KR20210142513A - Pharmaceutical Composition comprising Sorbaria kirilowii extract for Preventing or Treating Inflammatory disease as active ingredient - Google Patents

Abstract

The present invention relates to a composition for the prevention, alleviation or treatment of inflammatory diseases containing a Sorbaria sorbifolia extract as an active component, wherein the Sorbaria sorbifolia extract has an excellent anti-inflammatory effect, thereby being able to be widely used as an alleviation, prevention, or therapeutic agent for various inflammatory diseases.

Description

Pharmaceutical composition comprising Sorbaria kirilowii extract for Preventing or Treating Inflammatory disease as active ingredient}

The present invention relates to a composition for the prevention, improvement or treatment of inflammatory diseases, and the like, comprising an extract of Jomshitan tree as an active ingredient.

Inflammation refers to the pathological condition of an abscess formed by the invasion of external infectious agents (bacteria, fungi, viruses, and various types of allergens). Specifically, when external bacteria invade and proliferate in a specific tissue, the leukocytes of the living body recognize it and actively attack the proliferated external bacteria. At the same time, the cell debris of the invading bacteria killed by the white blood cells melts into the invaded tissue to form an abscess. The treatment of abscesses caused by inflammation can be promoted through anti-inflammatory action, which means using antibacterial agents to suppress the growth of invading bacteria or to activate macrophages that phagocytose the foreign substances accumulated during the abscess to digest the foreign substances. and promoting the treatment of inflammation, such as enhancing the function of excreted macrophages.

In general, the inflammatory reaction is a biological defense reaction process for repairing and regenerating damage caused by invasion that causes organic changes in cells or tissues of the living body. This works. While the inflammatory response normally induced by external invading bacteria is a defense system to protect the living body, when an abnormally excessive inflammatory response is induced, various diseases appear. These diseases are called inflammatory diseases. The inflammatory disease is a disease in which various inflammatory mediators secreted from target cells activated by external stimuli amplify and sustain inflammation, thereby threatening the life of the human body. The inflammatory disease is acute bronchitis, chronic bronchitis, acute bronchiolitis, chronic bronchiolitis, These include sepsis, septic shock, acute respiratory distress syndrome, multiple organ failure and chronic obstructive pulmonary disease.

The onset of many inflammation-related diseases is associated with the activation of macrophages and the excessive production of inflammation-related factors. Representative inflammation-related factors include IL-1β and TNF-α. In addition, among the inducing substances of the inflammatory response, LPS (lipopolysaccharide) is a substance constituting the cell surface of Gram-negative bacteria, interacts with immune cells such as leukocytes, and is involved in the regulation of inflammation-related cytokines.

On the other hand, it is not known whether Jomshtan tree is effective in the treatment of inflammatory diseases.

Korean Patent Registration 10-1705460

Accordingly, the present inventors have completed the present invention by confirming that the Jomshtang tree extract has a high therapeutic effect and few side effects, thereby having a therapeutic effect on inflammatory diseases.

Accordingly, an object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of inflammatory diseases comprising the extract of Jomshitan tree as an active ingredient.

Another object of the present invention is to provide a food composition for the prevention or improvement of inflammatory diseases comprising the extract of Jomshitan tree as an active ingredient.

However, the technical problem to be achieved by the present invention is not limited to the above-mentioned problems, and other problems not mentioned will be clearly understood by those skilled in the art from the following description.

In order to achieve the above object of the present invention, the present invention provides a pharmaceutical composition for the prevention or treatment of inflammatory diseases comprising a jomshitan tree extract as an active ingredient.

In addition, the present invention provides a composition for the prevention or improvement of inflammatory diseases comprising the extract of Jomshitan tree as an active ingredient.

In one embodiment of the present invention, the composition may be a food composition, but is not limited thereto.

In another embodiment of the present invention, the food composition may be a health functional food, but is not limited thereto.

In another embodiment of the present invention, the extract is water, an alcohol having 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate. , methylene chloride, hexane (hexane), cyclohexane (cyclohexane), petroleum ether (petroleum ether), subcritical fluid, and may be due to one or more solvents selected from the group consisting of supercritical fluid, but limited thereto it's not going to be

In another embodiment of the present invention, the inflammatory disease is gastritis, gastric ulcer, duodenal ulcer, inflammatory skin disease, allergic disease, Crohn's disease, irritable bowel syndrome, ulcerative colitis, inflammatory bowel disease, peritonitis, osteomyelitis , cellulitis, meningitis, encephalitis, pancreatitis, traumatic shock, bronchial asthma, cystic fibrosis, stroke, acute bronchitis, chronic bronchitis, acute bronchiolitis, chronic bronchiolitis, osteoarthritis, gout, spondyloarthropathy, ankylosing spondylitis, Reiter's syndrome, psoriasis Arthropathy, intestinal disease spondylitis, juvenile arthropathy, juvenile ankylosing spondylitis, reactive arthropathy, infectious arthritis, post-infectious arthritis, gonococcal arthritis, tuberculous arthritis, viral arthritis, fungal arthritis, syphilitic arthritis, Lyme disease, Arthritis associated with 'vasculitis syndrome', polyarteritis nodosa, vasculitis hypersensitivity, ALS, polymyalgia rheumatica, articular cell arteritis, calcium crystallization arthropathy, pseudogout, non-articular rheumatism, bursitis, tendinitis, epicondylitis (tennis) elbow), neuropathic joint disease (charco and joint), hemarthrosic, Henoch-Schenlein purpura, hypertrophic osteoarthropathy, multicentral reticulohistocytoma, surcoilosis, hemochromatosis, sickle cell disease and Other hemoglobinopathy, hyperproteinemia, hypogammaglobulinemia, familial mediterranean fever, Behat's disease, systemic lupus erythematosus, recurrent fever, psoriasis, multiple sclerosis, sepsis, septic shock, multiple organ dysfunction syndrome, acute respiratory distress syndrome , chronic obstructive pulmonary disease, acute lung injury, and broncho-pulmonary dysplasia may be at least one selected from the group consisting of, but is not limited thereto.

In another embodiment of the present invention, the Jomshitan tree extract may inhibit NF-κB activity, but is not limited thereto.

