. 예를 들어, 캡슐 형태의 건강기능 식품 중 경질 캡슐제는 통상의 경질 캡슐에본 발명에 따른 조성물을 부형제 등의 첨가제와 혼합 및 충진하여 제조할 수 있으며, 연질 캡슐제는 본 발명에 따른 조성물을 부형제 등의 첨가제와 혼합하고 젤라틴 등 캡슐기제에 충진하여 제조할 수 있다. 상기 연질 캡슐제는 필요에 따라 글리세린 또는 소르비톨 등의 가소제, 착색제, 보존제 등을 함유할 수 있다.The food composition of the present invention can be prepared and processed in the form of tablets, capsules, powders, granules, liquids, pills and the like. For example, among health functional foods in capsule form, hard capsules can be prepared by mixing and filling a composition according to the present invention with additives such as excipients in a conventional hard capsule, and soft capsules contain the composition according to the present invention. It can be prepared by mixing with additives such as excipients and filling in a capsule base such as gelatin. The soft capsule may contain a plasticizer such as glycerin or sorbitol, a coloring agent, a preservative, and the like, if necessary.
상기 부형제, 결합제, 붕해제, 활택제, 교미제, 착향제 등에 대한 용어 정의는 당업계에 공지된 문헌에 기재된 것으로 그 기능 등이 동일 내지 유사한 것들을 포함한다. 상기 식품의 종류에는 특별한 제한이 없으며, 통상적인 의미에서의 건강기능식품을 모두 포함한다.Definitions of terms for the excipients, binders, disintegrants, lubricants, corrigents, flavoring agents, etc. are described in literature known in the art, and include those having the same or similar functions. There is no particular limitation on the type of food, and it includes all health functional foods in the usual sense.
본 발명에서 용어 “예방”이란 본 발명에 따른 조성물의 투여로 질환의 억제 또는 지연시키는 모든 행위를 말한다. 본 발명에서 용어 “치료”는 본 발명에 따른 조성물의 투여로 질환의 증세가 호전되거나 이롭게 변경하는 모든 행위를 말한다. 본 발명에서 "개선"이란 본 발명의 조성물을 개체에 투여하거나 섭취시켜 질환의 나쁜 상태를 좋게 하는 모든 행위를 의미한다.In the present invention, the term "prevention" refers to all activities that inhibit or delay a disease by administering the composition according to the present invention. In the present invention, the term "treatment" refers to all activities that improve or beneficially change the symptoms of a disease by administering the composition according to the present invention. In the present invention, "improvement" means any action that improves the bad condition of a disease by administering or ingesting the composition of the present invention to a subject.
또한, 본 발명은 타이바질(Ocimum basilicum), 물엉겅퀴(Cirsium nipponicum(Maxim.) Makino) 및 눈개승마(Aruncus dioicus)로 이루어진 군에서 선택된 둘 이상의 추출물을 유효성분으로 포함하는 염증질환 예방 또는 개선용 화장료 조성물을 제공한다. In addition, the present invention Thai basil ( Ocimum basilicum ), water thistle ( Cirsium nipponicum (Maxim.) Makino) and snow riding horse ( Aruncus dioicus ) It provides a cosmetic composition for preventing or improving inflammatory diseases comprising two or more extracts selected from the group consisting of as an active ingredient.
상기 화장료 조성물은 스킨로션, 스킨소프너, 스킨토너, 아스트린젠트, 밀크로션, 모이스쳐로션, 영양로션, 마사지크림, 영양크림, 모이스쳐크림, 핸드크림, 파운데이션, 에센스, 영양에센스, 팩, 비누, 클렌징폼, 클렌징로션, 클렌징크림, 바디로션 및 바디클렌저로 이루어진 군에서 선택된 하나 이상의 제형을 갖는 것을 특징으로 하나, 이에 한정되는 것은 아니다.The cosmetic composition includes skin lotion, skin softener, skin toner, astringent, milk lotion, moisture lotion, nutrient lotion, massage cream, nutrient cream, moisture cream, hand cream, foundation, essence, nutrient essence, pack, soap, cleansing foam, It is characterized in that it has one or more formulations selected from the group consisting of cleansing lotion, cleansing cream, body lotion and body cleanser, but is not limited thereto.
구체적으로, 상기 화장료 조성물은 일반적인 유화 제형 및 가용화 제형으로 제조할 수 있다. 예컨대, 유연 화장수 또는 영양 화장수 등의 화장수; 훼이셜 로션, 바디로션 등의 유액; 영양 크림, 수분 크림, 아이 크림 등의 크림; 에센스; 화장연고; 스프레이; 젤; 팩; 선 스크린; 메이크업 베이스; 액체 타입, 고체 타입 또는 스프레이 타입 등의 파운데이션; 파우더; 클렌징 크림, 클렌징 로션, 클렌징 오일 등의 메이크업 제거제; 또는 클렌징 폼, 비누, 바디워시 등의 세정제로 제형화될 수 있으나 이에 한정되는 것은 아니다. 또한, 상기 피부 외용제는, 연고, 패치, 겔, 크림 또는 분무제로 제형화될 수 있으나 이에 한정되는 것은 아니다.Specifically, the cosmetic composition may be prepared in general emulsified and solubilized formulations. For example, cosmetic lotion, such as softening lotion or nourishing lotion; emulsions such as facial lotion and body lotion; creams such as nourishing creams, moisture creams, and eye creams; essence; cosmetic ointment; spray; gel; pack; sunscreen; makeup base; foundations such as liquid type, solid type or spray type; powder; makeup removers such as cleansing creams, cleansing lotions, and cleansing oils; Alternatively, it may be formulated as a cleanser such as a cleansing foam, soap, body wash, etc., but is not limited thereto. In addition, the external skin preparation may be formulated as an ointment, patch, gel, cream or spray, but is not limited thereto.
상기 화장료 조성물은 각각의 제형에 있어서 상기 필수성분 외에 제형의 종류 또는 사용 목적 등에 따라 본 발명에 따른 목적을 저해하지 않는 범위 내에서 다른 성분들이 적절히 배합될 수 있다.In addition to the essential components in each formulation, the cosmetic composition may be appropriately mixed with other components within a range that does not impair the purpose of the present invention depending on the type of formulation or purpose of use.
상기 화장료 조성물은 통상적으로 허용 가능한 담체를 포함할 수 있으며, 예컨대 유분, 물, 계면활성제, 보습제, 저급 알코올, 증점제, 킬레이트제, 색소, 방부제, 향료 등을 적절히 배합할 수 있으나, 이에 한정되는 것은 아니다.The cosmetic composition may include a conventionally acceptable carrier, and for example, oil, water, a surfactant, a moisturizer, a lower alcohol, a thickener, a chelating agent, a colorant, a preservative, a flavoring agent, and the like may be appropriately blended, but are not limited thereto. no.
상기 허용 가능한 담체는 제형에 따라 달리할 수 있다. 예컨대, 연고, 페이스트, 크림 또는 젤로 제형화될 때 담체 성분으로서 동물성 유, 식물성 유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오스 유도체, 폴리에틸렌글리콜, 실리콘, 벤토나이트, 실리카, 탈크, 산화아연 또는 이들의 추출물이 사용될 수 있다.The acceptable carrier may vary depending on the formulation. For example, animal oil, vegetable oil, wax, paraffin, starch, tracanthate, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide or any of these as a carrier component when formulated into an ointment, paste, cream or gel extracts may be used.
