WO2017023000A1 - Composition containing mixed medicinal herb extract for preventing, treating, or alleviating chronic inflammatory diseases - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for treating chronic inflammatory diseases and food composition for alleviating chronic inflammatory diseases, each composition containing, as an active ingredient, an extract of two or more herbal medicines of Inulae Flos, Schizonepetae Spica, and Magnolia Cortex. The composition of the present invention could be favorably used in the prevention or treatment of chronic inflammatory diseases since the mixed herbal medicinal extract inhibits the secretion of cytokines which influence immune diseases.

Description

 【Specification】

 [Name of invention]

 Composition for preventing, treating or ameliorating chronic inflammatory disease containing a common herbal extract

 The present invention relates to a composition for the prevention, treatment or amelioration of chronic inflammatory diseases comprising a common herbal extract, and more particularly, Inul ae Flos, Schi zonepetae Spi ca and Magnol ia Cortex It relates to a pharmaceutical composition for the treatment of chronic inflammatory diseases and a composition for improving food, comprising extracting at least two herbal drugs selected from the group consisting of) as an active ingredient.

Background Art

This application claims the priority of Korean Patent Application No. 10-2015-0108612 filed on July 31, 2015, the entirety of which is a reference of the present application. Chronic inflammation is a condition in which persistent inflammation occurs, which means that the immune system continues to respond inappropriately to certain exposures. In particular, autoimmune diseases are caused by attacking organs due to disturbances of the immune system. Chronic inflammation may be caused by single or multiple genetic factors or environmental factors such as food or smoke. Inflammation occurs and calms down, but continues and sometimes the treatment is not effective. Chronic inflammatory diseases cause significant damage to tissues of the body and, depending on their location, many problems occur. For example, chronic inflammation in the liver or digestive tract causes changes in the brain, causing fatigue, personality changes, organ dysfunction, and spreading inflammation throughout the body. Chronic Inflammatory Disease Psor i as is a pterygoid chronic dermatological disease, characterized by the rapid proliferation of keratinocytes and accompanying dermatitis. Lesions such as pustules can appear anywhere on the skin. Clinically, plaque psoriasis, mulberry psoriasis, pustulic psoriasis, inverted psoriasis, hemorrhagic psoriasis can be classified as. Psoriasis is caused by a combination of genetic and environmental factors, the cause of which is not clear. I, the changes in hormones, psychological aspects such as stress, trauma, infections such as tonsillitis, immunity, endocrine, inflammation, metabolic capacity, climate, dry skin, drugs, heredity, and the like. In addition, IL-17 and IL-6 secreted by T immune cells differentiated by hypersensitivity of the immune system or abnormal autoimmunity of patients due to chronic inflammatory diseases. Overexpression of cytokines such as TNF-Q causes inflammatory reactions in the skin, promotes hyperproliferation and division of keratinocytes and inhibits differentiation. These factors occur because the cells of the stratum corneum proliferate 6-7 times faster than normal cells, and the excessively and incompletely proliferated keratinocytes are piled up on the scales of white scales. Psoriasis is often accompanied by other diseases, so it is referred to as a companion disease. Psoriatic arthritis, cardiovascular disease, depression, Crohn's disease, etc. are frequently found in patients with psoriasis. Psoriatic arthritis is a painful chronic inflammatory disease with up to 30% of psoriasis patients with arthritis symptoms, 85% of which psoriasis precedes arthritis, and recent studies have shown that arthritis and psoriasis may occur simultaneously. The cause of the posterior relationship has not been reported yet. Severe psoriasis is also associated with obesity, hypertension, hyperlipidemia, and increased metabolic syndrome and mortality in type 2 diabetes mellitus, and an increased risk of cardiovascular disease, possibly due to the effects of chronic inflammatory responses on the endocrine system. There is this. In addition, psoriasis skin is not good aesthetics, and when it occurs in prominent parts such as hands and face, it has been reported that 40% of people with depression or anxiety have atrophy even in interpersonal relationships. In order to treat inflammatory diseases, especially psoriasis, the body's immune response is so diverse and complex that it weakens patients' immunity so that the overall immune system, including T cells, can function normally to remove unknown antigens present in the skin. It is important to maintain homeostas is by correcting factors and adjusting immunity to balance. However, the drugs used in the prior art simply suppress the immunity and cause various complications such as cancer and tuberculosis, or they are toxic and cannot be repaired for a long period of time. phenomenon) or treatment efficiency is low. Many patients with current chronic inflammatory disorders often do not achieve significant clinical improvement with current standard drug treatments. In addition, most drugs are expensive, the treatment process is long and complicated, and accompanied by various side effects, it is difficult to avoid recurrence even to some extent. Therefore, it is necessary to prescribe a new drug with fewer side effects and better treatment efficiency than existing drugs. When treating a disease using a composition using a natural extract, it is possible to obtain an excellent therapeutic effect while reducing or reducing side effects, the development of medicines using a natural extract has been proposed as an alternative.