In another embodiment of the present invention, the Jomshitan tree extract may be characterized in that it inhibits the activity of the Src protein, but is not limited thereto.

In another embodiment of the present invention, the Jomshitan tree extract may be characterized in that it inhibits nitric oxide (NO) production, but is not limited thereto.

In addition, the present invention provides a method for treating an inflammatory disease comprising administering to a subject a composition comprising an extract of Jomshitan tree as an active ingredient.

In addition, the present invention provides a use for preventing and/or treating inflammatory diseases of a composition comprising an extract of Jomshitan tree as an active ingredient.

In addition, the present invention provides a use for producing a medicament used in inflammatory diseases of the extract of Jomshtan tree.

The extract of Jomshtang tree according to the present invention has the effect of effectively reducing the generation of cytokines and corresponding reactants generated by the inflammatory reaction, as well as suppressing the expression of signaling proteins involved in immune activity. In addition, it is expected that the extract of Jomshitan tree as a natural product has no cytotoxicity, so that it can be widely used as an improvement, prevention, or therapeutic agent for various inflammatory diseases without side effects.

1 shows the inhibitory effect of Sorbaria kirilowii ethanol extract (hereinafter, Sk-EE) on the production of nitric oxide (NO) by the inflammatory substance lipopolysaccharide (hereinafter, LPS) in RAW 264.7 cells.
Figure 2 shows the inhibitory effect of NO production of Sk-EE in peritoneal macrophages obtained from ICR mouse induced inflammation with Thioglycollate (TG).
3 shows the results of confirming the cytotoxic effect of Sk-EE on RAW264.7 cells.
4 shows the results of confirming the cytotoxic effect of Sk-EE on peritoneal macrophage.
Figure 5 shows the effect of Sk-EE on the mRNA expression of nitric oxide synthase (iNOS), COX-2, and IL-1β, which are inflammation-related cytokines by LPS.
6 shows the effect of reducing the nuclear expression of p50, a protein constituting NF-κB, which is a transcription factor related to an inflammatory response, in the LPS-induced inflammatory state by Sk-EE.
7 shows the effect of Sk-EE on reducing intracellular NF-κB signaling proteins IKKα and IκBα phosphorylation.
8 shows the effect of Sk-EE on reducing the phosphorylation of Syk and Src proteins related to intracellular NF-κB signal transduction.
9 shows the effect of reducing Src phosphorylation by Sk-EE in the inflammatory response caused by Src overexpression.
Figure 10 shows the effect of Sk-EE binding to the inflammation-related protein Src to stabilize the protein structure, thereby inhibiting protein denaturation according to the increase in temperature.
11 shows the effect of reducing inflammation (bleeding) site by Sk-EE in an acute gastritis model using an ICR mouse.
12 shows the effect of reducing phosphorylation of the inflammation-related signaling protein IκBα by Sk-EE in an acute gastritis model using an ICR mouse.
13 is a schematic diagram showing the anti-inflammatory target signaling process of Sk-EE.
14 is a photograph showing the effect of recovering colon length by Sk-EE in a DSS (dextran sodium sulfate)-induced enteritis mouse model.
15 is a graph showing the quantitative measurement of colon length in a DSS (dextran sodium sulfate)-induced enteritis mouse model.

The present invention provides a composition for the prevention, improvement or treatment of inflammatory diseases comprising the extract of Jomshitan tree as an active ingredient.

Hereinafter, the present invention will be described in detail.

The jomsh tan tree of the present invention may preferably be a joms tan tree ( Sorbaria kirilowii ), but is not limited thereto.

The jomshitan tree is native to China and is a deciduous broad-leaved shrub planted for ornamental purposes around private houses and along roadsides. The stem grows straight to about 3~3m in height, and there is no hair on the whole. The stipule is long linear, 4~15mm long, hairless, and the tip is sharp or blunt. The leaves are alternate, pinnately compound. In the case of Korea, it is planted in the central region and is distributed in central and northern China. This species consists of 5-10 pairs of small leaves, and is distinguished from the stylus, which has star-shaped hairs as a whole, in that there are long hairs on the veins on the back side of the leaves.

The jomshitan tree of the present invention can be used as it is without damaging its original form, or after performing a pretreatment process in consideration of the process speed and process (manufacturing) efficiency intended by those skilled in the art, the pretreatment process includes, for example, Conventional screening, washing, cutting, powdering, drying, etc. steps may be performed.

The extract of Jomshtan tree of the present invention may be obtained from the whole plant or a part of the plant, for example, it may be obtained from a site comprising at least one selected from the group consisting of stems, roots, leaves, fruits, and flowers.

The extract of the Jomshtang tree of the present invention may be extracted by a known natural product extraction method. For example, it may be extracted with one or more solvents selected from the group consisting of water, an organic solvent having 1 to 6 carbon atoms, and a subcritical or supercritical fluid. The organic solvent having 1 to 6 carbon atoms is alcohol, acetone, ether, benzene, chloroform, ethyl acetate, and methylene chloride having 1 to 6 carbon atoms. chloride), hexane (hexane), cyclohexane (cyclohexane) and petroleum ether (petroleum ether) may be at least one selected from the group consisting of, but is not limited thereto.

The extract of Jomshtang tree of the present invention may be preferably extracted with ethanol, but is not limited thereto. As the extraction method, a conventional extraction method such as cold soaking, warm soaking, or heating using the solvent can be used.

The concentration of the ethanol is not limited, but for example, 30 to 100%, 40 to 100%, 50 to 100%, 60 to 100%, 70 to 100%, 80 to 100%, 90 to 100% or about 95 % ethanol extract.

The extract of the jomshtang tree of the present invention may be to inhibit NF-κB activity, but is not limited thereto.

The extract of the Jomshtang tree of the present invention may be to inhibit the activity of the Src protein, but is not limited thereto.

The extract of the jomshitan tree of the present invention may be to inhibit the production of nitric oxide (NO), but is not limited thereto.

