KR20220163842A - Extracellular vesicle derived from Lactobacillus acidophilus and use thereof - Google Patents

Abstract

The present invention relates to Lactobacillus acidophilus-derived vesicles and a use thereof and, more specifically, to a composition containing Lactobacillus acidophilus-derived vesicles as an active ingredient for alleviating, preventing, and treating inflammatory diseases and cancer. The vesicles are expected to be widely used as an alleviation, prevention, or treatment for various inflammatory diseases.

Description

Lactobacillus acidophilus-derived vesicle and use thereof {Extracellular vesicle derived from Lactobacillus acidophilus and use thereof}

The present invention relates to vesicles derived from Lactobacillus acidophilus and uses thereof, and more particularly, to a composition for preventing or treating inflammatory diseases comprising Lactobacillus acidophilus-derived vesicles as an active ingredient.

Entering the 21st century, the importance of acute infectious diseases, which were recognized as infectious diseases in the past, has decreased, while the disease pattern has changed to a major disease that determines the quality of life and human lifespan due to chronic diseases. The chronic disease is characterized by chronic inflammation accompanied by immune dysfunction, and various chronic inflammatory diseases and diseases such as cancer caused by this are becoming a major problem for public health.

Inflammation is a local or systemic defense mechanism against cell and tissue damage or infection. Inflammation is a direct response of humoral mediators that make up the immune system or by stimulating local or systemic effector systems. It is caused by a chain of biological reactions that occur. As a result of inflammation, cell death occurs, resulting in disease. Major inflammatory diseases include digestive diseases such as gastritis and inflammatory enteritis, oral diseases such as periodontitis, respiratory diseases such as asthma, chronic obstructive pulmonary disease (COPD), and rhinitis, skin diseases such as atopic dermatitis, hair loss, acne, and psoriasis, and degenerative diseases Musculoskeletal system inflammatory diseases such as arthritis, rheumatoid arthritis, gout, osteoporosis, and cancer are known.

Common compositions used to treat or prevent chronic inflammatory diseases are largely divided into steroid and non-steroidal compositions, most of which often accompany various side effects. Therefore, interest in inhibitors of TNF-α and IL-6, which are pro-inflammatory cytokines known to be the main causes of chronic inflammatory diseases, has recently increased.

However, development of drugs that can be effectively used for preventing and treating chronic inflammatory diseases by effectively inhibiting the expression of TNF-α and IL-6 while having low side effects is still insufficient.

Domestic registered patent 10-1620815

The inventors of the present invention completed the present invention by confirming that Lactobacillus acidophilus-derived vesicles have excellent TNF-α and IL-6 inhibitory effects as a result of intensive research to solve the above conventional problems.

Accordingly, an object of the present invention is to provide a composition for improving, preventing or treating inflammatory diseases comprising Lactobacillus acidophilus-derived vesicles as an active ingredient.

However, the technical problem to be achieved by the present invention is not limited to the above-mentioned problems, and other problems not mentioned can be clearly understood by those skilled in the art from the description below. will be.

In order to achieve the object of the present invention as described above, the present inventors cultured Lactobacillus acidophilus bacteria to isolate vesicles, and when administered orally to mice, the vesicles were found in the stomach, small intestine, large intestine, lung, liver, And distribution to organs such as the central nervous system was confirmed. In addition, it was confirmed that when the vesicles were treated with a mouse macrophage cell line, no inflammatory effect was shown, and the expression of TNF-α and IL-6, which were increased by vesicles derived from E. coli, which are inflammatory factors, was significantly suppressed.

Accordingly, the present invention provides a pharmaceutical composition for preventing or treating inflammatory diseases comprising Lactobacillus acidophilus-derived vesicles as an active ingredient.

In addition, the present invention provides a food composition for preventing or improving inflammatory diseases comprising Lactobacillus acidophilus-derived vesicles as an active ingredient.

In addition, the present invention provides an inhalant composition for preventing or treating inflammatory diseases, comprising Lactobacillus acidophilus-derived vesicles as an active ingredient.

In addition, the present invention provides a cosmetic composition for preventing or improving inflammatory diseases comprising Lactobacillus acidophilus-derived vesicles as an active ingredient.

