KR102872539B1 - Anti-inflammatory composition comprising extract of Hypholoma sublateritium as an active ingredient - Google Patents

Abstract

The present invention relates to an anti-inflammatory composition comprising a oyster mushroom extract as an active ingredient. More specifically, the composition can be used as a cosmetic composition by suppressing the production of nitric oxide (NO), prostaglandin E2 (PGE2), and inflammatory cytokines with reduced side effects, and can also be usefully used as a skin topical agent, a functional food composition, and tea.

Description

Anti-inflammatory composition comprising extract of Hypholoma sublateritium as an active ingredient

The present invention relates to an anti-inflammatory composition comprising a oyster mushroom extract as an active ingredient. More specifically, the composition relates to a composition capable of enhancing the inhibitory effect on the production of nitric oxide (NO), prostaglandin E2 (PGE2), and inflammatory cytokines with reduced side effects.

Inflammation is a localized defense response of the body in response to pathological conditions caused by physical trauma, harmful chemicals, infections by bacteria, fungi, and viruses, or irritants among the body's metabolic products. Inflammation is triggered by various inflammatory mediators produced by damaged tissues and migrating cells. During an inflammatory response, plasma accumulates at the site of inflammation, diluting the toxins secreted by bacteria. Blood flow increases, and symptoms such as erythema, pain, swelling, and fever accompany the response. Normally, the body uses the inflammatory response to neutralize or eliminate the pathogenic factor and regenerate damaged tissues to restore normal structure and function. However, if this fails, it can progress to a disease state such as chronic inflammation.

Recently, with the development of molecular biology, many studies on inflammatory responses at the molecular level are being conducted, and although various biochemical phenomena are involved in inflammatory responses, macrophages in particular are known to play an important role in inflammatory responses by producing nitric oxide (NO) and various inflammatory cytokines in response to chemical stimuli (Ito T., et al, Curr Drug Traget Inflamm Allergy, 2(3);257-265, 2003). Nitric oxide is synthesized from L-arginine by the action of nitric oxide synthase (NOS), and NOS exists in several isoforms. b/NOS (brain NOS) in the brain, nNOS (neuronal NOS) in the nervous system, and eNOS (endothelial NOS) in the vascular endothelium are always expressed at a certain level in the body, and the nitric oxide (NO) produced in small amounts by them plays an important role in maintaining homeostasis in the human body by performing various physiological reactions related to blood pressure regulation, neurotransmission, learning, and memory. In contrast, iNOS (induced NOS), whose expression is induced by a certain stimulus, produces excessive NO, and the nitric oxide (NO) produced excessively by iNOS reacts with superoxide to form peroxynitrite. It acts as a powerful oxidant and causes cell damage, thereby being involved in various pathological processes including inflammation and cancer (Gupta SC et al., Exp Biol Med., 236;658-671, 2011; Riehemann et al., FEBS Lett, 442;89-94, 1999; Stamler et al., Science, 358;1898-1902, 1992).

Meanwhile, cyclooxygenase (COX) has the function of COX and hydroperoxidase (HOX) activity, and synthesizes intermediates PGG ₂ and PGG ₂ from arachidonic acid, and produces PGE ₂ , PGF ₂ , PGD ₂ , prostacyclin, and thromboxane A2 (TxA2) from these compounds. There are two isoforms of COX. COX-1 is constantly expressed in most tissues and synthesizes prostaglandins (PGs) necessary for cell protection, whereas COX-2 is rapidly induced in response to inflammation and plays an important role in causing inflammation by producing PGE _2, etc. (Weisz A., Biochem. J., 316:209-215, 1996; (Miller MJ et al., Mediators of inflammation, 4:387-396, 1995; Appleton L. et al., Adv. Pharmacol., 35:27-28, 1996).

PGE ₂ is best known as an inflammatory mediator, and is involved in tissue damage by inducing the production of matrix metalloproteinases (MMPs) in inflammatory diseases such as periodontitis. Large amounts of PGE ₂ are produced in macrophages obtained from patients with rheumatoid arthritis, and the produced PGE ₂ plays an important role in the inflammatory response and tissue destruction mechanism in rheumatoid arthritis.

In addition to its well-known role as a mediator of inflammatory responses, PGE ₂ has also been shown in many studies in recent years to play a role as a regulator of immune responses by promoting Th2 type immune responses, suppressing Th1 type immune responses, inhibiting the production of inflammatory cytokines such as TNF-α, IL-1β, IL-8, and IL-12 in macrophages, and promoting the production of anti-inflammatory cytokines such as IL-10.

The expression of iNOS and COX-2, which induce excessive production of NO and PGs, which serve as mediators of this inflammatory response, is regulated by the nuclear transcription factor NF-κB. NF-κB is a nuclear protein of the Rel gene family, and exists in an inactive form in the cytoplasm by binding to I-κB. However, when I-κB kinase is activated by a certain factor, I-κB is detached through a phosphorylation process, thereby becoming activated. NF-κB, thus activated, moves to the nucleus and induces the gene expression of iNOS and COX-2, which induce an inflammatory response (Oh, G. T. et al., Atherosclerosis, 159(1):17-26, 2001).

Accordingly, the inventors of the present invention conducted research by using LPS (lipopolysaccharide), a bacterial endotoxin that can activate the transcription factor NF-κB and induce the expression of iNOS and COX-2, thereby inducing the secretion of various inflammation-regulating substances such as NO, inflammatory cytokines, and PGE ₂ .

These research results suggest that drugs that inhibit the production of NO or the production of inflammatory cytokines such as TNF-α and IL-6 may have the potential to be effective anti-inflammatory agents by inhibiting the expression of iNOS or COX-2.

Non-steroidal anti-inflammatory drugs (NSAIDS), which are currently widely used as anti-inflammatory agents, are known to cause serious side effects such as gastrointestinal disorders, liver disorders, and kidney disorders.

Therefore, it can be said that there is still a need for the development of new drugs that have anti-inflammatory activity, few side effects, and lasting effects.