상기 화장료 조성물은 파우더 또는 스프레이로 제형화될 때, 담체 성분으로서 락토스, 탈크, 실리카, 알루미늄히드록사이드, 칼슘 실케이트, 폴리아미드 파우더 또는 이들의 추출물이 사용될 수 있고, 스프레이의 경우 클로로플루오로히드로카본, 프로판, 부탄 또는 디메틸 에테르와 같은 추진제를 더 포함할 수 있다.When the cosmetic composition is formulated as a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder or extracts thereof may be used as a carrier component, and in the case of a spray, chlorofluorohydro It may further contain a propellant such as carbon, propane, butane or dimethyl ether.
상기 화장료 조성물은 용액 또는 유탁액으로 제형화될 때, 담체 성분으로서 용매, 용해화제, 또는 유탁화제가 사용될 수 있고, 예컨대 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 벤조에이트, 프로필렌 글리콜, 1,3-브틸글리콜 오일이 사용될 수 있고, 특히, 목화씨 오일, 땅콩 오일, 옥수수 배종 오일, 올리브 오일, 피마자 오일 및 참깨 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜 또는 소르비탄의 지방산 에스테르가 사용될 수 있다.When the cosmetic composition is formulated as a solution or emulsion, a solvent, solubilizing agent, or emulsifying agent may be used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil may be used, and in particular, cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, fatty acid esters of glycerol, polyethylene glycol or sorbitan may be used.
상기 화장료 조성물은 현탁액으로 제형화될 때, 담체 성분으로서 물, 에탄올 또는 프로필렌 글리콜과 같은 액상의 희석제, 에톡실화 이소스테아릴 알코올, 폴리옥시에틸렌 소르비톨 에스테르 및 폴리 옥시에틸렌 소르비탄 에스테르와 같은 현탁제, 미소결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가 또는 트라칸트 등이 사용될 수 있다.When the cosmetic composition is formulated as a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, an ethoxylated isostearyl alcohol, a suspending agent such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tracanth and the like can be used.
상기 화장료 조성물이 비누로 제형화될 때, 담체 성분으로서 지방산의 알칼리 금속 염, 지방산 헤미에스테르염, 지방산 단백질 히드롤리제이트, 이세티오네이트, 라놀린 유도체, 지방족 알코올, 식물성 유, 글리세롤, 당 등이 사용될 수 있다.When the cosmetic composition is formulated as a soap, alkali metal salts of fatty acids, fatty acid hemiester salts, fatty acid protein hydrolyzates, isethionates, lanolin derivatives, fatty alcohols, vegetable oils, glycerol, sugars, etc. can be used
상기 화장료 조성물은 최종 제품의 품질이나 기능에 따라 업계에서 통상적으로 사용되는 지방 물질, 유기용매, 용해제, 농축제, 겔화제, 연화제, 항산화제, 현탁화제, 안정화제, 발포제(foaming agent), 방향제, 계면활성제, 물, 이온형 또는 비이온형 유화제, 충전제, 금속이온봉 쇄제, 킬레이트화제, 보존제, 차단제, 습윤화제, 필수 오일, 염료, 안료, 친수성 또는 친유성 활성제, 화장품에 통상적으로 사용되는 임의의 다른 성분과 같은 화장품 학 또는 피부과학 분야에서 통상적으로 사용되는 보조제를 추가적으로 함유할 수 있다.The cosmetic composition includes fatty substances, organic solvents, solubilizers, thickeners, gelling agents, softeners, antioxidants, suspending agents, stabilizers, foaming agents, and fragrances commonly used in the industry depending on the quality or function of the final product. , surfactants, water, ionic or nonionic emulsifiers, fillers, sequestering agents, chelating agents, preservatives, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives commonly used in cosmetics It may additionally contain adjuvants commonly used in cosmetology or dermatology, such as any other ingredient.
다만, 상기 보조제 및 그 혼합 비율은 본 발명에 따른 화장료 조성물의 바람직한 성질에 영향을 미치지 않도록 적절히 선택할 수 있다.However, the adjuvant and its mixing ratio may be appropriately selected so as not to affect desirable properties of the cosmetic composition according to the present invention.
이하, 본 발명의 이해를 돕기 위하여 실시예를 들어 상세하게 설명하기로 한다. 다만 하기의 실시예는 본 발명의 내용을 예시하는 것일 뿐 본 발명의 범위가 하기 실시예에 한정되는 것은 아니다. 본 발명의 실시예는 당업계에서 평균적인 지식을 가진 자에게 본 발명을 보다 완전하게 설명하기 위해 제공되는 것이다.Hereinafter, examples will be described in detail to aid understanding of the present invention. However, the following examples are merely illustrative of the contents of the present invention, but the scope of the present invention is not limited to the following examples. The embodiments of the present invention are provided to more completely explain the present invention to those skilled in the art.
[제조예 1] 원료 추출물 제조[Preparation Example 1] Raw material extract preparation
각 약재의 단일 또는 중량 추출 비율 조건; 및 주정(에탄올) 농도(0, 30, 70, 100%)의 조건에 따라, 추출물을 제조하였다. 보다 상세하게는 먼저, 각 생약을 중량 비율별로 각 조건에 따라 혼합한 뒤, 물 또는 30 내지 100%의 주정을 추출 용매로 선택하여 추출하였다. 멀티 히팅맨틀(WiseTherm)을 사용하여 추출액의 온도를 70℃로 유지하고, 환류 냉각장치를 저온 항온수조(A&D)와 연결하여 환류 냉각 추출한 추출액을 진공회전농축기(EYELA)를 활용하여 45℃의 수욕 및 200 mbar 이하의 압력에서 감압 농축하여 최종적으로 각 조건의 추출물을 수득하였다. 각 약재의 단일 또는 혼합 추출 비율 조건은 표 1과 같다.Single or weight extraction ratio conditions of each medicinal material; And according to the conditions of alcohol (ethanol) concentration (0, 30, 70, 100%), extracts were prepared. More specifically, first, each herbal medicine was mixed according to each condition by weight ratio, and then extracted by selecting water or 30 to 100% alcohol as an extraction solvent. The temperature of the extract is maintained at 70℃ using a multi-heating mantle (WiseTherm), and the reflux cooling device is connected to a low-temperature constant temperature water bath (A&D) to reflux-cool the extracted extract into a 45℃ water bath using a vacuum rotary concentrator (EYELA). And concentrated under reduced pressure at a pressure of 200 mbar or less to finally obtain an extract of each condition. Table 1 shows the single or mixed extraction ratio conditions of each herb.
