Composition for treating or preventing inflammatory osteoarticular diseases comprising fermented mushroom mycelium extract as active ingredient

Abstract

The present invention relates to a composition for strengthening joints and cartilage, comprising a fermented mushroom mycelium extract as an active ingredient. The composition for preventing or treating inflammatory osteoarticular diseases, comprising a fermented extract of Auricularia auricula mycelium and Sparassis crispa mycelium, as an active ingredient, according to the present invention, has an anti-inflammatory effect and the effect of reducing joint swelling, and thus can be effectively used as a composition for protecting and strengthening joints and cartilage.

Description

Composition for treating or preventing inflammatory osteoarthritis, comprising mushroom mycelia extract fermentation as an active ingredient

[0001] The present invention relates to a composition for treating or preventing inflammatory osteoarthritis, which comprises a mushroom mycelia extract fermentation product as an active ingredient, and more specifically, to a composition for treating or preventing inflammatory osteoarthritis, which comprises a lactic acid bacteria fermentation product of a mixed mycelia of oyster mushroom mycelia, white oyster mushroom mycelia, and pine mushroom mycelia, as an active ingredient, and has a joint and cartilage strengthening or protective effect.

[0002] Arthritis is a disease caused by inflammation and pain in the joints. It includes osteoarthritis, rheumatoid arthritis, gout, and psoriatic arthritis, and 95% of arthritis patients suffer from degenerative arthritis. Degenerative arthritis is a disease in which local degenerative changes occur as the joint cartilage wears away. It is also called osteoarthritis. Degenerative arthritis is a representative degenerative disease closely related to aging, and approximately 10-15% of the general population suffers from degenerative arthritis, and in particular, 60-80% of the elderly population over 65 years of age suffer from degenerative arthritis.

[0003] Inflammatory osteoarthritis includes ankylosing spondylitis, psoriatic arthritis, osteoarthritis, periarthritis, tendonitis, tenosynovitis, and peritendinitis. Among them, osteoarthritis mainly occurs in middle-aged or elderly people. According to the National Health and Nutrition Examination Survey, the prevalence of osteoarthritis in Korea is 5.5% for men and 22.7% for women, which is more than four times higher in women than in men. The prevalence of osteoarthritis by age was 1.8% in men in their 50s, 8.1% in their 60s, and 10.9% in their 70s. In women, the prevalence increased rapidly with age, with 9.5% in their 50s, 25.3% in their 60s, and 41.5% in their 70s.

[0004] Although arthritis is not a fatal disease, it significantly reduces the quality of life due to joint pain and limited activity, and the financial burden, including individual and national medical expenses, increases depending on the purpose of treatment.

[0005] Osteoarthritis is a chronic degenerative disease in which articular cartilage is gradually damaged due to degeneration of the extracellular matrix that makes up the articular cartilage, resulting in inflammation and pain.

[0006] Osteoarthritis is particularly common in the elderly and typically affects one joint of the body through inflammation. The causes of osteoarthritis are closely related to aging and excess weight, and it occurs more frequently in women as they age. Early symptoms include stiffness and aching pain in one or two joints. If prolonged, it can lead to excessive bone formation around the joints and joint deformation. The mechanism of osteoarthritis involves increased production of proinflammatory cytokines and increased secretion of matrix metalloproteinases (MMPs), such as collagenase and stromelysin, which in turn damage the articular cartilage matrix.

[0007] Inflammation is a fundamental biological process, consisting of a dynamic complex of cellular and chemical reactions that occur in affected blood vessels and adjacent tissues in response to damage or abnormal stimuli caused by physical, chemical, or biological agents. The inflammatory process includes 1) local tissue reactions and resulting morphological changes; 2) destruction and removal of harmful substances; and 3) repair and healing responses. The so-called "cardinal symptoms" of inflammation are redness, heat (warmth), swelling, pain, and decreased or lost function. While all of these symptoms may be observed in certain cases, none are necessarily present all the time. Arthritis is an inflammatory disease characterized by inflammation of the joints, including the synovial tissue and membranes.

[0008] There are many types of arthritis, including, but not limited to, osteoarthritis (hypertrophic or degenerative arthritis), rheumatoid arthritis, infectious arthritis (e.g., tuberculosis, Lyme disease, rheumatic fever), septic arthritis, juvenile arthritis, and gouty arthritis. Elevated tissue levels of IL-1, IL-8, and TNF have been found in arthritis and other inflammatory conditions. In osteoarthritis, the cartilage lining the joints is gradually degraded by matrix metalloproteinases (MMPs), which are secreted into the synovial fluid by synovial lining cells in response to stimulation by various enzymes, particularly various proinflammatory cytokines, particularly IL-1 and TNF. Cartilage destruction by MMPs leads to a continuous inflammatory response, resulting in joint pain associated with osteoarthritis.

[0009] Rheumatoid arthritis is an inflammatory disease characterized by polyarthritis, with autoimmune mechanisms known to be the primary cause. Symptoms include inflammation of the synovial membrane, leading to the migration of macrophages, dendritic cells, T lymphocytes, and B lymphocytes into the synovial tissue. This results in increased synovial fluid, resulting in joint swelling and pain.

[0010] As inflammation persists, inflammatory synovial tissue hyperplasia occurs, destroying bone and cartilage, leading to joint structure deformation and impaired mobility. Rheumatoid arthritis is also known to increase the production of proinflammatory cytokines, which in turn increases the secretion of matrix metalloproteinases (MMPs), destroying collagen and proteoglycans that make up articular cartilage and damaging it.

[0011] Most arthritis treatments being developed using natural ingredients are based on natural extracts, and their precise mechanisms of action remain unknown. However, many are readily available as oral preparations, allowing for long-term use, and thus, active development is underway. Currently under development include injections of hyaluronic acid, which has similar components to synovial fluid, and supplements containing glucosamine and chondroitin. Furthermore, natural products used in established herbal medicine books such as Donguibogam are being developed.

[0012] The technical problem to be solved in the present invention is to provide a pharmaceutical composition for treating or preventing inflammatory osteoarthritis-related diseases.

[0013] Another technical problem to be solved in the present invention is to provide a functional food composition for preventing or improving inflammatory osteoarthritis-related diseases.

[0014] In addition, another technical problem to be solved in the present invention is to provide a method for preventing or treating inflammatory osteoarthritis using the composition.

[0015] In order to solve the above-mentioned technical problem, the present invention provides a pharmaceutical composition for preventing or treating inflammatory osteoarthritis, characterized in that it contains a mushroom mycelia extract fermentation product as an active ingredient.

[0016] Preferably, the mushroom mycelium is characterized by being at least one mushroom mycelium selected from the group consisting of oyster mushroom mycelium, white oyster mushroom mycelium, and chanterelle mushroom mycelium.

