Composition for preventing, improving, or treating inflammatory diseases, comprising extracellular vesicles derived from the genus Roseburia or Bifidobacterium

Abstract

The present invention relates to a composition for preventing, improving or treating inflammatory diseases, comprising extracellular vesicles (EVs) derived from Roseburia spp. strains and / or extracellular vesicles (EVs) derived from Bifidobacterium spp. strains. Not only have the EVs of the present invention been shown to be effective in improving inflammatory bowel disease (IBD), but it has also been confirmed that EVs derived from Roseburia strains or congenerally different strains corresponding to Bifidobacterium strains all exhibit strong anti-inflammatory effects. Therefore, the strains of the present invention and EVs derived therefrom can be provided as compositions for preventing, improving or treating various inflammatory diseases.

Description

[Technical field]

[0001] The present invention relates to a composition for preventing, ameliorating or treating an inflammatory disease, which comprises a Roseburia spp. strain, a Bifidobacterium spp. strain, or extracellular vesicles (EVs) derived therefrom.

[0002] The present invention claims priority based on Korean Patent Application No. 10-2022-0008554 filed on January 20, 2022 and Korean Patent Application No. 10-2023-0006807 filed on January 17, 2023, and all contents disclosed in the specification and drawings of said application are incorporated by reference into this application. [Background technology]

[0003] Background technology

[0004] Inflammation refers to the pathological state of abscesses formed by the invasion of external infectious agents (bacteria, fungi, viruses, and various types of allergy-inducing substances). Specifically, when external bacteria invade and grow in a specific tissue, the white blood cells of the body recognize this and actively attack the proliferated external bacteria. During this process, the corpses of white blood cells are accumulated in the tissue invaded by the bacteria, and the cell destruction products of the invading bacteria killed by the white blood cells are dissolved in the invaded tissue to form an abscess. The treatment of abscesses caused by inflammation is promoted through anti-inflammatory action, which means the action of suppressing the growth of invading bacteria using an antibacterial agent or promoting the function of macrophages that phagocytize foreign substances accumulated in the abscess and digest and excrete the foreign substances by activating the macrophages.

[0005] In general, an inflammatory response is a biological defense reaction process for repairing and regenerating damage caused by invasion that brings about matrix changes in cells and tissues of the living body, and this reaction process involves the action of local blood vessels, various tissue cells in body fluids, and immune cells. An inflammatory reaction normally induced by an external invading bacterium is a defense system for protecting the living body, whereas an abnormally excessive inflammatory reaction leads to various diseases, which are called inflammatory diseases. Inflammatory diseases may threaten the life of the human body by amplifying and sustaining inflammation due to various inflammatory mediators secreted from target cells activated by external stimuli. Inflammatory diseases include acute bronchitis, chronic bronchitis, acute bronchiolitis, chronic bronchiolitis, sepsis, septic shock, acute respiratory distress syndrome, multiple organ failure, and chronic obstructive pulmonary disease. Inflammatory bowel disease (IBD), one of the representative inflammatory diseases, causes chronic inflammation in the gastrointestinal tract and shows a chronic course, gradually progressing the disease, causing complications and reducing physical function (SY Na and W Moon, Korean J Gastroenterol Vol.71 No.2, 61-68). Inflammatory bowel disease is mainly classified into two forms, ulcerative colitis (UC) and Crohn's disease (CD). The clear pathogenesis of inflammatory bowel disease is unknown, and it is presumed to be caused by a complex interaction of genetic factors, stress, drug use, and other environmental factors. Therefore, it is a disease that is difficult to cure completely because each patient has a different cause, and the course and prognosis are also different.

[0006] The onset of many inflammation-related diseases is associated with the activation of macrophages and the resulting excessive production of inflammation-related factors, representative of which are IL-1β and TNF-α. Among the inducers of inflammatory responses, LPS (lipopolysaccharide) is a substance that constitutes the cell surface of gram-negative bacteria, and interacts with immune cells such as leukocytes and is involved in the regulation of inflammation-related cytokines. In particular, inflammatory cytokines such as IL-1β, TNF-α, and IL-6 play a crucial role in mediating the pathological progression of various infectious or inflammatory diseases. Such inflammatory cytokines are produced by several immune and inflammatory cells and contribute to recruiting additional inflammatory cells to the injury site, which causes tissue and organ damage. Summary of the Invention [Problem to be solved by the invention]

[0007] Detailed Description of the Invention technical challenges

[0008] The present inventors have made extensive efforts to find a drug having superior effects to conventional therapeutic agents for inflammatory diseases, and have confirmed that extracellular vesicles (EVs) derived from Roseburia spp. and Bifidobacterium spp. exhibit excellent preventive, ameliorative and therapeutic effects against inflammatory diseases, thereby completing the present invention.

[0009] Therefore, an object of the present invention is to provide a pharmaceutical composition for preventing or treating an inflammatory disease, comprising at least one selected from the group consisting of extracellular vesicles derived from Roseburia strains and extracellular vesicles derived from Bifidobacterium strains as an active ingredient.

[0010] Another object of the present invention is to provide a kit for preventing or treating an inflammatory disease, comprising at least one selected from the group consisting of extracellular vesicles derived from Roseburia strains and extracellular vesicles derived from Bifidobacterium strains as an active ingredient.

[0011] Yet another object of the present invention is to provide a method for preventing or treating an inflammatory disease, comprising administering to an individual in need thereof at least one selected from the group consisting of extracellular vesicles derived from Roseburia strains and extracellular vesicles derived from Bifidobacterium strains.

[0012] Yet another object of the present invention is to provide a food composition for preventing or ameliorating inflammatory diseases, comprising at least one selected from the group consisting of extracellular vesicles derived from Roseburia strains and extracellular vesicles derived from Bifidobacterium strains.

[0013] Yet another object of the present invention is to provide a feed composition for preventing or ameliorating inflammatory diseases, comprising at least one selected from the group consisting of extracellular vesicles derived from Roseburia strains and extracellular vesicles derived from Bifidobacterium strains.

[0014] However, the technical problems that the present invention aims to solve are not limited to the problems mentioned above, and other problems not mentioned will be clearly understood by a person having ordinary skill in the art to which the present invention pertains from the following description. [Means for solving the problem]

[0015] Problem solving means

[0016] The present invention provides a pharmaceutical composition for preventing or treating an inflammatory disease, comprising as an active ingredient at least one selected from the group consisting of extracellular vesicles derived from Roseburia spp. strains and extracellular vesicles derived from Bifidobacterium spp. strains.

[0017] The present invention also provides a food composition for preventing or ameliorating inflammatory diseases, comprising as an active ingredient at least one selected from the group consisting of extracellular vesicles derived from Roseburia strains and extracellular vesicles derived from Bifidobacterium strains.

[0018] The present invention also provides a feed composition for preventing or ameliorating inflammatory diseases, comprising as an active ingredient at least one selected from the group consisting of extracellular vesicles derived from Roseburia strains and extracellular vesicles derived from Bifidobacterium strains.

[0019] The present invention also provides a kit for preventing, ameliorating, or treating an inflammatory disease, comprising at least one selected from the group consisting of extracellular vesicles derived from a Roseburia strain and extracellular vesicles derived from a Bifidobacterium strain.

[0020] Furthermore, the present invention provides a method for preventing or treating an inflammatory disease, comprising administering to an individual in need thereof at least one selected from the group consisting of extracellular vesicles derived from a Roseburia strain and extracellular vesicles derived from a Bifidobacterium strain.

[0021] Furthermore, the present invention provides use of at least one selected from the group consisting of extracellular vesicles derived from Roseburia strains and extracellular vesicles derived from Bifidobacterium strains for the prevention or treatment of inflammatory diseases.

[0022] Furthermore, the present invention provides use of at least one selected from the group consisting of extracellular vesicles derived from a Roseburia strain and extracellular vesicles derived from a Bifidobacterium strain for the manufacture of a therapeutic agent for an inflammatory disease.

[0023] In one embodiment of the present invention, the extracellular vesicles may have an average diameter of 30 nm to 200 nm, but is not limited thereto.

[0024] In another embodiment of the present invention, the Roseburia strain-derived extracellular vesicles may be naturally or artificially secreted from a Roseburia strain, but is not limited thereto.

[0025] In yet another embodiment of the present invention, the extracellular vesicles derived from a Bifidobacterium strain may be naturally or artificially secreted from a Bifidobacterium strain, but are not limited thereto.

[0026] In yet another embodiment of the present invention, the Roseburia strain may be at least one selected from the group consisting of Roseburia intestinalis, Roseburia faecies, Roseburia hominis, and Roseburia inulinivorans, but is not limited thereto.

[0027] In yet another embodiment of the present invention, the Bifidobacterium strain may be at least one selected from the group consisting of Bifidobacterium longum, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium lactis, and Bifidobacterium adolescentis, but is not limited thereto.