In one embodiment of the present invention, the inflammatory disease may include the following diseases, but is not limited thereto:

one or more inflammatory skin diseases selected from the group consisting of eczema, atopic dermatitis, psoriasis, acne, alopecia, contact dermatitis, seborrheic dermatitis, and mastocytosis;

one or more inflammatory oral diseases selected from the group consisting of caries, gingivitis, and periodontitis;

Ulcerative colitis, Crohn's disease, intestinal Behcet's disease, indeterminate colitis, chronic gastritis, chronic peptic ulcer, celiac disease disease), and at least one inflammatory gastrointestinal disease selected from the group consisting of inflammatory bowel disease;

At least one inflammatory disease selected from the group consisting of non-alcoholic steatohepatitis (NASH), alcoholic steatohepatitis, liver cirrhosis, cholangitis, cholecystitis, and pancreatitis hepato-biliary-pancreatic disease;

one or more inflammatory nasal diseases selected from the group consisting of rhinitis, sinusitis, and sinusitis;

bronchitis, asthma, chronic obstructive pulmonary disease (COPD), emphysema, cystic fibrosis, pneumonia, chronic interstitial pneumonitis, bronchopulmonary dysplasia, and idiopathic pulmonary fibrosis ( at least one inflammatory lung disease selected from the group consisting of idiopathic pulmoniary fibrosis;

Hyperinsulinemia, dyslipidemia, arrhythmias, atherosclerosis, angina, metabolic syndrome, myocardial infartion, cardiomyopathy, stroke, ischemic heart disease, one or more inflammatory cardiovascular diseases selected from the group consisting of heart failure, thrombosis, and atherosclerosis;

Atony, muscular atrophy, muscular dystrophy, myasthenia, cachexia, Gout, sarcopenia, osteoporosis, Paget's disease, one or more inflammatory musculoskeletal disorders selected from the group consisting of rheumatoid arthritis and osteoarthritis;

Mild cognitive impairment, dementia, Alzheimer's disease, Parkinson's disease, Huntington's disease, Amyotrophic lateral sclerosis (ALS), Batten disease ), Kearns-Sayre syndrome (KSS), chronic progressive external ophthalmoplegia (CPEO), mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes (MELAS), MERRF syndrome ( myoclonic epilepsy with ragged-red fibers (MERRF), neurogenic weakness with ataxia and retinitis pigmentosa (NARP), Leigh syndrome (LS), mitochondrial recessive ataxia syndrome (MIRAS syndrome), anxiety disorder, post-traumatic stress disorder Post-traumatic stress disorder (PTSD), panic disorder, depression, autism spectrum disorders, attention deficit/hyperactivity disorder (ADHD), schizophrenia ), Dementia with Lewy Bodies (DLB), Multi-Infarct Dementia (MID), frontotemporal lobar degeneration (FTLD), Pick's disease, and cortical degeneration (Co rticobasal degeneration (CBD), progressive supranuclear palsy (PSP), age-related macular degeneration (AMD), dysosmia, deafness, and diabetic retinopathy ) At least one inflammatory central nervous system disease selected from the group consisting of; and

Proliferative vitreoretinopathy, macular degeneration, retinopathy pigmentosa, diabetic retinopathy, choroidal neovascularization, neovascular glaucoma, ischemic optic nerve disorder, retinopathy of prematurity, retinopathy of prematurity, epidemic keratoconjunctivitis, neovascular iris disease, postphakic fibroplasia, atopic At least one chronic inflammatory eye disease selected from the group consisting of keratitis, superior limbal keratitis, phase patch psoriatic keratitis, plyctenic keratoconjunctivitis, scleritis, and diabetic macular edema.

In another embodiment of the present invention, the inflammatory disease may be a disease mediated by TNF-α or IL-6, but is not limited thereto.

In another embodiment of the present invention, the vesicle may be naturally or artificially secreted from Lactobacillus acidophilus, but is not limited thereto.

In another embodiment of the present invention, the vesicles may have an average diameter of 10 to 300 nm, but is not limited thereto.

In another embodiment of the present invention, the vesicles may be isolated from a culture medium of Lactobacillus acidophilus or food cultured by adding Lactobacillus acidophilus, but is not limited thereto.

In addition, the present invention provides a drug delivery composition for treating diseases, comprising Lactobacillus acidophilus-derived vesicles as an active ingredient.

In one embodiment of the present invention, the disease may be one or more selected from the group consisting of gastric disease, intestinal disease, lung disease, liver disease, and central nervous system disease, but is not limited thereto.