[0017] The above white oyster mushroom mycelia and flower mushroom mycelia are characterized in that they are mixed in a ratio of 1:2 to 1:4 (w / w).

[0018] The above-mentioned oyster mushroom mycelia and pine mushroom mycelia are characterized in that they are mixed in a ratio of 1:2 to 1:4 (w / w).

[0019] Preferably, the mushroom mycelia extract is characterized in that it is prepared according to the following method:

[0020] (1) A step of mixing 30 to 50 parts by weight of glucose with 100 parts by weight of mushroom mycelium powder;

[0021] (2) a step of adding purified water in an amount of 9 to 11 times the weight of the mushroom mycelia to the mixture mixed in the above step (1) and extracting at 95 to 110°C for 12 to 48 hours; and

[0022] (3) A step of filtering the extract extracted in the above step (2).

[0023] Preferably, the mushroom mycelia extract fermentation product is characterized in that the mushroom mycelia extract is fermented using lactic acid bacteria.

[0024] The above lactic acid bacteria is characterized by using Weissella confusa WIKIM51 (Wilac D001) or Lactobacillus paracasei WiKim0171.

[0025] Preferably, the lactic acid bacteria is characterized by using a lactic acid bacteria culture solution.

[0026] The above lactic acid bacteria culture solution is characterized in that it is cultured according to the following method.

[0027] (1) Step for preparing MRS badge;

[0028] (2) a step of adding Weissella confusa WIKIM51 (Wilac D001) or Lactobacillus paracasei WiKim0171 to the medium; and

[0029] (3) A step of culturing at 35 to 40°C for 22 to 26 hours.

[0030] Preferably, the mushroom mycelia extract fermentation product is characterized in that it is manufactured according to the following method.

[0031] (1) A step of preparing mycelia of one or more strains selected from the group consisting of mycelia of oyster mushrooms, mycelia of white oyster mushrooms, and mycelia of pine mushrooms;

[0032] (2) A step of sterilizing the above strain mycelia under conditions of 1.2 atm and 120 to 125°C;

[0033] (3) OD of lactic acid bacteria 600 Step of inoculating at a concentration of 1-2% (v / v) under 0.3 conditions;

[0034] (4) Fermentation step at 35 to 40℃ for 45 to 50 hours;

[0035] (5) A step of filtering and concentrating the fermented product;

[0036] (6) Step of freeze-drying the concentrated fermented product and then powdering it.

[0037] Preferably, the inflammatory osteoarthritis is characterized by being at least one disease selected from the group consisting of ankylosing spondylitis, psoriatic arthritis, osteoarthritis, periarthritis of the shoulder joint, tendinitis, tenosynovitis and peritendinitis.

[0038] The present invention provides a functional food composition for preventing or improving inflammatory osteoarthritis, which contains a mushroom mycelia extract fermentation product as an active ingredient.

[0039] In addition, the present invention provides a feed composition for preventing or improving inflammatory osteoarthritis, which contains a mushroom mycelia extract fermentation product as an effective ingredient.

[0040] In addition, the present invention provides a method for preventing or treating inflammatory osteoarthritis, comprising a step of administering the composition to a subject other than a human.

[0041]

[0042] In this way, the composition for preventing or treating inflammatory osteoarthritis, which includes the mushroom mycelia extract fermentation product according to the present invention as an active ingredient, has an anti-inflammatory effect and an effect of improving joint swelling, and thus can be usefully used as a composition for protecting and strengthening joints and cartilage.

[0043] The following drawings attached to this specification illustrate preferred embodiments of the present invention and, together with the contents of the invention described above, serve to further understand the technical aspects of the present invention, and therefore, the present invention should not be interpreted as being limited to the matters described in such drawings.

[0044] Figure 1 illustrates an example of a process for obtaining a mushroom mycelia extract fermentation product from oyster mushroom mycelia or cauliflower mushroom mycelia.

[0045] Figure 2 shows the cytotoxicity of the mushroom mycelia extract fermentation product.

[0046] Figure 3 shows the anti-inflammatory efficacy of a mushroom mycelia extract fermented using Lactobacillus paracasei WiKim0171 in Raw 264.7 cells.

[0047] Figure 4 shows the anti-inflammatory efficacy of a mushroom mycelia extract fermented using Lactobacillus paracasei WiKim0171 in MH7A cells.

[0048] Figure 5 shows the anti-inflammatory efficacy of a mushroom mycelia extract fermented using Weissella confusa WIKIM51 (Wilac D001) in MH7A cells.

[0049] Figure 6 shows the results of observing the changes in growth parameters and serum biomarkers by fermented mushroom mycelia extract using Weissella confusa WIKIM51 (Wilac D001) in a collagen-induced arthritis (CIA) model.

[0050] Figure 7 shows the effect of improving arthritis symptoms by fermented mushroom mycelia extract using Weissella confusa WIKIM51 (Wilac D001) in a collagen-induced arthritis (CIA) model.

[0051] Figure 8 shows the effect of a mushroom mycelia extract fermented using Weissella confusa WIKIM51 (Wilac D001) on knee joint inflammation.

[0052] Figure 9 shows the results of body weight and joint swelling measurements using the RA model.

[0053]

[0054] Hereinafter, the present invention will be described in more detail.

[0055] The present invention provides a composition for treating and preventing inflammatory osteoarthritis, which comprises a fermented extract of mushroom mycelia as an active ingredient.

[0056] The above 'inflammatory osteoarthritis' refers to a general term for diseases occurring in bones and joints with inflammation as the main lesion, and may be any one selected from the group consisting of ankylosing spondylitis, psoriatic arthritis, osteoarthritis, periarthritis of the shoulder joint, tendinitis, tenosynovitis, and peritendinitis diseases, but is not limited thereto.

[0057] The above 'inflammatory disease' refers to a disease caused by a normal inflammatory response, and specifically includes, but is not limited to, gastritis, gastric ulcer, bronchitis, asthma, edema, hepatitis, nephritis, arteriosclerosis, inflammatory bowel disease (IBD), colitis, rheumatoid arthritis, osteoarthritis, arthritis-related diseases, cancer, and degenerative diseases.

[0058] The above mushroom mycelia extract fermentation product can be obtained by extracting mushroom mycelia and then fermenting them using lactic acid bacteria.

[0059] The mushroom mycelium used in the present invention may be one or more mushroom mycelium selected from the group consisting of oyster mushroom mycelium, white oyster mushroom mycelium, and pine mushroom mycelium, and is preferably characterized in that oyster mushroom mycelium and pine mushroom mycelium are used in combination.