[Detailed Description of the Invention]

 [Technical problem]

Substances that inhibit the function of IL-17, IL-6 and TNF-α include psoriasis, a chronic inflammatory immune disease, including psoriatic arthritis, rheumatoid arthritis, multiple sclerosis, systemic lupus erythematosus, inflammatory bowel disease, Crohn's disease, and ulcers It is known that it can have a therapeutic effect against colitis, ankylosing spondylitis, asthma, chronic obstructive pulmonary disease, periodontitis, and idiopathic inflammatory myopathy, but it is a therapeutic drug that inhibits IL-17, IL-6, and TNF-a in combination. Currently does not exist. Therefore, the present inventors studied natural materials that inhibit the activity of inflammatory cytokines IL-17, IL-6 and TNF-a in order to develop a composition for preventing and treating chronic inflammatory diseases. ), Two or more combination herbal extracts selected from Schizonepetae Spica and Magnolia Cortex have been found to have the effect of treating chronic inflammatory diseases. Therefore, an object of the present invention for the treatment of chronic inflammatory disease, characterized in that it comprises an extract of at least two herbal extracts selected from the group consisting of Inulae Flos, Schizonepetae .Spica and Magnolia Cortex. It is to provide a pharmaceutical composition. Another object of the present invention is to improve chronic inflammatory disease, characterized in that it comprises an extract of at least two herbal drugs selected from the group consisting of Inulae Flos, Schizonepetae Spica and Magnolia Cortex as an active ingredient It is to provide a composition for food. Other objects of the present invention are Inulae Flos, Schizonepetae Spica and The present invention provides a method for treating chronic inflammatory disease by administering an effective amount of an extract of two or more herbal drugs selected from the group consisting of Magnolia Cortex to an individual in need. Another object of the present invention is to prepare a preparation for the treatment of chronic inflammatory diseases comprising an extract of at least two herbal drugs selected from the group consisting of Inulae Flos, Schizonepetae Spica and Magnolia Cortex as an active ingredient. It is intended to provide a purpose for doing so.

Technical Solution

In order to achieve the above object, the present invention provides chronic inflammatory vaginal treatment, comprising at least two herbal extracts selected from the group consisting of Inulae Flos, Schizonepetae Spica, and Magnolia Cortex. It provides a pharmaceutical composition for the treatment of a hwan. In order to achieve the other object of the present invention, the present invention is characterized in that it comprises an extract of at least two herbal drugs selected from the group consisting of Inulae Flos, Schizonepetae Spica and Magnolia Cortex To provide a food composition for the improvement of chronic inflammatory diseases. In order to achieve another object of the present invention chronic inflammatory vaginal by administering to the subject in need of an extract of at least two herbal drugs selected from the group consisting of Inulae Flos, Schizonepetae Spica and Magnolia Cortex Provides a method of treating a ring. In order to achieve another object of the present invention, the preparation for the treatment of chronic inflammatory diseases comprising extracts of two or more herbal medicines selected from the group consisting of Inulae Flos, Schizonepetae Spica, and Magnolia Cortex as active ingredients. It provides a use for the preparation. Hereinafter, the present invention will be described in detail. The present invention is a pharmaceutical composition for the treatment of chronic inflammatory diseases, characterized in that it comprises an extract of at least two herbal drugs selected from the group consisting of Inulae Flos, Schizonepetae Spica and Magnolia Cortex. To provide. In the present invention, the above-mentioned extracts of sunkyeong, hyeonghyeong, hakbak has the effect of inhibiting the secretion of IL-17, IL-6 and TNF-CI cytokine, especially their combinations of the inhibition of cytokine secretion compared to the extract alone It was confirmed that there is a synergistic effect. In addition, it was confirmed that the extract containing two or more of the herbal medicines has an excellent therapeutic effect in an animal model of psoriasis. As described above, the therapeutic effect of chronic inflammatory diseases, particularly psoriasis, which has a composition containing extracts of two or more herbal medicines selected from among ablation, mold opening and thickening is disclosed for the first time in the present invention. Inulae Flos is a plant that has traditionally been used as a medicinal herb. It is originated from the flowers of Japonica Thunberg or Guantin britannica Linne of the Compositae. Standard collection), Diameter is 10 ~ 15 and has a spherical shape. In oriental medicine, eclampsia has the effect of clay, expectoration, bronchospasm, diuresis, phlegm, gastrointestinal disorders, chest swelling, It has been used for indigestion, hiccups, etc. Schizonepetae Spica is a flowering plant of Schizonepeta tenui folia Briquet, a Lamitae plant. . It is attached with a small stem and sometimes a calyx tube with fruit, and you can see milky white short hairs on Jeonju, and it has the effect of haematyeo, epoch, and bleeding. Et al., Yeonglimsa, pp. 863-864, (1998). Magnoliae Cortex is a deciduous tree belonging to Magnolia iaceae, s ¾ (Magnolia ovobata Thunberg), {Magnol ia officinalis Rehder et