Accordingly, the inventors of the present invention sought to provide anti-inflammatory effects by including a natural extract that is safe and has minimal side effects. They confirmed that the extract of the oyster mushroom exhibited anti-inflammatory effects, and completed the present invention.

[Prior Art Literature]

[Patent Document]

(Patent Document 1) Republic of Korea Patent No. 10-0847103

The purpose of the present invention is to provide an anti-inflammatory composition containing an extract of the dogtooth mushroom as an active ingredient.

Another object of the present invention is to provide a cosmetic composition that includes a mushroom extract as an active ingredient, thereby minimizing side effects upon administration and taking, and exhibiting an anti-inflammatory effect.

Another object of the present invention is to provide a skin topical preparation that includes an extract of the dogwood mushroom as an active ingredient, thereby minimizing side effects upon administration and taking, and exhibiting an anti-inflammatory effect.

Another object of the present invention is to provide a functional food composition that includes a mushroom extract as an active ingredient, thereby minimizing side effects upon administration and taking, and exhibiting an anti-inflammatory effect.

Another object of the present invention is to provide a tea that contains an extract of the dogwood mushroom as an active ingredient, thereby minimizing side effects upon administration and taking, and at the same time exhibiting an anti-inflammatory effect.

To achieve the above purpose, an anti-inflammatory composition according to one embodiment of the present invention includes a oyster mushroom extract as an active ingredient.

The above composition exhibits an anti-inflammatory effect by inhibiting the production of nitric oxide (NO), prostaglandin E2 (PGE2) and inflammatory cytokines.

The above extract is extracted using an extraction solvent selected from the group consisting of water, lower alcohols having 1 to 6 carbon atoms, and mixtures thereof.

The above composition further comprises a natural extract selected from the group consisting of an extract of Ophiopogon japonicus, an extract of Magnolia chinensis, an extract of Atractylodes japonica, and a mixture thereof.

A cosmetic composition according to another embodiment of the present invention comprises the composition.

A skin external preparation according to another embodiment of the present invention comprises the composition.

A functional food composition according to another embodiment of the present invention comprises the composition.

Tea according to another embodiment of the present invention comprises the composition.

Hereinafter, the present invention will be described in more detail.

Unless otherwise defined, all technical terms used in the present invention have the following definitions and correspond to the meanings generally understood by those skilled in the art in the relevant fields of the present invention. Furthermore, while preferred methods and samples are described herein, similar or equivalent methods are also included within the scope of the present invention.

In the specification of the present invention, the plants are not limited in type, and can be used without limitation on their source, such as those grown or those purchased commercially.

In the specification of the present invention, the extract may be extracted using a method known in the art, and the method is not particularly limited. Alternatively, a commercially available extract may be used.

The above plant extract can be made from any part of the plant, and is not limited to the extraction site. The production of the above plant extract is not limited by the form of the plant, and the plant includes all parts that have undergone processing such as drying. For example, the plant may be the whole plant, root, stem, leaf, fruit, flower, shoot, branch, bark, sap, bud, and/or seed (seed) of the above-mentioned plant.

The term "extract" used herein has the meaning commonly used in the art as a crude extract as described above, but in a broader sense, it also includes fractions obtained by further fractionation of the extract. That is, the extract includes not only those obtained using the above-mentioned extraction solvent, but also those obtained by additionally applying a purification process thereto. For example, fractions obtained by passing the extract through an ultrafiltration membrane having a certain molecular weight cut-off value, separation by various chromatographies (designed for separation according to size, charge, hydrophobicity, or affinity), and other fractions obtained through various additional purification methods are also included in the extract of the present invention. In addition, it includes all extracts in the form of extracts themselves and extracts that can be formed using the extract, such as dilutions or concentrates of the extract, dried products obtained by drying the extract, adjusted or purified products of the extract, fermented products of the extract, or mixtures thereof. In addition, it includes juice obtained by directly pressing or crushing a plant and then filtering it. The above plant can be extracted as is or by processing. The above "processing" refers to a pharmaceutical technology that changes the original properties of a medicinal herb by processing the medicinal herb based on oriental medicine theory, and examples thereof include the "cho" (frying) method of roasting medicinal herbs, the "ja" (roasting) method of roasting medicinal herbs with a certain amount of liquid auxiliary ingredients so that the auxiliary ingredients permeate the medicinal tissue, and the "jeung" (steaming) method of adding and mixing liquid auxiliary ingredients according to the processing regulations for each medicinal herb and heating and steaming in a suitable container, or steaming until it reaches a certain degree and then drying. In addition, it can refer to a product extracted using an extraction solvent from a strain itself, a fragment of a strain, a culture thereof, a dead body thereof, or a mixture thereof.

The term "fraction" used in the present invention means a result obtained by performing fractionation to separate a specific component or a specific group of components from a mixture containing various components.

As used herein, when a part is said to "include" a component, this does not mean that it excludes other components, but rather that it may include other components, unless otherwise specifically stated.

The term "improvement" as used herein means any action that at least reduces the severity of a symptom, for example, a parameter associated with the alleviation or treatment of a condition.

The term "about" as used herein means an amount, level, value, number, frequency, percentage, dimension, size, amount, weight, or length that varies by about 30, 25, 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1% relative to a reference amount, level, value, number, frequency, percentage, dimension, size, amount, weight, or length.

In addition, the food composition may include a health functional food. In this specification, "health functional food" refers to a food group that has been given added value by using physical, biochemical, or bioengineering techniques to enable the function of the food to function and express for a specific purpose, or a food that has been designed and processed to sufficiently express the bioregulatory function of the food composition, such as regulating the biological defense rhythm, disease prevention, and recovery, in the body. It has a more active health maintenance or promotion effect than general foods, and health supplement food refers to a food for the purpose of health supplementation. In some cases, the terms functional food, health food, and health supplement food are used interchangeably.

An anti-inflammatory composition according to one embodiment of the present invention comprises a oyster mushroom as an active ingredient.

