WO2019168327A1 - Nanovesicles derived from micrococcus bacteria and use thereof - Google Patents

Abstract

The present invention relates to vesicles derived from Micrococcus bacteria and a use thereof. The present inventors have confirmed experimentally that the vesicles were significantly reduced in samples of patients with gastric cancer, pancreatic cancer, bile duct cancer, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, variant angina, chronic obstructive pulmonary disease (COPD), dementia, and diabetes compared to a normal person, and that, when vesicles separated from the strain were injected, the secretion of an inflammatory mediator by pathogenic vesicles such as vesicles derived from Escherichia coli was remarkably inhibited. Therefore, the vesicles derived from Micrococcus bacteria according to the present invention may be usefully used for the purpose of developing a method for diagnosing gastric cancer, pancreatic cancer, bile duct cancer, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, variant angina, COPD, dementia, or diabetes and a composition for preventing, alleviating, or treating the diseases.

Description

Microvesicles-derived nanovesicles and uses thereof

The present invention relates to a microvesicle-derived nano-vesicles and uses thereof, more specifically, gastric cancer, pancreatic cancer, cholangiocarcinoma, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy using nano-vesicles derived from the micrococcus bacteria , Atrial fibrillation, heteroangular angina, chronic obstructive pulmonary disease (COPD), a method for diagnosing dementia or diabetes, and the like, and a composition for preventing, ameliorating or treating the disease, including the vesicles.

In the 21st century, acute infectious diseases, previously recognized as infectious diseases, have become less important, while chronic diseases with immune dysfunctions caused by the incompatibility of human and symbiotic microorganisms determine the quality of life and human life. The disease pattern has changed. As the refractory chronic disease of the 21st century, cancer, cardiovascular disease, chronic lung disease, metabolic disease, and neuro-psychiatric disease are major diseases that determine human life and quality of life, which is a major problem in public health. The intractable chronic disease is characterized by chronic inflammation accompanied by abnormal immune function caused by the causative factor.

The microorganisms symbiotic to the human body reaches 100 trillion times more than human cells, and the number of genes of microorganisms is known to be more than 100 times the number of human genes. Microbiota (microbiota or microbiome) refers to a microbial community including microbes, archae and eukarya that exist in a given settlement.

The symbiotic bacteria in the body and the bacteria present in the environment secrete nanometer-sized vesicles to exchange information such as genes, low molecular weight compounds, and proteins with other cells. The mucous membrane forms a physical protective film that particles larger than 200 nanometers (nm) in size can't pass through. If the bacteria are symbiotic bacteria, the mucosa cannot pass through the mucous membrane, but the bacteria-derived vesicles are 100 nanometers in size or less. It passes through epithelial cells through the mucous membrane and is absorbed by our body. Locally secreted bacterial-derived vesicles are absorbed through the epithelial cells of the mucosa to induce local inflammatory responses, as well as vesicles passing through the epithelial cells are systemically absorbed through the lymphatic vessels and distributed to each organ. Regulates immune and inflammatory responses For example, vesicles derived from pathogenic Gram-negative bacteria, such as Eshcherichia coli , locally cause colitis and, when absorbed into blood vessels, promote systemic inflammatory and blood coagulation through vascular endothelial inflammatory responses. In addition, when insulin is absorbed into muscle cells, it causes insulin resistance and diabetes. On the other hand, vesicles derived from beneficial bacteria can control the disease by controlling immune and metabolic abnormalities caused by pathogenic vesicles.

The immune response to factors such as vesicles derived from bacteria causes a Th17 immune response characterized by the secretion of Interleukin (IL) -17 cytokines, which secrete IL-6 upon exposure to bacterial vesicles, This induces a Th17 immune response. Inflammation by the Th17 immune response is characterized by neutrophil infiltration, and tumor necrosis factor-alpha (TNF-α), which is secreted from inflammatory cells such as macrophages in the process of inflammation, plays an important role. In charge.

Micrococcus bacteria are Gram-positive bacteria belonging to the micrococcus family, and are widely distributed in nature such as water, dust, and soil. Among them, Micrococc us luteus is known to produce riboflavin when grown in toxic organic pollutants such as pyridine, and absorb ultraviolet light through the lutein pigment. Micrococcus bacteria are isolated from dairy products and beer, grow in dry or high salt environments, do not form spores, but are known to survive long-term at refrigerated temperatures such as refrigerators. However, it has not been reported that micrococcus bacteria secrete extracellular vesicles, and there have been no reports on the application and diagnosis of cancer, cardiovascular disease, allergic-chronic lung disease, dementia, or metabolic disease. .

The present inventors earnestly researched to solve the above-mentioned conventional problems, gastric cancer, pancreatic cancer, cholangiocarcinoma, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, heteroangular angina, In the samples from patients with chronic obstructive pulmonary disease (COPD), dementia, and diabetic patients, it was confirmed that the contents of microcacus-derived vesicles were significantly reduced. In addition, when the vesicles are isolated from micrococcus luteus bacteria belonging to the bacteria of the micrococcus and treated with macrophages, it was confirmed that significantly inhibit the secretion of IL-6 and TNF-α by pathogenic vesicles, based on the present invention Was completed.

Accordingly, the present invention provides a method for providing information for the diagnosis of gastric cancer, pancreatic cancer, cholangiocarcinoma, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, angina, chronic obstructive pulmonary disease (COPD), dementia or diabetes It aims to do it.

In addition, the present invention is a gastric cancer, pancreatic cancer, cholangiocarcinoma, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, angina pectoris, atopic dermatitis, psoriasis, acne, hair loss Another object is to provide a composition for preventing, improving or treating macular degeneration, chronic obstructive pulmonary disease (COPD), dementia, or diabetes.

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

In order to achieve the object of the present invention as described above, the present invention comprises the following steps, gastric cancer, pancreatic cancer, cholangiocarcinoma, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, dysplastic angina, chronic obstructive pulmonary Provides informational methods for diagnosing disease (COPD), dementia, or diabetes:

(a) extracting DNA from extracellular vesicles isolated from normal and subject samples;

(b) performing PCR (Polymerase Chain Reaction) on the extracted DNA using a primer pair prepared based on the gene sequence present in the 16S rDNA, and then obtaining each PCR product; And

(c) Gastric cancer, pancreatic cancer, bile duct cancer, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrium when the content of extracellular vesicles derived from bacteria of Micrococcus is lower than that of normal people through quantitative analysis of the PCR products. Classification as fibrillation, angina, chronic obstructive pulmonary disease (COPD), dementia, or diabetes.