Wilson) or decay lobe G¾ g77o // s officinalis Rehder et Wilson var. bi loba Rehder et Wilson's stem bark (Korea Pharmacopoeia). In the traditional medicine, headache, chi It has been used for pain, sinusitis, and lowering blood pressure (Korean Medicinal Plant Book, 1993, 179-180, Academy Book). Thicken in the present invention means that derived from Magnol ia Cortex. The composition of the present invention is a composition comprising an extract of two or more herbal medicines selected from the group consisting of Inul ae Flos, Sch i zonepetae Spi ca and Magno ia Cortex. That is, the composition of the present invention may include extracts of two herbal medicines, such as sunkwa and hyeonghyeong, sunwha and hokwa, or hyunghwa and hobak as an active ingredient, and extracts of three herbal medicines, sunwha, hyunghwa and hohakwa It may be included as an active ingredient. The composition comprising the extract of two or more herbal medicines may be a mixture of two or more herbal extracts alone, or may be a complex extract extracted after mixing two or more herbal medicines. In the present specification, all of the combination or complex extract of the single extract may be collectively referred to as a combination extract. The composition of the present invention is not limited to the mixing mode of two or more herbal extracts containing as an active ingredient. There is no limitation on the form or property of the extract, it may be a solution, a concentrate, or may be a solid or powder from which the solvent used to prepare the extract is removed. The composition of the present invention may be a mixture of the sunkyeo, hyeonggeul and the thick extract alone in a weight ratio of 1: 0.1-1: 10: 0.1-10. When the composition of the present invention comprises a single extract of two herbal medicines (ie, when the extract of the sundae and mold opening, the sundae and huak, or the extract of hyungwa and huak), two herbal extracts are each 1: 0. 1 to 10 can be mixed in weight ratio. In addition, the composition of the present invention may be extracted after mixing the thickening, mold dog and thick foil on a dry weight basis in a weight ratio of 1: 0.1-1: 10: 1.10. When the composition of the present invention includes a composite extract of two herbal medicines, the two herbal medicines may be mixed and then extracted at a weight ratio of 1: 0 Λ˜10. The herbal extract in the present invention may be extracted with any one solvent selected from the group consisting of water, alcohol having 1 to 6 carbon atoms, nucleic acid, ethyl acetate and a mixed solvent thereof. Preferably, the extract of the present invention may be extracted with a solvent selected from water, alcohols having 1 to 6 carbon atoms and mixed solvents thereof, or an aqueous lower alcohol solution having 1 to 6 carbon atoms. The concentration of the aqueous alcohol solution is 10 to 10 (v / v), preferably 70 to 100¾) (v / v). The extraction solvent of the present invention may be most preferably an aqueous solution of 9 (v / v) of ethane. The extraction solvent in the present invention may be prepared by using about 3 to 30 times, preferably about 5 to 20 times, the dry herbal weight. Extraction of the herbal medicine of the present invention can be carried out at a temperature of 20 to 11 (C, preferably 60 to 90 ^° C. Extraction of the herbal medicine of the present invention is about 1 to 48 hours, preferably about 4 to 24 hours Extraction may be repeated one or more times In one embodiment of the present invention has been extracted twice every four hours The extraction method in the present invention can be used without limitation, if known in the art. ， For example, but not limited to cold soaking, hot water extraction, ultrasonic extraction, reflux cooling extraction, etc. Preferably, reflux extraction may be performed. In addition, an aqueous solution of an extraction solvent, such as alcohol, 4-7 times the weight of the crude ingredient is added to the residue, and the mixture is reextracted for 4 hours under reflux and extraction conditions, filtered, and mixed with the previous filtrate. In the present invention, a method of re-extracting after the first extraction is adopted, and thus, the extraction efficiency can be increased by mixing the two extractions and the filtrate obtained after each extraction. The extract of is not limited to the number of extractions, as described above, the combined filtrate after the second extraction is concentrated under reduced pressure at 40 to 60 ^° C to remove the solvent remaining in the extract, and the concentration of the concentrated concentrate is reduced to 25 After azeotropic concentration of 2 to 3 times with water of 50 to 50 times, and then suspended in a homogeneous solution by adding the same amount of water again, the final extract may be further purified or dried by lyophilization method to remove the solvent to solidify In the present inventors, the extract of sclerosis, mold opening, and gourd extract is used for splenocytes, macrophages, and the like. It was confirmed that there is an effect of inhibiting the secretion of inflammatory cytokines IL-17, IL-6 and TNF-α in immune cells. Therefore, it can be seen that the single extract of the herbal medicine has an effect of alleviating or treating the symptoms of immune diseases or chronic inflammatory diseases caused or mediated by the cytokines through such immune regulation. In particular, IL-17 augments aberrant cell proliferation and activity of psoriasis-inducing keratinocytes, so the extracts of acupuncture, glands and thickenings can treat psoriasis and psoriasis-related diseases. have. In addition, the inhibitory effect of IL-17, IL-6 and TNF-α secretion of herbal extracts was more remarkable when two or more herbal extracts were mixed than when the individual herbal extracts were treated. It can be seen that the synergistic effect of inflammatory cytokine inhibition of each herbal extract alone can be obtained through the composition comprising two or more herbal extracts according to the present invention. In other words, by administering the herbal extract according to the present invention it can be seen that more significant synergistic therapeutic effect can be expected for chronic inflammatory diseases such as psoriasis. In addition, in the present invention, the therapeutic effect of psoriasis, a chronic inflammatory disease of the complex extract of herbal medicine, was confirmed in a mouse model of psoriasis. Combined extracts of gnarled and thickened or gnarled and shaped dogs effectively reduced ear and ear tissue epidermal thickness, inhibited inflammatory cell infiltration, and psoriasis-related biomarkers in mouse models of psoriasis induced by IL-23 It was observed that the expression levels of keratin 17, IL-17, IL-23, TNF-α and IL-6 were reduced. In vivo, the composition comprising two or more herbal medicines according to the present invention as an active ingredient has the effect of alleviating and improving the symptoms of immune diseases or chronic inflammatory diseases caused or mediated by psoriasis or IL-23 through the immunomodulatory mechanism. Confirmed. Therefore, it can be expected that the composition of the present invention has the effect of preventing, alleviating or treating chronic inflammatory diseases mediated or aggravated by IL-17, IL-6, IL-23, TNF-α and the like. In particular, inflammatory cytokines such as IL-17, IL-6, and TNF-a also affect abnormal proliferation and activity of keratinocytes, which are important for psoriasis, and thus the composition of the present invention prevents and improves psoriasis and related symptoms. Or it can be seen that there is an excellent effect in the treatment. Accordingly, the composition of the present invention is a pharmaceutical composition for treating or preventing chronic inflammatory diseases, wherein the chronic inflammatory diseases are preferably psoriasis, psoriatic arthritis, rheumatoid arthritis, multiple sclerosis, systemic lupus erythematosus, inflammatory bowel disease, It may be a disease selected from the group consisting of Crohn's disease, ulcerative colitis, ankylosing spondylitis, asthma, chronic obstructive pulmonary disease, periodontitis and idiopathic inflammatory myopathy. The pharmaceutical composition of the present invention may be variously formulated according to the route of administration by a method known in the art together with a pharmaceutically acceptable carrier to produce the immunomodulation and inflammation inhibitory effect of the aforementioned herbal extract. 'Pharmaceutically acceptable' means a non-toxic composition which, when administered to humans, does not inhibit the action of the active ingredient and does not normally cause allergic reactions such as gastrointestinal disorders, dizziness or similar reactions. Say. Such carriers include all kinds of solvents, dispersion media, oil-in-water or water-in-oil emulsions, aqueous compositions, liposomes, microbeads and microsomes. The route of administration may be administered orally or parenterally. Parenteral administration methods include, but are not limited to, intravenous, intramuscular, intraarterial, intramedullary, intradural, cardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual or rectal administration Can be. In the case of oral administration of the pharmaceutical composition of the present invention, the pharmaceutical composition of the present invention may be a powder, granule, tablet, pill, dragee cap, liquid, gel, etc. according to methods known in the art together with a suitable oral carrier. And can be formulated in the form of syrups, suspensions, wafers and the like. Examples of suitable carriers include sugars and corn starch, wheat starch, rice starch and potato starch, including lactose, dextrose, sucrose, sorbet, mannitol, xylly, erytholi and maltyl, etc. Layered agents such as cellulose, gelatin, polyvinylpyrrolidone, and the like, including starch, cellulose, methyl cellulose, sodium carboxymethyl cellulose, hydroxypropylmethyl cellulose, and the like. In some cases, crosslinked polyvinylpyridone, agar, alginic acid or sodium alginate may be added as a disintegrant. Furthermore, the pharmaceutical composition may further include an anticoagulant, a lubricant, a humectant, a perfume, an emulsifier, and an antiseptic. In addition, when administered parenterally, the pharmaceutical compositions of the present invention may be formulated according to methods known in the art in the form of injections, transdermal and nasal inhalants together with suitable parenteral carriers. Such injections must be sterile and protected from contamination of microorganisms such as bacteria and fungi. Examples of suitable carriers for injection include, but are not limited to, solvents or dispersions comprising water, ethanol, polyols (e.g. glycerin, propylene glycol and liquid polyethylene glycols), mixtures thereof and / or vegetable oils May be a medium. More preferably, the suitable ^bearing, body is a Hanks solution, Ringer's solution, triethanolamine contains a PBS (phosphate buf fered sal ine) or injectable sterile water, 10% ethanol, 40% propylene glycol and 5% deck host rose and The same isotonic solution round can be used. In order to protect the injection from microbial contamination, various antibacterial and antifungal agents such as parabens, chlorobutane, phenol, sorbic acid, thimerosal, and the like may be further included. In addition, the injection may in most cases further comprise an isotonic agent such as sugar or sodium chloride. In the case of transdermal administrations, ointments, creams, lotions, gels, external preparations, pasta preparations, liniment preparations, aerosol preparations and the like are included. In the above, transdermal administration means that the pharmaceutical composition is locally administered to the skin such that an effective amount of the active ingredient contained in the pharmaceutical composition is delivered into the skin. For example, it may be administered in a way that the ^"state of the pharmaceutical composition of the invention was made of the scanning light coming from the skin formulations it as thin needles of 30 gauge terminals (pr ck i) or applied directly to the skin. These formulations are described in the prescription, generally known in pharmaceutical chemistry (Remington's Pharmaceut i Cal Science,-15th Edition, 1975, Mack Publ i Shing Company, East on, Pennsylvani a). In the case of inhaled dosages, the extracts used according to the invention can be prepared using pressurized packs or by means of suitable propellants, for example dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. It can be delivered conveniently from the nebulizer in the form of an aerosol spray. In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve to deliver a metered amount. For example, gelatin capsules and cartridges for use in inhalers or blowers may be formulated to contain a compound and a powdered mixture of suitable powder based such as lactose or starch. Other pharmaceutically acceptable carriers may be referred to those described in Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, East on, PA, 1995. The pharmaceutical compositions according to the invention may comprise one or more complete agents (e.g. saline or PBS), carbohydrates (e.g. glucose, mannose, sucrose or dextran), antioxidants, bacteriostatic agents, chelating agents (Eg, EDTA or glutathione), adjuvants (eg, aluminum hydroxide), suspending agents, thickening agents, and / or preservatives. The pharmaceutical compositions of the invention can be formulated using methods known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal. The pharmaceutical composition of the present invention can be administered in combination with known compounds that are effective in preventing or treating chronic inflammatory diseases. In another aspect, the present invention for the food for the improvement of chronic inflammatory diseases, characterized in that it comprises an extract of at least two herbal drugs selected from the group consisting of Inulae Flos, Schizonepetae Spica and Magnolia Cortex as an active ingredient To provide a composition. The composition of the present invention has the effect of improving the chronic inflammatory disease as demonstrated in the embodiment of the present invention. Chronic inflammatory diseases of the food composition of the present invention is preferably psoriasis, psoriatic arthritis, rheumatoid arthritis, multiple sclerosis, systemic lupus erythematosus, inflammatory bowel disease, Crohn's disease, ulcerative colitis, ankylosing spondylitis, asthma, chronic obstructive It may be selected from the group consisting of lung disease, periodontitis and idiopathic inflammatory myopathy. The food composition of the present invention includes all forms such as functional food, nutritional supplement, health food and food additives. Food compositions of this type are known in the art. It may be prepared in various forms according to conventional methods. For example, the health food may be prepared by drinking the composition itself in the form of tea, juice and drink, or ingested by granulation, encapsulation and powdering. Functional foods also include beverages (including alcoholic beverages), fruits and processed foods (e.g. canned fruit, canned food, jams, marmalade, etc.), fish, meat, and processed foods (e.g. ham, sausage corned beef, etc.); Breads and noodles (e.g. udon, soba, ramen, spaghetti, macaroni, etc.), fruit juices, various drinks, cookies, malts, dairy products (e.g. butter, cheese, etc.), edible vegetable oils, margarine, vegetable protein, retort food It may be prepared by adding extracts to frozen foods, various seasonings (eg, soy sauce, sauce, etc.). In addition, in order to use the composition of the present invention in the form of a food additive, it can be prepared in powder or concentrate form. A preferred content of the extract in the food composition of the present invention may be 0.001 to 50% based on the total weight of the food composition, preferably contained in the range of 0.1 to 50%. The invention is well illustrated in the Examples.