In one embodiment of the present invention, as a result of verifying the activity of the jomshitan tree extract, the extract not only inhibits the production of NO, which is a macrophage-derived cytotoxic substance without cytotoxicity (see Examples 3 and 4),

Inhibits NF-κB-related inflammation-related cytokines iNOS, IL-1β and COX-2 gene expression (see Example 5),

It significantly reduces the nuclear expression level of p50, a component protein of NF-κB, as well as reduces phosphorylation of IκBα and IKKα, and critically inhibits phosphorylation of Syk and Src, which are known as high-level factors among NF-κB signaling proteins. It was confirmed to decrease (see Example 7),

It was confirmed that Src phosphorylation was decreased by Sk-EE in the inflammatory response caused by Src overexpression (see Example 8),

It was confirmed that the degree of bleeding due to acute gastritis was significantly lower in the gastric wall of the mice pre-injected with Sk-EE (see Example 10),

It was confirmed that the effect of recovering enteritis of the mice injected with Sk-EE was excellent compared to the negative control group and the positive control group administered with sulfasalazine (see Example 11).

Accordingly, it is expected that the Jomshtan tree extract can be applied to the prevention, improvement or treatment of various inflammatory diseases. The inflammatory disease is gastritis, gastric ulcer, duodenal ulcer, inflammatory skin disease, allergic disease, Crohn's disease, irritable bowel syndrome, ulcerative colitis, inflammatory bowel disease, peritonitis, osteomyelitis, cellulitis, meningitis, encephalitis, pancreatitis, Trauma-induced shock, bronchial asthma, cystic fibrosis, stroke, acute bronchitis, chronic bronchitis, acute bronchiolitis, chronic bronchiolitis, osteoarthritis, gout, spondyloarthropathies, ankylosing spondylitis, Reiter's syndrome, psoriatic arthropathy, enteric spondylitis, juvenile spondylitis Arthropathy, juvenile ankylosing spondylitis, reactive arthropathy, infectious arthritis, post-infectious arthritis, gonococcal arthritis, tuberculous arthritis, viral arthritis, fungal arthritis, syphilitic arthritis, Lyme disease, arthritis associated with 'vasculitis syndrome', nodular Polyarteritis, hypersensitivity vasculitis, Lou Gehrig's granulomatosis, polymyalgia rheumatica, articular cell arteritis, calcium crystal deposition arthropathy, pseudo gout, non-articular rheumatism, bursitis, tendinitis, epicondylitis (tennis elbow), neuropathic joint disease ( charco and joint), hemarthrosic, Henoch-Schenlein purpura, hypertrophic osteoarthropathy, multicentral reticulocytoma, surcoilosis, hemochromatosis, sickle cell disease and other hemoglobinopathy, hyperproteinemia, hypo Gammaglobulinemia, familial Mediterranean fever, Behat's disease, systemic lupus erythematosus, recurrent fever, psoriasis, multiple sclerosis, sepsis, septic shock, multiple organ dysfunction syndrome, acute respiratory distress syndrome, chronic obstructive pulmonary disease disease), acute lung injury, and broncho-pulmonary dysplasia may be at least one selected from the group consisting of, but is not limited thereto.

In the present specification, the inflammatory disease consists of gastritis, gastric ulcer, gastric ulcer enteritis, duodenal ulcer, gastric cancer, intestinal obstruction, acute gastrointestinal bleeding, Crohn's disease, irritable bowel syndrome, inflammatory bowel disease, ulcerative colitis and functional dyspepsia. It may be a gastrointestinal disease including one or more selected from the group, but is not limited thereto.

The content of the extract of Jomshtan tree in the composition of the present invention can be appropriately adjusted according to the symptoms of the disease, the degree of progression of the symptoms, the condition of the patient, etc., for example, 0.0001 to 99.9% by weight, or 0.001 to 50, based on the total weight of the composition. It may be a weight %, but is not limited thereto. The content ratio is a value based on the dry amount from which the solvent is removed.

The pharmaceutical composition according to the present invention may further include suitable carriers, excipients and diluents commonly used in the preparation of pharmaceutical compositions. The excipient may be, for example, at least one selected from the group consisting of a diluent, a binder, a disintegrant, a lubricant, an adsorbent, a humectant, a film-coating material, and a controlled-release additive.

The pharmaceutical composition according to the present invention can be prepared according to a conventional method according to a conventional method, such as powders, granules, sustained-release granules, enteric granules, liquids, eye drops, elsilic, emulsions, suspensions, alcohols, troches, fragrances, and limonaade. , tablets, sustained release tablets, enteric tablets, sublingual tablets, hard capsules, soft capsules, sustained release capsules, enteric capsules, pills, tinctures, soft extracts, dry extracts, fluid extracts, injections, capsules, perfusates, Warnings, lotions, pasta, sprays, inhalants, patches, sterile injection solutions, or external preparations such as aerosols can be formulated and used, and the external preparations are creams, gels, patches, sprays, ointments, warning agents , lotion, liniment, pasta, or cataplasma.

Carriers, excipients and diluents that may be included in the pharmaceutical composition according to the present invention include lactose, dextrose, sucrose, oligosaccharide, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

In the case of formulation, it is prepared using diluents or excipients, such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants.

Corn starch, potato starch, wheat starch, lactose, sucrose, glucose, fructose, di-mannitol, precipitated calcium carbonate, synthetic aluminum silicate, phosphoric acid as additives for tablets, powders, granules, capsules, pills, and troches according to the present invention Calcium monohydrogen, calcium sulfate, sodium chloride, sodium hydrogen carbonate, purified lanolin, microcrystalline cellulose, dextrin, sodium alginate, methyl cellulose, sodium carboxymethyl cellulose, kaolin, urea, colloidal silica gel, hydroxypropyl starch, hydroxypropyl methyl excipients such as cellulose, 1928, 2208, 2906, 2910, propylene glycol, casein, calcium lactate, and primogel; Gelatin, gum arabic, ethanol, agar powder, cellulose acetate phthalate, carboxymethylcellulose, calcium carboxymethylcellulose, glucose, purified water, sodium caseinate, glycerin, stearic acid, sodium carboxymethylcellulose, sodium methylcellulose, methylcellulose, microcrystalline cellulose, dextrin , hydroxycellulose, hydroxypropyl starch, hydroxymethylcellulose, purified shellac, starch powder, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinyl alcohol, polyvinylpyrrolidone, etc. Hydroxypropyl methylcellulose, corn starch, agar powder, methylcellulose, bentonite, hydroxypropyl starch, sodium carboxymethylcellulose, sodium alginate, calcium carboxymethylcellulose, calcium citrate, sodium lauryl sulfate, silicic anhydride, 1-hydroxy Propylcellulose, dextran, ion exchange resin, polyvinyl acetate, formaldehyde treated casein and gelatin, alginic acid, amylose, guar gum, sodium bicarbonate, polyvinylpyrrolidone, calcium phosphate, gelled starch, gum arabic, disintegrants such as amylopectin, pectin, sodium polyphosphate, ethyl cellulose, sucrose, magnesium aluminum silicate, di-sorbitol solution, light anhydrous silicic acid; Calcium stearate, magnesium stearate, stearic acid, hydrogenated vegetable oil, talc, lycopodite, kaolin, petrolatum, sodium stearate, cacao butter, sodium salicylate, magnesium salicylate, polyethylene glycol 4000,6000, liquid paraffin, hydrogenated soybean oil (Lubri) wax), aluminum stearate, zinc stearate, sodium lauryl sulfate, magnesium oxide, macrogol, synthetic aluminum silicate, silicic anhydride, higher fatty acid, higher alcohol, silicone oil, paraffin oil, polyethylene glycol fatty acid ether, starch, sodium chloride, A lubricant such as sodium acetate, sodium oleate, dl-leucine, light anhydrous silicic acid; may be used.