Also. The present invention includes the following steps, gastric cancer, pancreatic cancer, cholangiocarcinoma, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, dysplastic angina, chronic obstructive pulmonary disease (COPD), dementia, or diagnosing method of diabetes Provides:

(a) extracting DNA from extracellular vesicles isolated from normal and subject samples;

(b) performing PCR (Polymerase Chain Reaction) on the extracted DNA using a primer pair prepared based on the gene sequence present in the 16S rDNA, and then obtaining each PCR product; And

(c) Gastric cancer, pancreatic cancer, bile duct cancer, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrium when the content of extracellular vesicles derived from bacteria of Micrococcus is lower than that of normal people through quantitative analysis of the PCR products. Determination of defibrillation, angina, chronic obstructive pulmonary disease (COPD), dementia, or diabetes.

In one embodiment of the invention, the sample in step (a) may be blood.

In another embodiment of the present invention, the primer pair in step (b) may be a primer of SEQ ID NO: 1 and SEQ ID NO: 2.

In addition, the present invention, including the micro-caustic bacteria-derived vesicles as an active ingredient, gastric cancer, pancreatic cancer, cholangiocarcinoma, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, angina, atopic dermatitis, psoriasis, acne, A pharmaceutical composition for preventing or treating hair loss, macular degeneration, chronic obstructive pulmonary disease (COPD), dementia, or diabetes is provided.

In one embodiment of the invention, the pharmaceutical composition may comprise an ophthalmic composition.

In addition, the present invention, including the micro-caustic bacteria-derived vesicles as an active ingredient, gastric cancer, pancreatic cancer, cholangiocarcinoma, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, angina, atopic dermatitis, psoriasis, acne, Provided is a food composition for preventing or improving hair loss, macular degeneration, chronic obstructive pulmonary disease (COPD), dementia, or diabetes.

In addition, the present invention, including the micro-caustic bacteria-derived vesicles as an active ingredient, gastric cancer, pancreatic cancer, cholangiocarcinoma, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, angina, atopic dermatitis, psoriasis, acne, Prevention of hair loss, macular degeneration, chronic obstructive pulmonary disease (COPD), dementia, or diabetes or Provided is a therapeutic inhalant composition.

The present invention also provides a cosmetic composition for the prevention or improvement of atopic dermatitis, psoriasis, acne or hair loss, including a micro-caustic bacteria-derived vesicles as an active ingredient.

In addition, the present invention comprises the step of administering to the subject a pharmaceutical composition comprising a microcacus-derived bacterial vesicles as an active ingredient, gastric cancer, pancreatic cancer, cholangiocarcinoma, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial Provided are methods of preventing or treating fibrillation, angina, atopic dermatitis, psoriasis, acne, hair loss, macular degeneration, chronic obstructive pulmonary disease (COPD), dementia, or diabetes.

In addition, the present invention is microbial vesicle-derived vesicles, gastric cancer, pancreatic cancer, cholangiocarcinoma, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, angina pectoris, atopic dermatitis, psoriasis, acne, hair loss, macular degeneration, Provided for the prevention or treatment of chronic obstructive pulmonary disease (COPD), dementia, or diabetes.

In one embodiment of the present invention, the vesicles may have an average diameter of 10 to 200 nm.

In another embodiment of the present invention, the vesicles may be secreted naturally or artificially from bacteria of the micrococcus.

In another embodiment of the present invention, the micro-caucus bacteria-derived vesicles may be micrococcus luteus-derived vesicles.

The present inventors confirmed that intestinal bacteria are not absorbed into the body, but in the case of bacterial-derived vesicles, they are absorbed into the body through epithelial cells, distributed systemically, and excreted in vitro through the kidneys, liver, and lungs. Bacterial-derived vesicle metagenome analysis in the gastric cancer, pancreatic cancer, bile duct cancer, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, dysplastic angina, COPD, dementia, and diabetes It was confirmed that the vesicle-derived microorganisms in the blood were significantly reduced compared to normal people. In addition, micrococcus luteus, a species of micrococcus bacteria, was cultured in vitro to separate vesicles, and when administered to inflammatory cells in vitro, it was observed that significantly inhibiting the secretion of inflammatory mediators caused by pathogenic vesicles. Micro-caustic bacteria-derived vesicles according to the invention is gastric cancer, pancreatic cancer, bile duct cancer, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, dysplastic angina, chronic obstructive pulmonary disease (COPD), diagnostic method for diabetes or diabetes And it is expected that it can be usefully used in the composition for the prevention, improvement or treatment for the above diseases.

Figure 1a is a picture of the distribution of bacteria and vesicles by time after oral administration of bacteria and bacteria-derived vesicles (EV) to the mouse, Figure 1b is 12 hours after oral administration, blood, kidney Figure shows the distribution of bacteria, vesicles and vesicles in the body, liver and various organs.

Figure 2 is a result of comparing the distribution of bacteria-derived vesicles of the micrococcus after performing a bacterial-derived vesicle metagenome analysis present in gastric cancer patients and normal blood.

Figure 3 is a result of comparing the distribution of bacteria-derived vesicles of the micrococcus after performing a bacterial-derived vesicle metagenome analysis present in pancreatic cancer patients and normal blood.

Figure 4 is a result of comparing the distribution of bacteria-derived vesicles of the genus Micrococcus after analyzing the bacteria-derived vesicles metagenome present in patients with bile duct cancer.

Figure 5 is a result of comparing the distribution of bacteria-derived vesicles of the micrococcus after performing a bacterial-derived vesicle metagenome analysis present in breast cancer patients and normal blood.

Figure 6 is a result of comparing the distribution of bacteria-derived vesicles of the genus Micrococcus after analyzing the bacteria-derived vesicles metagenome present in ovarian cancer patients and normal blood.

7 is a result of comparing the distribution of bacteria-derived vesicles of the micrococcus after analyzing the bacteria-derived vesicles metagenome present in the bladder cancer patients and normal blood.