Hypholoma sublateritium is a brown-rotting mushroom that grows in clusters on dead stumps of broad-leaved trees such as alder and oak in late autumn. The cap is hemispherical, 3–10 cm in diameter, and the stipe is usually 5–10 cm long. The stipe becomes hollow when mature, and the cap and stem are distinct. The surface of the cap is sticky and tile-colored when moist, and the periphery of the cap is initially covered with a thin, whitish fibrous membrane, which later turns light brown. The wrinkles on the back of the cap are dense and yellowish-white at first, and the spores are oval, measuring 5.5–8 × 3–4 ㎛. Hypholoma sublateritium is native to the northern hemisphere, including Korea, Japan, and China, and is known to have excellent pharmacological properties, including anticancer properties.

In the present invention, all parts of natural plants can be used, and there is no limitation on the extraction part. The production of the plant extract is not limited by the form of the plant, and the plant includes all plants that have undergone processing such as drying.

The present invention provides an anti-inflammatory composition characterized by including an extract of a oyster mushroom having the above-described characteristics.

The extraction method in the present invention is not particularly limited, and extraction can be performed according to methods commonly used in the art. Non-limiting examples of the extraction methods include solvent extraction, hot water extraction, ultrasonic extraction, filtration, and reflux extraction, and these may be performed alone or in combination of two or more methods.

In the present invention, the type of extraction solvent used for extraction is not particularly limited, and any solvent known in the art can be used. In the present invention, the extract of the oyster mushroom is extracted using an extraction solvent selected from the group consisting of water, lower alcohols having 1 to 6 carbon atoms, or mixed solvents thereof. In addition, non-limiting examples of the extraction solvent include water; lower alcohols having 1 to 6 carbon atoms such as methanol, ethanol, propyl alcohol, and butyl alcohol; polyhydric alcohols such as glycerin, butylene glycol, and propylene glycol; hydrocarbon solvents such as methyl acetate, ethyl acetate, acetone, benzene, hexane, diethyl ether, and dichloromethane; or mixtures thereof. Specifically, water, lower alcohols, 1,3-butylene glycol, and ethyl acetate may be used alone or in combination of two or more thereof. At this time, when two or more types of solvents are mixed and used, the mixing ratio between the solvents is not particularly limited.

Specifically, in order to manufacture a oyster mushroom extract, a step of washing oyster mushrooms; a step of drying after washing; a step of crushing oyster mushrooms after drying; a step of leaching the crushed product using an organic solvent; a step of drying the sample after leaching; a step of leaching using water; and a step of leaching are included, thereby obtaining a oyster mushroom extract.

For extraction, the sample may be left to leach in the extraction solvent for 1 to 72 hours, more specifically for 24 to 48 hours.

In the present invention, the extraction may be performed using a solvent of 1 to 100 times the weight, specifically 1 to 50 times the weight, and more specifically 2 to 20 times the weight of the plant dried material, at an extraction temperature of 10 to 80°C, specifically 15 to 50°C, for an extraction time of 2 hours to 30 days, specifically 12 hours to 18 days, and may include a process of obtaining a liquid crude extract by performing continuous extraction once to five times, including the dried and crushed material.

The extract extracted using the organic solvent may further include a step of performing fractionation using the organic solvent, and the method for preparing the extract may be a conventional extraction method in the art, such as an ultrasonic extraction method, an leaching method, and a reflux extraction method.

The above reflux extraction method is performed using 10 to 30 g of a pulverized natural product, 1 to 3 hours of reflux time, and 50 to 100% of alcohol having 1 to 6 carbon atoms or water, based on 100 mL of water or alcohol having 1 to 6 carbon atoms. More specifically, the method is performed using 10 to 20 g of a pulverized natural product, 1 to 2 hours of reflux time, and 70 to 90% of alcohol having 1 to 4 carbon atoms or water, based on 100 mL of water or alcohol having 1 to 6 carbon atoms.

The above leaching method is carried out at 15 to 30°C for 24 to 72 hours, and uses water or 50 to 100% alcohol having 1 to 6 carbon atoms as the extraction solvent. More specifically, it is carried out at 20 to 25°C for 30 to 54 hours, and the extraction solvent is water or 70 to 80% alcohol having 1 to 6 carbon atoms.

The above ultrasonic extraction method is carried out at 30 to 50°C for 0.5 to 2.5 hours, and the extraction solvent is water or 50 to 100% alcohol having 1 to 6 carbon atoms. Specifically, the extraction is carried out at 40 to 50°C for 1 to 2.5 hours, and the extraction solvent is water or 70 to 80% alcohol having 1 to 6 carbon atoms.

In the present invention, the extract may be used as is after filtering the particles using, for example, nylon, filter paper, etc. to remove floating solid particles, or filtering using a freeze filtration method, etc., or drying it using freeze drying, hot air drying, spray drying, etc.

The above liquid crude extract can be separated from the dried plant fragments by a method such as vacuum filtration, and then subjected to a process of concentration or drying. For example, the above liquid crude extract can be a concentrate obtained by concentrating under reduced pressure at 20 to 100°C, preferably 30 to 70°C, using a vacuum rotary concentrator, or the above liquid extract can be dried to obtain a powdered extract. The concentrated or powdered extract can be used as needed by being dissolved in water, alcohol, DMSO (dimethyl sulfoxide), or a mixed solvent thereof.

After obtaining the extract or fraction, additional methods such as concentration or freeze-drying can be used.

In the present invention, the fractionation method for obtaining the above fraction is not particularly limited and may be performed according to a method commonly used in the art. A non-limiting example of the above fractionation method includes a method of obtaining a fraction from a plant extract obtained by extracting the plant by treating the extract with a predetermined solvent.

The type of solvent used to obtain the fraction in the present invention is not particularly limited, and any solvent known in the art can be used. Non-limiting examples of the fractionation solvent include polar solvents such as water and alcohol; and non-polar solvents such as hexane, ethyl acetate, chloroform, dichloromethane, and butanol. These may be used alone or in combination of two or more. When using alcohol among the fractionation solvents, an alcohol having 1 to 6 carbon atoms can be used.