[0028] In another embodiment of the invention, the inflammatory disease is selected from the group consisting of gastritis, gastric ulcer, duodenal ulcer, inflammatory skin disease, allergic disease, irritable bowel syndrome, ulcerative colitis, inflammatory bowel disease, peritonitis, osteomyelitis, cellulitis, meningitis, encephalitis, pancreatitis, trauma-induced shock, bronchial asthma, cystic fibrosis, stroke, acute bronchitis, chronic bronchitis, acute bronchiolitis, chronic bronchiolitis, osteoarthritis, gout, spondyloarthropathy, ankylosing spondylitis, Reiter's syndrome, psoriatic arthropathy, enteropathy, spondylitis. , juvenile arthropathy, juvenile ankylosing spondylitis, reactive arthropathy, infectious arthritis, post-infectious arthritis, gonococcal arthritis, tuberculous arthritis, viral arthritis, fungal arthritis, syphilitic arthritis, Lyme disease, arthritis associated with vasculitis syndromes, polyarteritis nodosa, hypersensitivity vasculitis, Rugeric's granulomatosis, polymyalgia rheumatica, articular cell arteritis, calcium crystal deposition arthropathy, caustic gout, non-rheumatoid arthritis, mucositis, tenosynovitis, epicondylitis, neuropathic arthropathy (charco and joint, hemarthrosic, Henoch-Scholein purpura, hypertrophic osteoarthritis, multicentric reticulohistiocytosis, sarcoilosis, hemochromatosis, sickle cell disease, hemochromatosis, hyperlipoproteinemia, hypogammaglobulinemia, familial Mediterranean fever, Behart's disease, systemic lupus erythematosus, relapsing fever, psoriasis, multiple sclerosis, sepsis, septic shock, multiple organ dysfunction syndrome, acute respiratory distress syndrome, chronic obstructive pulmonary disease, acute lung injury, and broncho-pulmonary dysplasia, but is not limited thereto.

[0029] In yet another embodiment of the present invention, the inflammatory bowel disease may be at least one selected from the group consisting of ulcerative colitis, Crohn's disease, Behcet's disease, collagenous colitis, lymphocytic colitis, ischemic colitis, hemorrhagic rectal ulcer, pouchitis, and transformation colitis, but is not limited thereto.

[0030] In yet another embodiment of the present invention, the composition according to the present invention may have at least one effect selected from the group consisting of, but not limited to:

[0031] i) Reduction of intestinal epithelial cell barrier damage

[0032] ii) Reduced infiltration of immune cells into intestinal tissue

[0033] iii) Suppression of decrease in goblet cell number

[0034] iv) Inhibition of cecal weight loss

[0035] v) Inhibition of decrease in intestinal length

[0036] vi) Suppression of inflammatory cytokine production

[0037] vii) Promoting anti-inflammatory cytokine production

[0038] viii) Increased mucus secretion from goblet cells

[0039] ix) enhancement of dense junctions in intestinal epithelial cells; and

[0040] x) Increase in the number of Bifidobacteria in the intestines

[0041] In yet another embodiment of the present invention, the food composition may be, but is not limited to, a functional health food composition. Effect of the Invention

[0042] Effect of the Invention

[0043] The present invention relates to a composition for preventing, improving or treating inflammatory diseases, comprising extracellular vesicles (EVs) derived from Roseburia spp. and / or Bifidobacterium spp. In an animal model of inflammatory bowel disease, the extracellular vesicles of the present invention were shown to reduce damage to the intestinal epithelial cell barrier and infiltration of immune cells, maintain the number of goblet cells, inhibit the loss of cecal weight and intestinal length caused by enteritis, inhibit the secretion of inflammatory cytokines through changes in the intestinal flora, and restore mucus secretion from goblet cells and the tight junction ability of intestinal epithelial cells. In particular, it was confirmed that EVs derived from congenerally heterologous strains corresponding to Roseburia spp. or Bifidobacterium spp. all exhibit strong anti-inflammatory effects. In other words, the extracellular vesicles derived from Roseburia or Bifidobacterium strains according to the present invention can effectively suppress inflammation and improve tissue damage and functional abnormalities caused by inflammation, and therefore can be provided as a composition for preventing, improving and / or treating various inflammatory diseases.

[0044] BRIEF DESCRIPTION OF THE DRAWINGS [Brief description of the drawings]

[0045] [Figure 1] Figure 1 shows ultrathin section TEM (left) and cryo-TEM (right) images of extracellular vesicles (EVs) derived from Roseburia intestinalis (Ri-EVs).

[0046] [Diagram 2] Figure 2 shows the results of measuring the size and surface charge of Ri-EV.

[0047] [Figure 3a] Figure 3a shows the particle concentration of Ri-EV extracted from R. intestinalis.

[0048] [Figure 3b] Figure 3b shows the particle size measurements of Roseburia intestinalis-derived extracellular vesicles (Ri-EVs) and Bifidobacterium longum-derived EVs (Bl-EVs).

[0049] [Figure 4] FIG. 4 is a schematic diagram of an experiment using an animal model of dextransulfate sodium salt colitis grade (DSS)-induced inflammatory bowel disease (IBD).

[0050] [Diagram 5] FIG. 5 shows intestinal tissues stained with H&E (Hematoxilin-Eosin) and AB (Alcian blue) to observe the barrier of intestinal epithelial cells in the control group (control), DSS-induced IBD animal model (DSS), R. intestinalis administration group (Ri-Cell), and Ri-EV administration group (Ri-EV).

[0051] [Figure 6a] Figures 6a to 6d show the results of pathological analysis of intestinal tissues in the control group, DSS-induced IBD animal model, R. intestinalis-treated group, and Ri-EV-treated group, including histological scores and the number of goblet cells (Figure 6a), cecum weight (Figure 6b), colon length (Figure 6c), and disease activity index (Figure 6d). [Figure 6b] Same as above [Figure 6c] Same as above [Figure 6d] Same as above

[0052] [Figure 7a]Figures 7a and 7b show the results of measuring the expression levels of IL (Interleukin)-1β (Figure 7a), Muc-2 (Mucin2), and ZO-1 (Zonula occludens-1) in intestinal tissue (Figure 7b) in the control group, DSS-induced IBD animal model, R. intestinalis-treated group, and Ri-EV-treated group to confirm changes in intestinal inflammation levels, mucus protein levels, and dense junctions due to administration of R. intestinalis or Ri-EV. [Figure 7b] Same as above

[0053] [Figure 8a] Figures 8a and 8b show the results of measuring the intestinal microbial community (Figure 8a) and Shannon diversity (Figure 8b) in the control group, DSS-induced IBD animal model, R. intestinalis-treated group, and Ri-EV-treated group to confirm changes in the intestinal microbial community due to administration of R. intestinalis or Ri-EV. [Figure 8b] Same as above

[0054] [Figure 9] FIG. 9 shows the abundance of Bifidobacterium in the control group, the DSS-induced IBD animal model, the R. intestinalis administration group, and the Ri-EV administration group.

[0055] [Figure 10] Figure 10 shows the results of treating mouse macrophages with LPS to induce an inflammatory environment, and then measuring the amount of nitric oxide (NO) produced, the expression levels of inflammatory cytokines (TNF-α, IL-6), and the expression levels of anti-inflammatory cytokines (IL-10) after treatment with EVs derived from congeneric but different Roseburia strains (R. intestinalis (Ri), R. hominis (Rho), R. faecies (Rfa), and R. inulinivorans (Rin)).

[0056] [Figure 11]Figure 11 shows the results of measuring the amount of nitric oxide production to confirm the anti-inflammatory effect of treatment with EVs derived from Roseburia sp. strains (R. intestinalis, R. faecies, R. hominis, and R. inulinivorans) or EVs derived from Bifidobacterium sp. strains (B. bifidum, B. breve, B. lactis, B. adolescentis, and B. longum) after inducing an inflammatory environment by treating the cells with LPS in an in vitro environment mimicking Kuffer cells. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0057] Best Mode for Carrying Out the Invention

[0058] The present invention will be described in detail below with reference to examples to aid in understanding the present invention. However, the following examples are merely illustrative of the contents of the present invention, and the scope of the present invention is not limited to the following examples. The examples of the present invention are provided to more completely explain the present invention to those having average knowledge in the art.

[0059] The present invention relates to a pharmaceutical composition for preventing or treating inflammatory diseases, comprising extracellular vesicles derived from Roseburia spp. strains and extracellular vesicles derived from Bifidobacterium spp. strains as active ingredients, and has been completed by confirming that various Roseburia spp. strains, Bifidobacterium spp. strains and / or extracellular vesicles derived therefrom not only exhibit a strong anti-inflammatory effect but also improve tissue damage and functional abnormalities caused by inflammation. Therefore, the extracellular vesicles of the present invention may be used as a composition for preventing, improving and / or treating various inflammatory diseases.

[0060] The composition according to the present invention may contain at least one selected from the group consisting of Roseburia spp. strains, their homogenates, culture broths, extracellular vesicles, and mixtures thereof, and Bifidobacterium spp. strains, their homogenates, culture broths, extracellular vesicles, and mixtures thereof. In one embodiment of the present invention, the composition according to the present invention may not contain a bacterial strain.

[0061] In the present invention, "strains of the genus Roseburia" include strains that are already known in the art as strains belonging to the genus Roseburia, or strains that will be newly known in the future. Roseburia is a Gram-positive bacterium that inhabits the human colon and is known to have the ability to decompose sugar and produce butyric acid. The Roseburia strain according to the present invention includes any strain that belongs to the genus Roseburia, and is not limited to a specific type, but may be at least one selected from the group consisting of Roseburia intestinalis, Roseburia faecies, Roseburia hominis, Roseburia inulinivorans, and Roseburia cecicola. Among these, Roseburia intestinalis is the first microorganism isolated from human feces and is known to produce metabolic products in the intestine to maintain energy homeostasis.