In another embodiment of the present invention, the central nervous system disease is mild cognitive impairment, dementia, Alzheimer's disease, Parkinson's disease, Huntington's disease, Lou Gehrig's disease Amyotrophic lateral sclerosis (ALS), Batten disease, Kearns-Sayre syndrome (KSS), chronic progressive external ophthalmoplegia (CPEO), mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes (MELAS), myoclonic epilepsy with ragged-red fibers (MERRF), neurogenic weakness with ataxia and retinitis pigmentosa (NARP), Leigh syndrome (LS), MIRAS syndrome (mitochondrial recessive ataxia syndrome), anxiety disorder, post-traumatic stress disorder (PTSD), panic disorder, depression, autism spectrum disorders, attention deficit hyperactivity syndrome ADHD, schizophrenia, Dementia with Lewy Bodies (DLB), Multi-Infarct Dementia (MID), frontotemporal lobar degeneration (FTLD) ), Pick's disease, corticobasal degeneration (CBD), progressive supranuclear palsy (PSP), age-related macular degeneration (AMD), dysosmia, hearing It may be at least one selected from the group consisting of disability (deafness), and diabetic retinopathy (diabetic retinopathy), but is not limited thereto.

In addition, the present invention provides a method for preventing or treating an inflammatory disease comprising administering to a subject a composition containing Lactobacillus acidophilus-derived vesicles as an active ingredient.

In addition, the present invention provides a use of a composition comprising Lactobacillus acidophilus-derived vesicles as an active ingredient for preventing or treating inflammatory diseases.

In addition, the present invention provides a use of a composition comprising Lactobacillus acidophilus-derived vesicles as an active ingredient for producing a drug for treating inflammatory diseases.

In addition, the present invention provides a method for preventing, improving or treating inflammatory skin diseases, comprising administering to a subject a composition containing Lactobacillus acidophilus-derived vesicles as an active ingredient.

In addition, the present invention provides a composition containing Lactobacillus acidophilus-derived vesicles as an active ingredient for preventing, improving or treating inflammatory skin diseases.

In addition, the present invention provides a use of a composition comprising Lactobacillus acidophilus-derived vesicles as an active ingredient for producing a drug for treating inflammatory skin diseases.

In addition, the present invention provides a method for delivering a drug for treating a disease comprising administering to a subject a composition containing vesicles derived from Lactobacillus acidophilus as an active ingredient.

In addition, the present invention provides a use of a composition containing Lactobacillus acidophilus-derived vesicles as an active ingredient to deliver a drug for treating diseases.

In addition, the present invention provides a drug delivery system comprising vesicles derived from Lactobacillus acidophilus.

When the Lactobacillus acidophilus-derived vesicles according to the present invention are orally administered, they are absorbed through the stomach, small intestine, and large intestine, move into the blood through the lymphatic vessels, and are then distributed to organs such as the lungs, liver, and brain. In addition, since the vesicle can effectively suppress the expression of TNF-α, IL-6, etc., which are mediators that cause inflammatory diseases by pathogenic factors, it is expected that it can be widely used as an improvement, prevention, or treatment for various inflammatory diseases. do.

1 is a diagram illustrating the absorption, distribution, and excretion patterns of Lactobacillus acidophilus-derived vesicles over time after oral administration of vesicles derived from Lactobacillus acidophilus according to an embodiment of the present invention.
FIG. 2 is a diagram illustrating the absorption, distribution, and excretion of Lactobacillus acidophilus-derived vesicles orally administered to the stomach, small intestine, and large intestine over time according to an embodiment of the present invention.
FIG. 3 is a diagram illustrating absorption, distribution, and excretion into the lungs and liver over time after orally administering Lactobacillus acidophilus-derived vesicles according to an embodiment of the present invention.
Figure 4 is a view of analyzing the absorption, distribution, and excretion patterns in the central nervous system over time by orally administering Lactobacillus acidophilus-derived vesicles according to an embodiment of the present invention.
5 is a diagram illustrating absorption, distribution, and excretion into the heart and kidney over time after oral administration of Lactobacillus acidophilus-derived vesicles according to an embodiment of the present invention.
Figures 6a and 6b show the vesicles derived from Escherichia coli (E. coli EV) and vesicles derived from Lactobacillus acidophilus for the secretion of TNF-α and IL-6, which are inflammatory mediators, in inflammatory cells according to an embodiment of the present invention. It is a figure showing the result of comparing the secretion effect.
7a and 7b are Lactobacillus acidophilus-derived vesicles for the secretion of TNF-α and IL-6 by Escherichia coli-derived vesicles (E. coli EV), which are pathogenic factors in inflammatory cells according to an embodiment of the present invention. It is a drawing showing the result of evaluating the treatment effect of.