The liquid additives according to the present invention include water, dilute hydrochloric acid, dilute sulfuric acid, sodium citrate, monostearate sucrose, polyoxyethylene sorbitol fatty acid esters (Twinester), polyoxyethylene monoalkyl ethers, lanolin ethers, Lanolin esters, acetic acid, hydrochloric acid, aqueous ammonia, ammonium carbonate, potassium hydroxide, sodium hydroxide, prolamine, polyvinylpyrrolidone, ethyl cellulose, sodium carboxymethyl cellulose, and the like can be used.

In the syrup according to the present invention, a sucrose solution, other sugars or sweeteners may be used, and if necessary, a fragrance, colorant, preservative, stabilizer, suspending agent, emulsifying agent, thickening agent, etc. may be used.

Purified water may be used in the emulsion according to the present invention, and if necessary, an emulsifier, preservative, stabilizer, fragrance, etc. may be used.

Suspending agents such as acacia, tragacantha, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, microcrystalline cellulose, sodium alginate, hydroxypropylmethylcellulose, 1828, 2906, and 2910 may be used in the suspending agent according to the present invention, If necessary, surfactants, preservatives, stabilizers, colorants, and fragrances may be used.

Injectables according to the present invention include distilled water for injection, 0.9% sodium chloride injection, ring gel injection, dextrose injection, dextrose + sodium chloride injection, PEG (PEG), lactated ring gel injection, ethanol, propylene glycol, non-volatile oil-sesame oil , solvents such as cottonseed oil, peanut oil, soybean oil, corn oil, ethyl oleate, isopropyl myristate, and benzene benzoate; Solubilizing aids such as sodium benzoate, sodium salicylate, sodium acetate, urea, urethane, monoethylacetamide, butazolidine, propylene glycol, tweens, nijeongtinamide, hexamine, and dimethylacetamide; Weak acids and their salts (acetic acid and sodium acetate), weak bases and their salts (ammonia and ammonium acetate), organic compounds, proteins, buffers such as albumin, peptone, gum; isotonic agents such as sodium chloride; Stabilizers such as sodium bisulfite (NaHSO ₃ ) carbon dioxide gas, sodium metabisulfite (Na ₂ S ₂ O ₃ ), sodium sulfite (Na ₂ SO ₃ ), nitrogen gas (N ₂ ), ethylenediaminetetraacetic acid; sulphating agents such as sodium bisulfide 0.1%, sodium formaldehyde sulfoxylate, thiourea, disodium ethylenediaminetetraacetate, acetone sodium bisulfite; analgesic agents such as benzyl alcohol, chlorobutanol, procaine hydrochloride, glucose, and calcium gluconate; suspending agents such as SiMC sodium, sodium alginate, Tween 80, and aluminum monostearate.

The suppository according to the present invention includes cacao fat, lanolin, Witepsol, polyethylene glycol, glycerogelatin, methyl cellulose, carboxymethyl cellulose, a mixture of stearic acid and oleic acid, Subanal, cottonseed oil, peanut oil, palm oil, cacao butter + Cholesterol, Lecithin, Lanet Wax, Glycerol Monostearate, Tween or Span, Imhausen, Monolene (Propylene Glycol Monostearate), Glycerin, Adeps Solidus, Butyrum Tego -G), Cebes Pharma 16, Hexalide Base 95, Cotomar, Hydroxote SP, S-70-XXA, S-70-XX75 (S-70-XX95), Hydro Hydrokote 25, Hydrokote 711, Idropostal, Massa estrarium, A, AS, B, C, D, E, I, T, Massa-MF, Masupol, Masupol-15, Neosupostal-N, Paramound-B, Suposiro (OSI, OSIX, A, B, C, D, H, L), Suppository IV type (AB, B, A, BC, BBG, E, BGF, C, D, 299), supostal (N, Es), Wecobi (W, R, S, M, Fs), tester triglyceride base (TG-95, MA, 57) and The same mechanism may be used.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid preparations include at least one excipient in the extract, for example, starch, calcium carbonate, sucrose ) or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate talc are also used.

Liquid formulations for oral administration include suspensions, internal solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.

The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is determined by the type, severity, drug activity, and type of the patient's disease; Sensitivity to the drug, administration time, administration route and excretion rate, treatment period, factors including concurrent drugs and other factors well known in the medical field may be determined.

The pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered single or multiple. In consideration of all of the above factors, it is important to administer an amount that can obtain the maximum effect with a minimum amount without side effects, which can be easily determined by a person skilled in the art to which the present invention pertains.

The pharmaceutical composition of the present invention may be administered to an individual by various routes. All modes of administration can be envisaged, for example, oral administration, subcutaneous injection, intraperitoneal administration, intravenous injection, intramuscular injection, paraspinal (intrathecal) injection, sublingual administration, buccal administration, rectal insertion, vaginal It can be administered according to internal insertion, ocular administration, ear administration, nasal administration, inhalation, spraying through the mouth or nose, skin administration, transdermal administration, and the like.