[0062] In the present invention, "Bifidobacterium spp. or the genus Bifidobacterium strain" includes strains that are already known in the art as belonging to the genus Bifidobacterium, or strains that will be newly known in the future. Bifidobacteria are Gram-positive anaerobic bacteria, and are one of the major types of bacteria that make up the mammalian gastrointestinal microbial community. Some Bifidobacteria are also used as lactic acid bacteria. The Bifidobacterium strain according to the present invention is not limited to any particular species as long as it is a member of the genus Bifidobacterium, but is preferably at least one selected from the group consisting of Bifidobacterium longum, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium lactis, and Bifidobacterium adolescentis. Among these, Bifidobacterium longum is also known as bifidobacteria and is known to be effective in inhibiting the proliferation of Escherichia coli and enhancing intestinal motility and defecation.

[0063] In the present invention, the term "strain" includes not only live bacteria obtained from a culture such as a culture medium, but also any processed form of the strain known to those skilled in the art, including, but not limited to, crushed bacteria, dried products, frozen products, etc. In the present specification, the term "strain" may be used interchangeably with "bacteria", "cells", etc.

[0064] In the present invention, the term "disrupted product" refers to a product obtained by disrupting the cell walls of a strain by chemical treatment or by applying physical force to the strain.

[0065] In the present invention, the term "culture solution" refers to the entire medium containing the strain, its metabolites, excess nutrients, etc., obtained by culturing the strain for a certain period of time in a medium that can supply nutrients so that the strain can grow and survive in a test tube. The culture solution refers to a culture solution obtained by removing the bacteria from the culture solution obtained by culturing the strain. Meanwhile, the liquid from the culture solution from which the bacteria have been removed is also called "supernatant", and can be obtained by leaving the culture solution for a certain period of time and taking only the upper liquid excluding the part that has sunk to the bottom, removing the bacteria by filtration, or centrifuging the culture solution to remove the lower precipitate and taking only the upper liquid. The "bacterial cells" refers to the strain itself of the present invention, and includes the strain itself separated and selected from a biological sample or the like, or the strain separated from the culture solution in which the strain was cultured. The bacterial cells can be obtained by centrifuging the culture solution to take the part that has sunk to the bottom, or by leaving the culture solution for a certain period of time and removing the upper liquid since the bacterial cells sink to the bottom of the culture solution due to gravity. The culture medium may be referred to as a "culture supernatant," a "conditioned medium," or a "conditioned medium."

[0066] The culture medium may include, but is not limited to, a culture medium itself obtained by culturing a strain, an extract thereof, a concentrate thereof, or a freeze-dried product thereof, or a culture supernatant obtained by removing the strain from the culture medium, an extract thereof, a concentrate thereof, or a freeze-dried product thereof.

[0067] The culture medium may be one obtained by culturing the strain of the present invention in an appropriate medium (e.g., RCM (reinforced clostridial medium) medium) at any temperature of 10° C. to 50° C., 10° C. to 40° C., 20° C. to 50° C., 20° C. to 40° C., or 30° C. to 40° C. for a certain period of time, for example, 4 to 50 hours, 4 to 40 hours, 4 to 30 hours, 4 to 20 hours, or 10 to 20 hours, but is not limited thereto. The culture medium and culture conditions for culturing the strain of the present invention can be appropriately selected or modified by a person of ordinary skill in the art.

[0068] In one embodiment, the culture supernatant of the present invention is obtained by subjecting the above-mentioned strain culture solution to the step of removing the strain by a method such as centrifugation or filtration.

[0069] In the present invention, the term "concentrate" refers to the above-mentioned culture medium itself, or a culture medium obtained by concentrating a supernatant obtained by a method such as centrifugation or filtration.

[0070] In the present invention, the term "extract" means something extracted from the above-mentioned culture solution or a concentrate thereof, and may include an extract, a dilution or concentrate of the extract, a dried product or freeze-dried product obtained by drying the extract, or a crude or purified product thereof, or a fraction thereof.

[0071] In the present invention, the term "vesicle" refers to a particle secreted from a cell and released into the extracellular space, and may include a number of different species, such as exosomes, ectosomes, microvesicles, microparticles, and exosome-like vesicles. Extracellular vesicles (EVs) can reflect the state of the source cell (donor cell) from which they are secreted, and can exhibit various biological activities depending on which cell they are secreted from, and can play an important role in intercellular interactions by transferring genetic material and proteins between cells. In addition, cell-derived substances including the vesicles can cause disease or stimulate immune cells to fight disease, and can help decompose and absorb substances that humans cannot digest through the metabolic process of microorganisms. The vesicles are membrane-structured vesicles that are divided into an inside and an outside, contain plasma membrane lipids, plasma membrane proteins, nucleic acids, and cytoplasmic components, and may be smaller than the original cell. The term "extracellular vesicles" may be used interchangeably with "extracellular vesicles" and may also be simply called "endoplasmic reticulum" or "vesicles."

[0072] In particular, the vesicles derived from Gram-positive bacteria such as Roseburia strains and Bifidobacterium strains of the present invention may contain, in addition to proteins and nucleic acids, components of bacterial cell walls such as peptidoglycan and lipoteichoic acid.

[0073] In one embodiment of the present invention, the extracellular vesicles derived from a Roseburia strain may have a single lipid membrane structure, but is not limited thereto.

[0074] The vesicles according to the present invention may be produced during in vitro culture of the strains. Specifically, the vesicles according to the present invention may be naturally secreted from the strains of Roseburia and / or Bifidobacterium or may be artificially produced (isolated) by heat treatment, pressure treatment, etc., and may have a diameter of 30-200 nm, 30-150 nm, 30-100 nm, 30-80 nm, 40-200 nm, 50-200 nm, 50-100 nm, or 50-80 nm. In one embodiment of the present invention, the sizes of EVs derived from R. intestinalis and EVs derived from B. longum were measured using a zeta-sizer and were found to have an average particle size of about 76 nm (FIG. 1).

[0075] Furthermore, the extracellular vesicles according to the present invention may have a surface charge of -20 to 5 mV, -20 to 0 mV, -10 to 0 mV, -5 to 0 mV, -10 to -1 mV, or -5 to -1 mV, but are not limited thereto.

[0076] The vesicles may be isolated by subjecting a culture solution containing a Roseburia sp. strain and / or a Bifidobacterium sp. strain to at least one method selected from the group consisting of centrifugation, ultra-high speed centrifugation, extrusion, sonication, cell lysis, homogenization, freeze-thaw, electroporation, mechanical disintegration, chemical treatment, filtration through a filter, gel filtration chromatography, free-flow electrophoresis, and capillary electrophoresis, and may further include steps such as washing to remove impurities and concentrating the obtained vesicles.

[0077] The composition according to the present invention can have preventive, ameliorative, and / or therapeutic effects on inflammatory diseases.

[0078] As an example, the composition according to the present invention may have at least one effect selected from the following:

[0079] i) Reduction of intestinal epithelial cell barrier damage

[0080] ii) Reduced infiltration of immune cells into intestinal tissue

[0081] iii) Suppression of decrease in goblet cell number

[0082] iv) Inhibition of cecal weight loss

[0083] v) Inhibition of decrease in intestinal length

[0084] vi) Suppression of inflammatory cytokine production

[0085] vii) Promoting anti-inflammatory cytokine production

[0086] viii) Increased mucus secretion from goblet cells

[0087] ix) enhancement of dense junctions in intestinal epithelial cells; and

[0088] x) Increase in the number of Bifidobacteria in the intestines

[0089] That is, the preventive, ameliorative and / or therapeutic effects of the composition according to the present invention on inflammatory diseases may be exerted by the effects i) to x) above.

[0090] Regarding the effects of the composition of the present invention on i) reducing damage to intestinal epithelial cell barrier, ii) reducing immune cell infiltration, and iii) suppressing the decrease in the number of goblet cells, an experiment using an inflammatory bowel disease (IBD) animal model in one embodiment of the present invention showed that the IBD animal model had damaged intestinal epithelial cell barrier, infiltrated immune cells, and reduced the number of goblet cells responsible for mucus production compared to the normal control group, while the Ri-EV administration group of the present invention showed improved damage to the intestinal epithelial cell barrier, reduced infiltration of immune cells, and maintained the number of goblet cells at a similar level to the normal control group. In particular, the histological score due to the onset of IBD when Ri-EV was administered was significantly reduced, confirming that intestinal damage and functional abnormalities due to IBD were improved (Figures 5 and 6a).

[0091] Regarding the composition of the present invention, iv) suppression of the decrease in cecum weight and v) suppression of the decrease in intestinal length, in another embodiment of the present invention, the cecum weight and intestinal length were significantly decreased due to intestinal inflammation in an IBD animal model compared to a normal control group, but the decrease in cecum weight and intestinal length was suppressed in the R. intestinalis or Ri-EV administration group, and it was shown that the extracellular vesicles have a better effect of recovering / improving the cecum and intestine, especially compared to the strain itself (Figures 6b and 6c). The disease activity index was also reduced when R. intestinalis or Ri-EV was administered, and it was confirmed that the disease activity index was the lowest in the Ri-EV administration group (Figure 6d). The above results indicate that the extracellular vesicles according to the present invention can improve histological changes in the cecum and intestine caused by intestinal inflammation.