Specifically, it may be an extract obtained by removing foreign substances from a mushroom extract through washing and drying, and extracting the extract with water, alcohol having 1 to 6 carbon atoms, or a mixed solvent thereof, or an extract obtained by sequentially applying the above solvents to a sample.

By including the above-mentioned dogwood extract as an effective ingredient, the anti-inflammatory effect of the composition of the present invention can be enhanced.

Preferably, the composition of the present invention may additionally include a natural extract selected from the group consisting of an extract of Ophiopogon japonicus, an extract of Rhizoma japonica, an extract of Atractylodes japonica, and a mixture thereof.

The anti-inflammatory composition of the present invention comprises 20 to 300 parts by weight of an Ophiopogon japonicus extract, 20 to 300 parts by weight of a Rhizome chinensis extract, and 20 to 300 parts by weight of a Cornus officinalis extract, with respect to 100 parts by weight of a Hazel mushroom extract, and preferably 20 to 200 parts by weight of an Ophiopogon japonicus extract, 20 to 200 parts by weight of a Rhizome chinensis extract, and 20 to 200 parts by weight of a Cornus officinalis extract. More preferably, it may comprise 50 to 200 parts by weight of an Ophiopogon japonicus extract, 50 to 200 parts by weight of a Rhizome chinensis extract, and 50 to 200 parts by weight of a Cornus officinalis extract. When mixed and used within the above content range, an excellent anti-inflammatory effect can be exhibited due to the interaction between the components.

The above Liriopis Tuber is the bulge of the root of Liriope platyphylla Wang et Tang or Ophiopogon japonicus Ker-Gawler, a perennial herb belonging to the lily family (Liliaceae). Its active ingredients include β-sitosterol, stigmasterol, β-sitosterol-β-D-glucoside, steroidal saponins, isoflavonoids, oligosaccharides, and polysaccharides. Liriopis Tuber is a medicinal plant mainly distributed in China, Taiwan, and Korea, and has been traditionally used in Oriental medicine to treat coughs, phlegm, asthma, and neurodegenerative diseases. Also called Maekdong (麥冬), Mundong (門冬), Chondong (寸冬), Bulsacho (不死草), and Yeongyeoncho (沿階草), it is a perennial herb of the lily family (Liliaceae). Its purple flowers are used as ornamentals, and the swollen root is used for food and medicinal purposes. In Oriental medicine, Ophiopogon japonicus is said to have the effects of relaxing the five internal organs, brightening the eyes, relieving dry mouth, and stopping coughs. Existing scientific studies have reported that Ophiopogon japonicus has the following effects: enhancing immunity, anti-inflammatory action, suppressing IgM antibody production, lowering blood sugar, promoting coronary blood flow, reducing lipids, anticancer action, reducing stress, improving memory, improving heart muscle contractility, and sedative action.

Galla rhois is a nest created by the Galla aphid (Aphis chinensis J. Bell) on the leaves of Rhus javanica L., a member of the Anacardiaceae family. It is distributed throughout Korea. It is also called Munhap (文蛤) or Baekchungchang (白蟲倉). The exterior of the Galla rhois is gray-brown with light hairs, and is about 3-7 cm long, 2-5 cm wide, and 2 mm thick. It is hard but easily crushed. The interior is usually empty or sometimes contains gray-white dead insects and secretions, and sometimes has a foul odor. The collected Galla rhois are dried in the sun, boiled, or steamed before use. The main ingredient is pyrogallol tannin (50-70%), and also contains gallic acid, gallic acid, resin, fat, and starch. It has antimicrobial effects on dysentery and Pseudomonas aeruginosa, and its medicinal effects are as an astringent, antidiarrheal, and hemostatic, and it is used for diarrhea, phlegm, diabetes, hematochezia, and anemia, and it has liver function protection and antioxidant effects. In a clinical report, an effective reaction was observed when 15 grams of the above drug was mixed with Gaja (訶子) for digestive bleeding, and it has been reported to be effective in treating ulcerative colitis, radiation proctitis, spermatorrhea, night sweats due to pulmonary tuberculosis, hemoptysis, dysentery, spontaneous sweating, diabetes, esophageal cancer, hemorrhoids, burns, premature ejaculation, night sweats, diarrhea in children, and enteritis prevalent in autumn.

The above-mentioned Rubus sorbifolius var. myriadenus is a deciduous broadleaf shrub that grows 1 to 2 m tall and has dark red glandular hairs and hook thorns, with dense glandular hairs growing all over the young branches. The leaves are alternate and pinnately compound, with 3 to 5 leaflets, but the lower ones have 5 to 7 leaflets, broadly lanceolate, gradually tapering to a point, rounded at the base, with double serrations on the margins, and hairless except for the leaf veins. The flowers are white and bloom in April, borne in compound corymbs at the ends of branches, and have glandular and soft hairs. The fruit is an edible berry, ovate, and ripens to a white color.

In the cosmetic composition of the present invention, the natural material is preferably contained in an amount of 0.001 to 80 parts by weight based on the total weight of the cosmetic composition, and may preferably be contained in an amount of 0.01 to 50 parts by weight. More preferably, it may be contained in an amount of 0.01 to 30 parts by weight. However, the present invention is not necessarily limited thereto.

The composition according to one embodiment of the present invention exhibits anti-inflammatory activity through the inhibition of nitric oxide (NO), prostaglandin E2 (PGE2) and inflammatory cytokines.

Inflammation is one of the defense responses of living tissues to external stimuli. When an inflammatory response occurs, various pro-inflammatory mediators are produced, which clinically cause symptoms such as redness, fever, swelling, pain, and dysfunction.

The inflammatory response begins with various signaling substances secreted from immune cells that recognize antigens, and these signaling substances include inflammatory cytokines.