In the present invention, when Lactobacillus acidophilus-derived vesicles were orally administered, they were absorbed through the stomach, small intestine, and large intestine, and then absorbed into blood vessels through the lymphatic vessels, confirming that they were distributed to organs such as the lungs, liver, and brain. In addition, it was confirmed that the secretion of inflammatory mediators that cause inflammatory diseases caused by pathogenic factors was effectively inhibited, thereby exhibiting remarkable therapeutic effects in inflammatory diseases.

Accordingly, the present invention relates to a composition for preventing, improving or treating inflammatory diseases comprising Lactobacillus acidophilus-derived vesicles as an active ingredient.

Hereinafter, the present invention will be described in detail.

In the present invention, “Lactobacillus acidophilus” is a rod-shaped lactobacillus of Lactobacillus, which is a kind of probiotic. It is known to act as a decrease in vitamin B group synthesis.

In the present invention, “extracellular vesicle” or “vesicle” refers to a nano-sized membrane structure secreted by various bacteria, for example, endotoxin (lipopolysaccharide), toxic protein and vesicles or outer membrane vesicles (OMVs) derived from gram-negative bacteria such as Escherichia coli, which also have bacterial DNA and RNA, and peptidoglycan, a component of bacterial cell walls, in addition to proteins and nucleic acids. and vesicles derived from gram-positive bacteria, such as micrococcus bacteria that also contain lipoteichoic acid.

In the present invention, the vesicle is a generic term for all membrane structures naturally secreted or artificially produced from Lactobacillus acidophilus. The vesicles are obtained by centrifugation, ultra-high-speed centrifugation, high-pressure treatment, extrusion, sonication, cell lysis, homogenization, freeze-thaw, electroporation, mechanical disintegration, chemical treatment, filter Separation may be performed using at least one method selected from the group consisting of filtration by filtration, gel filtration chromatography, free-flow electrophoresis, and capillary electrophoresis. In addition, processes such as washing to remove impurities and concentration of the obtained vesicles may be further included.

The method of separating vesicles from the Lactobacillus acidophilus culture broth or fermented food of the present invention is not particularly limited as long as vesicles are included. For example, vesicles can be separated using methods such as centrifugation, ultra-high-speed centrifugation, filter filtration, gel filtration chromatography, free-flow electrophoresis, or capillary electrophoresis, and combinations thereof, and also removal of impurities. It may further include a process of washing for, concentration of the obtained vesicles, and the like.

The vesicles of the present invention may be isolated from a culture medium of Lactobacillus acidophilus or food prepared by adding Lactobacillus acidophilus, and may be naturally or artificially secreted from Lactobacillus acidophilus. , but is not limited thereto.

In the present invention, the vesicles isolated by the above method have an average diameter of 10-1000 nm, 10-900 nm, 10-800 nm, 10-700 nm, 10-600 nm, 10-500 nm, 10-400 nm, 10 ~300 nm, 10-200 nm, 10-100 nm, 10-90 nm, 10-80 nm, 10-70 nm, 10-60 nm, 10-50 nm, 10-40 nm, or 20-40 nm It may be, but is not limited thereto.

The term "comprising as an active ingredient" used in the present invention means containing an amount sufficient to achieve the efficacy or activity of the Lactobacillus acidophilus-derived vesicles.

In the present invention, the inflammatory disease is selected from the group consisting of eczema, atopic dermatitis, psoriasis, acne, alopecia, contact dermatitis, seborrheic dermatitis, and mastocytosis one or more inflammatory skin diseases;

one or more inflammatory oral diseases selected from the group consisting of caries, gingivitis, and periodontitis;

Ulcerative colitis, Crohn's disease, intestinal Behcet's disease, indeterminate colitis, chronic gastritis, chronic peptic ulcer, celiac disease disease), and at least one inflammatory gastrointestinal disease selected from the group consisting of inflammatory bowel disease;