The pharmaceutical composition of the present invention is determined according to the type of drug as an active ingredient along with several related factors such as the disease to be treated, the route of administration, the patient's age, sex, weight, and the severity of the disease.

In the present invention, "individual" means a subject in need of treatment for a disease, and more specifically, human or non-human primates, mice, rats, dogs, cats, horses, cattle, etc. It may be a mammal of, but is not limited thereto.

In the present invention, "administration" means providing a given composition of the present invention to a subject by any suitable method.

In the present invention, "prevention" means any action that inhibits or delays the onset of a desired disease, and "treatment" means that the desired disease and metabolic abnormalities are improved or It means any action that is advantageously changed, and "improvement" means any action that reduces a parameter related to a desired disease, for example, the degree of a symptom by administration of the composition according to the present invention.

In addition, the present invention provides a food composition comprising the extract of Jomshitan tree as an active ingredient.

In addition, the Jomshtan tree extract may be added to food for the purpose of improving inflammatory diseases.

In the present invention, food includes functional food and health functional food.

When the extract of Jomshtan tree of the present invention is used as a food additive, the extract of Jomshtan tree may be added as it is or used together with other foods or food ingredients, and may be appropriately used according to a conventional method. The mixed amount of the active ingredient may be appropriately determined according to the purpose of use (prevention, health or therapeutic treatment). In general, in the production of food or beverage, the extract of Jomshitan tree of the present invention may be added in an amount of 15% by weight or less, or 10% by weight or less based on the raw material. However, for health and hygiene purposes or health control In the case of long-term ingestion for the purpose of

There is no particular limitation on the type of the food. Examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, drinks, There are alcoholic beverages and vitamin complexes, and includes all health functional foods in the ordinary sense.

The health beverage composition according to the present invention may contain various flavoring agents or natural carbohydrates as additional ingredients, like a conventional beverage. The above-mentioned natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. As the sweetener, natural sweeteners such as taumatine and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like can be used. The proportion of the natural carbohydrate is generally about 0.01-0.20 g, or about 0.04-0.10 g per 100 mL of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, Carbonating agents used in carbonated beverages, etc. may be contained. In addition, the composition of the present invention may contain fruit for the production of natural fruit juice, fruit juice beverage, and vegetable beverage. These components may be used independently or in combination. The proportion of these additives is not critical, but is generally selected in the range of 0.01-0.20 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, preferred examples are presented to help the understanding of the present invention. However, the following examples are only provided for easier understanding of the present invention, and the contents of the present invention are not limited by the following examples.

Example 1. Experimental Preparation

Sorbaria kirilowii 95% ethanol extract (hereinafter, Sk-EE extract) was purchased from Korea Plant Extracts Bank (Cheongju, Korea).

Example 2. Cell culture method

Murine-derived macrophage line RAW264.7 cells were cultured in a 100 mm cell culture dish using RPMI 1640 medium containing penicillin (100 IU/ml) and streptomycin (100 μg/ml) and 10% FBS. It was cultured in an incubator maintained at a density of 37° C. and 5% CO _{2 .}

HEK293 cells, a human cell line, were cultured in a 100 mm cell culture dish at a density of 70-80% using DMEM medium containing penicillin (100 IU/ml) and streptomycin (100 μg/ml) and 5% FBS at 37°C. , incubated in an incubator maintaining 5% CO _{2 .}

Example 3. Measurement of cytotoxic substance NO (nitric oxide) production ability

The peritoneal macrophages isolated from the mouse macrophage line RAW264.7 and ICR mice were treated with RPMI 1640 medium containing penicillin (100 IU/ml) and streptomycin (100 μg/ml) and 10% FBS. Thus, at a concentration of 1×10 ⁵ cell/ml, 37° C., 5% CO ₂ was cultured for 24 hours in an incubator maintained. Then, the normal group was treated with DMSO as a control group, and 50 μl of each of Sk-EE was treated at 4 times the concentrations of 12.5, 25, 50, 100, and 200 μg/ml. After 30 minutes of incubation, 1 μg/ml LPS was applied to the positive control group and the extract-treated group 4 times, and the normal group was treated with 50 μg of the medium, and then cultured for 24 hours. Then, 100 μl of the culture solution from each well was transferred to a new 96 well plate, and absorbance was measured at 540 nm using _{Griess solution (0.5% naphthylethylenediamine dihydrochloride, 5% sulfanilamide, 25% H 3} PO _{4 ).} A calibration curve was prepared using sodium nitrite (0-100 μM), a standard material.

As a result, as shown in FIGS. 1 and 2 , it was confirmed that the Sk-EE extract inhibits the generation of nitric oxide (NO), a product of an inflammatory reaction by LPS and TG (Thioglycollate), in a concentration-dependent manner. did.

Example 4. Confirmation of cell viability

It was analyzed using MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphinyltetrazolium bromide) assay method. RAW264.7 cells (1 × 10 ⁶ cell/ml) and intraperitoneal macrophages were treated with Sk-EE at the indicated dose at 37° C. for 24 hours, and cultured in a _{CO 2 incubator.} Then, 10 μl MTT solution (stock concentration: 5 mg/ml) was added and an additional reaction was induced for 3 hours. For completion of the reaction and dissolution of formazan crystal, 100 μl MTT stopping solution (10% sodium dodecyl sulfate in 0.01M HCl) was additionally added to each well. Cell viability was calculated through the OD value obtained by measuring the absorbance at 570 nm of the amount of MTT reduced to formazan.

As a result, as shown in FIGS. 3 and 4 , it was confirmed that the cytotoxic effect of Sk-EE on macrophages and peritoneal macrophages was not significant. That is, it was confirmed that the Sk-EE extract effectively inhibited the inflammatory response without cytotoxicity.

Example 5. Confirmation of expression level of inflammation-related cytokines using RT-PCR

RAW264.7 cells were treated with RPMI 1640 medium containing penicillin (100 IU/ml) and streptomycin (100 μg/ml) and 10% FBS in a 12-well cell culture dish at ^{a concentration of 1 X 10 5} cells/ml. 37 ℃, 5% CO ₂ Incubated in an incubator maintained for 24 hours. Sk-EE was treated at a target concentration of 100 μg/ml and 200 μg/ml, respectively, and incubated for 30 minutes after processing 100 μl. Thereafter, 100 μl of 1 μg/ml of LPS was treated and incubated for 6 hours.