[0060] The above-mentioned wood ear mushroom (Auricularia auricula-judae) is a mushroom in the Auriculariaceae family, growing in colonies on old trees and dry branches of deciduous trees from spring to fall, and is distributed worldwide. The fruiting body is 3 to 12 cm in diameter, bell-shaped to ear-shaped, gelatinous, and the spore layer is densely covered with fine hairs and yellowish brown to red. Wood ear mushrooms contain abundant nutrients, and the main components include 21 types of free amino acids, ergosterol, 6 types of fatty acids, 12 types of trace metal elements, vitamins B1, B2, D, niacin, glycerol, mannitol, free sugars, fungal sugars, beta-glucan as an anti-tumor component, and gluconoxylomannan as an anti-inflammatory and cholesterol-lowering component. The main functions of oyster mushrooms include nourishment by providing nutrients, stimulating appetite, and regulating human health. Among mushrooms, oyster mushrooms are known to have a very high anti-tumor inhibition rate, and they are also known to improve the health of the five internal organs, stop dysentery and hematochezia, and purify the blood, helping with skin beauty and treating adult diseases such as anemia, high blood pressure, and arteriosclerosis. They also contain a large amount of natural gelatin, which promotes excretion and is effective in relieving constipation. They are also reported to have anti-thrombotic and memory-enhancing properties.

[0061] The Tremella fuciformis, an edible mushroom distributed in Korea, China, and other countries, has a pure white fruiting body. It grows on dead trees or branches of various broad-leaved trees in summer and fall. It has been confirmed that the fruiting bodies or mycelia of the Tremella fuciformis contain large amounts of amino acids that form collagen, such as glycine, proline, hydroxyproline, lysine, and hydroxylysine, in the form of glycoproteins. However, since the molecular weight of the glycoprotein extracted from the fruiting bodies or mycelia of the Tremella fuciformis is very large, exceeding 1,700,000 Da, it cannot be considered that the glycoprotein extracted from the fruiting bodies or mycelia of the Tremella fuciformis has the structure of animal collagen or fish collagen. Moreover, it has been confirmed that the fruiting bodies or mycelia of the Tremella fuciformis contain not only the amino acids that form collagen, but also a large amount of sugars.

[0062] The above-mentioned flower mushroom (Sparassis crispa) is a mushroom in the Sparassis family of the Basidiomycete order. The fruiting body is white to brown and resembles numerous wavy petals. It is highly sensitive to its surroundings, and its growing conditions, including temperature, humidity, and lighting, are demanding, making it difficult to encounter in the wild. Consequently, the flower mushroom is called a gift from God and a miracle. The beta-glucan contained in the flower mushroom is known to activate the immune function of normal cells, inhibit the proliferation and recurrence of cancer cells, and have the activity of reducing blood sugar and cholesterol.

[0063] In the present invention, "mycelium" is a general term referring to a densely entangled growth of hyphae (singular: hypha, plural: hyphae), which are observed in eukaryotic fungi and prokaryotic actinobacteria. Mushroom mycelia are an aggregate of hyphae and are the vegetative organs of fungi. The fruiting bodies of fungi, commonly called mushrooms, are also composed entirely of mycelia, except for the hymenium. These mycelia have the property of self-assembly, in which biopolymers assemble themselves under appropriate environmental conditions to form specific higher-order structures.

[0064] In the present invention, the mushroom mycelium is characterized by using one or more mushroom mycelia selected from the group consisting of oyster mushroom mycelia, white oyster mushroom mycelia, and pine mushroom mycelia, and using them by extracting each extract.

[0065] According to one embodiment of the present invention, the mushroom mycelia extract is characterized in that it is prepared according to the following method.

[0066] (1) A step of mixing 30 to 50 parts by weight of glucose with 100 parts by weight of mushroom mycelium powder;

[0067] (2) A step of adding purified water in an amount of 9 to 11 times the weight of the mushroom mycelia to the mixture mixed in the above step (1) and extracting at 95 to 100°C for 2 to 4 hours; and

[0068] (3) A step of filtering the extract extracted in the above step (2).

[0069] The composition for treating and preventing inflammatory osteoarthritis of the present invention is characterized in that it contains, as an active ingredient, a fermented extract obtained by mixing and then fermenting one or more mushroom mycelia extracts selected from the group consisting of oyster mushroom mycelia, white oyster mushroom mycelia, and pine mushroom mycelia.

[0070] According to one embodiment of the present invention, it is preferable to mix the mycelial extract of the oyster mushroom and the mycelial extract of the pine mushroom in a ratio of 1:2 to 1:4 (v / v), particularly in a ratio of 1:3 (v / v).

[0071] According to one embodiment of the present invention, it is preferable to mix the white oyster mushroom mycelia extract and the flower mushroom mycelia extract in a ratio of 1:2 to 1:4 (v / v), particularly in a ratio of 1:3 (v / v).

[0072] According to one embodiment of the present invention, a mixture of one or more mushroom mycelia extracts selected from the group consisting of the oyster mushroom mycelia, the white oyster mushroom mycelia, and the pine mushroom mycelia can be cultured with lactic acid bacteria to obtain an extract fermentation product.

[0073] The above-mentioned lactic acid bacteria are Gram-positive, lactic acid-producing organisms with various health benefits. Major strains may include probiotics, live microorganisms that, when administered in appropriate amounts, provide health benefits to the host.

[0074] The above lactic acid bacteria include at least one lactic acid bacterium of the genus Lactobacillus selected from L. acidophilus, L. casei, L. gasseri, L. delbrueckii ssp. bulgaricus, L. helveticus, L. fermentum, L. paracasei, L. plantarum, L. reuteri, L. rhamnosus, and L. salivarius; It may be at least one lactic acid bacterium selected from the group consisting of Lactococcus lactis (Lc. lactis); Enterococcus faecium (E. faecium) and Enterococcus faecalis (E. faecalis); Streptococcus thermophilus (S. thermophilus); and at least one lactic acid bacterium of the genus Bifidobacterium selected from B. bifidum, B. breve, B. longum and B. animalis spp. lactis.

[0075] Among the various probiotics mentioned above, the Lachnospiraceae family is the most well-known and widely used in the probiotics industry. Lactobacillaceae strains are involved in fermentation, regulating microbial growth and enzymatically converting food components to produce beneficial end products such as organic acids, bacteriocins, and peptides. Weissella, a genus within the Lactobacillaceae family, is known to exhibit beneficial effects on the host, including antibacterial, anti-inflammatory, and anticancer activities.

[0076] The present invention is characterized in that Weissella confusa WIKIM51 (Wilac D001) or Lactobacillus paracasei WiKim0171 is used as lactic acid bacteria.

[0077] The above Weissella confusa WIKIM51 (Wilac D001) is a lactic acid bacterium derived from dandelion kimchi and has completed the U.S. Food and Drug Administration (FDA) New Drug Indication (NDI) registration. In animal testing, its effectiveness has been confirmed, including suppressing fat synthesis, promoting fat breakdown, and improving body fat levels and blood lipid levels.