The above inflammatory cytokines may be IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, TNF-α, TNF-β, TGF-β (transforming growth factor-β), MIG (CXCL9) or interferons (IFNs), and more preferably IL-6, IL-1β, TNF-α.

These inflammatory cytokines no longer increase when the antigen is removed. However, if the inflammatory response is abnormally high, it can damage normal cells. Therefore, if abnormal cytokine secretion and excessive activation persist, inflammatory diseases can develop.

In particular, harmful stimuli such as lipopolysaccharide (LPS), inflammatory factors, and radiation are known to over-activate the human immune system, causing immune cells such as macrophages to excessively induce inflammatory mediators such as tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1), IL-6, IL-1β, prostaglandins, leukotrienes, and nitric oxide (NO), which in turn cause inflammatory diseases, tissue transplant rejection, autoimmune diseases, diabetes, and other immune diseases, as well as the death of nerve cells.

Nitric oxide (NO), one of the inflammatory mediators, is produced in endothelial cells and macrophages under normal conditions and exhibits various physiological activities in addition to cell killing and bactericidal effects. In particular, it is known to play a role in maintaining blood pressure homeostasis through the relaxation of vascular endothelial cells. Stimuli such as LPS, inflammatory factors, and radiation exposure in particular induce the production of iNOS (inducible nitric oxide synthetase), which continuously produces a large amount of nitric oxide (NO) for 4 to 6 hours. The excessive amount of NO produced in this way causes the diseases described above.

Prostaglandins, such as PGE2, PGF2a, and PGI2, are hormones derived from arachidonic acid and involved in various physiological activities. Their production is regulated by cyclooxygenase (COX). Nonsteroidal anti-inflammatory drugs (NSAIDs), such as aspirin and indomethacin, which inhibit COX activity and thus prostaglandin synthesis, are known to be effective in treating arthritis.

In addition, TNF-α is produced by macrophages and fibroblasts activated by BCG, LPS, and inflammatory factors, and induces cell death, production of prostaglandins, and cytokines such as IL-1, IL-6, and IL-1β. TNF-α, which was discovered as a tumor necrosis factor, has been reported to be related to autoimmune diseases such as rheumatoid arthritis and organ transplant rejection, allergic inflammatory diseases such as asthma and atopic dermatitis, and acute immune diseases such as sepsis and acute liver disease. Accordingly, active research is being conducted on the development of compounds that inhibit TNF-α synthesis. In addition, according to a recent report, mice with a defective TNF-α receptor gene are resistant to lethal toxicity by LPS, and mice administered TNF-α antibodies have the effect of preventing arthritis and LPS toxicity.

The present applicant has confirmed that a composition containing the extract of the above-mentioned Hae-am-mulberry as an active ingredient can exhibit an anti-inflammatory effect by inhibiting the production of nitric oxide (NO), prostaglandin E2 (PGE2) and inflammatory cytokines, and that the composition can be used not only as a cosmetic composition, but also as a skin external preparation, a functional food composition and tea.

The term “moisturizer” used in the present invention means a moisturizer used by a method conventional in the art.

The above cosmetic composition further comprises a moisturizer, and the complex composition may be included in a ratio of 0.01 to 0.5 times the amount of the moisturizer added. The moisturizer may be included in an amount of 0.01 to 20 parts by weight, preferably 0.001 to 10 parts by weight, based on the total weight of the cosmetic composition. The moisturizer may include glycerin, 1,3 butylene glycol, dilauroyl glutamate lysine, or a non-toxic salt thereof.

The cosmetic composition of the present invention may contain, but is not limited to, not only the above-described complex herbal extract, but also auxiliary agents commonly used in cosmetic compositions, such as hydrophilic or lipophilic gelling agents, hydrophilic or lipophilic active agents, preservatives, antioxidants, solvents, fragrances, fillers, blocking agents, pigments, deodorants, and dyes.

The above cosmetic composition may further include, but is not limited to, one or more selected from a surfactant, a pH regulator, a swelling agent, a buffer, a preservative, a fragrance, a moisturizer, a viscosity regulator, an aliphatic alcohol, and a vitamin.

The above surfactant may be used without particular limitation as long as it is commonly used in the art, and for example, among surfactants known in the art, such as cationic surfactants, anionic surfactants, nonionic surfactants, amphoteric surfactants, or mixtures thereof, may be appropriately selected and used so as to achieve the effects of the present invention.

More specifically, the surfactant may be an alkyl sulfate (AS), an alkyl ether sulfate (AES), a sodium alkane sulfonate (SAS), an alkyl trimethyl ammonium halide, an alkyl pyridinium halide, an alkyl imidazolium halide, a polyethylene oxide-based surfactant, a polyvinyl-based water-soluble polymer, and the like, and non-limiting examples thereof include sodium dodecyl sulfate (SDS), sodium octylbenzene sulfonate (NaOBS), sodium dodecylbenzene sulfonate (SDBS), ammonium lauryl sulfate (ALS), and Triton It may be at least one selected from X-100 (Triton X-100) and polyvinyl pyrrolidone (PVP, polyvinyl pyrrolidone), but is not limited thereto.

The above pH adjusting agent is for adjusting the pH of the aqueous dye composition, and any agent commonly used in the art may be used without particular limitation, and any one or more selected from sodium hydroxide, potassium hydroxide, triethanolamine, citric acid, and sodium citrate may be used.

The above swelling agent may be used without particular limitation as long as it is commonly used in the art, and any one or more selected from urea, thiourea, urea peroxide, polyoxymethylene urea, hydroxyethyl urea, ammonia, and salts thereof may be used.

The above buffering agent may be used without particular limitation as long as it is commonly used in the art, and any one or two or more selected from salicylic acid, sodium salicylate, lactic acid, and sodium lactate may be used.

The above preservative may be used without particular limitation as long as it is commonly used in the art, and may include at least one selected from among methylparaben, propylparaben, phenoxyethanol, methyl p-hydroxybenzoate, and ethyl p-hydroxybenzoate, and the above fragrance may be used without particular limitation as long as it is commonly used in the art, and may be selected from natural fragrances or artificial synthetic fragrances.