실시예Example 타이바질Thai basil 눈개승마Snow Riding 물엉겅퀴water thistle 추출용매extraction solvent
1One 1One 　 　 물water
22 　 1One 　 물water
33 　 　 1One 물water
44 1One 1One 　 물water
55 1One 22 　 물 water
66 1One 44 　 물water
77 1One 66 　 물water
88 22 1One 　 물water
99 44 1One 　 물water
1010 66 1One 　 물 water
1111 1One 　 1One 물water
1212 1One 　 22 물water
1313 1One 　 44 물water
1414 1One 　 66 물 water
1515 22 　 1One 물water
1616 44 　 1One 물water
1717 66 　 1One 물water
1818 　 1One 1One 물water
1919 　 22 1One 물 water
2020 　 44 1One 물water
2121 　 66 1One 물water
2222 　 1One 22 물water
2323 　 1One 44 물water
2424 　 1One 66 물water
2525 1One 1One 1One 물water
2626 22 1One 1One 물water
2727 44 1One 1One 물water
2828 66 1One 1One 물water
2929 1One 22 1One 물water
3030 1One 44 1One 물water
3131 1One 66 1One 물water
3232 1One 1One 22 물water
3333 1One 1One 44 물water
3434 1One 1One 66 물water
항염증효능 검정에 사용된 시약인 lipopolysaccharide(LPS), 3-(4,5-dime thylthiazol-2-yl)- 2,5- diphenyltetrazolium(MTT) 및 dimethyl sulfoxide(DMSO)는 Sigma(St. Louis, MO, USA)에서 구입하였고, fetal bovine serum(FBS) 및 antibiotics는 Gibco/BRL(Eggenstein, Germany)에서 구입하여 사용하였으며, 1차 및 2차 antibody는 BD Bioscience(San Diego, USA), Cayman(Ann Arbor, USA) 및 Zymed(South San Francisco, USA)에서 각각 구입하였다. Tumor necrosis factor (TNF)-α, IL-1β 및 IL-6의 ELISA kit는 Pierce endogen(Rockford, IL., USA)에서 구입하였다. 그 외의 기타 시약은 특급 시약을 사용하였으며, UV/vis spectrophotometer(Hitachi, Japan), centrifuge (Hitachi, Japan), ELISA reader (Bio Rad, Japan) 등을 사용하여 측정하였다.Reagents used in the anti-inflammatory efficacy assay, lipopolysaccharide (LPS), 3-(4,5-dime thylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT), and dimethyl sulfoxide (DMSO) were purchased from Sigma (St. Louis, USA). MO, USA), fetal bovine serum (FBS) and antibiotics were purchased from Gibco/BRL (Eggenstein, Germany), and primary and secondary antibodies were purchased from BD Bioscience (San Diego, USA) and Cayman (Ann Arbor, USA) and Zymed (South San Francisco, USA), respectively. Tumor necrosis factor (TNF)-α, IL-1β, and IL-6 ELISA kits were purchased from Pierce Endogen (Rockford, IL., USA). For other reagents, special grade reagents were used, and measurements were performed using a UV/vis spectrophotometer (Hitachi, Japan), centrifuge (Hitachi, Japan), ELISA reader (Bio Rad, Japan), and the like.
[[ 실험예Experimental example 1] 항염증 효능 평가 1] Evaluation of anti-inflammatory efficacy
1) 항염증 효과 측정을 위한 세포배양1) Cell culture for measuring anti-inflammatory effect
Murine macrophage 세포주인 Raw 264.7 cells은 ATCC(American Type Culture Collection)에서 구입하였으며, Dulbecco's modified Eagle's medium(DMEM)에 10% fetal bovine serum(FBS), 100U/mL penicillin 및 100μg/mL streptomycin을 혼합한 배지를 사용하여 37℃, 5% CO2 incubator에서 배양하였다. 실험을 할 때는, 80%의 confluency와 20회 이하 passages 조건을 준수하였고, 실험 전 12시간은 FBS를 제거한 배지로 세포를 배양시켰다.Raw 264.7 cells, a murine macrophage cell line, were purchased from ATCC (American Type Culture Collection), and Dulbecco's modified Eagle's medium (DMEM) was mixed with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μg/mL streptomycin. 37 ℃, 5% CO 2 was cultured in an incubator. During the experiment, the conditions of 80% confluency and less than 20 passages were observed, and the cells were cultured in a medium without FBS for 12 hours before the experiment.
2) 일산화질소(Nitric Oxide) 측정2) Measurement of Nitric Oxide
일산화질소(Nitric Oxide, 이하 NO라 함) 측정은 세포의 상층액에서의 NO의 량을 아질산염(nitrite)과 질산염(nitrate)으로서 측정을 하였다. 아질산염에서 질산염으로는 산화된 후의 안전한 형태는 그리스시약(griess reagent; Sigma, USA)을 사용하여 측정하였다. 1×106개의 세포를 24시간 동안 배양하여 안정화 후에 normal군을 제외한 모든 well에 lipopolysaccharide(LPS) 500μM을 넣어서 자극시켰다. 1시간 후에 각각의 well에 실시예 1 내지 34의 타이바질, 눈개승마 및 물엉겅퀴 단일 또는 복합추출물을 200μg/mL 농도로 처리한 후, 24시간 인큐베이션 시켰다. NO 생성량은 시간별로 상층액을 모아 그리스시약(griess reagent)으로 20분간 차광시켜 반응시킨 후에 540㎚에서 흡광도로 측정하였다. NO 저해율은 하기 식에 따라 계산하였다.Nitric Oxide (hereinafter referred to as NO) was measured by measuring the amount of NO in the supernatant of cells as nitrite and nitrate. The safe form after oxidation from nitrite to nitrate was measured using a grease reagent (Sigma, USA). After stabilization by culturing 1×10 6 cells for 24 hours, 500 μM of lipopolysaccharide (LPS) was added to all wells except for the normal group to stimulate them. After 1 hour, the single or combined extracts of Examples 1 to 34 of Examples 1 to 34 were treated with 200 μg/mL concentration of Thai basil, snow sage, and water thistle, and then incubated for 24 hours. The amount of NO production was measured by absorbance at 540 nm after collecting the supernatant over time and reacting by blocking light with a grease reagent for 20 minutes. The NO inhibition rate was calculated according to the following formula.
[수학식 1][Equation 1]
Figure PCTKR2022019994-appb-img-000001
Figure PCTKR2022019994-appb-img-000001
NO 저해율을 측정한 결과, 모든 실시예에서 30% 이상의 NO 억제능을 보였고, 그 중 실시예 29~31이 뛰어난 NO 억제능을 보였으며, 특히 실시예 30이 86.9%로 가장 높은 NO 억제능을 보이는 것을 확인하였다(표 2).As a result of measuring the NO inhibition rate, it was confirmed that all examples showed NO inhibition ability of 30% or more, and among them, Examples 29 to 31 showed excellent NO inhibition ability, and in particular, Example 30 showed the highest NO inhibition ability at 86.9%. (Table 2).