8 is a result of comparing the distribution of bacteria-derived vesicles of the micrococcus after analyzing the bacteria-derived vesicles metagenome present in myocardial infarction patients and normal blood.

9 is a result of comparing the distribution of microbial bacteria-derived vesicles after performing a microorganism-derived vesicle metagenome analysis present in cardiomyopathy patients and normal blood.

10 is a result of comparing the distribution of bacteria-derived vesicles of the micrococcus after analyzing the bacteria-derived vesicles metagenome present in atrial fibrillation patients and normal blood.

11 is a result of comparing the distribution of bacteria-derived vesicles of the micrococcus after analyzing the bacteria-derived vesicles metagenome present in patients with heteroangular angina and normal blood.

12 is a result of comparing the distribution of the bacterial-derived vesicles of the micrococcus after performing the bacterial-derived vesicle metagenome analysis present in patients with chronic obstructive pulmonary disease (COPD) and normal blood.

Fig. 13 shows the results of comparing the distribution of bacterial-derived vesicles of the micrococcus after analyzing the bacterial-derived vesicles metagenome present in dementia patients and normal blood.

14 is a result of comparing the distribution of bacteria-derived vesicles belonging to the micrococcus after analyzing the bacteria-derived vesicles metagenome present in diabetic patients and normal blood.

15 is a pathogenic effect of IL-6 and TNF-α secretion of inflammatory mediators by treating microcacus luteus-derived vesicles in macrophages (Raw264.7 cells) in order to evaluate the inflammation-inducing effect of microcaus luteus-derived vesicles. Compared to the vesicle E. coli vesicles (E. coli EV).

16 is a pre-treatment of micrococcus luteus-derived vesicles before treatment with Escherichia coli vesicles (E. coli EV) to evaluate the anti-inflammatory and immunomodulatory effects of micrococcus luteus-derived vesicles. The results of evaluation of the effect on the secretion of IL-6 and TNF-α.

The present invention relates to vesicles derived from bacteria of the micrococcus and their use.

The present inventors have found that the microcacus-derived vesicles from the genome of the micrococcus have gastric cancer, pancreatic cancer, bile duct cancer, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, dysplastic angina, and chronic obstructive pulmonary disease (COPD). Significant reductions were found in clinical samples of patients with diabetes, dementia, and diabetes, confirming that the disease could be diagnosed. In addition, the vesicles were isolated and characterized from micrococcus luteus, which showed gastric cancer, pancreatic cancer, bile duct cancer, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, dysplastic angina, atopic dermatitis, macular degeneration, and chronic It was confirmed that it can be used as a composition for preventing, improving or treating diseases such as obstructive pulmonary disease (COPD), dementia, and diabetes.

Accordingly, the present invention includes the following steps, gastric cancer, pancreatic cancer, bile duct cancer, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, dysplastic angina pectoris, chronic obstructive pulmonary disease (COPD), dementia, or diabetes mellitus Provide informational methods for diagnosis:

(a) extracting DNA from extracellular vesicles isolated from normal and subject samples;

(b) performing PCR (Polymerase Chain Reaction) on the extracted DNA using a primer pair prepared based on the gene sequence present in the 16S rDNA, and then obtaining each PCR product; And

(c) Gastric cancer, pancreatic cancer, bile duct cancer, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrium when the content of extracellular vesicles derived from bacteria of Micrococcus is lower than that of normal people through quantitative analysis of the PCR products. Classification as fibrillation, angina, chronic obstructive pulmonary disease (COPD), dementia, or diabetes.

As used herein, the term "diagnosis" in the broad sense means to determine the actual condition of the patient's disease in all aspects. The content of the judgment is the name of the disease, the etiology, the type of disease, the seriousness, the detailed mode of the condition, the presence or absence of complications, and the prognosis. Diagnosis in the present invention is the presence and disease of gastric cancer, pancreatic cancer, cholangiocarcinoma, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, dysplastic angina, chronic obstructive pulmonary disease (COPD), dementia, and / or diabetes To judge the level and so on.

As used herein, the term "nanovesicle" or "vesicle" refers to a structure of nanoscale membranes secreted by various bacteria. Gram-negative bacteria-derived vesicles or outer membrane vesicles (OMVs) contain toxic proteins, bacterial DNA and RNA as well as lipopolysaccharides, and gram-positive bacteria-derived vesicles. In addition to proteins and nucleic acids, it also contains peptidoglycan and lipoteichoic acid, which are components of bacterial cell walls. In the present invention, nanovesicles or vesicles are naturally secreted or artificially produced by micrococcus bacteria, and have a spherical shape and have an average diameter of 10 to 200 nm.

The term "metagenome" used in the present invention, also referred to as "gunoelectric", refers to the sum total of the genome including all viruses, bacteria, fungi, etc. in an isolated area such as soil, animal intestine, mainly culture It is used as a concept of genome explaining the identification of many microorganisms at once using sequencer to analyze microorganisms that are not. In particular, the metagenome does not refer to one genome or genome, but to a kind of mixed dielectric as the genome of all species of one environmental unit. This is a term from the point of view of defining a species in the course of the evolution of biology in terms of functional species as well as various species that interact with each other to create a complete species. Technically, rapid sequencing is used to analyze all DNA and RNA, regardless of species, to identify all species in one environment, and to identify interactions and metabolism.

In the present invention, the sample may be blood, but is not limited thereto.

As another embodiment of the present invention, the present invention comprises a micro-caustic bacteria-derived vesicles as an active ingredient, gastric cancer, pancreatic cancer, bile duct cancer, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, dysplastic angina, atopic dermatitis It provides a composition for preventing, treating or improving psoriasis, acne, hair loss, macular degeneration, chronic obstructive pulmonary disease (COPD), dementia, or diabetes. The composition includes food, inhalants, cosmetics and pharmaceutical compositions.

In addition, the food composition in the present invention includes a health functional food composition, the pharmaceutical composition may include an ophthalmic composition, but is not limited thereto.

As used herein, the term "prevention" refers to gastric cancer, pancreatic cancer, cholangiocarcinoma, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, angina pectoris, atopic dermatitis, psoriasis, by administration of the composition according to the invention Means any action that inhibits or delays the development of acne, hair loss, macular degeneration, chronic obstructive pulmonary disease (COPD), dementia, and / or diabetes.