[0092] Regarding the effects of the composition of the present invention on vi) suppressing inflammatory cytokine secretion, vii) promoting anti-inflammatory cytokine production, viii) promoting mucus secretion in goblet cells, and ix) promoting dense junctions in intestinal epithelial cells, in another embodiment of the present invention, it was confirmed that the expression of IL-1β, a proinflammatory cytokine, was increased in an IBD animal model compared to a normal control group, and the expression of Muc-2 and ZO-1 associated with mucus proteins and dense junctions was decreased, thereby increasing intestinal inflammation and weakening mucus secretion and dense junctions. Meanwhile, the R. intestinalis or Ri-EV administration group showed a decrease in the expression of IL-1β and an increase in the expression of Muc-2 and ZO-1. In particular, it was shown that the anti-inflammatory effect in the intestine, the ability to secrete mucus, and the ability to restore dense junctions were more excellent in the extracellular vesicles than in the bacterial strains. In addition, extracellular vesicles were isolated from various congeneric and heterogeneous Roseburia and Bifidobacterium strains and their anti-inflammatory effects were confirmed. As a result, it was confirmed that all of the extracellular vesicles suppress the production of inflammatory factors and promote the production of anti-inflammatory cytokines, thereby exerting excellent anti-inflammatory effects (Figures 10 and 11).

[0093] Regarding the effect of the composition of the present invention on increasing the number of intestinal Bifidobacteria, in another embodiment of the present invention, a significant change in the intestinal microbial community, such as an increase in the Proteobacteria and Deferribacteres phylum, was observed in the IBD animal model compared to the control group (FIGS. 8a and 8b). In the R. intestinalis or Ri-EV administration group, the number of Bifidobacteria, which is known to contribute to the alleviation of IBD, increased, and in particular, the number of Bifidobacteria increased significantly in the Ri-EV administration group compared to the R. intestinalis administration group (FIG. 9). This shows that administration of the Roseburia strain-derived extracellular vesicles according to the present invention improves IBD by inducing changes in the intestinal flora through the growth induction of Bifidobacteria in the intestinal environment of IBD, thereby suppressing the secretion of inflammatory cytokines and restoring the mucus secretion of goblet cells and the tight junction ability of intestinal epithelial cells.

[0094] In summary, the composition according to the present invention can reduce damage to the intestinal epithelial cell barrier and infiltration of immune cells into the intestine caused by inflammation, maintain the number of goblet cells at a normal level, inhibit the loss of cecum weight and intestinal length caused by intestinal inflammation, inhibit the secretion of inflammatory cytokines through changes in the intestinal flora, and restore mucus secretion of goblet cells and the tight junction ability of intestinal epithelial cells, and therefore may be used for the prevention, amelioration, and / or treatment of inflammatory diseases.

[0095] On the other hand, the composition according to the present invention may be used for the prevention, amelioration, and / or treatment of "inflammatory diseases". In the present invention, the inflammatory disease includes, without limitation, any inflammation-related disease or disease that can be ameliorated / treated by suppressing inflammation, and is not limited to a specific type, but is preferably gastritis, gastric ulcer, duodenal ulcer, inflammatory skin disease, allergic disease, Crohn's disease, irritable bowel syndrome, ulcerative colitis, inflammatory bowel disease, peritonitis, osteomyelitis, cellulitis, meningitis, encephalitis, pancreatitis, trauma-induced shock, bronchial asthma, cystic fibrosis, stroke, acute bronchitis, chronic bronchitis, acute bronchiolitis, chronic bronchiolitis, osteoarthritis, gout, spondyloarthropathy, ankylosing spondylitis, Reiter's syndrome, psoriatic arthropathy, intestinal spondylitis, juvenile arthropathy, juvenile ankylosing spondylitis, reactive arthropathy, infection Arthritis of the genital tract, post-infectious arthritis, gonococcal arthritis, tuberculous arthritis, viral arthritis, fungal arthritis, syphilitic arthritis, Lyme disease, arthritis associated with vasculitis syndromes, polyarteritis nodosa, hypersensitivity vasculitis, Rugeric's granulomatosis, polymyalgia rheumatica, articular cell arteritis, calcium crystal deposition arthropathy, caustic gout, non-rheumatoid arthritis, mucositis, tenosynovitis, epicondylitis (tennis elbow), neuropathic arthropathy (charco and joint, hemarthrosic, Henoch-Scholein purpura, hypertrophic osteoarthropathy, multicentric reticulohistiocytosis, sarcoilosis, hemochromatosis, sickle cell disease and other hemochromatopathies, hyperlipoproteinemia, hypogammaglobulinemia, familial Mediterranean fever, Behart's disease, systemic lupus erythematosus, relapsing fever, psoriasis, multiple sclerosis, sepsis, septic shock, multiple organ dysfunction syndrome, acute respiratory distress syndrome, chronic obstructive pulmonary disease, acute lung injury, and broncho-pulmonary dysplasia.

[0096] In the present invention, the term "inflammatory bowel disease (IBD)" refers to a disease that causes chronic inflammation in the gastrointestinal tract. Inflammatory bowel disease refers to a disease in which chronic inflammation occurs in the intestine, and the main symptoms are recurrent abdominal pain, diarrhea, bloody stool, weight loss, etc. Specifically, it is a common name for a chronic digestive disease in which inflammation that occurs in the intestine continues for 6 months or more and becomes chronic. Therefore, it may include infectious enteritis such as bacterial, viral, amoebic, and tuberculous enteritis, ischemic enteritis, and radiation enteritis. In the present invention, inflammatory bowel disease includes without limitation any disease related to intestinal inflammation or a disease that can be improved / treated by suppressing inflammation, and is not limited to a specific type, but may be at least one selected from the group consisting of ulcerative colitis, Crohn's disease, Behcet's disease, collagenous colitis, lymphocytic colitis, ischemic colitis, hemorrhagic rectal ulcer, pouchitis, and transformation colitis.

[0097] Preferably, the inflammatory bowel disease according to the present invention may be selected from ulcerative colitis (UC), Crohn's disease (CD), and Behcet's disease. "Ulcerative colitis" refers to a chronic inflammatory disease in the large intestine, which can be further divided into proctitis, left-sided colitis, and extensive colitis depending on the extent of the disease. "Crohn's disease" refers to a disease in which inflammation occurs throughout the entire digestive tract from the mouth to the anus. However, the inflammatory sites are not always continuous, and may be present separately in various locations. About one-third of Crohn's disease patients develop inflammation only in the small intestine, one-third develop inflammation only in the large intestine, and the remaining one-third develop chronic inflammation in both the large intestine and the small intestine. In particular, inflammation most often occurs at the site where the end of the small intestine meets the large intestine. Ulcerative colitis is characterized by inflammation occurring only in the mucosal layer of the intestine, whereas Crohn's disease is characterized by an inflammatory reaction that affects all layers of the barrier, including the mucosal layer, submucosal layer, muscular layer, and serosal layer. "Behçet's disease" is a chronic, recurrent systemic disease in which inflammation affects various organs, including the skin, mucous membranes, eyes, intestines, joints, urogenital tract, and nervous system. In 3-5% of patients with Behçet's disease, inflammation occurs in the small intestine or large intestine, most commonly at the site where the end of the small intestine joins the large intestine.

[0098] The content of the strain, lysate, culture medium and / or extracellular vesicles in the composition of the present invention can be appropriately adjusted depending on the symptoms of the disease, the degree of progression of the symptoms, the condition of the patient, etc., and may be, for example, 0.0001 to 99.9% by weight, or 0.001 to 50% by weight based on the total composition weight, but is not limited thereto. The content ratio is a value based on the dry amount after removing the solvent.

[0099] The pharmaceutical composition according to the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions, such as at least one selected from the group consisting of diluents, binders, disintegrants, lubricants, adsorbents, moisturizers, film-coating materials and controlled release additives.

[0100] The pharmaceutical composition according to the present invention may be formulated by a conventional method into the form of a powder, granule, sustained-release granule, enteric-coated granule, liquid, eye drop, elucic, emulsion, suspension, spirit, troche, aromatic perfume, lemonade, tablet, sustained-release tablet, enteric-coated tablet, sublingual tablet, hard capsule, soft capsule, sustained-release capsule, enteric-coated capsule, pill, tincture, soft extract, dry extract, fluid extract, injection, capsule, perfusion solution, plaster, lotion, paste, spray, inhalant, patch, sterile injection, or external preparation such as aerosol, and the external preparation may have the form of a cream, gel, patch, spray, ointment, plaster, lotion, liniment, paste, or cataplasm.

[0101] Carriers, excipients and diluents that may be included in pharmaceutical compositions according to the invention include lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginates, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, amorphous cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil.

[0102] When the formulation is made, it is usually prepared by using a diluent or excipient such as a filler, extender, binder, wetting agent, disintegrant, surfactant, etc.