In one embodiment of the present invention, 200 g of scallop, mold opening, and thick foil were added twice each for 4 hours under reflux cooling extraction conditions by adding 4 L of 95% ethanol, and then the extraction solution was filtered with a filtering device and decompressed. Ethanol was removed by the apparatus to obtain a single extract. In another embodiment of the present invention, the effect of a single extract and a mixed extract prepared through the above process on the secretion of inflammatory cytokines was confirmed in the splenic cells of ICR mice and RAW264.7 cells, which are mouse macrophage lines. Splenocytes were stimulated with IL-23 and measured for IL-17. Macrophages were stimulated with LPS (l ipopolysacchar i de) and measured for IL-6 and TNF. The single extract of each herbal medicine also had the effect of inhibiting the secretion of the inflammatory cytokines, and especially when two or more single extracts were combined, the cytokine inhibitory effect was observed more remarkably. In another embodiment of the present invention was examined the content ratio to maximize the synergistic effect of inhibition of inflammation between herbal extracts. Compound extracts were prepared by mixing two or more herbal medicines selected from sunbok, gyeonghyeong and thick cakes in various weight ratios. The inhibitory effects of inflammatory cytokines such as IL-17, IL-6, and TNF-α were compared in splenocytes and macrophages according to the weight of herbal medicines. In another embodiment of the present invention, the therapeutic effect of the composition according to the present invention for chronic inflammatory diseases was confirmed in an animal model of psoriasis. Intradermal administration of IL-23 into the ear of C57BL / 6 mice every other day induces psoriasis symptoms. The expression levels of keratin 17, IL-17, IL-23, IL-6, and TNF-α, which are biomarkers of psoriasis, were measured. The complex extract was confirmed to have an effect of alleviating and reducing psoriasis symptoms in a mouse model of psoriasis to the extent that the tofacitinib positive control group. The present invention provides a method for treating chronic inflammatory disease by administering to a subject in need thereof an extract of at least two herbal drugs selected from the group consisting of Inulae Flos, Schizonepetae Spica, and Magnolia Cortex. do. The present invention provides a use for the preparation of a therapeutic agent for the treatment of chronic inflammatory diseases comprising an extract of at least two herbal drugs selected from the group consisting of Inulae Flos, Schizonepetae Spica and Magnolia Cortex. to provide. The term 'effective amount' of the present invention, when administered to a subject, refers to an amount that exhibits an effect of improving, treating and preventing chronic inflammatory diseases, and the term 'individual' refers to an animal, preferably a mammal, particularly an animal including a human. It may be a cell, tissue, organ, etc. derived from the animal. The subject may be a patient in need of treatment.