To measure regulation at the transcriptional level, total RNA was extracted using Trizol reagent. cDNA was prepared from the extracted total RNA using the First strand cDNA synthesis kit (Thermo scientific), and then the same amount of cDNA was amplified by PCR. The base sequences of the sense and antisense primers of the target protein used are shown in Table 1, and GAPDH was used as the control gene. For PCR amplification, use the Hipi PCR kit (Elpis biotech) for each experimental group cDNA, sense and antisense primers of target proteins, and control GAPDH primers with dNTP 250 μM, Tris-HCl (pH 8.3) 10 mM, KCl 50 mM, MgCl ₂ 1.5 It was performed in 20 μl of Hipi PCR kit containing mM. PCR was performed under the conditions of denaturation at 95 °C for 45 seconds, annealing at 55 °C for 45 seconds, and extension at 72 °C for 1 minute, and a total of 30 cycles were performed. DNA amplified by PCR was loaded on a 1% agarose gel and electrophoresed at 100V for 30 minutes, and the intensity of the DNA band fractionated in UV was measured.

Primers Gene Target Sequence SEQ ID NO: iNOS F CCC TTC CGA AGT TTC TGG CAG CAG C One R GGC TGT CAG AGC CTC GTG GCT TTG G 2 COX-2 F CAC TAC ATC CTG ACC CAC TT 3 R ATG CTC CTG CTT GAG TAT GT 4 IL-1β F CAG GAT GAG GAC ATG AGC ACC 5 R CTC TGC AGA CTC AAA CTC CAC 6 GADPH F CAA TGA ATA CGG CTA CAG CA 7 R AGG GAG ATG CTC AGT GTT GG 8

As a result, as shown in Figure 5, Sk-EE extract has the effect of reducing inflammation-related cytokines caused by LPS. Specifically, the Sk-EE extract has the effect of inhibiting nitric oxide synthase (iNOS) that produces NO as well as the inflammatory mediators Interleukin 1 beta, and COX-2 at the mRNA level. confirmed that there is.

Example 6. Evaluation of nuclear translocation of transcription factors

RAW264.7 cells were placed in a 10 cm cell culture dish containing RPMI 1640 medium containing penicillin (100 IU/ml) and streptomycin (100 μg/ml) and 10% FBS at a concentration of 1 X 10 ⁵ cells/ml. , 37 ℃, 5% CO ₂ Incubated in an incubator maintained for 24 hours.

After each treatment with 200 μg/ml of Sk-EE at a target concentration, incubated for 30 minutes. Then, after treatment with 1 μg/ml of LPS, the cells were scraped at each time point. After stopping the protein function with cold PBS, homogenization buffer (20 mM Tris-HCl pH8.0, 10 mM EGTA, 2 mM EDTA, 2 mM DTT, 1 mM PMSF, 25 μg/ml Aprotinin, 10 μg/ml Leupeptin) ), and then lysing the cells using a sonicator, followed by centrifugation at 8000 rpm, 4 °C for 15 minutes. After transferring the supernatant (cytosolic fraction), the pellet (nuclear fraction) was treated with homogenization buffer B (1% TritonX-100 in Homogenization buffer A) and vortexed. The obtained sample was subjected to SDS-PAGE and western blotting for target signal transduction process and protein expression analysis.

As a result, as shown in FIG. 6 , it was confirmed that the Sk-EE extract significantly reduced the nuclear expression level of p50, a protein constituting NF-κB, a transcription factor related to an inflammatory response by LPS.

Example 7. Confirmation of anti-inflammatory target protein and signal transduction process

RAW264.7 cells, a murine macrophage cell line, were 3 cm cells at a concentration of ^{2.5X10 5} cells/ml using RPMI 1640 medium containing penicillin (100 IU/ml) and streptomycin (100 μg/ml) and 10% FBS. It was pre-cultured in a culture dish. After 24 hours, the Sk-EE extract was pretreated at a target concentration of 200 μg/ml for 30 minutes, and then treated with LPS having a final concentration of 1 μg/ml to induce an inflammatory response, and after a certain time depending on the drug, the cells lysis buffer (20mM Tris-HCl, pH 7.4, 2mM EDTA, 2mM EGTA, 50mM glycerol phosphate, 1mM DTT, 2 μg/ml aprotinin, 2 μg/ml leupeptin, 1 μg/ml pepstatin, 50 μM PMSF, 1 mM benzamide , 2% Triton X-100, 10% glycerol, 0.1mM sodium vanadate, 1.6mM pervanadate, and 20mM NaF) were added to lyse the cells. The lysate was quantified using a concentration formula using BSA as a standard, followed by SDS-PAGE. Then, blotting was performed using a PVDF membrane, followed by blocking with 3% BSA. Antibodies specific for each of the target proteins p65, p50, LaminA/C, IKKα/β, IκBα, Syk, Src, β-actin, HA, and Myc (Cell Signaling Technology Danvers, MA, USA) was treated first, and then after washing, the secondary antibody was treated. After washing again, the ECL (Amersham, England) solution was evenly treated and photosensitized with an X-ray film.

As a result, as shown in FIG. 7 , it was confirmed that phosphorylation of intracellular NF-κB signaling proteins IκBα and IKKα was decreased by Sk-EE.

In addition, as shown in FIG. 8 , it was confirmed that the phosphorylation of Syk and Src, which are proteins related to intracellular NF-κB signal transduction, was decreased by Sk-EE.