[0078] The above Lactobacillus paracasei WiKim0171 is a lactic acid bacterium isolated from makgeolli. It is a gram-positive bacterium and a facultative anaerobe that can grow under both aerobic and anaerobic conditions. It is non-motile and its cells have a rod-shaped form. Catalase and indole production abilities are not observed. The growth temperature is 15 to 40°C, and the optimal growth temperature is 30°C. The growth pH is 4.0 to 8.5, and the optimal growth pH is 6.0 to 6.5.

[0079] According to one embodiment of the present invention, the present invention is characterized in that a lactic acid bacteria culture solution cultured according to the following method is used.

[0080] (1) Step for preparing MRS badge;

[0081] (2) a step of adding Weissella confusa WIKIM51 (Wilac D001) or Lactobacillus paracasei WiKim0171 to the medium; and

[0082] (3) A step of culturing at 35 to 40°C for 22 to 26 hours.

[0083] According to one embodiment of the present invention, a mixture of one or more mushroom mycelia extracts selected from the group consisting of oyster mushroom mycelia, white oyster mushroom mycelia, and pine mushroom mycelia is cultured using lactic acid bacteria, and a mushroom mycelia extract fermentation product can be obtained according to the following method:

[0084] (1) A step of preparing one or more mushroom mycelia extracts selected from the group consisting of oyster mushroom mycelia, white oyster mushroom mycelia, and pine mushroom mycelia;

[0085] (2) A step of sterilizing the above mixed extract under conditions of 1.2 atm and 120 to 125°C;

[0086] (3) OD of lactic acid bacteria 600 Step of inoculating at a concentration of 1-2% (v / v) under 0.3 conditions;

[0087] (4) Fermentation step at 35 to 40℃ for 45 to 50 hours;

[0088] (5) A step of filtering and concentrating the fermented product;

[0089] (6) Step of freeze-drying the concentrated fermented product and then powdering it.

[0090] The present invention provides a composition for treating and preventing inflammatory osteoarthritis, characterized in that it contains the above mushroom mycelia extract fermentation as an effective ingredient.

[0091] Hereinafter, the above mushroom mycelia extract fermentation product is referred to as ATJ-301.

[0092] According to one embodiment of the present invention, the composition is characterized by having an edema reducing effect.

[0093] According to one embodiment of the present invention, the composition is characterized by having an anti-inflammatory effect.

[0094] According to one embodiment of the present invention, the composition is characterized by having a protective and strengthening effect on joints and cartilage.

[0095] In the present invention, the “active ingredient” refers to an ingredient that exhibits the desired activity alone or can exhibit the activity together with a carrier that is inactive in itself.

[0096] According to one embodiment of the present invention, the mushroom mycelia extract fermentation is characterized in that it is included in an amount of 1 to 80 wt% based on the total weight of the composition. In this case, if the content is less than 1 wt%, the joint or cartilage protection and strengthening effect, which is the intended effect of the present invention, cannot be obtained, and if it exceeds 80 wt%, the effect is not proportional to the increase in content, which may be inefficient, and there is a problem that stability in the formulation is not secured.

[0097] In one embodiment of the present invention, the composition of the present invention may be a pharmaceutical composition for treating or preventing inflammatory osteoarthritis.

[0098] The composition of the present invention may additionally contain a suitable carrier, excipient or diluent that is commonly used in addition to the active ingredient.

[0099] The pharmaceutical composition according to the present invention may include a pharmaceutically acceptable carrier in addition to the mushroom mycelia extract fermentation product. The pharmaceutically acceptable carrier included in the pharmaceutical composition of the present invention is one commonly used in formulation, and includes, but is not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. In addition to the above components, the pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

[0100] In this specification, the term "prevention" means any action that inhibits or delays the progression of inflammatory osteoarthritis by administering the composition of the present invention.

[0101] As used herein, the term "treatment" means inhibition of the development of inflammatory osteoarthritis; alleviation of inflammatory osteoarthritis; and elimination of inflammatory osteoarthritis.

[0102] The pharmaceutical composition of the present invention can be administered orally or parenterally.

[0103] The appropriate dosage of the pharmaceutical composition of the present invention can be prescribed in various ways depending on factors such as the formulation method, administration method, patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and reaction sensitivity. The dosage of the pharmaceutical composition of the present invention is generally 0.001 to 1000 mg / kg, preferably 0.05 to 200 mg / kg, and more preferably 0.1 to 100 mg / kg for adults. In addition, in the case of external preparation, it is preferable to apply 1.0 to 6.0 ml once to 5 times a day for adults and continue for more than 1 month. However, the above dosage does not limit the scope of the present invention.

[0104] The pharmaceutical composition of the present invention can be manufactured in unit dosage form or can be manufactured by inserting it into a multi-dose container by formulating it using a pharmaceutically acceptable carrier and / or excipient according to a method that can be easily performed by a person of ordinary skill in the art. In this case, the formulation may be in the form of a solution, suspension, syrup, or emulsion in an oil or aqueous medium, or in the form of an extract, powder, granule, tablet, or capsule, and may additionally include a dispersant or stabilizer.

[0105] The composition of the present invention may be a functional food composition for improving or preventing inflammatory osteoarthritis.

[0106] The term "functional food" used in the present invention refers to a food manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc. using raw materials or ingredients that have functionality useful to the human body. Here, functionality means obtaining a useful effect for health purposes, such as regulating nutrients for the structure and function of the human body or physiological functions. The health functional food of the present invention can be manufactured by a method commonly used in the art, and can be manufactured by adding raw materials and ingredients commonly added in the art during the manufacturing process. In addition, unlike general drugs, it has the advantage of not having side effects that may occur with long-term use of drugs because it uses food as a raw material, and can be highly portable.

[0107] The food composition according to the present invention may additionally include not only the mushroom mycelia extract fermentation product as an effective ingredient, but also ingredients commonly added during food manufacturing, such as proteins, carbohydrates, fats, nutrients, seasonings, and flavoring agents.

[0108] Examples of the carbohydrates include monosaccharides such as glucose, fructose, etc.; disaccharides such as maltose, sucrose, oligosaccharides, etc.; and polysaccharides such as dextrin, cyclodextrin, etc., and common sugars and sugar alcohols such as xylitol, sorbitol, erythritol, etc. As flavoring agents, natural flavoring agents [thaumatin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.]) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used.

[0109] For example, when the food composition of the present invention is manufactured as a drink, in addition to the mushroom mycelia extract fermentation of the present invention, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, juice, Eucommia extract, jujube extract, licorice extract, etc. may be additionally included.

[0110] The composition of the present invention may be a feed composition for preventing or improving inflammatory osteoarthritis.