실시예Example NO NO 억제능inhibitory ability (( %% )) 실시예Example NO NO 억제능inhibitory ability (( %% ))
1One 35.735.7 1818 37.237.2
22 48.748.7 1919 45.745.7
33 37.837.8 2020 45.645.6
44 48.648.6 2121 47.547.5
55 48.948.9 2222 30.930.9
66 54.854.8 2323 38.838.8
77 52.152.1 2424 39.139.1
88 45.045.0 2525 50.250.2
99 44.144.1 2626 49.849.8
1010 47.947.9 2727 52.652.6
1111 43.543.5 2828 50.950.9
1212 43.843.8 2929 61.561.5
1313 40.540.5 3030 86.986.9
1414 42.542.5 3131 77.277.2
1515 40.540.5 3232 50.650.6
1616 44.544.5 3333 30.730.7
1717 45.745.7 3434 45.445.4
3) TNF-α 생성량 측정 3) Measurement of TNF-α production
1×106개의 세포를 24시간 동안 배양하여 안정화 후에 normal군을 제외한 모든 well에 lipopolysaccharide (LPS) 500μM을 넣어서 자극시켰다. 1시간 후에 각각의 well에 실시예 1 내지 34의 타이바질, 눈개승마 및 물엉겅퀴 단일 또는 복합추출물을 200μg/mL의 농도로 24시간 처리하였고, 세포 배양액을 모아 TNF-α enzyme immunoassay(EIA) kit (R&D Systems Inc. Minneapolis, MN, USA)를 사용하여 측정하였다. TNF-α 저해율은 하기 식과 같이 계산하였다.After stabilization by culturing 1×10 6 cells for 24 hours, 500 μM of lipopolysaccharide (LPS) was added to all wells except for the normal group to stimulate them. After 1 hour, the single or combined extracts of Examples 1 to 34 of Examples 1 to 34 in each well were treated for 24 hours at a concentration of 200 μg/mL, and the cell culture medium was collected and TNF-α enzyme immunoassay (EIA) kit (R&D Systems Inc. Minneapolis, MN, USA). The TNF-α inhibition rate was calculated according to the following formula.
[수학식 2][Equation 2]
Figure PCTKR2022019994-appb-img-000002
Figure PCTKR2022019994-appb-img-000002
TNF-α 저해율을 측정한 결과, 실시예 29~31이 높은 TNF-α 억제능을 보였고, 특히 실시예 30은 75.8%로 가장 높은 TNF-α 억제능을 보이는 것을 확인하였다(표 3).As a result of measuring the TNF-α inhibition rate, it was confirmed that Examples 29 to 31 showed high TNF-α inhibitory ability, and in particular, Example 30 showed the highest TNF-α inhibitory ability at 75.8% (Table 3).
실시예Example TNFTNF -α -α 억제능inhibitory ability (( %% )) 실시예Example TNFTNF -α -α 억제능inhibitory ability (( %% ))
1One 24.524.5 1818 11.011.0
22 36.236.2 1919 38.438.4
33 18.918.9 2020 35.535.5
44 38.638.6 2121 37.237.2
55 35.635.6 2222 7.47.4
66 48.548.5 2323 10.510.5
77 42.742.7 2424 11.511.5
88 27.627.6 2525 44.544.5
99 26.726.7 2626 42.542.5
1010 27.127.1 2727 38.338.3
1111 10.510.5 2828 35.635.6
1212 30.030.0 2929 55.655.6
1313 31.531.5 3030 75.875.8
1414 28.828.8 3131 50.250.2
1515 20.720.7 3232 13.213.2
1616 21.521.5 3333 14.914.9
1717 24.224.2 3434 22.422.4
4) 추출용매별 NO 및 TNF-α억제능 측정4) Measurement of NO and TNF-α inhibitory ability for each extraction solvent
실시예 25~34(3종 복합추출물) 조건에서 추출 용매를 달리하여 NO 및 TNF-α 저해율에 대하여 비교 실험을 진행하였다. 추출 용매 조건 및 3종 복합추출물 중량 비율은 표 4와 같이 설정하여 비교예 1~30을 제조하였다.In Examples 25 to 34 (three composite extracts), a comparative experiment was conducted for NO and TNF-α inhibition rate by changing the extraction solvent. The extraction solvent conditions and the weight ratio of the three composite extracts were set as shown in Table 4 to prepare Comparative Examples 1 to 30.
비교예comparative example 타이바질Thai basil 눈개승마Snow Riding 물엉겅퀴water thistle 추출용매extraction solvent NO
억제능 (%)NO
Inhibiting ability (%) 		TNF-α
억제능 (%)TNF-α
Inhibiting ability (%)
1One 1One 1One 1One 30% 에탄올30% ethanol 48.948.9 40.940.9
22 22 1One 1One 30% 에탄올30% ethanol 48.148.1 40.440.4
33 44 1One 1One 30% 에탄올30% ethanol 50.950.9 36.136.1
44 66 1One 1One 30% 에탄올30% ethanol 52.652.6 30.630.6
55 1One 22 1One 30% 에탄올30% ethanol 58.958.9 55.455.4
66 1One 44 1One 30% 에탄올30% ethanol 83.183.1 70.070.0
77 1One 66 1One 30% 에탄올30% ethanol 75.175.1 50.950.9
88 1One 1One 22 30% 에탄올30% ethanol 48.948.9 12.412.4
99 1One 1One 44 30% 에탄올30% ethanol 31.531.5 10.610.6
1010 1One 1One 66 30% 에탄올30% ethanol 40.540.5 23.423.4
1111 1One 1One 1One 70% 에탄올70% ethanol 48.748.7 38.438.4
1212 22 1One 1One 70% 에탄올70% ethanol 45.645.6 40.940.9
1313 44 1One 1One 70% 에탄올70% ethanol 49.749.7 34.634.6
1414 66 1One 1One 70% 에탄올70% ethanol 53.453.4 29.729.7
1515 1One 22 1One 70% 에탄올70% ethanol 61.261.2 52.252.2
1616 1One 44 1One 70% 에탄올70% ethanol 70.970.9 50.050.0
1717 1One 66 1One 70% 에탄올70% ethanol 68.868.8 50.150.1
1818 1One 1One 22 70% 에탄올70% ethanol 45.745.7 14.014.0
1919 1One 1One 44 70% 에탄올70% ethanol 33.433.4 11.411.4
2020 1One 1One 66 70% 에탄올70% ethanol 38.738.7 25.425.4
2121 1One 1One 1One 100% 에탄올100% ethanol 45.745.7 35.135.1
2222 22 1One 1One 100% 에탄올100% ethanol 40.940.9 37.837.8
2323 44 1One 1One 100% 에탄올100% ethanol 45.745.7 33.033.0
2424 66 1One 1One 100% 에탄올100% ethanol 50.550.5 24.824.8
2525 1One 22 1One 100% 에탄올100% ethanol 50.150.1 50.650.6
2626 1One 44 1One 100% 에탄올100% ethanol 70.670.6 50.650.6
2727 1One 66 1One 100% 에탄올100% ethanol 65.465.4 51.251.2
2828 1One 1One 22 100% 에탄올100% ethanol 46.846.8 22.422.4
2929 1One 1One 44 100% 에탄올100% ethanol 30.430.4 24.724.7
3030 1One 1One 66 100% 에탄올100% ethanol 35.135.1 20.120.1
추출용매 조건별 NO 및 TNF-α 저해율 측정 결과, 물 추출물(실시예 25~34)이 조건별 에탄올 추출물보다 우수한 NO 및 TNF-α 억제능을 보였고, 에탄올 추출물 중에서는 30% 에탄올 추출물(비교예 1~10)이 다른 에탄올 추출 조건보다 우수한 억제능을 보였으며, 비교예 6이 각각 83.1%, 70.0% 억제 활성을 보여 에탄올 추출물 중 가장 우수한 억제 활성을 보이는 것을 확인하였다(표 4).As a result of measuring the NO and TNF-α inhibition rate for each extraction solvent condition, the water extract (Examples 25 to 34) showed superior NO and TNF-α inhibitory ability than the ethanol extract for each condition, and among the ethanol extracts, the 30% ethanol extract (Comparative Example 1 ~ 10) showed better inhibitory activity than other ethanol extraction conditions, and Comparative Example 6 showed 83.1% and 70.0% inhibitory activity, respectively, showing the best inhibitory activity among ethanol extracts (Table 4).