As used herein, the term "treatment" refers to gastric cancer, pancreatic cancer, cholangiocarcinoma, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, angina pectoris, atopic dermatitis, psoriasis, by administration of the composition according to the invention It means any behavior that improves or beneficially changes the condition for acne, hair loss, macular degeneration, chronic obstructive pulmonary disease (COPD), dementia, or diabetes.

As used herein, the term “improvement” means any action that at least reduces the parameters associated with the condition being treated, for example, the extent of symptoms.

The vesicles are centrifuged, ultra-fast centrifugation, high pressure treatment, extrusion, sonication, cell lysis, homogenization, freeze-thaw, electroporation, mechanical decomposition, chemical treatment, filtration by a filter containing micrococcus bacteria. It can be separated using one or more methods selected from the group consisting of, gel filtration chromatography, pre-flow electrophoresis, and capillary electrophoresis. In addition, it may further include a process for washing to remove impurities, concentration of the obtained vesicles and the like.

The pharmaceutical composition according to the invention may comprise a pharmaceutically acceptable carrier. Such pharmaceutically acceptable carriers are conventionally used in the preparation, and include, but are not limited to, saline solution, sterile water, Ringer's solution, buffered saline, cyclodextrin, dextrose solution, maltodextrin solution, glycerol, ethanol, liposomes, and the like. If necessary, other conventional additives such as antioxidants and buffers may be further included. In addition, diluents, dispersants, surfactants, binders, lubricants and the like may be additionally added to formulate injectable formulations, pills, capsules, granules, or tablets such as aqueous solutions, suspensions, emulsions and the like. Suitable pharmaceutically acceptable carriers and formulations can be preferably formulated according to the individual components using methods disclosed in Remington's literature. The pharmaceutical composition of the present invention is not particularly limited in formulation, but may be formulated as an injection, inhalant, external skin preparation, ophthalmic, oral ingestion, and the like.

The pharmaceutical composition of the present invention can be administered orally or parenterally (eg, applied intravenously, subcutaneously, skin, nasal, airways) according to the desired method, and the dosage is determined by the condition and weight of the patient, disease Depending on the degree, drug form, route of administration, and time, it may be appropriately selected by those skilled in the art.

The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, the pharmaceutically effective amount means an amount sufficient to treat the disease at a reasonable benefit / risk ratio applicable to the medical treatment, and the effective dose level refers to the type of disease, the severity, the activity of the drug and the drug. Sensitivity, time of administration, route of administration and rate of release, duration of treatment, factors including concurrent use of drugs, and other factors well known in the medical arts. The composition according to the present invention may be administered as a separate therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be single or multiple doses. Taking all of the above factors into consideration, it is important to administer an amount that can obtain the maximum effect in a minimum amount without side effects, which can be easily determined by those skilled in the art.

Specifically, the effective amount of the pharmaceutical composition according to the present invention may vary depending on the age, sex and weight of the patient, and generally 0.001 to 150 mg, preferably 0.01 to 100 mg daily or every other day, per kg of body weight Or divided into 1 to 3 times a day. However, the dosage may be increased or decreased depending on the route of administration, the severity of obesity, sex, weight, age, etc., and the above dosage does not limit the scope of the present invention in any way.

The inhalant composition of the present invention may be added to the inhalant as it is, or used in combination with other ingredients, and may be suitably used according to conventional methods. The amount of the active ingredient to be mixed may be suitably determined depending on the purpose of use (prophylactic or therapeutic).

The food composition of the present invention includes a nutraceutical composition. The food composition according to the present invention may be used as it is, or may be used in combination with other foods or food ingredients, or may be appropriately used according to conventional methods. The mixing amount of the active ingredient can be suitably determined according to the purpose of use (prevention or improvement). Generally, in the preparation of food or beverages the compositions of the invention are added in amounts of up to 15% by weight, preferably up to 10% by weight relative to the raw materials. However, in the case of prolonged intake for health and hygiene purposes or health control purposes, the amount may be below the above range.

The food composition of the present invention, in addition to containing the active ingredient as an essential ingredient in the indicated ratio, there are no particular restrictions on other ingredients, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract, for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. . The proportion of the natural carbohydrate can be appropriately determined by the choice of those skilled in the art.

In addition to the above, the food composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese, chocolate), pectic acid and salts thereof, alginic acid and Salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. These components can be used independently or in combination. The proportion of such additives may also be appropriately selected by those skilled in the art.

The cosmetic composition of the present invention may include components commonly used in cosmetic compositions, as well as microbe decay-derived vesicles, such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances Adjuvants, and carriers.

In addition, the composition of the present invention may be used in addition to microvesicles-derived microvesicles, organic sunscreens that have been conventionally used as long as they do not impair the skin protection effect by reacting with microvesicles-derived microvesicles. Examples of the organic sunscreen include glyceryl pava, drometrizole trisiloxane, drometrizole, digaloyltrioleate, disodium phenyldibenzimidazole tetrasulfonate, diethylhexyl butamidotriazone, diethylamino Hydroxybenzoylhexylbenzoate, die-methoxycinnamate, a mixture of lowson and dihydroxyacetone, methylenebis-benzotriazolyltetramethylbutylphenol, 4-methylbenzylidene camphor, menthyl anthranilate, benzophenone -3 (oxybenzone), benzophenone-4, benzophenone-8 (dioxyphenbenzone), butylmethoxydibenzoylmethane, bisethylhexyloxyphenol methoxyphenyltriazine, synoxate, ethyldihydroxypropylpava, Octocrylene, ethylhexyldimethylpava, ethylhexylmethoxycinnamate, ethylhexylsalicylate, ethylhexyltrizone, isoamyl-p-methoxycinnamate, polysilicon-15 (dimethicodiethylbenzalmal Ronate), terephthalylidene dicamphor sulfonic acid and salts thereof, thy-salicylate and aminobenzoic acid (Pava) can be used.