Example 8. Confirmation of signal transduction process upon overexpression of target protein Src

HEK293T cells were cultured in DMEM culture media containing 5% Fetal Bovine Serum (FBS) in a 12-well cell culture plate at ^{a concentration of 2.5 X 10 5} cells/ml at 37 ° C. and 5% CO ₂ in an incubator maintained for 24 hours. did. Target DNA and control DNA (HA-Src, pcDNA) were transfected with PEI and cultured for 24 hours. The medium was changed to the same amount with DMEM 1640 culture medium containing penicillin (100 IU/ml) and streptomycin (100 μg/ml) and 5% FBS, and then 100 μg/ml, 200 μg/ml of Sk-EE was added. After treatment in each group, culture was performed again for 24 hours. After stopping protein function with cold PBS, cell lysis buffer (20mM Tris-HCl, pH 7.4, 2mM EDTA, 2mM EGTA, 50mM glycerol phosphate, 1mM DTT, 2 μg/ml aprotinin, 2 μg/ml leupeptin, 1 μg/ml pepstatin , 50 μM PMSF, 1 mM benzamide, 2% Triton X-100, 10% glycerol, 0.1 mM sodium vanadate, 1.6 mM pervanadate, and 20 mM NaF) to lyse the cells. SDS-PAGE and western blotting were performed to analyze the target protein expression level.

As a result, as shown in FIG. 9 , it was confirmed that Src phosphorylation was reduced by Sk-EE in the inflammatory response caused by Src overexpression.

From the results of Examples 6 and 7, the Sk-EE extract not only strongly inhibited IκBα, which directly binds to NF-κB, but also critically inhibited Src protein, which is known as the highest factor among NF-κB signaling proteins. It was confirmed that the effect was clear. The above results indicate that the Sk-EE extract acts as a target for the Src protein.

Example 9. Confirmation of the protein denaturation inhibitory effect of Sk-EE

HEK293T cells were cultured in DMEM culture media containing 5% Fetal Bovine Serum (FBS) in a 12-well cell culture plate at ^{a concentration of 2.5 X 10 5} cells/ml at 37 ° C. and 5% CO ₂ in an incubator maintained for 24 hours. did. Then, Myc-Syk, HA-Src plasmids were transfected through PEI. After 24 hours of incubation, Sk-EE (200 μg/ml) was treated with a fresh medium and incubated for 24 hours again. After removing the cells through trypsinization, the cells were collected by centrifugation at 1000 rpm for 5 minutes. Then, after washing 3 times with PBS, cell counting was performed. ^{After quantifying 1.5 X 10 6} cells in each group, the temperature was changed using a real-time PCR machine (43-64 ℃, 3 minutes at each temperature). After repeating the freezing-thawing process with liquid nitrogen three times, centrifugation was performed, and 5X sample buffer was added to the supernatant of each separated group. Protein expression was measured by Western blotting, and detection was carried out with ECL reagent.

As a result, as shown in FIG. 10 , it was confirmed that Sk-EE binds to the inflammation-related protein Src to stabilize the protein structure, and thus has an excellent effect of inhibiting protein denaturation according to temperature rise.

Example 10. Confirmation of effect of Sk-EE in acute gastritis model

Twenty-five 6-week-old ICR mice were divided into 5 groups of 5 in plastic cages each, acclimatized for 4 days, and fasted on day 0. Gastritis was induced by injection of HCl/EtOH into the remaining groups except for the Normal group, and one group was treated as a control group by injection of 0.5% CMC (carboxyl methyl cellulose), and the two groups were each 100% Sorbaria kirilowii ethanol extract (Sk-EE). mg/kg and 200 mg/kg were injected, and the last group was injected with 40 mg/kg of Ranitidine. After sacrifice, the stomach was removed and the extent of inflammation was quantified by measuring the size of the bleeding site due to gastritis on the wall of the stomach.

As a result, as shown in FIG. 11 , as a result of analyzing the gastric wall of the mice induced with gastritis, the degree of bleeding due to acute gastritis was significantly higher in the gastric wall of the mice pre-injected with Sk-EE compared to the control group. low was confirmed. The gastritis relief effect of Sk-EE increased in a concentration-dependent manner when comparing the 100 mg/ml treatment group and the 200 mg/ml treatment group. It was confirmed that the alleviation effect was shown.

In addition, as shown in FIG. 12 , the effect of reducing phosphorylation of the inflammation-related signaling protein IκBα by Sk-EE was confirmed in the acute gastritis model, and it was confirmed that the Jomshtan tree extract showed the potential as an effective pharmaceutical material.

Example 11. Confirmation of the effect of Sk-EE in the enteritis model

25 5-week-old C57BL/6 mice were divided into 5 groups in 5 plastic cages each, acclimatized for 4 days, and the substance was injected from day 0 to 10 days. The normal and positive groups were injected with 0.2 ml of 0.5% CMC by oral injection, and both groups were injected with Sorbaria kirilowii ethanol extract (Sk-EE) 100 mg/kg and 200 mg/kg 0.2 ml each. The other group was injected with 0.2 ml of 200 mg/kg of Sulfasalazine as a positive control. At the same time as the material injection from Day 3, DSS (dextran sodium sulfate) was dissolved in drinking water at a concentration of 0.5% and administered to induce enteritis in mice in each group except for the Normal group. After sacrificing the mice on Day 10, the colon was removed and the length was measured, and the extent of the decrease in the length of the intestine due to the induction of enteritis was quantified by the material.

As a result, as shown in FIGS. 14 and 15 , as a result of measuring the colon length of the mice induced with enteritis, the colon length of the mice administered Sk-EE was recovered compared to the DSS-administered negative control group, and the positive control group Compared with sulfasalazine, the colon length was elongated higher, indicating that the recovery effect of the Sk-EE extract showed a higher recovery effect of enteritis even compared to the positive control group.

Summarizing the above examples, the present inventors confirmed that the Sorbaria kirilowii ethanol extract effectively reduced the production of cytokines and corresponding reactants generated in the inflammatory reaction, and suppressed the expression of signaling proteins involved in immune activity. , These results show that Sorbaria kirilowii extract can be used in pharmaceuticals and health functional foods for the prevention, treatment or improvement of inflammatory diseases and materials for their development.

The above description of the present invention is for illustration, and those of ordinary skill in the art to which the present invention pertains can understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive.