[[ 실험예Experimental example 2] 2] 인도메타신indomethacin 유발 위궤양 gastric ulcer 랫트rat 모델에서의 in the model 항궤양anti-ulcer 효과 effect
1) 시험동물1) Test animals
7주령 암컷 흰쥐(Sprage-Dawley rats)를 코아텍에서 구입하였다. 상기 흰쥐를 쥐 우리에 넣고 온도 23±3℃, 상대습도 55±15%, 환기횟수 10∼20회/hr, 조명시간 12시간(오전 8시 점등∼오후 8시 소등) 및 조도 150∼300Lux로 설정한 방에서 사육하였으며, 일반적인 사료(normal laboratory diet)를 섭취하도록 하였다.Seven-week-old female rats (Sprage-Dawley rats) were purchased from Coretech. The rats were placed in a rat cage at a temperature of 23 ± 3 ° C, relative humidity of 55 ± 15%, ventilation frequency of 10 to 20 times / hr, lighting time of 12 hours (8:00 am to 8:00 pm off), and illumination of 150 to 300 Lux. They were reared in a set room and fed a normal laboratory diet.
2) 위염 억제율 평가2) Evaluation of gastritis inhibition rate
상기 실험용 SD랫트를 48시간 절식시킨 후에 타이바질, 눈개승마 및 물엉겅퀴 단일 또는 복합추출물을 각각 체중기준으로 10mL/kg으로 300mg/kg 경구 투여하고, 1시간 후에 인도메타신(indomethacin) 80㎎/㎏을 경구투여 하였다. 양성대조군으로는 위염치료제로 사용하는 의약품 원료인 애엽 에탄올 연조엑스를 경구투여 하였다. 인도메타신 투여 5시간째에 동물을 마취하여 치사시켜 위를 적출하고 세척한 후에 위 점막면을 디지털카메라로 촬영하였다. 디지털카메라로 촬영한 병변 발생 위 점막면에서 손상된 부위의 면적 및 위 전체 면적을 ImageJ software(NIH, Bethesda, MD)를 이용하여 분석하고, Ulcer index(%)와 위염 억제율을 산출하였다. Ulcer index(%)와 위염 억제율은 하기 식에 따라 계산하였다.After the experimental SD rats were fasted for 48 hours, single or combined extracts of Thai basil, snow sage, and water thistle were orally administered at 10 mL/kg based on body weight and 300 mg/kg, respectively, and 1 hour later, indomethacin 80 mg/kg kg was orally administered. As a positive control group, ethanol soft extract, which is a pharmaceutical raw material used as a gastritis treatment, was orally administered. At 5 hours after indomethacin administration, the animals were killed by anesthesia, and the stomach was removed and washed, and the mucosal surface of the stomach was photographed with a digital camera. The area of the damaged area and the total area of the stomach on the mucosal surface of the lesion-prone gastric mucosa photographed with a digital camera were analyzed using ImageJ software (NIH, Bethesda, MD), and the Ulcer index (%) and gastritis inhibition rate were calculated. Ulcer index (%) and gastritis inhibition rate were calculated according to the following formula.
[수학식 3][Equation 3]
Ulcer index(Ulcer index ( %% ) = 손상면적 / 전체면적 X 100) = damaged area / total area X 100
위염 억제율(Gastritis inhibition rate ( %% ) = [ 1 - () = [ 1 - ( 경구투여군의oral administration group ulcer index / ulcer index / 위염유발군의Gastritis provoking group ulcer index) ] X 100 ulcer index)] X 100
실시예Example 타이바질Thai basil 눈개승마Snow Riding 물엉겅퀴water thistle Ulcer index(%)Ulcer index (%) 위염 억제율Gastritis inhibition rate
위염 유발군gastritis-causing group 7.2±3.07.2±3.0 --
1One 1One 　 　 4.3 ±2.8*4.3±2.8* 40.440.4
22 　 1One 　 4.9 ±2.94.9±2.9 31.231.2
33 　 　 1One 4.9 ±1.54.9±1.5 31.531.5
2525 1One 1One 1One 2.6 ±3.2**2.6±3.2** 63.163.1
2929 1One 22 1One 1.7 ±1.0**1.7±1.0** 75.775.7
3030 1One 44 1One 1.0 ±0.8**1.0 ±0.8** 85.985.9
3131 1One 66 1One 2.3 ±1.2**2.3 ±1.2** 68.268.2
애엽 에탄올 연조엑스(양성 대조군)Ayeop ethanol softened extract (positive control) 2.4 ±1.7**2.4 ±1.7** 66.566.5
Ulcer index(%) 및 위염 억제율 측정 결과, 실시예 29~31이 양성 대조군보다 낮은 Ulcer index(%) 및 높은 위염 억제율을 보였고, 그 중 실시예 30이 가장 우수한 효과를 보이는 것을 확인하였다(표 5).As a result of measuring the Ulcer index (%) and gastritis inhibition rate, Examples 29 to 31 showed lower Ulcer index (%) and higher gastritis inhibition rate than the positive control group, and it was confirmed that Example 30 showed the most excellent effect (Table 5). ).
3) 위 점막 PEG2(Prostaglandin E2) 측정3) Gastric mucosal PEG2 (Prostaglandin E2) measurement
적출된 위의 일부는 일정한 무게로 잘라 RIPA buffer(Cell Signaling Technology, Danvers, MA, USA)에 녹여 단백질을 분리한 뒤 원심분리기로 4℃, 13,000 × g 조건에서 10분간 원심분리 하였고, 일정한 양의 단백질 용해액을 PGE2 ELISA kit(Cusabio Biotech, Wuhan, China)를 통해 측정하였다. A portion of the extracted stomach was cut to a certain weight, dissolved in RIPA buffer (Cell Signaling Technology, Danvers, MA, USA) to separate proteins, and then centrifuged for 10 minutes at 4℃ and 13,000 × g in a centrifuge. Protein lysates were measured using a PGE2 ELISA kit (Cusabio Biotech, Wuhan, China).
위 점막 PEG2 측정 결과, 실시예 30이 140.9 pg/mg/protein로 양성대조군을 포함한 모든 실험군 중에서 가장 높은 값을 보이는 것을 확인하였다(도 1). As a result of gastric mucosal PEG2 measurement, it was confirmed that Example 30 showed the highest value among all experimental groups including the positive control group at 140.9 pg/mg/protein (FIG. 1).
4) 위 조직내 항산화 효소 측정4) Measurement of antioxidant enzymes in gastric tissue
위 조직에서의 산화적 스트레스에 대한 보호기전을 확인하고자 대표적인 항산화 효소인 카탈라아제(catalase)와 SOD(superoxide dismutase)의 활성을 측정하였다. 액체질소로 동결된 위 조직 약 100㎎에 100mM sodium phosphate buffer(pH 7.4) 1㎖를 넣어 homogenizer(TissueLyser LT, Qiagen, USA)로 균질화한 후 균질액을 4℃, 4,000rpm에서 4분간 원심분리한 다음, 상층액을 두 개로 나누어 취하였다. 하나는 SOD assay kit(Biomax, korea)를 이용하여 제조사의 프로토콜을 따라 SOD 활성을 측정하였고, 다른 하나는 4℃, 10,000rpm에서 15분간 원심분리하여 상층액을 취한 후 CAT assay kit(Biomax, korea)를 이용하여 제조사 프로토콜에 따라 측정하였다.In order to confirm the protective mechanism against oxidative stress in gastric tissue, the activities of catalase and superoxide dismutase (SOD), which are representative antioxidant enzymes, were measured. 1 ml of 100 mM sodium phosphate buffer (pH 7.4) was added to about 100 mg of gastric tissue frozen in liquid nitrogen, homogenized with a homogenizer (TissueLyser LT, Qiagen, USA), and the homogenate was centrifuged at 4°C and 4,000 rpm for 4 minutes. Next, the supernatant was divided into two parts and taken. In one, SOD activity was measured using the SOD assay kit (Biomax, Korea) according to the manufacturer's protocol, and in the other, the supernatant was centrifuged at 4℃ and 10,000 rpm for 15 minutes, and then the CAT assay kit (Biomax, Korea) was used. ) was measured according to the manufacturer's protocol.