Examples of products to which the cosmetic composition of the present invention may be added include, for example, cosmetics such as astringent cosmetics, soft cosmetics, nourishing cosmetics, various creams, essences, packs, foundations, and the like, cleansing agents, soaps, treatments, and essences. Etc. Specific formulations of the cosmetic composition of the present invention include skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, essence, nutrition essence, pack, Formulations such as soaps, shampoos, cleansing foams, cleansing lotions, cleansing creams, body lotions, body cleansers, emulsions, lipsticks, makeup bases, foundations, press powders, loose powders, eye shadows and the like.

In one embodiment of the present invention, the bacteria and bacteria-derived vesicles orally administered to observe the body absorption, distribution, and excretion of the bacteria and vesicles in the body, in the case of bacteria is not absorbed through the intestinal membrane, the vesicles are administered 5 It was confirmed that it was absorbed within minutes and distributed systemically and excreted through the kidney, liver, and the like (see Example 1).

In another embodiment of the present invention, age and sex in gastric cancer, pancreatic cancer, cholangiocarcinoma, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, angina, chronic obstructive pulmonary disease (COPD), dementia, and diabetes patients Bacterial metagenome analysis was performed using vesicles isolated from the blood of healthy individuals. As a result, compared to the normal samples, gastric cancer, pancreatic cancer, cholangiocarcinoma, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, dysplastic angina pectoris, chronic obstructive pulmonary disease (COPD), dementia, and diabetes in clinical samples It was confirmed that microvesicle-derived vesicles were significantly reduced (see Examples 3 to 15).

In another embodiment of the present invention, micrococcus luteus strains were cultured to evaluate whether vesicles secreted therefrom exhibited immunomodulatory and anti-inflammatory effects. Later, by treating E. coli-derived vesicles, the inflammatory disease-causing factors, and evaluating the secretion of inflammatory mediators, it was confirmed that the micro-cuscus luteus-derived vesicles effectively suppressed IL-6 and TNF-α secretion by E. coli-derived vesicles ( See Example 18).

Hereinafter, preferred examples are provided to aid in understanding the present invention. However, the following examples are merely provided to more easily understand the present invention, and the contents of the present invention are not limited by the following examples.

EXAMPLE

Example  1. Analysis of absorption, distribution, and excretion of intestinal bacteria and bacterial-derived vesicles

In order to evaluate whether the intestinal bacteria and bacteria-derived vesicles are absorbed systemically through the gastrointestinal tract, experiments were performed as follows. Fluorescently labeled enterobacteriaceae and enteric bacteria-derived vesicles were administered to the gastrointestinal tract at doses of 50 μg, respectively, and the fluorescence was measured after 0, 5, 3, 6 and 12 hours. As a result of observing the whole image of the mouse, as shown in FIG. 1A, the bacteria were not absorbed systemically, but in the case of bacterial-derived vesicles, they were absorbed systemically 5 minutes after administration, and the bladder fluoresced 3 hours after administration. This was observed strongly, indicating that the vesicles were excreted by the urinary system. In addition, the vesicles were found to exist in the body until 12 hours of administration.

In addition, after the intestinal bacteria and enteric bacteria-derived vesicles are absorbed systemically, 50 μg of fluorescently labeled bacteria and bacteria-derived vesicles are administered in the same manner as described above in order to evaluate the infiltration into various organs. After 12 hours, blood, heart, lungs, liver, kidneys, spleen, fat and muscle were collected. As a result of fluorescence observed in the collected tissue, as shown in Figure 1b, the bacteria-derived vesicles were distributed in the blood, heart, lung, liver, spleen, fat, muscle, kidney, it can be seen that the bacteria are not absorbed.

Example 2 Bacterial-derived Vesicular Metagenome Analysis in Clinical Samples

Clinical samples such as blood were first placed in 10 ml tubes and the suspension was allowed to settle by centrifugation (3,500 x g, 10 min, 4 ° C.) and only the supernatant was transferred to a new 10 ml tube. After removing the bacteria and foreign substances using a 0.22㎛ filter, and transferred to centrifugal filters (centrifugal filters 50 kD) and centrifuged at 1500 x g, 4 ℃ for 15 minutes to discard the material smaller than 50 kD concentrated to 10 ml. Once again, the bacteria and foreign substances were removed using a 0.22㎛ filter, and the supernatant was discarded using ultrafast centrifugation for 3 hours at 150,000 xg and 4 ℃ with a Type 90ti rotor, and the pulverized pellet was physiologically It was dissolved in saline (PBS).

100 μl of the vesicles separated by the above method was boiled at 100 ° C. to let the internal DNA come out of the lipid and then cooled on ice for 5 minutes. And centrifuged at 10,000 x g, 4 ℃ 30 minutes to remove the remaining suspended solids and collected only the supernatant. The amount of DNA was quantified using Nanodrop. Thereafter, PCR was performed with the 16s rDNA primer shown in Table 1 to confirm whether the bacteria-derived DNA exists in the extracted DNA, and it was confirmed that the bacteria-derived gene exists in the extracted gene.

primer		order	SEQ ID NO:
16S rDNA	16S_V3_F	5'-TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGCCTACGGGNGGCWGCAG-3 '	One
	16S_V4_R	5'-GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGGACTACHVGGGTATCTAATCC-3	2

DNA extracted by the above method was amplified using the above 16S rDNA primers, followed by sequencing (Illumina MiSeq sequencer), and the results were outputted in a Standard Flowgram Format (SFF) file, using GS FLX software (v2.9). After converting the SFF file into a sequence file (.fasta) and a nucleotide qualityscore file, the credit rating of the lead was confirmed, and the window (20 bps) average base call accuracy was less than 99% (Phred score <20). . For operational taxonomy unit (OTU) analysis, clustering is performed according to sequence similarity using UCLUST and USEARCH. Genus is 94%, family is 90%, order is 85%, and steel ( class is 80% and phylum is clustered based on 75% sequence similarity and the phylum, class, order, family and genus levels of each OTU Sorting was performed, and bacteria with greater than 97% sequence similarity at the genus level were profiled (QIIME) using BLASTN and GreenGenes' 16S RNA sequence database (108,453 sequences).