[0103] Additives for the tablets, powders, granules, capsules, pills and lozenges according to the present invention include excipients such as corn starch, potato starch, wheat starch, lactose, sucrose, glucose, fructose, D-mannitol, precipitated calcium carbonate, synthetic aluminum silicate, calcium hydrogen phosphate, calcium sulfate, sodium chloride, sodium bicarbonate, purified lanolin, microcrystalline cellulose, dextrin, sodium alginate, methylcellulose, sodium carboxymethylcellulose, kaolin, urea, colloidal silica gel, hydroxypropyl starch, hydroxypropylmethylcellulose (HPMC) 1928, HPMC 2208, HPMC 2906, HPMC 2910, propylene glycol, casein, calcium lactate and Primozel, gelatin, gum arabic, ethanol, agar powder, cellulose acetate phthalate, carboxymethylcellulose, calcium carboxymethylcellulose, glucose, purified water, casein sodium, Binders such as cellulose, glycerin, stearic acid, sodium carboxymethylcellulose, sodium methylcellulose, methylcellulose, microcrystalline cellulose, dextrin, hydroxycellulose, hydroxypropyl starch, hydroxymethylcellulose, purified shellac, starch paste, hydroxypropyl cellulose, hydroxypropyl methylcellulose, polyvinyl alcohol, and polyvinylpyrrolidone may be used, and hydroxypropyl methylcellulose, corn starch, agar powder, methylcellulose, bentonite, hydroxypropyl starch, sodium carboxymethylcellulose, sodium alginate, calcium carboxymethylcellulose, calcium citrate, sodium lauryl sulfate, anhydrous silicic acid, 1-hydroxypropyl cellulose, dextran, ion exchange resins, polyvinyl acetate, formaldehyde-treated casein and gelatin, alginic acid, amylose, guar gum (Guargum), baking soda, polyvinylpyrrolidone, calcium phosphate, gelling starch, gum arabic, amylopectin, pectin, sodium polyphosphate, ethylcellulose, white sugar, magnesium aluminum silicate, D-sorbitol liquid, disintegrants such as hard anhydrous silicic acid, calcium stearate, magnesium stearate, stearic acid, hydrogenated vegetable oil, talc, lycopodium, kaolin, petrolatum, sodium stearate, cacao butter, sodium salicylate, magnesium salicylate, polyethylene glycol (PEG) 4000, PEG 6000, liquid paraffin, hydrogenated soybean oil (Lubri Lubricants such as wax, aluminum stearate, zinc stearate, sodium lauryl sulfate, magnesium oxide, Macrogol, synthetic aluminum silicate, anhydrous silicic acid, higher fatty acids, higher alcohols, silicone oil, paraffin oil, polyethylene glycol fatty acid ether, starch, sodium chloride, sodium acetate, sodium oleate, dl-leucine, and hard anhydrous silicic acid may be used.

[0104] Examples of additives that may be used in the liquid preparation according to the present invention include water, dilute hydrochloric acid, dilute sulfuric acid, sodium citrate, sucrose monostearate, polyoxyethylene sorbitol fatty acid esters (twin esters), polyoxyethylene monoalkyl ethers, lanolin ethers, lanolin esters, acetic acid, hydrochloric acid, aqueous ammonia, ammonium carbonate, potassium hydroxide, sodium hydroxide, prolamine, polyvinylpyrrolidone, ethyl cellulose, and sodium carboxymethylcellulose.

[0105] The syrup according to the present invention may contain a solution of white sugar, other sugars or sweeteners, and may contain flavorings, coloring agents, preservatives, stabilizers, suspending agents, emulsifiers, thickening agents, etc., as necessary.

[0106] The emulsion of the present invention may contain purified water, and may contain emulsifiers, preservatives, stabilizers, fragrances, etc., as required.

[0107] The suspension according to the present invention may contain a suspending agent such as acacia, tragacanth, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, microcrystalline cellulose, sodium alginate, hydroxypropylmethylcellulose (HPMC), HPMC1828, HPMC2906, or HPMC2910, and may also contain a surfactant, a preservative, a stabilizer, a coloring agent, or a fragrance, as necessary.

[0108] The injections according to the present invention may contain distilled water for injection, 0.9% sodium chloride injection, Ringer's injection, dextrose injection, dextrose + sodium chloride injection, PEG, lactated Ringer's injection, ethanol, propylene glycol, non-volatile oils - sesame oil, cottonseed oil, peanut oil, soybean oil, corn oil, solvents such as ethyl oleate, isopropyl myristate, and benzene benzoate, solubilizers such as sodium benzoate, sodium salicylate, sodium acetate, urea, urethane, monoethylacetamide, butazolidine, propylene glycol, twines, nidiotinamide, hexamine, and dimethylacetamide, weak acids and their salts (acetic acid and sodium acetate), and weak bases and their salts (ammonia and ammonium acetate). It may also contain buffers such as organic compounds, proteins, albumin, peptones, and gums, isotonicity agents such as sodium chloride, stabilizers such as sodium hydrogen sulfite (NaHSO3), carbon dioxide gas, sodium metabisulfite (Na2S2O5), sodium sulfite (Na2SO3), nitrogen gas (N2), and ethylenediaminetetraacetic acid, sulfating agents such as sodium bisulfide 0.1%, sodium formaldehyde sulfoxylate, thiourea, disodium ethylenediaminetetraacetate, and acetone sodium bisulfite, soothing agents such as benzyl alcohol, chlorobutanol, procaine hydrochloride, glucose, and calcium gluconate, and suspending agents such as sodium CMC, sodium alginate, Tween 80, and aluminum monostearate.

[0109] Suppositories according to the invention may contain any of the following ingredients: cocoa butter, lanolin, witepsol, polyethylene glycol, glycerogelatin, methylcellulose, carboxymethylcellulose, mixture of stearic and oleic acids, Subanal, cottonseed oil, peanut oil, coconut oil, cocoa butter + cholesterol, lecithin, lanet wax, glycerol monostearate, Tween or Span, Imhausen, Monolen (propylene glycol monostearate), glycerin, Adeps solidus, Buytyrum Tego-G, Cebes Pharma 16, etc. 16), Hexalide Base 95, Cotomar, Hydrocoat SP, S-70-XXA, S-70-XX75 (S-70-XX95), Hydrokote 25, Hydrocoat 711, Idropostal, Massa Estralium Bases such as estrarium, A, AS, B, C, D, E, I, T), Masa-MF, Maspol, Maspol-15, Neospostal-En, Paramound-B, Sporsilo (OSI, OSIX, A, B, C, D, H, L), suppository base type IV (AB, B, A, BC, BBG, E, BGF, C, D, 299), Spostal (N, Es), Wecoby (W, R, S, M, Fs), and Tezeta triglyceride base (TG-95, MA, 57) may be used.

[0110] The solid preparation for oral administration includes tablets, pills, powders, granules, capsules, etc., and such solid preparations are prepared by mixing the extract with at least one excipient, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium styrenate talc are also used.

[0111] Liquid preparations for oral administration include suspensions, liquid preparations, emulsions, syrups, etc., and may contain various excipients such as wetting agents, sweeteners, flavorings, preservatives, etc. in addition to water and liquid paraffin, which are commonly used simple diluents. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, etc.

[0112] The pharmaceutical composition according to the present invention is administered in a pharmacologic effective amount. In the present invention, the term "pharmacologic effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to any medical treatment, and the effective dose level may be determined by factors including the type and severity of the patient's disease, the activity of the drug, sensitivity to the drug, administration time, administration route and excretion rate, treatment duration, concurrently used drugs, and other factors well known in the medical field.

[0113] The pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or in multiple doses. Taking into consideration all of the above factors, it is important to administer an amount that provides maximum effect at a minimum amount without side effects, which may be easily determined by a person skilled in the art to which the present invention pertains.

[0114] The pharmaceutical compositions of the present invention may be administered to an individual by a variety of routes, including, for example, oral ingestion, subcutaneous injection, intraperitoneal administration, intravenous injection, intramuscular injection, paraspinal space (intradural) injection, sublingual administration, buccal administration, rectal insertion, vaginal insertion, ocular administration, aural administration, nasal administration, inhalation, spraying through the mouth or nose, dermal administration, transdermal administration, and the like, although all modes of administration are contemplated.

[0115] The pharmaceutical composition of the present invention is determined by the type of drug as an active ingredient, along with various related factors such as the disease to be treated, the administration route, the age, sex, weight and severity of the disease of the patient. Specifically, the effective amount of the composition according to the present invention may vary depending on the age, sex and weight of the patient, and is generally 0.001 to 150 mg, preferably 0.01 to 100 mg per kg of body weight, administered daily or every other day, or administered in 1 to 3 divided doses per day. However, since the amount may be increased or decreased depending on the administration route, severity of the disease, sex, weight, age, etc., the above-mentioned dosage does not limit the scope of the present invention in any way.

[0116] In the present invention, the term "individual" refers to a subject in need of treatment for a disease, and more specifically, refers to mammals such as humans or non-human primates, mice, rats, dogs, cats, horses, and cows.

[0117] In the present invention, "administration" means providing a given composition of the present invention to an individual by any suitable method.

[0118] In the present invention, "prevention" refers to any action that suppresses or delays the onset of a desired disease, "treatment" refers to any action that improves or beneficially alters the symptoms of a desired disease and associated metabolic disorders by administering a pharmaceutical composition according to the present invention, and "improvement" refers to any action that reduces parameters related to a target disease, such as the severity of symptoms, by administering a composition according to the present invention.