[0111] In the present invention, the term "feed" means any natural or artificial diet, meal, etc. or a component of said meal for eating, ingesting, and digesting by an animal or suitable therefor.

[0112] The type of the above feed is not particularly limited, and feed commonly used in the relevant technical field can be used. Non-limiting examples of the above feed include plant feed such as grains, roots, food processing by-products, algae, fiber, pharmaceutical by-products, oils, starches, meal, or grain by-products; and animal feed such as proteins, inorganic substances, oils, minerals, oils, single-cell proteins, zooplankton, or food. These may be used alone or in combination of two or more.

[0113] The above feed composition may include a feed additive. The feed additive of the present invention corresponds to a supplementary feed under the Feed Management Act.

[0114] The composition of the present invention is not particularly limited and can be applied to any subject for the purpose of preventing or improving inflammatory osteoarthritis. For example, it can be used in any non-human animal such as monkeys, dogs, cats, rabbits, guinea pigs, rats, mice, cows, sheep, pigs, goats, birds, fish, etc., and the composition can be administered parenterally, subcutaneously, intraperitoneally, intrapulmonary, and intranasally, and can be administered by any suitable method including intralesional administration if necessary for local treatment. The preferred dosage of the pharmaceutical composition of the present invention varies depending on the condition and body weight of the subject, the degree of the disease, the drug form, the route and duration of administration, but can be appropriately selected by those skilled in the art. For example, it can be administered orally, rectally, or by intravenous, intramuscular, subcutaneous, intrauterine, intradural, or intracerebrovascular injection, but is not limited thereto.

[0115] Another aspect of the present invention provides a method for preventing or treating inflammatory osteoarthritis, comprising administering to a subject a composition of the present invention.

[0116] In the present invention, the term "subject" means any animal other than humans that has developed or may develop an inflammatory osteoarthritis, and by administering the composition of the present invention to a subject suspected of having an inflammatory osteoarthritis, the subject can be effectively treated.

[0117] As used herein, the term "administration" refers to introducing the composition of the present invention into a subject suspected of having inflammatory osteoarthritis through any suitable method. The route of administration may be oral or parenteral, as long as it can reach the target tissue, and is as described above.

[0118] In this way, the composition for preventing or treating inflammatory osteoarthritis, which includes the mushroom mycelia extract fermentation product according to the present invention as an active ingredient, has an anti-inflammatory effect and an effect of improving joint swelling, and thus can be usefully used as a composition for protecting and strengthening joints and cartilage.

[0119] To facilitate understanding of the present invention, examples and other embodiments will be described in detail below. However, the embodiments according to the present invention may be modified in various ways, and the scope of the present invention should not be construed as being limited to the following examples. These examples are provided to more fully explain the present invention to those of average skill in the art.

[0120]

[0121] <Manufacturing Example 1> Manufacturing of lactic acid bacteria culture solution

[0122] Weissella confusa WIKIM51 (Wilac D001) or Lactobacillus paracasei WiKim0171 was added to MRS medium and cultured at 38°C for 24 hours to obtain a lactic acid bacteria culture solution.

[0123] Weissella confusa WiKim51 (W. confusa) (99.73% identity; accession number LC063164.1) strain was obtained from Pharmsville Co., Ltd. (Seoul, Republic of Korea). 16S rRNA sequencing was performed at Macrogen (Seoul, Republic of Korea) for strain identification.

[0124] W. confusa WiKim51 was cultured in Man, Rogosa, and Sharpe (MRS) medium (BD Co., Franklin Lakes, NJ, USA) at 38°C for 24 h to obtain cultures of Weissella confusa WIKIM51 (Wilac D001) and Lactobacillus paracasei WiKim0171. All strains were subcultured at least three times before use.

[0125]

[0126] <Example 1> Production of mushroom mycelia extract fermentation product

[0127] Auricularia auricula-judae mycelia powder (Mush&, Jeonju, South Korea) or Tremella fuciformis and Sparassis crispa mycelia powder (Mush&, Jeonju, South Korea) were mixed in a ratio of [1:3 w / w], dissolved in distilled water [1:10 w / v], and reflux extracted at 100°C for 6 hours. The resulting extract was mixed with minimal medium (glucose [1:25 w / v], peptone [1:100 w / v], sodium acetate 3H2O [1:200 w / v], magnesium sulfate 7H2O [1:10,000 w / v], manganese sulfate 4H2O [1:20,0000 w / v], Tween 80 [1:100 v / v], ammonium citrate [1:500 w / v], potassium hydrogen phosphate [1:500 w / v]) and then autoclaved (121°C, 15 min). At this time, the mixture of Auricularia auricula-judae mycelia and Sparassis crispa mycelia was named 'mycelial extract (ME)'.

[0128] Afterwards, the W. confusa WiKim51 (WC) culture broth was centrifuged at 10,800 × g for 3 min (VS-180Cfi, Vision Scientific Co., Daejeon, South Korea) and washed twice with phosphate-buffered saline (PBS). The optical density of the harvested bacterial pellet at 600 nm was adjusted to 0.3, and then added to ME (1:50 v / v) and cultured for 48 h. Samples were prepared at 0, 24, and 48 h of fermentation, and plated on MRS agar medium (Kisan Bio) to evaluate bacterial growth. The ME fermented with WC for 24 h was designated 'WCF-ME'. The WCF-ME was then filtered through Whatman filter paper-Grade 3 (6 μm) (WhatmanTM, Maidstone, United Kingdom), lyophilized, and stored at -80°C.

[0129]

[0130] <Example 2> Analysis of components of mushroom mycelia extract fermentation product

[0131] The mushroom mycelia extract fermentation product prepared in Example 3 above was analyzed.

[0132] The short-chain fatty acid (SCFA) content of ME and WCF-ME was measured by gas chromatography / mass spectrometry (GC / MS) using a slightly modified method of Kim and Jang. Specifically, 50 mg of ME or WCF-ME sample was mixed with 100 μL crotonic acid, 50 μL HCl, and 200 μL ether, homogenized, and centrifuged at 1000 ×g for 10 min. The supernatant was transferred to a vial, and 16 μL N-tert-butyldimethylsilyl-N-methyltrifluoroacetamide (Sigma-Aldrich, USA) was added. After mixing, the vial was sealed and heated at 80 °C for 20 min and then stored at room temperature for 48 h. The sample was analyzed by HP-5MS column (30 m, 0.25 mm, 0.25 μm) and 5973 network mass selective detector (Agilent Technologies, The SCFAs were injected into a 6890N network GC system (Agilent Technologies, California, USA) equipped with a helium (99.9999% purity) carrier gas at a flow rate of 1.2 mL / min. The head pressure was 97 kPa, and the split ratio was 20:1. The inlet and transfer line temperatures were 250 °C and 260 °C, respectively. The following temperature program was used: 60 °C (3 min), 60–120 °C (5 °C / min), and 120–300 °C (20 °C / min). 1 μL of sample was injected, and the analysis time was 30 min. SCFA concentrations were quantified by comparison with the peak areas of the standards.