The above moisturizer can be used without any particular limitation as long as it is commonly used in the art, and commonly used ingredients including glycols and oils such as polyethylene glycol can be used.

The viscosity modifier may be used without particular limitation as long as it is commonly used in the art, and may include one or more selected from polystearate, sodium carboxymethyl cellulose, etc., and the fatty alcohol may be used without particular limitation as long as it is commonly used in the art, and may be used by selecting one or more fatty alcohols having 16 to 22 carbon atoms, such as cetyl alcohol, stearyl alcohol, or behenyl alcohol, and the vitamin may be used to supplement nutrition to hair.

Meanwhile, the cosmetic composition according to the present invention may contain water in addition to the aforementioned ingredients. The water may be purified water, such as ion-exchanged water or distilled water. The content is not particularly limited, but may be any amount sufficient to sufficiently dissolve or disperse the aforementioned components.

Cosmetics prepared from the cosmetic composition of the present invention can be prepared in the form of general emulsified formulations and solubilized formulations. Cosmetics prepared in emulsified formulations include nourishing toners, creams, and essences, while cosmetics prepared in solubilized formulations include emollient toners.

In addition to the composition of the present invention, the above cosmetic may be manufactured in the form of an auxiliary agent for topical or systemic application commonly used in the art by containing a dermatologically acceptable medium or base.

Suitable cosmetic formulations may be, for example, solutions, gels, solid or paste anhydrous products, emulsions obtained by dispersing an oil phase in an aqueous phase, suspensions, microemulsions, microcapsules, microgranules or ionic (liposomes) or non-ionic vesicular dispersions, creams, toners, lotions, powders, ointments, sprays or concealer sticks. They may also be prepared in the form of foams or aerosol compositions further containing compressed propellants.

Products to which the cosmetic composition of the present invention can be added include, for example, cosmetics such as astringent toner, emollient toner, nourishing toner, various creams, essences, packs, foundations, and the like, as well as cleansing agents, cleansers, soaps, treatments, or beauty solutions. Specific formulations include, but are not limited to, formulations such as skin lotions, skin softeners, skin toners, astringents, lotions, milk lotions, moisture lotions, nourishing lotions, massage creams, nourishing creams, moisture creams, hand creams, essences, nourishing essences, packs, soaps, shampoos, cleansing foams, cleansing lotions, cleansing creams, body lotions, body cleansers, milky lotions, pressed powders, loose powders, or eye shadows.

The composition according to one embodiment of the present invention can be used as an external skin agent.

When used as the above skin external preparation, it may additionally contain adjuvants commonly used in the field of dermatology, such as fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic or nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, humectants, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles, or any other ingredients commonly used in skin external preparations. In addition, the above ingredients may be introduced in amounts commonly used in the field of dermatology.

When the above extract is provided in the form of a skin external preparation, it may be manufactured and used as a medicine or quasi-drug in the form of a skin external preparation such as a cream, gel, patch, spray, ointment, ointment, lotion, liniment, paste or cataplasma, but is not limited thereto.

The effective amount of the plant extract included in the composition of the present invention may vary depending on the form in which the composition is manufactured, the method by which the compound is applied to the skin, and the length of time it remains on the skin. For example, when the composition is manufactured as a pharmaceutical formulation, the plant extract may be included in a higher concentration than when the composition is manufactured as a cosmetic product that is routinely applied to the skin. Accordingly, the daily dosage is 0.1 to 100 mg/kg, preferably 30 to 80 mg/kg, and more preferably 50 to 60 mg/kg, based on the amount of the plant extract, and may be administered 1 to 6 times a day.

In another embodiment of the present invention, the functional food comprises the functional health food composition.

The above food additives refer to components that can be added to food for preservation purposes, and can be appropriately selected and used by those skilled in the art as they are added in the manufacture of health functional foods of each formulation. Examples of food additives include, but are not limited to, various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, binders and fillers, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, and carbonating agents used in carbonated beverages.

Specifically, the health functional food refers to a food product manufactured by adding the plant extract to food materials such as beverages, teas, spices, gums, and confectionery, or by manufacturing it in the form of capsules, powders, suspensions, etc., and means that when consumed, it brings about a specific health effect, but unlike general drugs, it has the advantage of not having side effects that may occur with long-term use of drugs because it is made from food as a raw material.

The above food may contain food additives that are food-scientifically acceptable, and may further contain suitable carriers, excipients, and diluents commonly used in the manufacture of health functional foods.

Each of the above-described components included in the food composition according to the present invention may be included in the food composition of the present invention within a range that does not exceed the maximum usage amount stipulated in the food safety standards of each country.

The composition may include additional ingredients commonly used in food compositions to improve odor, taste, sight, etc. For example, it may include vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, pantothenic acid, etc. In addition, it may include minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), etc. In addition, it may include amino acids such as lysine, tryptophan, cysteine, valine, etc. There is no limitation on the form that the health functional food of the present invention may take, and it may include all foods in the conventional sense, and may be used interchangeably with terms known in the art such as functional food.

In addition, the health functional food of the present invention can be manufactured by mixing other appropriate auxiliary ingredients and known additives that can be included in foods according to the selection of a person skilled in the art. Examples of foods to which the health functional food of the present invention can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and the food extract of the present invention can be manufactured by adding it to juice, tea, jelly, and juice manufactured using the food extract of the present invention as a main ingredient. It also includes foods used as feed for animals.

When the food formulation is a beverage, it may contain various flavoring agents or natural carbohydrates as additional ingredients, just like conventional beverages. The natural carbohydrates mentioned above 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 a sweetener, a natural sweetener such as thaumatin and stevia extract, or a synthetic sweetener such as saccharin and aspartame can be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g, per 100 mL of the composition of the present invention.