The ^first treatment of the invention'inclusively refers to ameliorating the symptoms of a chronic inflammatory disease, which may include treating, substantially preventing, and / or ameliorating the condition, from a chronic inflammatory disease It includes, but is not limited to, alleviating, healing or preventing one symptom or most of the symptoms that result. Advantageous Effects

 Therefore, the present invention provides a composition for the prevention, treatment or improvement of chronic inflammatory diseases comprising a mixed herbal extract. The composition of the present invention is effective in preventing and treating chronic inflammatory diseases by effectively inhibiting the secretion of inflammatory cytokines such as IL-17, IL-6, TNF-Q in immune cells. In addition, since the composition of the present invention contains an herbal extract as an active ingredient, side effects on the human body are extremely smaller than those of chemically synthesized drugs.

[Brief Description of Drawings]

 Figure 1 is a composite extract of algae and mold in the animal model of psoriasis (Example <3-1>, weight ratio 1: 6), composite extract of algae and thickening (Example <3-2>, weight ratio 2: 1 ) And the effect of tofacitinib (positive control) on ear thickness.

 Figure 2 is a composite extract of algae and molds in animal models of psoriasis (Example <3-1>, weight ratio 1: 6). It is a representative slide photograph showing the effect on the epidermal thickness in the ear tissue when the complex extract of the ablation and thickening (Example <3-2>, weight ratio 2: 1) and tofacitinib (positive control) was treated.

 Figure 3 is a composite extract of gnarling and mold of psoriasis in the animal model of psoriasis (Example <3-1>, weight ratio 1: 6), composite extract of gnarling and thickening (Example <3-2>, weight ratio 2: 1 ) And tofacitinib (positive control) are graphs showing the degree of inflammatory cell invasion in the ear tissue. Inflammatory cell invasion was observed by optical microscope after H & E staining to evaluate the degree of inflammatory cell invasion. Asymptomatic 0, mild 1, moderate 2, severe 3

 Fig. 4a shows a composite extract of algae and molds in an animal model of psoriasis (Example <3-1>, weight ratio 1: 6), a composite extract of algae and thick skin (Example <3-2>, weight ratio 2: 1 ) And tofacitinib (positive control) showed mRNA expression in keratin 17 and IL-17 genes in your tissues compared to the PBS control group.