[0133] Table 1 below shows the effective ingredients of mushroom mycelia extract fermentation.

[0134]

[0135]

[0136] (1) White oyster mushroom mycelia indicator component

[0137] · Ingredient name: Limonene

[0138] · Molecular formula: C 10 H 16

[0139] · Molecular weight: 136.23 g / mol

[0140] · CAS No.: 138-86-3

[0141] · Chemical formula 1

[0142]

[0143]

[0144] Limonene, the aforementioned limonene, is an aliphatic hydrocarbon classified as a cyclic monoterpene and a volatile oil component primarily found in the peels of citrus fruits. It is primarily used industrially as a fragrance in cosmetics, foods, and solvents, and is known to be a low-toxic chemical. It has been reported to suppress stress and inflammation and promote skin cell regeneration.

[0145] Assay method: Phenomenex Kinetex C 18 HPLC analysis was performed at 25°C using a column (100 mm * 2.1 mm, particle size 1.7 μm). 1 μl of sample was injected, and the mobile phase was 78% acetonitrile and 22% distilled water. The analysis was performed at a flow rate of 0.65 mL / min for 1.2 minutes, and the wavelength was analyzed at 210 nm using a PDA detector.

[0146]

[0147] (2) Indicator component of oyster mushroom mycelia

[0148] · Ingredient name: β-Glucan

[0149] · Molecular formula: (C6H 10 O5) n

[0150] · Molecular weight: (180.16 g / mol) n

[0151] · Chemical formula 2

[0152]

[0153]

[0154] Beta-glucans (β-Glucans) constitute a group of β-D-glucose polysaccharides that occur naturally in the cell walls of cereals, bacteria, and fungi, and possess significantly different physicochemical properties depending on their source. Typically, beta-glucans form a linear backbone with 1-3 beta-glycosidic linkages, but they vary in molecular mass, solubility, viscosity, branching structure, and gelling properties, resulting in a variety of physiological effects in animals.

[0155] Analytical method: The β-Glucan Assay Kit (Yeast and Mushroom) (K-YBGL) was purchased from NEOGEN (Michigan, USA). The experimental method was based on the kit usage protocol provided by NEOGEN and the β-glucan analysis method in the Health Functional Food Codex of the Ministry of Food and Drug Safety.

[0156]

[0157] (3) Indicator component of flower mushroom mycelia

[0158] · Ingredient name: Fraxinellone

[0159] · Molecular formula: C 14 H 16 O3

[0160] · Molecular weight: 232.27 g / mol

[0161] · CAS No.: 228808-62-0

[0162] · Chemical formula 3

[0163]

[0164]

[0165] Fraxinelone, a substance found primarily in the root bark of perennial plants like Rutaceae, is known for its insecticidal activity. It has also been reported to have various health benefits, including liver protection, anticancer effects, anti-inflammation, immunomodulation, and relief of inflammatory arthritis.

[0166] Method: Thermo Accucorea QC 18 HPLC analysis was performed at 40℃ using a column (150 mm * 2.1 mm). 2 μl of sample was injected, and the mobile phase was 0.1% formic acid aqueous solution 95%, acetonitrile 5% for 0 to 20 minutes, 100% acetonitrile for 20 to 23 minutes, and 0.1% formic acid aqueous solution 70%, acetonitrile 30% from 23 minutes. The analysis was performed at a flow rate of 0.4 mL / min for 23 minutes, and the wavelength was analyzed at 260 nm with a PDA detector.

[0167] · Ingredient name: Veratric acid

[0168] · Molecular formula: C9H 10 O4

[0169] · Molecular weight: 182.17 g / mol

[0170] · CAS No.: 93-07-2

[0171] · Chemical formula 4

[0172]

[0173] Veratric acid, a type of benzoic acid identified as a metabolite of certain plants, is known to exhibit antibacterial and anti-inflammatory activities. It has also been reported to suppress LPS-induced inflammatory damage by inhibiting the NF-κB signaling pathway.

[0174] Analysis method: HPLC analysis was performed at 25℃ using a YMC-Pack ODS AM-303 column (250 mm * 4.6 mm). The mobile phase used was 0.1% acetic acid aqueous solution (A) and 0.1% acetic acid acetonitrile solution (B) with a sample injection volume of 20 μl. The conditions were 92% A and 8% B for 0 to 2 minutes, 90% A and 10% B for 2 to 27 minutes, 10% A and 90% B for 27 to 50 minutes, 100% B for 50 to 60 minutes, and 92% A and 8% B from 63 minutes onwards at a flow rate of 1.0 mL / min for 60 minutes, and the wavelength was analyzed at 280 nm with a PDA detector.

[0175] · Ingredient name: Sparassol

[0176] · Molecular formula: C 10 H 12 O4

[0177] · Molecular weight: 196.29 g / mol

[0178] · CAS No.: 520-43-4

[0179] Chemical Formula 5

[0180]

[0181]

[0182] Sparasol is a substance produced by the chanterelle mushroom during its growth process and is known to have antibiotic and antifungal properties.

[0183] Analytical method: HPLC analysis was performed at 25℃ using an ACE 5 C18-HL column (250 mm * 4.6 mm). The mobile phase used was 0.1% acetic acid aqueous solution and methanol with a sample injection volume of 10 μl. The conditions were 0 to 60 minutes, 50% 0.1% acetic acid aqueous solution, 50% methanol, and 100% methanol after 60 minutes, at a flow rate of 1.0 mL / min for 60 minutes, and the wavelength was analyzed at 266 nm with a PDA detector.

[0184] · Ingredient name: Methyl orsellinate

[0185] · Molecular formula: C9H 10 O4

[0186] · Molecular weight: 182.17 g / mol

[0187] · CAS No.: 3187-58-4

[0188] Chemical formula 6

[0189]

[0190]

[0191] Methyl orcelinate is a substance produced during the growth process of the flower mushroom, similar to Sparassol. It is an antibiotic substance in which the methoxy group at carbon 4 of Sparassol is replaced with a hydroxy group, and their structures are very similar.

[0192] Analysis method: Simultaneously analyzed using the same method as Sparasol.