실시예Example 타이바질Thai basil 눈개승마Snow Riding 물엉겅퀴water thistle SOD activity SOD activity
(U/g of protein)(U/g of protein) 		CatalaseCatalase activity activity
(U/g of protein)(U/g of protein)
VehicleVehicle 68.3±3.868.3±3.8 11.2±4.011.2±4.0
1One 1One 　 　 80.6±5.580.6±5.5 19.5±2.219.5±2.2
22 　 1One 　 82.1±5.182.1±5.1 20.5±3.420.5±3.4
33 　 　 1One 83.7±4.083.7±4.0 16.8±2.616.8±2.6
2525 1One 1One 1One 97.5±7.197.5±7.1 21.5±3.521.5±3.5
2929 1One 22 1One 99.1±3.799.1±3.7 22.4±4.022.4±4.0
3030 1One 44 1One 116.9±5.7116.9±5.7 30.4±3.530.4±3.5
3131 1One 66 1One 104.5±4.6104.5±4.6 25.5±1.525.5±1.5
양성대조군 : 애엽 에탄올 연조엑스Positive control group: Ayeop ethanol soft-textured extract 100.1±6.2100.1±6.2 22.1±0.522.1±0.5
카탈라아제(catalase) 및 SOD(superoxide dismutase)의 활성 측정 결과, 실시예 25, 29, 30 및 31에서 양성대조군과 비슷한 SDO 및 카탈라아제 활성을 나타냈고, 특히 실시예 30은 SOD 및 카탈라아제 활성 모두에서 양성대조군을 포함한 모든 실험군 중 가장 높은 활성을 보이는 것을 확인하였다(표 6). As a result of measuring the activities of catalase and SOD (superoxide dismutase), Examples 25, 29, 30 and 31 showed SDO and catalase activities similar to those of the positive control group, and in particular, Example 30 was a positive control group in both SOD and catalase activities. It was confirmed that it showed the highest activity among all experimental groups including (Table 6).
5) 위 조직내 항염증 인자 측정5) Measurement of anti-inflammatory factors in gastric tissue
항염증 효과를 확인하기 위해, 위 점막 조직 내 항염증 인자인 TNF-α 및 IL-6를 측정하였다. 액체질소로 동결된 위 조직으로부터 세포질을 얻기 위해 100mM Tris-HCl(pH 7.4), 5mM Tris-HCl(pH 7.5), 2mM MgCl2, 15mM CaCl2, 1.5M sucrose 및 0.1M DTT protease inhibitor cocktail을 첨가한 buffer A를 넣고 tissue grinder(Bio Spec Product, USA)로 분쇄한 후 10% NP-40 용액을 첨가하였다. 아이스 위에서 20분간 정치시킨 후 12,000 rpm으로 2분간 원심분리 하여 세포질을 포함하고 있는 상층액을 분리하였다. 이 상층액을 이용하여 TNF-α 및 IL-6 enzyme immunoassay(EIA) kit (RayBiotech, Norcross, GA, USA)로 측정하였다. 양성대조군으로 애엽 에탄올 연조엑스를 사용하였다.To confirm the anti-inflammatory effect, anti-inflammatory factors TNF-α and IL-6 in gastric mucosal tissue were measured. 100mM Tris-HCl (pH 7.4), 5mM Tris-HCl (pH 7.5), 2mM MgCl 2 , 15mM CaCl 2 , 1.5M sucrose and 0.1M DTT protease inhibitor cocktail were added to obtain cytoplasm from gastric tissue frozen in liquid nitrogen. After adding one buffer A and grinding with a tissue grinder (Bio Spec Product, USA), a 10% NP-40 solution was added. After standing on ice for 20 minutes, the supernatant containing cytoplasm was separated by centrifugation at 12,000 rpm for 2 minutes. TNF-α and IL-6 enzyme immunoassay (EIA) kit (RayBiotech, Norcross, GA, USA) were used to measure the supernatant. As a positive control group, ethanol soft extract was used.
TNF-α 측정 결과, 실시예 25, 29, 30 및 31이 양성대조군보다 낮은 TNF-α 값을 보였고, 그 중 실시예 30이 20.3pg/mg/protein으로 가장 낮은 값을 보이는 것을 확인하였다(도 2).As a result of TNF-α measurement, it was confirmed that Examples 25, 29, 30, and 31 showed lower TNF-α values than the positive control group, and among them, Example 30 showed the lowest value at 20.3 pg/mg/protein (Fig. 2).
IL-6 측정 결과, 실시예 29, 30 및 31이 양성대조군보다 낮은 IL-6 값을 보였고, 그 중 실시예 30이 107.7pg/mg/protein으로 가장 낮은 값을 보이는 것을 확인하였다(도 3).As a result of the IL-6 measurement, it was confirmed that Examples 29, 30, and 31 showed lower IL-6 values than the positive control group, and among them, Example 30 showed the lowest value at 107.7 pg/mg/protein (FIG. 3) .
[[ 실험예Experimental example 3] 피부보호 효과 측정 3] Measurement of skin protection effect
1) 피부보호 효과 확인을 위한 세포배양 1) Cell culture to confirm skin protection effect
인간피부 섬유아세포주인 CCD986sk는 ATCC(American Type Culture Collection)에서 구입하였으며, Iscove Modified Dulbecco Media(IMDM)에 10% fetal bovine serum(FBS)를, 100U/mL penicillin 및 100μg/mL streptomycin을 혼합한 배지를 사용하여 37℃, 5% CO2 incubator에서 배양하였다. 실험을 할 때는, 80%의 confluency와 10회 이하 passages 조건을 준수하여 실험 시, 실험 전 12시 간은 FBS를 제거한 배지로 세포를 배양시켰다.CCD986sk, a human skin fibroblast cell line, was purchased from ATCC (American Type Culture Collection), and Iscove Modified Dulbecco Media (IMDM) was mixed with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μg/mL streptomycin. 37 ℃, 5% CO 2 was cultured in an incubator. When performing the experiment, the cells were cultured in a medium without FBS for 12 hours before the experiment in compliance with the conditions of 80% confluency and less than 10 passages.