Example 3 Analysis of Bacterial-Derived Vesicular Metagenome in Blood of Gastric Cancer Patients

In the method of Example 2, the blood of 66 gastric cancer patients and 198 normal blood patients whose age and sex were matched were extracted from the vesicles in the blood and subjected to metagenomic analysis. The distribution of vesicles was evaluated. As a result, it was confirmed that microvesicle-derived microvesicles were significantly reduced in the blood of gastric cancer patients compared to normal blood (see Table 2 and FIG. 2).

blood	Control		Stomach cancer		t-test
Taxon	Mean	SD	Mean	SD	p-value	Ratio
g__Micrococcus	0.0083	0.0115	0.0029	0.0051	<0.0001	0.35

Example  4. Bacterial-derived vesicles from pancreatic cancer patients Metagenome  analysis

In the method of Example 2, 176 bloods of pancreatic cancer patients and 271 bloods of normal age matched with age and sex were extracted from the vesicles in the blood and subjected to metagenomic analysis, followed by micrococcus-derived bacteria. The distribution of vesicles was evaluated. As a result, it was confirmed that microvesicle-derived microvesicles were significantly reduced in the blood of pancreatic cancer patients compared to normal blood (see Table 3 and FIG. 3).

blood	Control		Pancreatic cancer		t-test
Taxon	Mean	SD	Mean	SD	p-value	Ratio
g__Micrococcus	0.0059	0.0101	0.0029	0.0042	<0.0001	0.49

Example  5. Blood Bacteria Derived from Bile Duct Cancer Patients Metagenome  analysis

In the blood of 79 patients with cholangiocarcinoma patients and blood of 259 normal people whose age and sex were matched by the method of Example 2, genes were extracted from vesicles present in the blood and subjected to metagenomic analysis. The distribution of vesicles was evaluated. As a result, it was confirmed that the vesicle-derived bacteria of the micrococcus significantly decreased in the blood of the bile duct cancer patients compared to the normal blood (see Table 4 and FIG. 4).

blood	Control		Bile duct cancer		t-test
Taxon	Mean	SD	Mean	SD	p-value	Ratio
g__Micrococcus	0.0058	0.0101	0.0031	0.0107	0.04	0.54

Example  6. Blood-derived vesicles derived from breast cancer patients Metagenome  analysis

By the method of Example 2, the genes were extracted from the vesicles in the blood of 96 breast cancer patients and 192 normal blood patients whose ages and genders were matched. The distribution of vesicles was evaluated. As a result, it was confirmed that microbe decay-derived vesicles significantly reduced in the blood of breast cancer patients compared to normal blood (see Table 5 and Figure 5).

blood	Control		Breast cancer		t-test
Taxon	Mean	SD	Mean	SD	p-value	Ratio
g__Micrococcus	0.0118	0.0164	0.0037	0.0061	<0.0001	0.31

Example  7. Blood Bacteria Derived from Ovarian Cancer Patients Metagenome  analysis

In the method of Example 2, 137 bloods of ovarian cancer patients and 139 bloods of normal age and sex matched were extracted from the vesicles in the blood and subjected to metagenomic analysis, followed by bacteria of the micrococcus. The distribution of the derived vesicles was evaluated. As a result, it was confirmed that microvesicle-derived vesicles are significantly reduced in the blood of ovarian cancer patients compared to normal blood (see Table 6 and FIG. 6).

blood	Control		Ovarian Cancer		t-test
Taxon	Mean	SD	Mean	SD	p-value	Ratio
g__Micrococcus	0.0115	0.0185	0.0019	0.0034	<0.0001	0.17

Example  8. Vesicle-derived vesicles from bladder cancer patients Metagenome  analysis

The blood of 91 bladder cancer patients and blood of 176 normal people whose age and sex were matched by the method of Example 2 were extracted from the vesicles in the blood and subjected to a metagenome analysis. The distribution of bacterial derived vesicles was evaluated. As a result, it was confirmed that microvesicle-derived microvesicles were significantly reduced in the blood of bladder cancer patients compared to normal blood (see Table 7 and FIG. 7).

blood	Control		Bladder cancer		t-test
Taxon	Mean	SD	Mean	SD	p-value	Ratio
g__Micrococcus	0.0059	0.0120	0.0024	0.0024	0.0006	0.41

Example  9. Blood Bacteria Derived from Myocardial Infarction Patients Metagenome  analysis

In the method of Example 2, blood was collected from 57 patients with myocardial infarction and 163 normal blood patients whose ages and genders were matched. The distribution of the derived vesicles was evaluated. As a result, it was confirmed that microvesicle-derived microvesicles are significantly reduced in the blood of myocardial infarction patients compared to normal blood (see Table 8 and FIG. 8).

blood	Control		Myocardial infarction		t-test
Taxon	Mean	SD	Mean	SD	p-value	Ratio
g__Micrococcus	0.0105	0.0165	0.0017	0.0043	<0.0001	0.16

Example  10. Blood Bacteria Derived from Cardiomyopathy Patients Metagenome  analysis

In the method of Example 2, the genes were extracted from the vesicles in the blood and 163 normal people whose age and sex were matched with the blood of 72 cardiomyopathy patients, and then subjected to metagenomic analysis. The distribution of the derived vesicles was evaluated. As a result, it was confirmed that microbe decay-derived vesicles significantly reduced in the blood of cardiomyopathy patients compared to normal blood (see Table 9 and Figure 9).

blood	Control		Cardiomyopathy		t-test
Taxon	Mean	SD	Mean	SD	p-value	Ratio
g__Micrococcus	0.0105	0.0165	0.0033	0.0052	<0.0001	0.32

Example  11. Atrial fibrillation vesicles derived from blood bacteria Metagenome  analysis

In the blood of 34 patients with atrial fibrillation and blood of 62 healthy subjects whose age and gender were matched by the method of Example 2, genes were extracted from vesicles present in the blood, and a metagenome analysis was performed. The distribution of the derived vesicles was evaluated. As a result, it was confirmed that microvesicle-derived vesicles are significantly reduced in blood of atrial fibrillation patients compared to normal blood (see Table 10 and FIG. 10).

blood	Control		Atrial fibrillation		t-test
Taxon	Mean	SD	Mean	SD	p-value	Ratio
g__Micrococcus	0.0066	0.0113	0.0017	0.0016	0.001	0.26

Example  12. Patients with heterozygous angina  Blood Bacteria-Derived Vesicles Metagenome  analysis