[0193]

[0194] <Test Example 1> Cytotoxicity evaluation using CCK-8 assay

[0195] Cytotoxicity was measured using the Cell Counting Kit-8 (CCK-8) assay (K1018, APExBIO Corporation, USA). MH7A cells were seeded at 3 × 10 per well in a 48-well plate. 4 Cells were seeded at a density of 100 μg / mL. After 24 h of culture, cells were pretreated with ME or WCF-ME at various concentrations (100, 200, 300, 500, 750, and 1000 μg / mL) and cultured for 24 h (37°C, 5% CO2). Subsequently, cells were treated with 200 μL of 10% CCK-8 solution and cultured in the dark for 2 h. Finally, absorbance was measured at 450 nm using an Epoch microplate spectrophotometer (BioTek, VT, USA), and the relative rate of cell proliferation was calculated.

[0196] Figure 2 shows the cytotoxicity of mushroom mycelia extract fermentation. Here, (a) bacterial growth of W. confusa WiKim51 in growth medium supplemented with ME for 48 h; (b) acetic acid content of ME and WCF-ME; (c) viability of MH7A cells pretreated with various concentrations (0, 100, 200, 300, 500, 750, 1000 μg / mL) of ME and (d) WCF-ME. As shown in Figure 2, ME was not toxic to bacterial growth, and WC-fermented ME (WCF-ME) showed an increase in acetate concentration after 24 h.

[0197]

[0198] <Test Example 2> In vitro anti-inflammatory evaluation

[0199] Anti-inflammatory efficacy experiments were performed using Raw 264.7 cells and MH7A cells.

[0200] Gene expression of inflammation-related biomarkers was measured by RT-qPCR. MH7A cells were seeded at 2 × 10 per well in 6-well plates. 5Cells were seeded at respective cell densities. After 24 h of culture, cells were pretreated with ME (750 μg / mL) and WCF-ME (750 μg / mL) diluted in DMEM and incubated for 18 h (37°C, 5% CO2). To induce an inflammatory response, 10 ng / mL TNFα (PeproTech, USA) diluted in DMEM was added and incubated for 6 h (37°C, 5% CO2). RNA was extracted from cells using TRIzol reagent (Thermo Fisher Scientific) according to the manufacturer's protocol. Final RNA concentration and purity were measured by UV absorbance using a NanoDrop spectrophotometer (BioTek, VT, USA). cDNA was synthesized using a cDNA reverse transcription kit (Thermo Fisher Scientific). Reverse transcription quantitative real-time PCR (RT-qPCR) was performed using a CFX96 Real-Time PCR Detection System (Bio-Rad, Hercules, CA, USA). Target gene expression was quantified using the 2X Real-Time PCR Smart Mix (SolGent, Daejeon, Korea). The mRNA expression level of each target gene was analyzed and normalized relative to the GAPDH family gene.

[0201] Figure 3 shows the anti-inflammatory efficacy of a mushroom mycelia extract fermented using Lactobacillus paracasei WiKim0171 in Raw 264.7 cells. Here, L001 is a mushroom mycelia extract fermented using a culture medium of the Lactobacillus paracasei WiKim0171 strain. As shown in Figure 3, it was confirmed that the mushroom mycelia extract fermented significantly suppressed the expression rate of inflammatory cytokines.

[0202] Figure 4 shows the anti-inflammatory efficacy of a mushroom mycelia extract fermented with Lactobacillus paracasei WiKim0171 in MH7A cells. Evaluation of MMPs, inflammatory cytokines, and HO-1 using MH7A cells revealed that the anti-inflammatory efficacy was further enhanced when the extract was fermented with L001.

[0203] Figure 5 shows the anti-inflammatory efficacy of mushroom mycelia extract fermented with Weissella confusa WIKIM51 (Wilac D001) in MH7A cells. Here, the effects of ME and WCF-ME on the gene expression of inflammation-related markers (e) MMP3, (f) MMP13, (g) IL1B, and (h) IL6 in MH7A cells treated with TNFα. As shown in Figure 5, ME and WCF-ME reduced the expression of inflammation-related genes (MMP3, MMP13, IL1B) increased by TNFα induction, and in particular, WCF-ME also reduced IL6 gene expression.

[0204]

[0205] <Test Example 3> Effect of reducing joint swelling

[0206] The effects of WC, ME, and WCF-ME on body weight, serum biomarkers, and joint destruction in the collagen-induced arthritis (CIA) model were observed.

[0207] Seven-week-old Wistar rats were divided into six groups (Con, CIA, MTX, WC, ME, WCF-ME) and immunized twice. Immunization was administered subcutaneously using type II collagen and Freund's adjuvant. After adaptation, each group received oral gavage for 4 weeks, and body weight, food intake, paw thickness, and arthritis severity were measured.

[0208] Serum analysis

[0209] Serum TNFα was measured using an ELISA kit (K0331196, Komabiotech, Korea) and PGE2 (KGB004B, R&D Systems, USA).

[0210] Pathological analysis

[0211] At the end of the experiment, pathological analysis of the hind paw joints of each rat was performed. The hind paw joints of each group were fixed in 10% (v / v) formaldehyde, decalcified (RDO Gold, Apex Engineering Products Corporation, USA), and embedded in paraffin blocks using standard techniques. Sections of 4–5 μm thickness were then prepared and stained with H&E. Digital photomicrographs were taken from representative areas using an EasyScan Pro6, Motic slide scanner (Hong Kong).

[0212] CIA treatment induced weight loss and decreased food intake, indicating the induction of inflammation. ME, WC, and WCF-ME treatments did not affect weight loss or food intake.

[0213] Figure 6 shows the results of observing the changes in growth parameters and serum biomarkers by the mushroom mycelia extract fermented using Weissella confusa WIKIM51 (Wilac D001) in the collagen-induced arthritis (CIA) model. Here, (a) body weight; (b) food intake; (c) water intake; (d) serum levels of TNFα and (e) PGE2 are shown. As shown in Figure 6, in the collagen-induced arthritis (CIA) model, CIA-induced serum TNFα was normalized by WCF-ME treatment, and PGE2 was reduced by WC and WCF-ME treatments. ME had no effect on either serum marker. CIA-induced paw edema and deformation were significantly reduced by WC and WCF-ME from day 21 after immunization, but ME had no effect. That is, serum inflammatory markers TNFα and PGE2 increased after CIA injection, and WCF-ME and MTX (comparator drug) treatment normalized TNFα, while WC and WCF-ME reduced PGE2.

[0214] These results suggest that MTX, WC, and WCF-ME have joint protective effects by inhibiting CIA-induced cartilage destruction, synovial proliferation, and pannus formation.

[0215]

[0216] <Experimental Example 4> Observation of the effect on knee joint inflammation

[0217] Using primers as shown in Table 2 below

[0218] The expression of inflammation-related genes (IL6, IL1B, MMP3, MMP13) in the knee joint was evaluated using RT-qPCR.