2) IL-6(Interleukin 6) 생성량 측정2) Measurement of IL-6 (Interleukin 6) production
전염증성 사이토카인인 IL-6를 측정하기 위해, 1×106개의 세포를 24시간동안 배양하여 안정화 후에 normal군을 제외한 모든 well에 TNF-a 20ng/ml을 넣어서 자극시켰다. 1시간 후에 각각의 well에 타이바질, 눈개승마 및 물엉겅퀴 단일 또는 복합추출물을 200μg/mL의 농도로 처리한 후, 24시간 인큐베이션 시켰다. IL-6 생성량 측정은 세포 배양액을 모아 IL-6 enzyme immunoassay(EIA) kit(R&D Systems Inc. Minneapolis, MN, USA)를 이용하여 제조사 프로토콜에 따라 측정하였다.In order to measure IL-6, a pro-inflammatory cytokine, 1×10 6 cells were cultured for 24 hours and then stimulated by adding 20 ng/ml of TNF-a to all wells except for the normal group. After 1 hour, each well was treated with single or combined extracts of 200 μg/mL of Thai basil, snow sage, and water thistle, and then incubated for 24 hours. The amount of IL-6 produced was measured by collecting the cell culture medium and using an IL-6 enzyme immunoassay (EIA) kit (R&D Systems Inc. Minneapolis, MN, USA) according to the manufacturer's protocol.
IL-6 생성량 측정 결과, 실시예 30이 81.8%로 무처리군을 포함한 모든 실험군 중에서 가장 적은 IL-6 생성량을 보이는 것을 확인하였다(도 4).As a result of measuring the amount of IL-6 production, it was confirmed that Example 30 showed the lowest amount of IL-6 production among all experimental groups including the untreated group at 81.8% (FIG. 4).
3) MMP-1(matrix metalloproteinase-1) 생성량 측정3) Measurement of MMP-1 (matrix metalloproteinase-1) production
피부보호 효과를 확인하기 위하여, 피부 콜라겐 분해효소인 MMP-1(matrix metalloproteinase-1) 분비량을 측정하였다. 1×106개의 세포를 24시간 동안 배양하여 안정화 후에 normal군을 제외한 모든 well에 TNF-a 20ng/ml을 넣어서 자극시켰다. 1시간 후에 각각의 well에 타이바질, 눈개승마 및 물엉겅퀴 단일 또는 복합추출물을 200μg/mL의 농도로 처리한 후, 24시간 인큐베이션 시켰다. 세포 배양액을 모아 Human MMP1 ELISA Kit(Abcam, Cambridge, MA, USA)를 이용하여 제조사 프로토콜에 따라 측정하였다.In order to confirm the skin protection effect, the amount of MMP-1 (matrix metalloproteinase-1) secretion, which is a skin collagen degrading enzyme, was measured. After stabilization by culturing 1×10 6 cells for 24 hours, 20 ng/ml of TNF-a was added to all wells except for the normal group to stimulate them. After 1 hour, each well was treated with single or combined extracts of 200 μg/mL of Thai basil, snow sage, and water thistle, and then incubated for 24 hours. The cell culture medium was collected and measured using the Human MMP1 ELISA Kit (Abcam, Cambridge, MA, USA) according to the manufacturer's protocol.
MMP-1 측정 결과, 실시예 30이 101.3%로 가장 적은 MMP-1 생성량을 보이는 것을 확인하였다(도 5).As a result of MMP-1 measurement, it was confirmed that Example 30 showed the smallest amount of MMP-1 production at 101.3% (FIG. 5).
이상으로 본 발명의 특정한 부분을 상세히 기술한 바, 당업계의 통상의 지식을 가진 자에게 있어서 이러한 구체적인 기술은 단지 바람직한 구현 예일 뿐이며, 이에 본 발명의 범위가 제한되는 것이 아닌 점은 명백하다. 따라서, 본 발명의 실질적인 범위는 첨부된 청구항과 그의 등가물에 의하여 정의된다고 할 것이다.Having described specific parts of the present invention in detail above, it is clear that these specific techniques are only preferred embodiments for those skilled in the art, and the scope of the present invention is not limited thereto. Accordingly, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.
본 발명의 범위는 후술하는 특허 청구범위에 의하여 나타내어지며, 특허 청구범위의 의미 및 범위 그리고 그 균등개념으로부터 도출되는 모든 변경 또는 변형된 형태가 본 발명의 범위에 포함되는 것으로 해석되어야 한다.The scope of the present invention is indicated by the following claims, and all changes or modifications derived from the meaning and scope of the claims and equivalent concepts thereof should be construed as being included in the scope of the present invention.

Claims (11)

  1. 타이바질(Ocimum basilicum), 물엉겅퀴(Cirsium nipponicum(Maxim.) Makino) 및 눈개승마(Aruncus dioicus)로 이루어진 군에서 선택된 둘 이상의 추출물을 유효성분으로 포함하는 염증질환 예방 또는 치료용 약학 조성물.Thai basil ( Ocimum basilicum ), water thistle ( Cirsium A pharmaceutical composition for the prevention or treatment of inflammatory diseases, comprising two or more extracts selected from the group consisting of nipponicum (Maxim.) Makino) and snow horse riding ( Aruncus dioicus ) as an active ingredient.
  2. 청구항 1에 있어서, 상기 추출물은 The method according to claim 1, wherein the extract
    (1) 타이바질 : 눈개승마 = 1 : (1~6); (1) Thai basil: Snow riding = 1: (1~6);
    (2) 타이바질 : 물엉겅퀴 = 1 : (2~6);(2) Thai basil: water thistle = 1: (2~6);
    (3) 물엉겅퀴 : 타이바질 = 1 : (2~6);(3) Water thistle : Thai basil = 1 : (2~6);
    (4) 물엉겅퀴 : 눈개승마 = 1 : (2~6) 및(4) Water thistle: riding snow = 1: (2~6) and
    (5) 타이바질 : 물엉겅퀴 : 눈개승마 = (1~6) : (1~6) : 1로 이루어진 군에서 선택된 하나의 중량 비율로 추출한 것을 특징으로 하는 염증질환 예방 또는 치료용 약학 조성물.(5) Thai basil: Water thistle: Snow sage = (1 to 6): (1 to 6): A pharmaceutical composition for preventing or treating inflammatory diseases, characterized by being extracted in a weight ratio selected from the group consisting of 1.
  3. 청구항 1 또는 2에 있어서, 상기 추출물은 물, (C1~C4) 알콜 또는 이의 혼합용매로 추출한 것을 특징으로 하는 염증질환 예방 또는 치료용 약학 조성물.The pharmaceutical composition for preventing or treating inflammatory diseases according to claim 1 or 2, wherein the extract is extracted with water, (C1-C4) alcohol or a mixed solvent thereof.