<110> Research & Business Foundation SUNGKYUNKWAN UNIVERSITY <120> Pharmaceutical Composition comprising Sorbaria kirilowii extract for Preventing or Treating Inflammatory disease as active ingredient <130> MP20-049P1 <150> KR 10-2020-0058851 <151> 2020-05-18 <160> 8 <170> KoPatentIn 3.0 <210> 1 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> iNOS forward primer <400> 1 cccttccgaa gtttctggca gcagc 25 <210> 2 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> iNOS reverse primer <400> 2 ggctgtcaga gcctcgtggc tttgg 25 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> COX-2 forward primer <400> 3 cactacatcc tgacccactt 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> COX-2 reverse primer <400> 4 atgctcctgc ttgagtatgt 20 <210> 5 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> IL-1beta forward primer <400> 5 caggatgagg acatgagcac c 21 <210> 6 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> IL-1beta reverse primer <400> 6 ctctgcagac tcaaactcca c 21 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> GADPH forward primer <400> 7 caatgaatac ggctacagca 20 <210> 8 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> GADPH reverse primer <400> 8 agggagatgc tcagtgttgg 20

Claims (10)

A pharmaceutical composition for the prevention or treatment of inflammatory diseases comprising an extract of Jomshitan tree as an active ingredient.
According to claim 1,
The extract is water, alcohol having 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane (hexane), cyclohexane (cyclohexane), petroleum ether (petroleum ether), subcritical fluid, and characterized in that the extract by one or more solvents selected from the group consisting of supercritical fluid, inflammatory disease prevention or treatment pharmaceutical enemy composition.
According to claim 1,
The inflammatory disease is gastritis, gastric ulcer, duodenal ulcer, inflammatory skin disease, allergic disease, Crohn's disease, irritable bowel syndrome, ulcerative colitis, inflammatory bowel disease, peritonitis, osteomyelitis, cellulitis, meningitis, encephalitis, pancreatitis, Trauma-induced shock, bronchial asthma, cystic fibrosis, stroke, acute bronchitis, chronic bronchitis, acute bronchiolitis, chronic bronchiolitis, osteoarthritis, gout, spondyloarthropathies, ankylosing spondylitis, Reiter's syndrome, psoriatic arthropathy, enteric spondylitis, juvenile spondylitis Arthropathy, juvenile ankylosing spondylitis, reactive arthropathy, infectious arthritis, post-infectious arthritis, gonococcal arthritis, tuberculous arthritis, viral arthritis, fungal arthritis, syphilitic arthritis, Lyme disease, arthritis associated with 'vasculitis syndrome', nodular Polyarteritis, hypersensitivity vasculitis, Lou Gehrig's granulomatosis, polymyalgia rheumatica, articular cell arteritis, calcium crystal deposition arthropathy, pseudo gout, non-articular rheumatism, bursitis, tendinitis, epicondylitis (tennis elbow), neuropathic joint disease ( charco and joint), hemarthrosic, Henoch-Schenlein purpura, hypertrophic osteoarthropathy, reticulocytoma multicentre, surcoilosis, hemochromatosis, sickle cell disease and other hemoglobinopathy, hyperproteinemia, hypo Gammaglobulinemia, familial Mediterranean fever, Behat's disease, systemic lupus erythematosus, recurrent fever, psoriasis, multiple sclerosis, sepsis, septic shock, multiple organ dysfunction syndrome, acute respiratory distress syndrome, chronic obstructive pulmonary disease disease), acute lung injury (acute lung injury) and bronchial lung dysplasia (broncho-pulmonary dysplasia) characterized in that at least one selected from the group consisting of, the prevention or treatment of an inflammatory disease pharmaceutical composition.
According to claim 1,
The Jomshitan tree extract is a pharmaceutical composition for the prevention or treatment of inflammatory diseases, characterized in that it inhibits NF-κB activity.
According to claim 1,
The Jomshitan tree extract is a pharmaceutical composition for the prevention or treatment of inflammatory diseases, characterized in that it inhibits the activity of the Src protein.
According to claim 1,
The Jomshitan tree extract is a pharmaceutical composition for the prevention or treatment of inflammatory diseases, characterized in that it suppresses NO (Nitric oxide) production.
A food composition for the prevention or improvement of inflammatory diseases comprising the extract of Jomshitan tree as an active ingredient.
8. The method of claim 7,
The food composition is a health functional food, characterized in that the food composition.
8. The method of claim 7,
The extract is water, alcohol having 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane (hexane), cyclohexane (cyclohexane), petroleum ether (petroleum ether), subcritical fluid, and a food composition characterized in that the extract by one or more solvents selected from the group consisting of supercritical fluid.
8. The method of claim 7,
The inflammatory disease is gastritis, gastric ulcer, duodenal ulcer, inflammatory skin disease, allergic disease, Crohn's disease, irritable bowel syndrome, ulcerative colitis, inflammatory bowel disease, peritonitis, osteomyelitis, cellulitis, meningitis, encephalitis, pancreatitis, Trauma-induced shock, bronchial asthma, cystic fibrosis, stroke, acute bronchitis, chronic bronchitis, acute bronchiolitis, chronic bronchiolitis, osteoarthritis, gout, spondyloarthropathies, ankylosing spondylitis, Reiter's syndrome, psoriatic arthropathy, enteric spondylitis, juvenile spondylitis Arthropathy, juvenile ankylosing spondylitis, reactive arthropathy, infectious arthritis, post-infectious arthritis, gonococcal arthritis, tuberculous arthritis, viral arthritis, fungal arthritis, syphilitic arthritis, Lyme disease, arthritis associated with 'vasculitis syndrome', nodular Polyarteritis, hypersensitivity vasculitis, Lou Gehrig's granulomatosis, polymyalgia rheumatica, articular cell arteritis, calcium crystal deposition arthropathy, pseudo gout, non-articular rheumatism, bursitis, tendinitis, epicondylitis (tennis elbow), neuropathic joint disease ( charco and joint), hemarthrosic, Henoch-Schenlein purpura, hypertrophic osteoarthropathy, reticulocytoma multicentre, surcoilosis, hemochromatosis, sickle cell disease and other hemoglobinopathy, hyperproteinemia, hypo Gammaglobulinemia, familial Mediterranean fever, Behat's disease, systemic lupus erythematosus, recurrent fever, psoriasis, multiple sclerosis, sepsis, septic shock, multiple organ dysfunction syndrome, acute respiratory distress syndrome, chronic obstructive pulmonary disease disease), acute lung injury (acute lung injury) and bronchopulmonary dysplasia (broncho-pulmonary dysplasia) characterized in that at least one selected from the group consisting of, a food composition.

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