[0119] The present invention also provides a kit for preventing, ameliorating, or treating an inflammatory disease, comprising the composition according to the present invention.

[0120] The kit according to the present invention is not limited to a specific form as long as it can achieve the purpose of preventing, improving, or treating an inflammatory disease, and may include any components and devices for producing the composition according to the present invention (e.g., culturing a strain, isolating extracellular vesicles, etc.), storing, administering, etc. In addition, the kit may include an instruction manual that describes the characteristics, usage method, storage method, dosage, etc. of the composition of the present invention.

[0121] The present invention also provides a food composition for preventing or improving an inflammatory disease, comprising at least one selected from the group consisting of extracellular vesicles derived from a Roseburia strain and extracellular vesicles derived from a Bifidobacterium strain as an active ingredient. The food composition includes a functional health food composition.

[0122] When the strain, lysate, culture solution, and / or extracellular vesicles of the present invention are used as food additives, the strain, extracellular vesicles, etc. may be added as is or together with other foods or food ingredients, or may be used appropriately by a conventional method. The content of the active ingredient in the food composition may be appropriately determined depending on the purpose of use (prevention, improvement, or therapeutic treatment). The amount of the active ingredient mixed may be appropriately determined depending on the purpose of use (prevention, health, or therapeutic treatment). In general, when producing a food or beverage, the strain, extracellular vesicles, etc. of the present invention may be added in an amount of 15% by weight or less, or 10% by weight or less, based on the raw material. However, in the case of long-term intake for the purpose of health and hygiene or health regulation, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount greater than the above range.

[0123] The type of the food is not particularly limited. Examples of the food to which the substance can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, drinking water, tea, energy drinks, alcoholic beverages, and vitamin complexes, and include all health functional foods in the usual sense.

[0124] The health drink composition according to the present invention may contain various flavorings or natural carbohydrates as additional ingredients, like ordinary drinks. The above-mentioned natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As sweeteners, natural sweeteners such as thaumatin and stevia extract, and synthetic sweeteners such as saccharin and aspartame may be used. The proportion of the natural carbohydrates is generally about 0.01 to 0.20 g, or about 0.04 to 0.10 g per 100 mL of the composition of the present invention.

[0125] In addition to the above, the 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, carbonation agents used in carbonated drinks, etc. In addition, the composition of the present invention 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 combination. The proportion of such additives is not very important, but is generally selected from the range of 0.01 to 0.20 parts by weight per 100 parts by weight of the composition of the present invention.

[0126] In this specification, the term "health functional food" is the same as food for special health use (FoSHU), and refers to food with high medical and therapeutic effects that is processed to efficiently exhibit bioregulatory functions in addition to providing nutrients. The food may be manufactured in various forms such as tablets, capsules, powders, granules, liquids, pills, etc. to obtain useful effects in preventing or improving inflammatory diseases.

[0127] The health functional food of the present invention can be manufactured by a method commonly used in the art, and may be manufactured by adding raw materials and ingredients commonly added in the art. Also, unlike general medicines, it has the advantage of being free of side effects that may occur when taking medicines for a long period of time, as it is made from food as a raw material, and is highly portable.

[0128] The present invention also provides a feed composition for preventing or ameliorating inflammatory diseases, comprising as an active ingredient at least one selected from the group consisting of extracellular vesicles derived from Roseburia strains and extracellular vesicles derived from Bifidobacterium strains.

[0129] The term "feed" in the present invention refers to any natural or artificial diet, meal, etc., or components of said meal, which are intended or suitable for an animal to eat, ingest, and digest, and the feed containing the graphene nanoparticles of the present invention as an active ingredient can be produced in various forms of feed known in the art, and specifically may include concentrated feed, roughage, and / or special feed.

[0130] The amount of the bacterial strain, homogenate, culture medium, and / or extracellular vesicles of the present invention contained in the feed composition of the present invention may vary depending on the purpose and conditions of use of the feed, and may be, for example, 0.01 to 100 wt %, more specifically 1 to 80 wt %, based on the total weight of the livestock feed composition, but is not limited thereto.

[0131] When the composition is produced as a feed additive, the composition may be produced in the form of a highly concentrated liquid (20 to 90%), powder, or granules. The feed additive may further contain any one or more of organic acids such as citric acid, fumaric acid, adipic acid, lactic acid, malic acid, etc., phosphates such as sodium phosphate, potassium phosphate, acid pyrophosphate, and polyphosphates (polymerized phosphates), and natural antioxidants such as polyphenols, catechins, α-tocopherol, rosemary extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, and phytic acid. When produced as a feed, the composition may be formulated in the form of a normal feed and may contain normal feed ingredients.

[0132] The feed and feed additives may further include grains such as crushed or crushed wheat, oats, barley, corn, and rice, vegetable protein feeds such as feeds mainly composed of rapeseed, soybeans, and sunflowers, animal protein feeds such as blood meal, meat and bone meal, bone meal, and fish meal, sugar, and dairy products such as various types of milk powder and whey powder, and may further include nutritional supplements, digestion and absorption enhancers, growth promoters, and the like.

[0133] The feed additives may be administered to animals alone or in combination with other feed additives in an edible carrier. The feed additives may also be readily administered to animals as a top dressing, or mixed directly into these animal feeds, or in an oral dosage form separate from the feed. When the feed additives are administered separately from the animal feed, they may be combined with pharma- ceutically acceptable edible carriers and made into immediate or sustained release formulations, as is well known in the art. These edible carriers may be solid or liquid, such as corn starch, lactose, sucrose, soybean flakes, peanut oil, olive oil, sesame oil, and propylene glycol. When solid carriers are used, the feed additives may be in the form of tablets, capsules, powders, lozenges, or lozenges or top dressings in microdispersible form. When liquid carriers are used, the feed additives may be in the form of gelatin soft capsules, or syrups, suspensions, emulsions, or solutions.

[0134] The feed and feed additives may also contain auxiliary agents, such as preservatives, stabilizers, wetting agents, emulsifiers, solution promoters, etc. The feed additives may be used by acupuncture, spraying, or mixing and adding to animal feed.

[0135] The feed or feed additive of the present invention may be applied to the diets of many animals including mammals, poultry and fish.

[0136] The mammals may include pigs, cows, horses, sheep, rabbits, goats, rodents, and laboratory rodents such as mice, hamsters, and guinea pigs, as well as pet animals (e.g., dogs and cats). The poultry may include chickens, turkeys, ducks, geese, pheasants, and quails. The fish may include carp, crucian carp, and trout, but are not limited thereto.

[0137] MODE FOR CARRYING OUT THEINVENTION

[0138] In order to facilitate understanding of the present invention, the present invention will be described in detail with reference to the following examples. However, the following examples are merely illustrative of the contents of the present invention, and the scope of the present invention is not limited to the following examples. The examples of the present invention are provided to more completely explain the present invention to those having average knowledge in the art.

[0139] (blank) EXAMPLES

[0140] [Example]

[0141] Example 1. Extraction and size analysis of extracellular vesicles (EVs) (Ri-EVs, Bl-EVs) from Roseburia intestinalis or Bifidobacterium longum

[0142] To extract EVs from R. intestinalis and B. longum, R. intestinalis KCTC 15746 and B. longum KCTC 3249 were cultured in reinforced clostridial medium (MB cell, MB-R1602) in an anaerobic chamber at 37°C for 18 hours. The culture was centrifuged at 2000×g for 20 minutes, and the supernatant was collected and 0.22μm filter was used to remove cell debris and waste products from the culture medium. EVs were extracted from the culture through a 300kDa molecular weight TFF (tangential flow filtration) membrane filter-based TFF system, diluted with phosphate-buffered saline (PBS), purified, and finally, R. intestinalis-derived EVs (Ri-EVs) and B. longum-derived EVs (Bl-EVs) dispersed in PBS were extracted.

[0143] Next, the physicochemical properties of Ri-EVs were confirmed using electron microscopes (Ultrathin-section TEM, Cryo-TEM) and nanoparticle tracking analyzers. As a result, it was confirmed through ultrathin-section TEM that R. intestinalis produces EVs during in vitro culture, and through Cryo-TEM that the EVs have a single lipid membrane structure (Figure 1). The extracted Ri-EVs showed a size of 76 ± 10 nm and a surface charge of -4 ± 1 mV (Figure 2), with a molecular weight of 4.8 × 10 10 ±5.9×10 7 The particles were extracted at a concentration of 10 ...

[0144] Furthermore, the size of Ri-EVs and Bl-EVs was measured using a zeta-sizer and confirmed to have an average particle size of 75 nm and 58 nm, respectively (Figure 3b).

[0145] (blank)

[0146] Example 2. Evaluation of the anti-inflammatory effect of Ri-EV administration in an animal model of DSS (dextran sulfate sodium salt colitis grade)-induced inflammatory bowel disease (IBD)

[0147] An IBD animal model was created by providing normal C57BL / 6 mice with drinking water containing 2.5% [v / v] DSS, which is known to induce IBD, for one week (Figure 4). For the experimental group, R. intestinalis strain or Ri-EV extracted in Example 1 was orally administered at a total protein level of 200 μg / mouse / day on days 1, 3, 5, and 7 during the one-week period during which DSS was injected. Mice were sacrificed on day 10 of the experiment to confirm the anti-inflammatory effect of the EV administration. Normal mice not administered DSS served as the control group.