  4. 청구항 1에 있어서, 상기 염증질환은 급성 및 만성 위염, 소화성궤양, 십이지장염, 위궤양, 장염, 염증성 장질환, 궤양성 대장염, 크론병, 간염, 기관지염, 알레르기성 비염, 천식, 만성폐쇄성폐질환, 접촉성 피부염, 알레르기성 피부염, 아토피성 피부염, 아나필락시스, 지루성 피부염, 전신성 홍반성 루푸스, 경피증, 구내염, 퇴행성 과절염, 류마티스성 관절염, 신경염, 당뇨병성 망막증, 망막염, 황반 변성, 포도막염, 결막염, 강직성 척추염, 골관절염, 신염, 신장염, 쇼그렌 증후군, 자가 면역 췌장염, 치주질환, 만성 골반 염증 질환, 자궁내막염, 비염, 편도염, 중이염, 인후염, 방광염 및 만성 전립선염으로 이루어진 군에서 선택된 하나 이상인 것을 특징으로 하는 염증질환 예방 또는 치료용 약학 조성물.The method according to claim 1, wherein the inflammatory disease is acute and chronic gastritis, peptic ulcer, duodenitis, gastric ulcer, enteritis, inflammatory bowel disease, ulcerative colitis, Crohn's disease, hepatitis, bronchitis, allergic rhinitis, asthma, chronic obstructive pulmonary disease, Contact dermatitis, allergic dermatitis, atopic dermatitis, anaphylaxis, seborrheic dermatitis, systemic lupus erythematosus, scleroderma, stomatitis, degenerative arthritis, rheumatoid arthritis, neuritis, diabetic retinopathy, retinitis, macular degeneration, uveitis, conjunctivitis, ankylosing At least one selected from the group consisting of spondylitis, osteoarthritis, nephritis, nephritis, Sjogren's syndrome, autoimmune pancreatitis, periodontal disease, chronic pelvic inflammatory disease, endometritis, rhinitis, tonsillitis, otitis media, sore throat, cystitis and chronic prostatitis A pharmaceutical composition for preventing or treating inflammatory diseases.
  5. 청구항 1에 있어서, 상기 추출물은 IL-6(Interleukin 6) 및 TNF-α(tumor necrosis factor-α) 활성 억제를 통한 항염 효과가 있는 것을 특징으로 하는 염증질환 예방 또는 치료용 약학 조성물.The pharmaceutical composition for preventing or treating inflammatory diseases according to claim 1, wherein the extract has an anti-inflammatory effect by inhibiting IL-6 (Interleukin 6) and TNF-α (tumor necrosis factor-α) activity.
  6. 청구항 1에 있어서, 상기 추출물은 카탈라아제(catalase) 및 SOD(superoxide dismutase) 활성 억제를 통한 항산화 효과가 있는 것을 특징으로 하는 염증질환 예방 또는 치료용 약학 조성물.The pharmaceutical composition for preventing or treating inflammatory diseases according to claim 1, wherein the extract has an antioxidant effect through inhibition of catalase and superoxide dismutase (SOD) activity.
  7. 청구항 1에 있어서, 상기 추출물은 IL-6(Interleukin 6) 및 MMP-1(matrix metalloproteinase-1) 활성 억제를 통한 피부 보호 효과가 있는 것을 특징으로 하는 염증질환 예방 또는 치료용 약학 조성물.The pharmaceutical composition for preventing or treating inflammatory diseases according to claim 1, wherein the extract has a skin protection effect by inhibiting IL-6 (Interleukin 6) and MMP-1 (matrix metalloproteinase-1) activity.
  8. 타이바질(Ocimum basilicum), 물엉겅퀴(Cirsium nipponicum(Maxim.) Makino) 및 눈개승마(Aruncus dioicus)로 이루어진 군에서 선택된 둘 이상의 추출물을 유효성분으로 포함하는 염증질환 예방 또는 개선용 건강기능식품 조성물.Thai basil ( Ocimum basilicum ), water thistle ( Cirsium nipponicum (Maxim.) Makino) and snow horse riding ( Aruncus dioicus ) A health functional food composition for preventing or improving inflammatory diseases comprising two or more extracts selected from the group consisting of as active ingredients.
  9. 청구항 8에 있어서, 상기 건강기능식품 조성물은 이너뷰티 푸드(inner beauty food)인 것을 특징으로 하는 염증질환 예방 또는 개선용 건강기능식품. The health functional food for preventing or improving inflammatory diseases according to claim 8, wherein the health functional food composition is an inner beauty food.
  10. 타이바질(Ocimum basilicum), 물엉겅퀴(Cirsium nipponicum(Maxim.) Makino) 및 눈개승마(Aruncus dioicus)로 이루어진 군에서 선택된 둘 이상의 추출물을 유효성분으로 포함하는 염증질환 예방 또는 개선용 화장료 조성물.Thai basil ( Ocimum basilicum ), water thistle ( Cirsium A cosmetic composition for preventing or improving inflammatory diseases comprising two or more extracts selected from the group consisting of nipponicum (Maxim.) Makino) and snow horse riding ( Aruncus dioicus ) as an active ingredient.
  11. 청구항 10에 있어서, 상기 화장료 조성물은 스킨로션, 스킨소프너, 스킨토너, 아스트린젠트, 밀크로션, 모이스쳐로션, 영양로션, 마사지크림, 영양크림, 모이스쳐크림, 핸드크림, 파운데이션, 에센스, 영양에센스, 팩, 비누, 클렌징폼, 클렌징로션, 클렌징크림, 바디로션 및 바디클렌저로 이루어진 군에서 선택된 하나 이상의 제형을 갖는 것을 특징으로 하는 염증질환 예방 또는 개선용 화장료 조성물.The cosmetic composition of claim 10, wherein the cosmetic composition is skin lotion, skin softener, skin toner, astringent, milk lotion, moisture lotion, nutrient lotion, massage cream, nutrient cream, moisture cream, hand cream, foundation, essence, nutrient essence, pack, A cosmetic composition for preventing or improving inflammatory diseases, characterized in that it has at least one formulation selected from the group consisting of soap, cleansing foam, cleansing lotion, cleansing cream, body lotion and body cleanser.
PCT/KR2022/019994 2022-01-26 2022-12-09 Composition for preventing or treating inflammatory diseases comprising composite extract of ocimum basilicum, cirsium nipponicum (maxim.) makino, and aruncus dioicus as active ingredient WO2023146123A1 (en)

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Citations (5)

* Cited by examiner, † Cited by third party
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KR20120009554A (en) * 2010-07-19 2012-02-02 웅진코웨이주식회사 Cosmetic composition comprising mixed herbal extracts having anti-oxidant activity and anti-allergic activity
JP2013514994A (en) * 2009-12-18 2013-05-02 エクソドス ライフ サイエンシーズ リミテッド パートナーシップ Methods and compositions for treating skin inflammation
KR101869353B1 (en) * 2017-08-23 2018-06-21 재단법인 제주테크노파크 Novel Compound Isolated from Aruncus dioicus and Anti-inflammation Composition Using the Same
KR20210087406A (en) * 2020-01-02 2021-07-12 주식회사 엘지생활건강 Compositions Containing Plant Extracts
KR20210087405A (en) * 2020-01-02 2021-07-12 주식회사 엘지생활건강 Compositions Containing Plant Extracts

Patent Citations (5)

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Publication number Priority date Publication date Assignee Title
JP2013514994A (en) * 2009-12-18 2013-05-02 エクソドス ライフ サイエンシーズ リミテッド パートナーシップ Methods and compositions for treating skin inflammation
KR20120009554A (en) * 2010-07-19 2012-02-02 웅진코웨이주식회사 Cosmetic composition comprising mixed herbal extracts having anti-oxidant activity and anti-allergic activity
KR101869353B1 (en) * 2017-08-23 2018-06-21 재단법인 제주테크노파크 Novel Compound Isolated from Aruncus dioicus and Anti-inflammation Composition Using the Same
KR20210087406A (en) * 2020-01-02 2021-07-12 주식회사 엘지생활건강 Compositions Containing Plant Extracts
KR20210087405A (en) * 2020-01-02 2021-07-12 주식회사 엘지생활건강 Compositions Containing Plant Extracts

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