In the method of Example 2, the blood of 80 patients with heterocystic angina and 80 normal blood whose age and sex were matched were extracted from the vesicles in the blood and subjected to metagenomic analysis. The distribution of the derived vesicles was evaluated. As a result, it was confirmed that microvesicle-derived vesicles are significantly reduced in blood of patients with heteroangular angina compared with normal blood (see Table 11 and FIG. 11).

blood	Control		Angina		t-test
Taxon	Mean	SD	Mean	SD	p-value	Ratio
g__Micrococcus	0.0065	0.0151	0.0028	0.0045	0.03	0.43

Example  13. COPD Patient  Blood Bacteria-Derived Vesicles Metagenome  analysis

In the method of Example 2, 205 bloods of COPD patients and 231 bloods of normal and matched age and sex were extracted from the vesicles in the blood and subjected to metagenomic analysis. The distribution of vesicles was evaluated. As a result, it was confirmed that microvesicle-derived microvesicles were significantly reduced in blood of COPD patients compared to normal blood (see Table 12 and FIG. 12).

blood	Control		COPD		t-test
Taxon	Mean	SD	Mean	SD	p-value	Ratio
g__Micrococcus	0.0125	0.0121	0.0010	0.0017	<0.0001	0.08

Example  14. Vesicle-derived vesicles from dementia patients Metagenome  analysis

In the method of Example 2, the genes were extracted from the vesicles present in the blood of 67 people with dementia and 70 blood of normal people who matched their age and sex. The distribution of vesicles was evaluated. As a result, it was confirmed that microbe decay-derived vesicles significantly reduced in the blood of dementia patients compared to normal blood (see Table 13 and Figure 13).

blood	Control		dementia		t-test
Taxon	Mean	SD	Mean	SD	p-value	Ratio
g__Micrococcus	0.0031	0.0087	0.0012	0.0027	0.0003	0.36

Example  15. Diabetic blood bacteria-derived vesicles Metagenome  analysis

In the method of Example 2, the blood of 61 diabetic patients and 122 normal blood of age and sex matched were extracted from the vesicles present in the blood and subjected to metagenomic analysis, followed by micrococcus bacteria. The distribution of vesicles was evaluated. As a result, it was confirmed that microvesicle-derived vesicles are significantly reduced in the blood of diabetic patients compared to normal blood (see Table 14 and FIG. 14).

blood	Control		diabetes		t-test
Taxon	Mean	SD	Mean	SD	p-value	Ratio
g__Micrococcus	0.0093	0.0154	0.0001	0.0001	<0.0001	0.01

Example  16. Micrococcus Luteus  Separation of Vesicles from Culture

Based on the above embodiment, after culturing the micro base Lactococcus Proteus (M. luteus) strains were characterized by separating the vesicles thereof. Micrococcus luteus was subcultured after incubating in de Man-Rogosa and Sharpe (MRS) medium until the absorbance (OD ₆₀₀ ) was 1.0 to 1.5 in an aerobic chamber at 37 ° C. Then, the culture medium supernatant containing the strain was recovered and centrifuged at 10,000 g, 4 ° C. for 20 minutes to remove the strain, filtered through a 0.22 μm filter, and the filtered supernatant was filtered using a 100 kDa Pellicon 2 Cassette filter membrane ( Merck Millipore, US) using a MasterFlex pump system (Cole-Parmer, US) was concentrated to a volume of 50 ml through microfiltration (microfiltration). Then, the concentrated supernatant was once again filtered through a 0.22 μm filter and protein was quantified using BCA (Bicinchoninic acid) assay, and the following experiments were performed on the obtained vesicles.

Example 17. Inflammatory Effect of Micrococcus Luteus-derived Vesicles

To investigate the effects of inflammatory mediators (IL-6, TNF-α) secretion of micrococcus luteus-derived vesicles in inflammatory cells, micrococcus luteus-derived vesicles were collected in raw 264.7 cells, a mouse macrophage. 1, 10 μg / ml), followed by ELISA.

More specifically, incubated for 12 hours by treating various concentrations of micrococcus Luteus-derived vesicles in which Dulbeco's Modified Eagle's Medium (DMEM) serum-free medium was added to Raw 264.7 cells dispensed 4 x 10 ⁴ in 48-well cell culture plates. It was. Then, the cell culture solution was collected in a 1.5 ml tube, centrifuged at 3000 g for 5 minutes, the supernatant was collected and stored at -80 ° C, followed by ELISA.

In order to perform ELISA, the capture antibody was diluted in phosphate buffered saline (PBS), and 50 μl was dispensed into 96 well polystyrene plates according to the working concentration, followed by reaction at 4 ° C. overnight. I was. After washing three times with 100 μl of PBST (PBS containing 0.05% tween-20), 100 μl of RD (PBS containing 1% BSA) solution was dispensed and blocked for 1 hour at room temperature. 50 μl of the sample and standard were dispensed according to the concentration and reacted at room temperature for 2 hours. Then, after washing three times with 100 μl of PBST, the detection antibody was diluted in RD and 50 μl aliquots were adjusted according to the working concentration, and reacted at room temperature for 2 hours.

Then, after washing three times with 100 μl of PBST, Strpetavidin-HRP (R & D system, USA) was diluted 1/40 in RD, 50 μl were dispensed and reacted at room temperature for 20 minutes. After washing twice, 50 μl of TMB substrate (SurModics, USA) was dispensed, and when color development progressed after 5 to 20 minutes, 50 μl of 1 M sulfuric acid solution was dispensed to stop the reaction and the SpectraMax M3 microplate reader (Molecular Devices, USA) and absorbance at 450 nm.

As a result, as shown in Figure 15, when treated with macrophage cells derived from micrococcus luteus, it was confirmed that the secretion of inflammatory mediators is significantly lower than Escherichia coli-derived vesicles (E. coli EV).

Example 18 Anti-inflammatory Effects of Microcaucus Luteus-derived Vesicles

Based on the results of Example 17, in order to determine the anti-inflammatory effect of micrococcus luteus-derived vesicles on inflammatory mediator secretion from inflammatory cells, micrococcus luteus-derived vesicles were concentrated in Raw 264.7 cells. After treatment with (0.1, 1, 10 μg / ml), the secretion amount of inflammatory mediators (IL-6, TNF-α, etc.) was measured by treatment with E. coli EV ( E. coli EV), an inflammatory pathogenic vesicle.