[0148] First, for the pathological analysis of intestinal tissue, mouse intestines were excised, fixed in 4% neutral buffered formalin, embedded in paraffin, and then 4 μm-thick tissue sections were prepared. The tissue sections were stained with H&E (Hematoxilin-Eosin) and AB (Alcian blue), and then observed under an optical microscope.

[0149] Next, cecum weight and intestinal length associated with intestinal inflammation were measured.

[0150] Next, to confirm the ability of Ri-EVs to regulate inflammation, mucus proteins, and dense junctions in the intestinal environment, the expression levels of IL (Interleukin)-1β, Muc-2 (Mucin 2), and ZO-1 (Zonula occludens-1) in the intestinal tissue obtained above were confirmed by qRT-PCR. After the intestinal tissue was dissolved in Trizol solution (Life Technologies, CA, USA), total RNA was isolated from the tissue using the chloroform-isopropanol method according to the manufacturer's analytical method. cDNA of the isolated samples was synthesized using a high-capacity cDNA synthesis kit (Invitrogen, CA, USA) and amplified using TaqMan gene-specific probes (Thermo Fisher Scientific) in a 7500 Real-Time PCR System (Applied Biosystems, CA, USA).

[0151] As a result, as shown in Figure 5, in the DSS-induced IBD animal model, compared to the control group not administered DSS, the intestinal epithelial cell barrier was damaged, immune cells infiltrated, and the number of goblet cells responsible for mucus production was reduced. On the other hand, in the Ri-EV administration group, the damage to the intestinal epithelial cell barrier and the infiltration of immune cells were reduced, and the number of goblet cells was maintained similar to that of the normal group. In particular, the Ri-EV administration group showed a significant decrease in the histological score due to the onset of IBD, confirming that IBD was improved (Figure 6a).

[0152] In addition, in the DSS-induced IBD animal model, the cecum weight and intestinal length associated with intestinal inflammation were significantly reduced compared to the control group, but the R. intestinalis or Ri-EV administration group suppressed the reduction in cecum weight and intestinal length, and in particular, the Ri-EV administration group significantly suppressed the reduction in cecum weight and intestinal length compared to the R. intestinalis administration group (Figures 6b and 6c). The disease activity index was also confirmed to be the lowest in the Ri-EV administration group (Figure 6d).

[0153] In addition, in the DSS-induced IBD animal model, the expression of IL-1β, an inflammatory cytokine that induces inflammation, was increased, and the expression of Muc-2 and ZO-1, which are associated with mucus proteins and dense junctions, was decreased, thereby confirming that intestinal inflammation was increased and mucus secretion and dense junctions were weakened. Meanwhile, the R. intestinalis or Ri-EV administration group showed a decrease in IL-1β expression (Figure 7a) and an increase in Muc-2 and ZO-1 expression (Figure 7b) in the IBD animal model. In particular, the Ri-EV administration group showed a significant decrease in IL-1β expression and an increase in Muc-2 and ZO-1 expression compared to the R. intestinalis administration group. The above results indicate that Ri-EV administration can not only effectively reduce intestinal inflammation, but also restore mucus secretion and dense junctions, and such effects are even better than those of the administration of the strain itself.

[0154] (blank)

[0155] Example 3. Evaluation of changes in gut flora by Ri-EV administration in DSS-induced IBD animal model

[0156] To confirm the change in the intestinal flora and the resulting anti-inflammatory effect caused by Ri-EV administration in an IBD animal model, 16S ribosomal RNA gene sequencing analysis was performed on fecal samples obtained from the DSS-induced IBD animal model in Example 2 using the QIAamp Power Fecal Pro DNA Kit (QIAGEN, Germany).

[0157] As a result, as shown in Figure 8a, significant changes in the gut microbial community, such as an increase in the Proteobacteria and Deferribacteres phylum, were observed in the DSS-induced IBD animal model compared to the control group not administered DSS. In addition, the Shannon diversity value in the Ri-EV administration group was significantly higher than in the other groups, confirming that various changes in the microbiota occurred in the samples (Figure 8b). In particular, as shown in Figure 9, the number of Bifidobacterium, which is known to contribute to the alleviation of IBD in changes in gut bacteria, increased in the R. intestinalis or Ri-EV administration groups, and was significantly higher in the Ri-EV administration group than in the R. intestinalis administration group. These results suggest that Ri-EV administration can improve IBD by suppressing the secretion of inflammatory cytokines through changes in the intestinal flora by inducing the growth of Bifidobacterium in the intestinal environment of IBD, and by restoring mucus secretion from goblet cells and the tight junction ability of intestinal epithelial cells.

[0158] (blank)

[0159] Example 4. Analysis of anti-inflammatory activity of extracellular vesicles derived from Roseburia spp. strains

[0160] In this example, in addition to the R. intestinalis EVs extracted through the process of Example 1, it was confirmed whether EVs from other Roseburia strains could also exert a therapeutic effect on IBD.

[0161] Specifically, extracellular vesicles were extracted from Roseburia intestinalis (Ri) and three other Roseburia strains (Roseburia faecies (Rfa, KCTC 15739), Roseburia hominis (Rho, KCTC 5845), and Roseburia inulinivorans (Rin, DSM 16841)) and their anti-inflammatory activities were analyzed. EVs were extracted using the same procedure as in Example 1.

[0162] To evaluate the anti-inflammatory activity of each EV, the amount of nitric oxide (NO) produced, an indicator of inflammation, was measured, and the expression levels of inflammatory and anti-inflammatory cytokines were also measured. The results are shown in Figure 10.

[0163] Specifically, the amount of nitric oxide produced was measured using mouse macrophages. Mouse macrophage Raw 264.7 cells were cultured in RPMI1640 culture medium containing 10% fetal bovine serum (FBS) and 1% antibiotics (100 U / ml penicillin and 100 μg / ml streptomycin) at 37°C in the presence of 5% CO2. Then, 5 × 10 5 The cells were dispensed at a concentration of 1 ml per well and incubated at 37°C for 24 hours in a CO2 incubator. To induce inflammation, the wells were treated with medium containing 10 μg / ml lipopolysaccharide (LPS) and incubated for an additional 4 hours. The vesicles were added to the medium at a total protein weight of 2 mg and incubated at 37°C for 16 hours. Then, 50 μl of the well supernatant was mixed with 50 μl of Griess reagent and reacted at room temperature for 10 minutes, and the amount of nitric oxide produced was confirmed by measuring the absorbance at 540 nm using a plate reader.

[0164] The expression levels of inflammatory and anti-inflammatory cytokines were also measured using LPS-treated Raw264.7 cells. Raw264.7 cells were treated with LPS in the same manner as above, and 2 mg of the vesicles (based on the total protein amount) were treated and then cultured at 37°C for 16 hours. The protein expression levels of TNF-α, IL-6, and IL-10 in the cells were then measured by absorbance at 450 nm using an ELISA kit (R&D Systems, US) according to the manufacturer's protocol.

[0165] As a result, as shown in Figure 10, when the extracellular vesicles derived from the same but different species of Roseburia strains were treated with inflammation-induced mouse macrophages, it was confirmed that the amount of nitric oxide produced was significantly reduced in the remaining extracellular vesicles derived from Roseburia strains, except for Rin-derived EVs (Rin-EVs). In addition, in terms of the expression levels of inflammatory and anti-inflammatory cytokines, it was confirmed that all the extracellular vesicles derived from Roseburia strains used as experimental groups (hereinafter referred to as Ri-EVs, Rho-EVs, Rfa-EVs, and Rin-EVs) not only effectively reduced TNF-α and IL-6, but also increased IL-10, compared to the control group.

[0166] The anti-inflammatory activity analysis of the Roseburia strains among different species of the same genus demonstrated that all extracellular vesicles derived from all Roseburia strains used in the experiment exhibited excellent anti-inflammatory effects and therefore could be used as a composition for preventing, ameliorating or treating inflammatory bowel disease.

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[0168] Example 5. Analysis of anti-inflammatory activity of extracellular vesicles derived from Roseburia spp. and Bifidobacterium spp. strains

[0169] Following Example 4, it was confirmed whether EVs derived from strains belonging to the genus R. intestinalis or B. longum could exert an anti-inflammatory effect.

[0170] Specifically, four species of Roseburia spp. strains (Roseburia faecies (Rfa, KCTC 15739), Roseburia hominis (Rho, KCTC 5845), and Roseburia inulinivorans (Rin, DSM 16841)) and five species of Bifidobacterium spp. strains (Bifidobacterium bifidum (KCTC 3281), Bifidobacterium breve (KCTC 3220), Bifidobacterium lactis (KCTC 5854), Bifidobacterium alesentis (KCTC 5854), B ... Extracellular vesicles were extracted from each of A. adolescentis (KCTC 3216) and then analyzed for their anti-inflammatory activity. EVs were extracted in the same manner as in Example 1.