Figure 4b is a composite extract of algae and mold in the animal model of psoriasis (Example <3-1>, weight ratio 1: 6), composite extract of algae and thickening (Example <3-2>, weight ratio 2: 1 ) And tofacitinib (positive control) showed mRNA expression levels of IL-23, TNF-α, and IL-6 genes in your tissues compared to PBS controls. [Form for implementation of invention]

Hereinafter, the present invention will be described in detail. ^'

 However, the following examples are merely to illustrate the present invention, the content of the present invention is not limited to the following embodiments.

<Example 1>

 Preparation of Herbal Extracts

<ι-ι> Preparation of Roots Alone Extract

Inulae Flos was washed with water to remove contaminants and completely dried. The _{200 g} of the prepared flounder was reflux-cooled twice with 4 _L of 95% (v / v) ethanol solution for 4 hours and then extracted. Concentration under reduced pressure after filtration. After azeotropically concentrating azeotropically by adding 0.2 L of water while most solvents were evaporated, the mixture was repeated twice, suspended in a homogeneous manner, and then lyophilized to obtain 26 g of a powdered extract.

<1-2> Preparation of a single open extract

 The mold dog (Schizonepetae Spi ca) was washed with water to remove impurities and completely dried. 200 g of the molds thus prepared were refluxed twice with 4 L of 95% (v / v) ethane twice in an aqueous solution for 4 hours, and the extract was filtered and concentrated under reduced pressure. 0.2L of water was removed while most solvents were evaporated. After the azeotropic concentration was added, the mixture was repeated twice, suspended in a homogeneous amount by adding the same amount of water again, and then lyophilized to obtain 10 g of a mold extract in powder form.

<1-3> Preparation of Thick Single Extract

The hump (Magnol ia Cortex) was washed with water to remove any contaminants and allowed to dry completely. 200 g of the prepared thin foil was reflux-cooled twice with 4 L of 95% (v / v) ethanol solution for 4 hours, and the extract was filtered and concentrated under reduced pressure. 0.2L of water was added and most of the solvent was evaporated, and then azeotropically concentrated. Then, the mixture was repeated twice, suspended in a homogeneous solution, and then lyophilized to obtain 34 g of a powdery extract. <Example 2>

 Gymnastics mixture of herbal extracts

<2-1> Preparation of mixed extract of loosening and mold

 The sundae monocotyledonous extract prepared in Example <1-1> and the mold alone extract prepared in Example <1-2> were mixed at a weight ratio of 1: 1.

<2-2> Combined extract manufacturing of loosening and thickening

 The sunkalone extract prepared in Example <1-1> and the hulk extract alone prepared in Example <1-3> were mixed at a weight ratio of 1: 1.

<2-3> Preparation of a mixed extract of a mold and a thick foil

 The mold alone extract prepared in Example <1-2> and the extract of hupak alone prepared in Examples <1-3> were mixed at a weight ratio of 1: 1.

<Example 3>

 Gymnastics of herbal medicine extract

<3-1> Preparation of loosening and open complex extract

Liberation and mold opening were combined in a weight ratio of 1:15, 1: 9, 1: 6, 1: 3, 1: 1, 1.6: 1. To the mixture thus prepared was added 9 (v / v) ethane aqueous solution, reflux cooled for 4 hours, filtered and the filtrate was collected. For residue, 95% (v / v) ethanol aqueous solution was added and 80 ^{° C} for 4 hours. The filtrate obtained by reheating extraction at C was combined with the filtrate previously collected and concentrated under reduced pressure.

<3-2> Preparation of thinning and thick composite extract

Trivia and thickening were combined in weight ratios of 3: 1, 2: 1, 1: 1, 1: 2, 1: 3. 95% (v / v) ethane was added to the mixture, and the mixture was refluxed and cooled for 4 hours, and the filtrate was collected and collected. For residue, 9 (v / v) ethane was added to the mixture for 4 hours. The filtrate obtained by reheating extraction at 80 ^° C. was concentrated under reduced pressure by mixing with the previously collected filtrate. <3-3> Preparation of Hyungae and Thubark Composite Extracts

The mold and thickening were mixed in a weight ratio of 3: 1, 1: 1, 1: 1.6. 95% (v / v) ethanol aqueous solution was added to the mixture thus prepared, reflux-cooled extraction was performed for 4 hours, the filtrate was collected by collecting the filtrate, and the residue was added with 95% (v / v) ethanol aqueous solution at 80 ^{° C} for 4 hours. The filtrate obtained by reheating extraction at C was combined with the filtrate previously collected and concentrated under reduced pressure.

<3-4> Preparation of loosening, mold opening and thick extract

Trichophysis, mold opening and thickening were combined at a weight ratio of 3: 1: 1. 95% (v / v) ethanol aqueous solution was added to the mixture thus prepared, reflux-cooled for 4 hours, filtered and the filtrate was collected and the residue was collected for 4 hours by adding 95% (v / v) ethanol aqueous solution. The filtrate obtained by reheating extraction at 80 ^° C. was concentrated under reduced pressure by mixing with the previously collected filtrate.

<Example 4>

 Pharmacological Activity Experiments of Herbal Extracts

<4-1> Analysis of the effect of the herbal extracts on the secretion of IL-17-induced IL-23 alone extract of the herbal preparations prepared in <Example 1> and prepared in <Example 2> and <Example 3> The effect of mixed herbal extracts on the secretory activity of IL-17 was confirmed in splenocytes of mice.