In addition to the above, the food formulation may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, the food formulation may contain fruit pulp for the production of natural fruit juices, fruit juice drinks, and vegetable drinks. These ingredients may be used independently or in mixtures. The proportion of these additives is not particularly important, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

When the above plant extract is used as a food additive, the plant extract can be added as is or used together with other foods or food ingredients, and can be used appropriately according to a conventional method. The amount of the active ingredient mixed can be appropriately determined depending on the purpose of use (prevention, health, or therapeutic treatment). Generally, when manufacturing a food or beverage, the composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on the raw material. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount can be below the above range, and since there is no problem in terms of safety, the active ingredient can also be used in an amount exceeding the above range.

When the food composition of the present invention is a beverage composition, there is no particular limitation on the other components other than containing the extract as an essential component in the indicated ratio, and various flavoring agents or natural carbohydrates, etc. may be contained as additional components like a typical beverage. Examples of the above-mentioned natural carbohydrates are monosaccharides such as glucose, fructose, etc.; disaccharides such as maltose, sucrose, etc.; and polysaccharides such as dextrin, cyclodextrin, etc., and typical sugars, and sugar alcohols such as xylitol, sorbitol, erythritol, etc. As flavoring agents other than those described above, natural flavoring agents (thaumatin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) may be used, but may not be limited thereto. The ratio of the above-mentioned natural carbohydrates may generally be in the range of about 1 to 20 g per 100 ml of the composition of the present invention, but may not be limited thereto.

Meanwhile, the functional food referred to in the present invention includes health supplements, which are foods manufactured or processed using specific ingredients as raw materials or by extracting, concentrating, refining, mixing, etc. specific ingredients contained in food raw materials for the purpose of health supplementation, and other foods designed and processed so that the food ingredients can sufficiently exert their bioregulatory functions, such as biodefense, biorhythm regulation, disease prevention, and recovery, on the body, and include all foods that also have functions related to disease prevention and disease recovery.

Tea is an example of a functional food, and there is no particular limitation on the type of the functional food. Examples of functional foods to which the composition of the present invention can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and may include all health functional foods in the conventional sense, and may include foods used as feed for animals. In addition to the above, the health functional food composition of the present invention may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, and the like. In addition, it may contain fruit pulp for the production of natural fruit juice, fruit juice drinks, and vegetable drinks.

The present invention provides a composition that exhibits an anti-inflammatory effect by including a oyster mushroom as an active ingredient, and can provide an anti-inflammatory composition with reduced side effects by including a natural product as an active ingredient. Furthermore, the composition can be used not only as a cosmetic composition, but also as a skin topical agent, a functional food composition, and tea.

Figure 1 shows the results of cytotoxicity evaluation according to one embodiment of the present invention.
Figure 2 shows the results of cytotoxicity evaluation according to one embodiment of the present invention.
Figure 3 shows the results of an experiment to inhibit PGE2 production by a composition according to one embodiment of the present invention.
Figure 4 shows the results of an experiment to inhibit NO production by a composition according to one embodiment of the present invention.
Figure 5 shows the results of an experiment to inhibit the production of inflammatory cytokines by a composition according to one embodiment of the present invention.
Figure 6 shows the results of an experiment to inhibit the production of inflammatory cytokines by a composition according to one embodiment of the present invention.

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily implement them. However, the present invention may be implemented in various different forms and is not limited to the embodiments described herein.

[Manufacturing Example 1: Manufacturing of Natural Products]

1. Preparation of dogwood mushrooms

Wash and dry the dogwood mushrooms, crush them, add distilled water as an extraction solvent in a ratio of 1:10 (w:v), and then completely soak them. Afterwards, the extraction was repeated three times for 2 hours each while refluxing at 80℃. The extract was filtered through Whatman No. 2 filter paper. The filtrate was concentrated under reduced pressure at 60℃ to prepare a powder sample and used in the experiment. Hot water extraction, concentration, and filtration were performed to prepare a Hae-am mushroom extract (HE).

2. Production of other natural products

In the same manner as the above-mentioned extract of the dogtooth mushroom (HE), extracts of Ophiopogon japonicus (LE), extracts of the magnolia berry (GE), and extracts of the atractylodes japonica (RE) were prepared.

3. Production of complex natural products

The above-mentioned extracts of the dogtooth mushroom, the extract of Ophiopogon japonicus, the extract of the magnolia berry, and the extract of the fruit of the Chinese chive were mixed in the weight ranges shown in Table 1 below to prepare a composite extract.

M1 M2 M3 M4 M5 M6 M7 M8 M9 HE 100 100 100 100 100 100 100 100 100 LE - 100 - - 20 50 100 200 300 GE - - 100 - 20 50 100 200 300 RE - - - 100 20 50 100 200 300

(Unit: parts by weight)

[Manufacturing Example 2: Cell Culture]

RAW 264.7 cells were purchased from the Korea Cell Line Bank (Korea) and cultured in 10% (v/v) fetal bovine serum (FBS), Dulbecco's modified Eagle medium containing 100 U/ml penicillin and 100 U/ml streptomycin The cells were cultured at 37°C and 5% CO2 (v/v) using Eagle's medium.

[Experimental Example 1: Cytotoxicity Evaluation]

The cytotoxicity of the extract of the mushroom (HE) on the RAW 264.7 cells was measured using the MTT reduction assay.

Cells were treated with various concentrations of oyster mushroom extract (HE). Afterwards, the medium was removed and MTT solution (Sigma-Aldrich Chemical Co.) 0.5 mg/ml was distributed to each well and reacted at 37°C. After incubation for 3 hours, the supernatant was removed. Blue formazan crystals were dissolved by adding DMS for 10 min. The absorbance per well was measured at a wavelength of 540 nm using an enzyme-linked immunosorbent assay (ELISA) plate reader (Dynatech Laboratories, Chantilly, VA, USA).