More specifically, 1 × 10 ⁵ Raw 264.7 cells were dispensed into 24-well cell culture plates, and then cultured in DMEM complete medium for 24 hours. Then, the culture supernatant was collected in a 1.5 ml tube, centrifuged at 3000 g for 5 minutes, the supernatant was collected and stored at 4 ° C., followed by ELISA analysis. As a result, as shown in Figure 14, when pre-treated with micrococcus luteus vesicles, it was confirmed that IL-6 and TNF-α secretion by E. coli-derived vesicles is significantly suppressed. This means that microcacus luteus-derived vesicles can effectively suppress the occurrence of inflammation induced by pathogenic vesicles such as E. coli-derived vesicles.

The foregoing description of the present invention is intended for illustration, and it will be understood by those skilled in the art that the present invention may be easily modified in other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are exemplary in all respects and not restrictive.

Micro-caustic bacteria-derived vesicles according to the present invention is gastric cancer, pancreatic cancer, cholangiocarcinoma, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, dysplastic angina, chronic obstructive pulmonary disease (COPD), diagnosis of dementia or diabetes As well as the method and can be used as a composition for preventing, improving or treating the above diseases, it is expected to be usefully used in the related medical, beauty and food industries.

Claims (15)

1. Providing information for the diagnosis of gastric cancer, pancreatic cancer, cholangiocarcinoma, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, angina, chronic obstructive pulmonary disease (COPD), dementia, or diabetes, comprising the following steps Way:

   (a) extracting DNA from extracellular vesicles isolated from normal and subject samples;

   (b) performing PCR (Polymerase Chain Reaction) on the extracted DNA using a primer pair prepared based on the gene sequence present in the 16S rDNA, and then obtaining each PCR product; And

   (c) Gastric cancer, pancreatic cancer, bile duct cancer, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrium when the content of extracellular vesicles derived from bacteria of Micrococcus is lower than that of normal people through quantitative analysis of the PCR products. Classification as fibrillation, angina, chronic obstructive pulmonary disease (COPD), dementia, or diabetes.
2. The method of claim 1,

   The sample in step (a) is characterized in that the blood, information providing method.
3. Micro Rhodococcus (Micrococcus) in bacteria containing the resulting package to the active ingredient, gastric cancer, pancreatic cancer, bile duct cancer, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, variant angina, atopic dermatitis, psoriasis, acne, alopecia, A pharmaceutical composition for preventing or treating at least one disease selected from the group consisting of macular degeneration, chronic obstructive pulmonary disease (COPD), dementia, and diabetes.
4. The method of claim 3,

   The vesicles are characterized in that the average diameter of 10 to 200 nm, pharmaceutical composition.
5. The method of claim 3,

   The vesicles are characterized in that they are secreted naturally or artificially from bacteria of the genus Micrococcus , pharmaceutical composition.
6. The method of claim 3,

   The micrococcus bacteria-derived vesicles are characterized in that the micrococcus luteus- derived vesicles, pharmaceutical compositions.
7. Micro Rhodococcus (Micrococcus) in bacteria containing the resulting package to the active ingredient, gastric cancer, pancreatic cancer, bile duct cancer, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, variant angina, atopic dermatitis, psoriasis, acne, alopecia, Macular degeneration, chronic obstructive pulmonary disease (COPD), dementia, and food composition for preventing or ameliorating one or more diseases selected from the group consisting of diabetes.
8. The method of claim 7, wherein

   The vesicles are characterized in that the average diameter of 10 to 200 nm, food composition.
9. The method of claim 7, wherein

   The vesicles are characterized in that they are secreted naturally or artificially from bacteria of the genus Micrococcus , food composition.
10. The method of claim 7, wherein

    The micrococcus bacteria-derived vesicles are characterized in that the micrococcus luteus- derived vesicles, food composition.
11. Stomach cancer, pancreatic cancer, cholangiocarcinoma, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, angina pectoris, atopic dermatitis, psoriasis, acne, hair loss, including vesicles derived from micrococcus bacteria as an active ingredient Inhalation composition for preventing or treating at least one disease selected from the group consisting of macular degeneration, chronic obstructive pulmonary disease (COPD), dementia, and diabetes.
12. Cosmetic composition for preventing or ameliorating at least one disease selected from the group consisting of atopic dermatitis, psoriasis, acne, and hair loss, including a micrococcus-derived microbial vesicle as an active ingredient.
13. A method of diagnosing gastric cancer, pancreatic cancer, bile duct cancer, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, angina, chronic obstructive pulmonary disease (COPD), dementia, or diabetes, comprising the following steps:

    (a) extracting DNA from extracellular vesicles isolated from normal and subject samples;

    (b) performing PCR (Polymerase Chain Reaction) on the extracted DNA using a primer pair prepared based on the gene sequence present in the 16S rDNA, and then obtaining each PCR product; And

    (c) Gastric cancer, pancreatic cancer, bile duct cancer, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrium when the content of extracellular vesicles derived from bacteria of Micrococcus is lower than that of normal people through quantitative analysis of the PCR products. Determination of defibrillation, angina, chronic obstructive pulmonary disease (COPD), dementia, or diabetes.
14. Stomach cancer, pancreatic cancer, cholangiocarcinoma, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, dysplastic angina, comprising administering to the subject a pharmaceutical composition comprising a microcacus-derived vesicle as an active ingredient Atopic dermatitis, psoriasis, acne, hair loss, macular degeneration, chronic obstructive pulmonary disease (COPD), dementia and diabetes, one or more methods of preventing or treating diseases selected from the group consisting of.
15. Bacterial-derived vesicles of the micrococcus, stomach cancer, pancreatic cancer, cholangiocarcinoma, breast cancer, ovarian cancer, bladder cancer, myocardial infarction, cardiomyopathy, atrial fibrillation, dysplastic angina, atopic dermatitis, psoriasis, acne, hair loss, macular degeneration, chronic obstructive pulmonary disease ( COPD), dementia and diabetes for use in the prevention or treatment of one or more diseases.

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