At least one inflammatory disease selected from the group consisting of non-alcoholic steatohepatitis (NASH), alcoholic steatohepatitis, liver cirrhosis, cholangitis, cholecystitis, and pancreatitis hepato-biliary-pancreatic disease;

one or more inflammatory nasal diseases selected from the group consisting of rhinitis, sinusitis, and sinusitis;

bronchitis, asthma, chronic obstructive pulmonary disease (COPD), emphysema, cystic fibrosis, pneumonia, chronic interstitial pneumonitis, bronchopulmonary dysplasia, and idiopathic pulmonary fibrosis ( at least one inflammatory lung disease selected from the group consisting of idiopathic pulmoniary fibrosis;

Hyperinsulinemia, dyslipidemia, arrhythmias, atherosclerosis, angina, metabolic syndrome, myocardial infartion, cardiomyopathy, stroke, ischemic heart disease, one or more inflammatory cardiovascular diseases selected from the group consisting of heart failure, thrombosis, and atherosclerosis;

Atony, muscular atrophy, muscular dystrophy, myasthenia, cachexia, Gout, sarcopenia, osteoporosis, Paget's disease, one or more inflammatory musculoskeletal disorders selected from the group consisting of rheumatoid arthritis and osteoarthritis;

Mild cognitive impairment, dementia, Alzheimer's disease, Parkinson's disease, Huntington's disease, Amyotrophic lateral sclerosis (ALS), Batten disease ), Kearns-Sayre syndrome (KSS), chronic progressive external ophthalmoplegia (CPEO), mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes (MELAS), MERRF syndrome ( myoclonic epilepsy with ragged-red fibers (MERRF), neurogenic weakness with ataxia and retinitis pigmentosa (NARP), Leigh syndrome (LS), mitochondrial recessive ataxia syndrome (MIRAS syndrome), anxiety disorder, post-traumatic stress disorder Post-traumatic stress disorder (PTSD), panic disorder, depression, autism spectrum disorders, attention deficit/hyperactivity disorder (ADHD), schizophrenia ), Dementia with Lewy Bodies (DLB), Multi-Infarct Dementia (MID), frontotemporal lobar degeneration (FTLD), Pick's disease, and cortical degeneration (Co rticobasal degeneration (CBD), progressive supranuclear palsy (PSP), age-related macular degeneration (AMD), dysosmia, deafness, and diabetic retinopathy ) At least one inflammatory central nervous system disease selected from the group consisting of; and

Proliferative vitreoretinopathy, macular degeneration, retinopathy pigmentosa, diabetic retinopathy, choroidal neovascularization, neovascular glaucoma, ischemic optic nerve disorder, retinopathy of prematurity, retinopathy of prematurity, epidemic keratoconjunctivitis, neovascular iris disease, postphakic fibroplasia, atopic It may be at least one selected from the group consisting of one or more chronic inflammatory eye diseases selected from the group consisting of keratitis, limbal keratitis, phase patch psoriatic keratitis, plyctenic keratoconjunctivitis, scleritis, and diabetic macular edema, but is not limited thereto.

In the present invention, tumor necrosis factor α (TNF-α) is an intracellular mediator of immune responses produced by various cells including activated macrophages and monocytes. Responses elicited by TNF-α are initiated through interaction with two distinct TNF-α cell surface receptors, TNFαR1 and TNFαR2. TNF-α binds to these cell surface receptors and induces activation of transcription factors such as nuclear factor κB (NFκB), which regulates the expression of various immune and inflammatory response genes. Upon binding of TNF-α, the TNF-α receptor interacts with various intracellular signal translation proteins through its cytoplasmic domain. There are a variety of disease states associated with regulatory pathways controlled by TNF-α binding. In certain instances, TNF-α binding triggers an inflammatory response and ultimately results in a disease state. Therefore, it would be desirable to develop means to prevent diseases associated with TNF-α receptor binding. In particular, there is a need to identify methods to prevent activation of the inflammatory response that may be initiated by TNF-α activation. On the other hand, IL-23, IL-17, and IL-6 secreted by T immune cells differentiated by hypersensitivity of the immune system due to chronic inflammatory disease or autoimmune abnormality of patients. Excessive expression of cytokines such as TNF-α causes an inflammatory response.

The content of the vesicles in the composition of the present invention can be appropriately adjusted according to the symptoms of the disease, the progress of the symptoms, the condition of the patient, etc., for example, 0.0001 to 99.9% by weight, or 0.001 to 50% by weight based on the total weight of the composition. It may, but is not limited thereto. The content ratio is a value based on the dry amount after removing the solvent.