Referring to Figure 1, it was confirmed that there was no cytotoxicity on RAW 264.7 cells at a concentration of 500 μg/ml or less of the extract of the dogwood mushroom (HE), and Figure 2 shows the cytotoxicity on RAW 264.7 cells treated with 50 μg/ml of standard diesel particulate matter (SRM 1650b, hereinafter referred to as PM2.5). It was confirmed that administration of less than 500 μg/ml of the extract of the dogtooth mushroom (HE) to RAW 264.7 cells did not have a significant adverse effect on cell survival.

The anti-inflammatory effect of the following extract of the dogwood mushroom (HE) at a concentration of 500 μg/ml was confirmed, and 500 μg/ml was also used as the standard when manufacturing the complex extract (M2 to M9).

[Experimental Example 2: Experiment to inhibit the production of NO, PGE2, IL-6, and IL-1β]

RAW 264.7 cells were treated with 200 μg/ml and 500 μg/ml of Helianthus annuus extract (HE) for 1 h, followed by 50 μg/ml for 24 h. Stimulated with PM2.5. NO levels in the medium were assessed by measuring the amount of nitrite using Griess reagent (Sigma-Aldrich Chemical Co.). 100 μL of cell-conditioned medium was mixed with an equal volume of Griess reagent for 10 min.

Absorbance was measured at 540 nm using an ELISA reader and compared to a sodium nitrite (NaNO2) standard curve. Calculated.

Additionally, to investigate the level of PGE2, cultures were cultured using an ELISA kit (R & D Systems Inc., Minneapolis, MN, USA). The supernatant was collected and evaluated. The absorbance was measured at a wavelength of 450 nm using an ELISA reader.

The results of the above experiment, as shown in Figs. 3 and 4, showed that PM2.5 treatment significantly increased the emission of NO and PGE2 compared to the untreated control group, but this increase was significantly reduced as the treatment concentration of the extract of the oyster mushroom (HE) increased.

In addition, the effect of the extract of the mushroom (HE) on the production and expression of inflammatory cytokines increased by PM2.5 treatment is shown in Fig. 5 and 6. Specifically, after stimulation by PM2.5, proinflammatory cytokines including IL-6 and IL-1β released into the culture supernatant A significant increase in the amount of cytokines was confirmed. However, it was confirmed that the production of proinflammatory cytokines by PM2.5 was significantly suppressed by pretreatment with the extract of Hae-am-mushroom (HE).

[Experimental Example 3: Experiment on the Inhibition of NO, PGE2, IL-6, and IL-1β Production by Compound Extracts]

An experiment was conducted in the same manner as the experiment that confirmed the inhibitory effect of the above-mentioned extract of the dogwood mushroom (HE) on the production of NO, PGE2, IL-6, and IL-1β.

In the cytotoxicity results, it was confirmed that there was no cytotoxicity in RAW 264.7 cells even when VE, RE, and SE were treated at the same concentration (500 μg/ml), and the same no cytotoxicity was confirmed when treated with the mixed complex.

Afterwards, experiments were conducted on the inhibitory effects on the production of NO, PGE2, IL-6, and IL-1β.

The case where M1 was treated at a concentration of 500 μg/ml was set as index 5, and the cases where M2 to M9 were treated at a concentration of 500 μg/ml were evaluated as indexes.

The above index was evaluated from 1 to 10, and the higher the number, the better the effect of inhibiting the production of NO, PGE2, IL-6, and IL-1β.

The results are shown in Table 2 below.

M1 M2 M3 M4 M5 M6 M7 M8 M9 Inhibition of NO and PGE2 production 5 6 6 6 4 8 9 8 4 Inhibition of inflammatory cytokine (IL-6, IL-1β) production 5 5 6 6 3 7 9 8 4

(Unit: Index)

According to the above experimental results, it was confirmed that M2 to M9, used as a mixed composition, showed an equally excellent NO and PGE2 production inhibition effect compared to M1, and also showed an excellent inflammatory cytokine production inhibition effect.

More specifically, it can be confirmed that the above effect is maximized in the range of M6 to M8.

[Experimental Example 4: Sensory Evaluation]

When the composition containing the above-mentioned extract of the dogwood mushroom (HE) and each mixed extract was provided as a functional food composition, the sensory properties for flavor and taste according to each combination were evaluated.

Specifically, M1, an extract of the dogwood mushroom (HE), and each of the mixed extracts M2 to M9 were prepared as tea beverages, and a sensory evaluation was conducted on 20 adult men and women.

The sensory evaluation was evaluated on a scale of 1 to 10 based on the preference for aroma and taste, and the average index was determined. The higher the number, the better the symbolism.

The specific results are shown in Table 3 below.

M1 M2 M3 M4 M5 M6 M7 M8 M9 incense 5 6 5 6 4 8 9 8 3 taste 5 5 6 6 3 9 9 8 4 Comprehensive symbol map 5 5.5 5.5 6 3.5 8.5 9 8 3.5

(Unit: Index)

Looking at the results in Table 3 above, it can be confirmed that the preference for flavor and taste is improved in M2 to M9 used as a mixed composition compared to M1.

More specifically, it can be confirmed that the above effect is maximized in the range of M6 to M8.

Although the preferred embodiments of the present invention have been described in detail above, the scope of the present invention is not limited thereto, and various modifications and improvements made by those skilled in the art using the basic concept of the present invention defined in the following claims also fall within the scope of the present invention.

Claims (7)

Contains extracts of dogwood, lily of the valley, magnolia, and holly as active ingredients
Anti-inflammatory composition. In the first paragraph,
The above composition exhibits an anti-inflammatory effect by inhibiting the production of nitric oxide (NO), prostaglandin E2 (PGE2) and inflammatory cytokines.
Anti-inflammatory composition. In the first paragraph,
The above extract is extracted using an extraction solvent selected from the group consisting of water, lower alcohols having 1 to 6 carbon atoms, and mixtures thereof.
Anti-inflammatory composition. Comprising a composition according to any one of claims 1 to 3
Cosmetic composition. Comprising a composition according to any one of claims 1 to 3
Topical skin preparation. Comprising a composition according to any one of claims 1 to 3
Functional food composition. Comprising a composition according to any one of claims 1 to 3
Tea.