Experimental animals (ICR mice) were euthanized by C0 ₂ anesthesia or cervical dislocation method.

The abdomen was cleaned with 70% ethanol, opened using surgical scissors, spleens were removed, peripheral tissues were removed, and placed in PBS (Phosphate buffered saline) solution. After washing the spleen twice with PBS, the spleen was transferred to a culture dish, sliced into 1 ~ 2 dragons with a scalpel blade, and the cells were separated using a cell strainer and a syringe. The separated solution at the bottom of the strainer was transferred to a microfluid and centrifuged at 800 g for 3 minutes. The supernatant was removed, ACK lysis buffer (Amnion ium-Chlor i de-Potassium lysis buffer) was added 5m and the pellets were redispersed and incubated for 4 to 5 minutes. PBS 40rn.e was added and centrifuged at 800 g for 3 minutes. The supernatant was again removed, 5m RPMI-1640 complete let media was added and gently redispersed. Since sal Clumps were removed by filtration with strainer, and then the number of cells was ^^ and seeded in 100, 96 wel l plate at a concentration of 6 x loVwel l. After incubation for 1 hour after treatment of the test material, and incubated for 72 hours after treatment with 10ng / IL-23-. The supernatant was collected and used immediately or stored at -70 ° C. ELISA assay was performed using R & D IL-17 (Cat No. DY421) analysis system and was performed according to the manufacturer's recommended method. Cytokine inhibition rate excluded effects by cytotoxicity. The experimental results are shown in the IL-17 column of Table 1 and Table 2 as the efficiency (inhibition rate,%) of each herbal extract to inhibit the secretion of IL-17 in mouse spleen cells. First, as shown in Table 1, the effects of the single extract of each herbal prepared in <Example 1> and the herbal extracts prepared in <Example 2> was compared. The single extract of each herb also showed the effect of inhibiting the secretion of IL-17, but it was confirmed that the effect of inhibiting the secretion of IL-17 was further improved when the mixed extract was treated more than the single extract of the herb. In other words, by extracting each herbal extract, a synergistic effect of inhibiting the secretion of the inflammatory cytokine IL-17 can be obtained. Next, as shown in Table 2, in order to determine the most preferable composition ratio of the herbal extracts were compared the inhibitory effect of IL-17 secretion of the herbal extracts prepared in <Example 3>. As a result, the combined extracts of sunbyeong and hyeonggae were superior to the inhibitory effect of IL-17 especially when the two herbal medicines were mixed at a weight ratio of 1: 6. Or when mixed at a weight ratio of 2: 1, the most effective. In addition, the extracts of hyunggae and hoobak showed the best inhibitory effect of IL-17 when the two herbal medicines were mixed in a weight ratio of 3: 1.

<4-2> Effect of Herbal Extracts on IL-6 and TNF- Secretion Induced by LPS Treatment

 Effects of the extracts of the herbal extract prepared in <Example 1> and the herbal extracts prepared in <Example 2> and <Example 3> on the IL-6 and TNF-α secretion activity RAW It was confirmed in 264.7 cells.

 RAW 264.7 cells were cultured in DMEM medium containing 10% FBS and W penicillin / streptomycin. On the day of the experiment, cells were collected with a scraper and then hemocytometer

Measure the number of cells using a hematocytometer and place 5xl () ⁴ cel l / wel 1 on a 96 wel l plate. Seeding was carried out. This was incubated for 24 hours, and the herbal sample treatment was completely dissolved in DMSOCDimethyl Sulfoxide using serum-free medium as a conditional medium. After 1 hour, LPS (l ipolypolysaccharide) was added. Treated to 100ng / and incubated for 24 hours. After culture for 24 hours, the culture medium supernatant of rat macrophages was collected using a rat IL-6 and TNF-α immunoassay kit. Collected medium supernatant is diluted in serum-free medium and the baseline concentration is 15.625-1000 pg / m. The diluted solution is added to 96 wel l piate and reacted at room temperature for 2 hours. After reaction, the solution was washed five times with washing solution, and the reaction was performed at room temperature for 1 hour after the addition of each antibody conjugate. After the reaction, the solution was washed 7 times with the washing solution, the substrate was added and reacted at room temperature for 30 minutes. The reaction was stopped with the stop solution and the absorbance was measured at 450 nm. Immunoassay kits of IL-6 and TNF-α were used for mouse EL ISA kit (BD science), respectively. First, as shown in Table 1, the effect of inhibiting the secretion of IL-6 and TNF-α between the extracts of the herbal preparations prepared in <Example 1> and the herbal extracts prepared in <Example 2> was compared. In addition, the extracts of each herb showed the effect of inhibiting the secretion of IL-6 and TNF-a, but the combination of the extracts treated with the combination extract of IL-6 and TNF-a further improved the effect of inhibiting the secretion of IL-6 and TNF-a. Confirmed. In other words, when the extracts of each herb are treated in combination, it can be seen that they produce a synergistic effect of inhibiting the secretion of inflammatory cytokines IL-6 and TNF-a. In order to determine the most preferable composition ratio of the herbal extract as shown in Table 2, the secretion inhibitory effect of IL-6 and TNF-a of the herbal complex extract prepared in <Example 3> was compared. As a result, the combined extracts of sunbyeong and hyeonggae showed high inhibitory effect of IL-6 at most weight ratios, and the inhibitory effect of TNF-a on complex ratios of 1: 1 and 1.6: 1 The effect appeared to be good. On the other hand, the combined extracts of the abalone and hubac were found to have high inhibitory effects on the secretion of IL-6 and TNF-a at most weight ratios. On the other hand, in the case of complex extracts of hyunghwa and hobak, the secretion inhibitory effect of IL-6 or TNF-a was lower than that of the complex extracts of sundae.