[0171] To evaluate the anti-inflammatory activity of each EV, an in vitro environment replicating Kupffer cells, one of the cells that compose liver tissue, was created using mouse macrophages, Raw264.7 cells, and the amount of nitric oxide (NO) produced was measured, as shown in Figure 11. Specifically, mouse macrophages, Raw264.7 cells, were cultured at 37°C in the presence of 5% CO2 in RPMI1640 culture medium containing 10% fetal bovine serum (FBS) and 1% antibiotics (100 U / ml penicillin and 100 μg / ml streptomycin). Then, 5 × 10 cells were cultured in a 12-well plate. 5The cells were dispensed at a concentration of 1 ml per well and incubated in a CO2 incubator at 37°C for 24 hours. To induce inflammation, the wells were treated with medium containing 10 μg / ml lipopolysaccharide (LPS) and incubated for an additional 4 hours. The vesicles were added to the medium at a concentration of 200 μg of total protein and incubated at 37°C for 16 hours. Then, 50 μl of the well supernatant and 50 μl of Griess reagent were mixed and reacted at room temperature for 10 minutes, and the amount of nitric oxide produced was confirmed by measuring the absorbance at 540 nm using a plate reader.

[0172] As a result, it was confirmed that LPS-induced nitric oxide production was significantly and effectively reduced in all groups administered extracellular vesicles derived from Roseburia strains, which were used as experimental groups, as shown in Figure 11. It was also confirmed that nitric oxide production was effectively reduced in the group administered extracellular vesicles derived from Bifidobacterium strains, and it was confirmed that the ability of Bbr-EV and Bla-EV to suppress nitric oxide production was regulated at a total protein amount of 200μg or 50μg.

[0173] The above results indicate that EVs from the same genus, different strains of the genus Roseburia or Bifidobacterium, have excellent anti-inflammatory effects. In other words, the above results confirm that extracellular vesicles derived from Roseburia and Bifidobacterium strains can be used to prevent, improve, or treat various inflammatory diseases through their strong anti-inflammatory effects.

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[0175] As described in the above examples, the extracellular vesicles derived from Roseburia and non-Bifidobacterium strains of the present invention were shown to reduce damage to the intestinal epithelial cell barrier and infiltration of immune cells due to inflammation, maintain the number of goblet cells, inhibit the reduction in cecal weight and intestinal length due to enteritis, inhibit the secretion of inflammatory cytokines through changes in the intestinal flora, and restore mucus secretion from goblet cells and the tight junction ability of intestinal epithelial cells. In particular, it was confirmed that extracellular vesicles derived from various congeneric and heterogeneous strains of Roseburia or Bifidobacterium all exhibit excellent anti-inflammatory effects. Therefore, the extracellular vesicles derived from Roseburia and Bifidobacterium strains of the present invention not only exhibit a strong anti-cancer effect but also improve histological damage and loss of function due to inflammation, and therefore can be provided as a composition for the strong prevention, amelioration, and / or treatment of various inflammatory diseases.

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[0177] From the above description, a person skilled in the art to which the present invention belongs can understand that the present invention can be embodied in other specific forms without changing the technical idea or essential characteristics thereof. In this regard, the above-described embodiments should be understood to be illustrative and not limiting in all respects. The scope of the present invention should be interpreted as including all modifications or variations derived from the meaning and scope of the claims described below from the above detailed description, and their equivalent concepts. [Industrial Applicability]

[0178] Industrial Applicability

[0179] The present invention relates to a composition for preventing, improving or treating inflammatory diseases, comprising extracellular vesicles (EVs) derived from Roseburia spp. and / or Bifidobacterium spp. In an animal model of inflammatory bowel disease, the extracellular vesicles of the present invention have been shown to reduce damage to the intestinal epithelial cell barrier and infiltration of immune cells, maintain the number of goblet cells, inhibit the loss of cecal weight and intestinal length caused by enteritis, inhibit the secretion of inflammatory cytokines through changes in the intestinal flora, and restore mucus secretion from goblet cells and the tight junction ability of intestinal epithelial cells. In particular, it has been confirmed that EVs derived from congenerally heterologous strains corresponding to Roseburia spp. or Bifidobacterium spp. both exhibit strong anti-inflammatory effects. In other words, the extracellular vesicles derived from Roseburia or Bifidobacterium strains according to the present invention can effectively suppress inflammation and improve tissue damage and functional abnormalities caused by inflammation, and therefore can be provided as a composition for preventing, improving and / or treating various inflammatory diseases.

Claims

1. A pharmaceutical composition for preventing or treating an inflammatory disease, comprising as an active ingredient at least one selected from the group consisting of extracellular vesicles derived from Roseburia spp. strains and extracellular vesicles derived from Bifidobacterium spp. strains.

2. The pharmaceutical composition of claim 1, wherein the extracellular vesicles have an average diameter of 30 nm to 200 nm.

3. The extracellular vesicles derived from a Roseburia strain are naturally or artificially secreted from a Roseburia strain, The pharmaceutical composition according to claim 1, wherein the extracellular vesicles derived from a Bifidobacterium strain are naturally or artificially secreted from a Bifidobacterium strain.

4. 2. The pharmaceutical composition according to claim 1, wherein the Roseburia strain is at least one selected from the group consisting of Roseburia intestinalis, Roseburia faeces, Roseburia hominis, and Roseburia inulinivorans.

5. 2. The pharmaceutical composition according to claim 1, wherein the Bifidobacterium strain is at least one selected from the group consisting of Bifidobacterium longum, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium lactis, and Bifidobacterium adolescentis.

6. The inflammatory diseases include gastritis, gastric ulcer, duodenal ulcer, inflammatory skin diseases, allergic diseases, irritable bowel syndrome, ulcerative colitis, inflammatory bowel disease, peritonitis, osteomyelitis, cellulitis, meningitis, encephalitis, pancreatitis, trauma-induced shock, bronchial asthma, cystic fibrosis, stroke, acute bronchitis, chronic bronchitis, acute bronchiolitis, chronic bronchiolitis, osteoarthritis, gout, spondyloarthropathy, ankylosing spondylitis, Reiter's syndrome, psoriatic arthropathy, enteropathy spondylitis, juvenile arthropathy, juvenile ankylosing spondylitis, reactive arthropathy, infectious arthritis, post-infectious arthritis, gonococcal arthritis, tuberculous arthritis, viral arthritis, fungal arthritis, syphilitic arthritis, Lyme disease, arthritis associated with vasculitis syndrome, polyarteritis nodosa, hypersensitivity vasculitis, Ruge's syndrome, and the like.

2. The pharmaceutical composition according to claim 1, wherein the therapeutic agent is at least one selected from the group consisting of Rick's granulomatosis, polymyalgia rheumatica, articular cell arteritis, calcium crystal deposition arthropathy, caustic gout, non-rheumatoid arthritis, mucositis, tenosynovitis, epicondylitis, neurogenic arthropathy, hemarthrosis, Henoch-Scholein purpura, hypertrophic osteoarthropathy, multicentric reticulohistiocytosis, sarcoidosis, hemochromatosis, sickle cell disease, hemochromatosis, hyperlipoproteinemia, hypogammaglobulinemia, familial Mediterranean fever, Behart's disease, systemic lupus erythematosus, relapsing fever, psoriasis, multiple sclerosis, sepsis, septic shock, multiple organ dysfunction syndrome, acute respiratory distress syndrome, chronic obstructive pulmonary disease, acute lung injury, and bronchopulmonary dysplasia.

7. 7. The pharmaceutical composition of claim 6, wherein the inflammatory bowel disease is at least one selected from the group consisting of ulcerative colitis, Crohn's disease, Behcet's disease, collagenous colitis, lymphocytic colitis, ischemic colitis, hemorrhagic rectal ulcer, pouchitis, and transformation colitis.

8. The pharmaceutical composition according to claim 1, characterized in that it has at least one effect selected from the group consisting of the following: i) Reduction of intestinal epithelial cell barrier damage ii) Reduced intestinal tissue infiltration of immune cells iii) Suppression of decrease in number of goblet cells iv) Inhibition of reduction in cecum weight v) Inhibition of decrease in intestinal length vi) Suppression of inflammatory cytokine production vii) Promoting anti-inflammatory cytokine production viii) Increased mucus secretion from goblet cells ix) enhancement of dense junctions in intestinal epithelial cells; and x) Increase in the number of Bifidobacterium in the intestines

9. A kit for preventing or treating an inflammatory disease, comprising the pharmaceutical composition according to any one of claims 1 to 8.

10. A method for preventing or treating an inflammatory disease, comprising administering to an individual in need thereof at least one selected from the group consisting of extracellular vesicles derived from a Roseburia strain and extracellular vesicles derived from a Bifidobacterium strain.

11. A food composition for preventing or ameliorating an inflammatory disease, comprising as an active ingredient at least one selected from the group consisting of extracellular vesicles derived from a Roseburia strain and extracellular vesicles derived from a Bifidobacterium strain.

12. The food composition according to claim 11, wherein the food composition is a functional health food composition.

13. A feed composition for preventing or ameliorating an inflammatory disease, comprising as an active ingredient at least one selected from the group consisting of extracellular vesicles derived from a Roseburia strain and extracellular vesicles derived from a Bifidobacterium strain.

14. 2. Use of at least one selected from the group consisting of extracellular vesicles derived from a Roseburia strain and extracellular vesicles derived from a Bifidobacterium strain for the prevention or treatment of an inflammatory disease.

15. 1. Use of at least one selected from the group consisting of extracellular vesicles derived from a Roseburia strain and extracellular vesicles derived from a Bifidobacterium strain for the manufacture of a therapeutic agent for an inflammatory disease.

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