[0219]

[0220] As a result, Il6 and Mmp3 gene expression in the knee joint was normalized by all treatments, and Il1b gene expression was normalized only in the WC group. Mmp13 gene expression was normalized by WC and WCF-ME, but there was no significant difference in ME.

[0221] Figure 7 shows the effect of mushroom mycelia extract fermented with Weissella confusa WIKIM51 (Wilac D001) on improving arthritis symptoms in a collagen-induced arthritis (CIA) model. Here, (a) gross morphology (scale bar: 1 cm) and representative images of H&E-stained hind paw joints. (b) Magnification: 40x, scale bar: 1000 μm; (c) Magnification: 160x, scale bar: 1000 μm. The black box indicates the enlarged area in the corresponding figure below. (d) paw thickness. (e) arthritis score. The arthritis score was calculated as shown in Table 3 below.

[0222]

[0223]

[0224] <Experimental Example 5> Spearman's Correlation Analysis

[0225] The correlation between serum TNFα and PGE2 concentrations and IL6, IL1B, MMP3, and MMP13 gene expression in the knee joint was evaluated.

[0226] Figure 8 shows the effects of mushroom mycelia extract fermented using Weissella confusa WIKIM51 (Wilac D001) on knee joint inflammation. Here, gene expression in the knee joint (a) Il6, (b) Il1b, (c) Mmp3, and (d) Mmp13 is shown. (e) Serum TNFα and PGE2 concentrations; and Spearman correlation analysis results for knee joint Il6, Il1b, Mmp3, and Mmp13 gene expression.

[0227] As shown in Figure 8, Spearman's correlation analysis showed that serum PGE2 was proportional to Il6, Il1b, and Mmp13. Collectively, these results suggest that WC and WCF-ME improve CIA-induced RA symptoms by suppressing PGE2 activity and reduce Il6, Mmp3, and Mmp13 expression in the knee. Therefore, WC and WCF-ME are expected to have potential as novel RA therapeutics.

[0228]

[0229] <Experimental Example 5> In vivo RA model - Measurement of body weight and joint swelling

[0230] Figure 9 shows the results of body weight and joint swelling measurements using the RA model. The high-dose group, like the control group, showed no weight loss, and compared to the CIA group after arthritis induction, the high-dose group showed less joint swelling. The low-dose group showed similar increases in joint swelling as the CIA group.

[0231]

[0232] While specific aspects of the present invention have been described in detail above, it should be apparent to those skilled in the art that these specific descriptions are merely preferred embodiments and do not limit the scope of the present invention. Therefore, the substantial scope of the present invention is defined by the appended claims and their equivalents.

[0233]

[0234] In this way, the composition for preventing or treating inflammatory osteoarthritis, which includes the mushroom mycelia extract fermentation product according to the present invention as an active ingredient, has an anti-inflammatory effect and an effect of improving joint swelling, and thus can be usefully used as a composition for protecting and strengthening joints and cartilage.

Claims

1. A pharmaceutical composition for preventing or treating inflammatory osteoarthritis, characterized in that it contains a mushroom mycelia extract fermentation product as an effective ingredient.

2. In paragraph 1, A pharmaceutical composition for preventing or treating inflammatory osteoarthritis, characterized in that the mushroom mycelia are at least one mushroom mycelia selected from the group consisting of oyster mushroom mycelia, white oyster mushroom mycelia, and pine mushroom mycelia.

3. In paragraph 2, A pharmaceutical composition for preventing or treating inflammatory osteoarthritis, characterized in that the mushroom mycelia are a mixture of oyster mushroom mycelia and pine mushroom mycelia in a ratio of 1:2 to 1:4 (w / w).

4. In paragraph 1, A pharmaceutical composition for preventing or treating inflammatory osteoarthritis, characterized in that the above mushroom mycelia extract fermentation product is obtained by fermenting mushroom mycelia extract with lactic acid bacteria.

5. In paragraph 4, A pharmaceutical composition for preventing or treating inflammatory osteoarthritis, characterized in that the mushroom mycelia extract is prepared according to the following method: (1) A step of mixing 30 to 50 parts by weight of glucose with 100 parts by weight of mushroom mycelium powder; (2) a step of adding purified water in an amount of 9 to 11 times the weight of the mushroom mycelia to the mixture mixed in the step (1) and extracting at 95 to 100°C for 2 to 4 hours; and (3) A step of filtering the extract extracted in the above step (2).

6. In paragraph 4, A pharmaceutical composition for preventing or treating inflammatory osteoarthritis, characterized in that the lactic acid bacteria used are Weissella confusa WIKIM51 (Wilac D001) or Lactobacillus paracasei WiKim0171.

7. In paragraph 6, A pharmaceutical composition for preventing or treating inflammatory osteoarthritis, characterized in that the above lactic acid bacteria uses a lactic acid bacteria culture solution.

8. In paragraph 7, A pharmaceutical composition for preventing or treating inflammatory osteoarthritis, characterized in that the above lactic acid bacteria culture solution is cultured according to the following method: (1) Step for preparing MRS badge; (2) Step of adding Weissella confusa WIKIM51 (Wilac D001) or Lactobacillus paracasei WiKim0171 to the medium; and (3) A step of culturing at 35 to 40°C for 22 to 26 hours.

9. In paragraph 1, A pharmaceutical composition for preventing or treating inflammatory osteoarthritis, characterized in that the above mushroom mycelia extract fermentation product is manufactured according to the following method: (1) A step of mixing one or more mushroom mycelia extracts selected from the group consisting of oyster mushroom mycelia, white oyster mushroom mycelia, and chanterelle mushroom mycelia at a ratio of 1:2 to 1:4 (v / v); (2) A step of sterilizing the above mixed extract under conditions of 1.2 atm and 120 to 125°C; (3) OD of lactic acid bacteria 600 Step of inoculating at a concentration of 1-2% (v / v) with a 0.3 condition; (4) Fermentation step at 35 to 40℃ for 45 to 50 hours; (5) A step of filtering and concentrating the fermented product; (6) Step of freeze-drying the concentrated fermented product and then powdering it.

10. In paragraph 1, A pharmaceutical composition for preventing or treating inflammatory osteoarthritis, characterized in that the inflammatory osteoarthritis is at least one disease selected from the group consisting of ankylosing spondylitis, psoriatic arthritis, osteoarthritis, periarthritis of the shoulder joint, tendonitis, tenosynovitis, and peritendinitis.

11. A functional food composition for preventing or improving inflammatory osteoarthritis, characterized by containing a mushroom mycelia extract fermentation product as an effective ingredient.

12. A feed composition for preventing or improving inflammatory osteoarthritis, characterized in that it contains a mushroom mycelia extract fermentation product as an effective ingredient.

13. A method for preventing or treating inflammatory osteoarthritis, comprising administering a composition according to any one of claims 1 to 10 to a subject other than a human.

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