Table 11 Comparison of Cytokine Secretion Inhibitory Activity between Herbal Extracts and Mixed Extracts

Table 2

Cytokine Secretion Inhibitory Activity of the Medicinal Herbal Extracts

Therapeutic Effect of Herbal Extracts in Animal Models of Psoriasis

 Combined extracts of concoctions and ginsengs showing excellent synergistic effect of secretion of IL-17, IL-6 and TNF-α in the above examples (Examples <3-1>, weight ratio 1: 6) Using a complex extract (Example <3_2>, weight ratio 2: 1) of the psoriasis treatment effect in the body according to the method of Ma [J. Clin. Cell Immunol. 4: 6. (2013)].

Psoriasis was induced by intradermal administration of 500 ng IL-23 every other day into the ears of C57BL / 6 mice (6 weeks old Orient Bio). Tofacitinib, an oral drug, was used as a positive control and 10 rag / kg bid and 30 mg / kg bid were administered with reference to the literature. The administration method of the combined extract and thinning extract (Example <3-1>, weight ratio 1: 6) and the combined extract and thick extract (Example <3-2>, weight ratio 2: 1) are the same as the positive control group. The bid oral administration method was used and the dose was administered at 100 mg / kg bid and 200 mg / kg bid, respectively. The administration was performed 14 times for a total of 16 days, and the experiment was terminated at the start of 17 days. After the experiment was finished, the ear thickness of the mouse was measured, and ear tissues were extracted, and then epidermal thickness, inflammatory cell invasion, and mRNA expression analysis were performed. The results are shown in FIGS. 1 to 4. As shown in Fig. 1, the composite extract of the capillary scapulae and ginseng (Example <3-1>, weight ratio 1: 6) and the complex extract of the capillary and scabbard (Example <3-2>, weight ratio 2: 1 Psoriasis animals It can be seen that there is an ear thickness reduction effect in the model. In addition, when compared with the positive control tofacitinib in the ear thickness reduction portion, it can be confirmed that the non-inferiority. As shown in FIG. 2, the combined extract and mold opening extract (Example 3-1>, weight ratio 1: 6) and the combined extract and thick extract (Example <3-2>, weight ratio 2: 1) are tofacitinib. Compared to the tofacitinib-treated positive control group, psoriasis animal model has an effect of reducing epidermal thickness in the ear tissue. As shown in Fig. 3, the composite extract of the capillary linus and the mold of the capillary (Example <3-1>, the middle ring: ratio 1: 6) and the complex extract of the capillary lining and the thick leaf (Example <3-2>, weight ratio 2 : 1) can be found to reduce the infiltration of inflammatory cells in psoriasis animal model. In addition, it was confirmed that the efficacy of the combined extract of Hwahwa and hyeonggae (Example <3-1>, weight ratio 1: 6) is relatively better than the combined extract of Hwahwa and thick skin (Example <3-2>, weight ratio 2: 1). Can be. As shown in Figures 4a and 4b, the composite extract of the sunkwa and hyeongjeop (Example <3-1>, weight ratio 1: 6) and the complex extract of sunkwa and thick (Example <3-2>, weight ratio 2 : 1)) can be found to reduce the mRNA expression of keratin 17, IL-17, IL-23, TNF-α and IL-6, which are related to psoriasis biomarkers in psoriasis animal models. Compared with the positive control group, it can be confirmed that there is an effect of lowering the expression of keratin 17, IL-17, and IL-23 genes corresponding to a more psoriasis-specific mechanism.

Industrial Applicability

 As described above, the present invention provides a composition for preventing, treating or ameliorating chronic inflammatory disease, which comprises at least two herbal extracts selected from the group consisting of acidification, mold opening and thickening. The composition of the present invention can be effectively used to prevent and treat chronic inflammatory diseases by effectively inhibiting the activity of inflammatory cytokines.

Claims

[Range of request]

 [Claim 1]

 A pharmaceutical composition for the treatment of chronic inflammatory diseases, comprising extracts of two or more herbal drugs selected from the group consisting of Inulae Flos, Schizonepetae Spica, and Magnolia Cortex.

[Claim 2]

 The composition according to claim U, wherein the extract is extracted with any one solvent selected from the group consisting of water, alcohols having 1 to 6 carbon atoms, nucleic acids, ethyl acetate, and mixed solvents thereof.

[Claim 3]

 The method of claim 1, wherein the chronic inflammatory disease is psoriasis, psoriatic arthritis, rheumatoid arthritis, multiple sclerosis, systemic lupus erythematosus, inflammatory bowel disease, Crohn's disease, ulcerative colitis. Ankylosing spondylitis, asthma, chronic obstructive pulmonary disease, periodontitis, and idiopathic inflammatory myopathy.

[Claim 4]

 A composition for improving chronic inflammatory disease, comprising extracts of at least one herbal medicine selected from the group consisting of Inulae Flos, Schizonepetae Spica and Magnolia Cortex as active ingredients.

[Claim 5]

 A method of treating chronic inflammatory disease by administering an effective amount of an extract of two or more herbal drugs selected from the group consisting of Inulae Flos, Schizonepetae Spica, and Magnolia Cortex.

[Claim 6]

 Use for the preparation of a therapeutic agent for the treatment of chronic inflammatory diseases comprising an extract of at least two herbal drugs selected from the group consisting of Inulae Flos, Schizonepetae Spica and Magnolia Cortex.