The pharmaceutical composition according to the present invention may further include suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions. The excipient may be, for example, one or more selected from the group consisting of a diluent, a binder, a disintegrant, a lubricant, an adsorbent, a moisturizer, a film-coating material, and a controlled release additive.

The pharmaceutical compositions according to the present invention are powders, granules, sustained-release granules, enteric granules, solutions, eye drops, elsilic agents, emulsions, suspensions, spirits, troches, perfumes, and limonadese, respectively, according to conventional methods. , tablets, sustained-release tablets, enteric tablets, sublingual tablets, hard capsules, soft capsules, sustained-release capsules, enteric capsules, pills, tinctures, soft extracts, dry extracts, fluid extracts, injections, capsules, perfusate, It can be formulated and used in the form of external preparations such as warning agents, lotions, pasta agents, sprays, inhalants, patches, sterile injection solutions, or aerosols, and the external agents are creams, gels, patches, sprays, ointments, and warning agents. , lotion, liniment, pasta, or cataplasma may have formulations such as the like.

Carriers, excipients and diluents that may be included in the pharmaceutical composition according to the present invention include lactose, dextrose, sucrose, oligosaccharide, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

When formulated, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants.

Corn starch, potato starch, wheat starch, lactose, sucrose, glucose, fructose, di-mannitol, precipitated calcium carbonate, synthetic aluminum silicate, phosphoric acid as additives for tablets, powders, granules, capsules, pills, and troches according to the present invention Calcium monohydrogen, calcium sulfate, sodium chloride, sodium bicarbonate, purified lanolin, microcrystalline cellulose, dextrin, sodium alginate, methylcellulose, sodium carboxymethylcellulose, kaolin, urea, colloidal silica gel, hydroxypropyl starch, hydroxypropylmethyl Excipients such as cellulose (HPMC) 1928, HPMC 2208, HPMC 2906, HPMC 2910, propylene glycol, casein, calcium lactate, Primogel; Gelatin, gum arabic, ethanol, agar powder, cellulose phthalate acetate, carboxymethyl cellulose, calcium carboxymethyl cellulose, glucose, purified water, sodium caseinate, glycerin, stearic acid, sodium carboxymethyl cellulose, sodium methyl cellulose, methyl cellulose, microcrystalline cellulose, dextrin Binders such as hydroxycellulose, hydroxypropyl starch, hydroxymethylcellulose, purified shellac, starch arc, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinyl alcohol, and polyvinylpyrrolidone may be used, Hydroxypropyl Methyl Cellulose, Corn Starch, Agar Powder, Methyl Cellulose, Bentonite, Hydroxypropyl Starch, Sodium Carboxymethyl Cellulose, Sodium Alginate, Calcium Carboxymethyl Cellulose, Calcium Citrate, Sodium Lauryl Sulfate, Silicic Anhydride, 1-Hydroxy Propyl cellulose, dextran, ion exchange resin, polyvinyl acetate, formaldehyde-treated casein and gelatin, alginic acid, amylose, guar gum, sodium bicarbonate, polyvinylpyrrolidone, calcium phosphate, gelled starch, gum arabic, disintegrants such as amylopectin, pectin, sodium polyphosphate, ethyl cellulose, white sugar, magnesium aluminum silicate, di-sorbitol solution, and light anhydrous silicic acid; Calcium stearate, magnesium stearate, stearic acid, hydrogenated vegetable oil, talc, lycopod, kaolin, vaseline, sodium stearate, cacao butter, sodium salicylate, magnesium salicylate, polyethylene glycol 4000, PEG 6000, liquid paraffin, hydrogenated soybean oil ( Lubri wax), aluminum stearate, zinc stearate, sodium lauryl sulfate, magnesium oxide, macrogol, synthetic aluminum silicate, silicic anhydride, higher fatty acid, higher alcohol, silicone oil, paraffin oil, polyethylene glycol fatty acid ether, starch, sodium chloride , lubricants such as sodium acetate, sodium oleate, dl-leucine, light anhydrous silicic acid; may be used.

Additives for the liquid formulation according to the present invention include water, dilute hydrochloric acid, dilute sulfuric acid, sodium citrate, sucrose monostearate, polyoxyethylene sorbitol fatty acid esters (tween esters), polyoxyethylene monoalkyl ethers, lanolin ethers, Lanolin esters, acetic acid, hydrochloric acid, aqueous ammonia, ammonium carbonate, potassium hydroxide, sodium hydroxide, prolamine, polyvinylpyrrolidone, ethyl cellulose, sodium carboxymethyl cellulose, and the like may be used.

In the syrup according to the present invention, a solution of white sugar, other sugars, or a sweetener may be used, and aromatics, coloring agents, preservatives, stabilizers, suspending agents, emulsifiers, thickeners, etc. may be used as necessary.

Purified water may be used in the emulsion according to the present invention, and emulsifiers, preservatives, stabilizers, fragrances, etc. may be used as needed.

Suspension agents according to the present invention include acacia, tragacantha, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, microcrystalline cellulose, sodium alginate, hydroxypropylmethylcellulose (HPMC), HPMC 1828, HPMC 2906, HPMC 2910, etc. Agents may be used, and surfactants, preservatives, stabilizers, colorants, and fragrances may be used as needed.

Injections according to the present invention include distilled water for injection, 0.9% sodium chloride injection, IV injection, dextrose injection, dextrose + sodium chloride injection, PEG, lactated IV injection, ethanol, propylene glycol, non-volatile oil-sesame oil , solvents such as cottonseed oil, peanut oil, soybean oil, corn oil, ethyl oleate, isopropyl myristate, and benzene benzoate; solubilizing agents such as sodium benzoate, sodium salicylate, sodium acetate, urea, urethane, monoethylacetamide, butazolidine, propylene glycol, twins, nijuntinamide, hexamine, and dimethylacetamide; buffers such as weak acids and their salts (acetic acid and sodium acetate), weak bases and their salts (ammonia and ammonium acetate), organic compounds, proteins, albumins, peptones, and gums; tonicity agents such as sodium chloride; Stabilizers such as sodium bisulfite (NaHSO ₃ ) carbon dioxide gas, sodium metabisulfite (Na ₂ S ₂ O ₃ ), sodium sulfite (Na ₂ SO ₃ ), nitrogen gas (N ₂ ), ethylenediaminetetraacetic acid; Sulfating agents such as sodium bisulfide 0.1%, sodium formaldehyde sulfoxylate, thiourea, ethylenediamine disodium tetraacetate, acetone sodium bisulfite; analgesics such as benzyl alcohol, chlorobutanol, procaine hydrochloride, glucose, and calcium gluconate; Suspending agents such as Siemesis sodium, sodium alginate, Tween 80, aluminum monostearate may be included.

The suppository according to the present invention includes cacao butter, lanolin, witapsol, polyethylene glycol, glycerogelatin, methylcellulose, carboxymethylcellulose, a mixture of stearic acid and oleic acid, subanal, cottonseed oil, peanut oil, palm oil, cacao butter + Cholesterol, Lecithin, Lannet Wax, Glycerol Monostearate, Tween or Span, Imhausen, Monolen (Propylene Glycol Monostearate), Glycerin, Adeps Solidus, Buytyrum Tego-G -G), Cebes Pharma 16, Hexalide Base 95, Cotomar, Hydroxycote SP, S-70-XXA, S-70-XX75 (S-70-XX95), Hyde Hydrokote 25, Hydrokote 711, Idropostal, Massa estrarium (A, AS, B, C, D, E, I, T), Massa-MF, Masupol, Masupol-15, Neosupostal-N, Paramound-B, Suposiro (OSI, OSIX, A, B, C, D, H, L), suppository type IV (AB, B, A, BC, BBG, E, BGF, C, D, 299), Supostal (N, Es), Wecobi (W, R, S, M, Fs), testosterone triglyceride base (TG-95, MA, 57) and The same mechanism can be used.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations contain at least one excipient, for example, starch, calcium carbonate, sucrose, etc. ) or by mixing lactose and gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used.

Liquid preparations for oral administration include suspensions, solutions for oral administration, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, aromatics, and preservatives may be included. have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried formulations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspending agents.

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

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 single or multiple times. Considering all of the above factors, it is important to administer an amount that can obtain the maximum effect with the minimum amount without side effects, which can be easily determined by a person skilled in the art to which the present invention belongs.

The pharmaceutical composition of the present invention can be administered to a subject by various routes. All modes of administration can be envisaged, eg oral administration, subcutaneous injection, intraperitoneal administration, intravenous injection, intramuscular injection, paraspinal space (intrathecal) injection, sublingual administration, buccal administration, intrarectal insertion, vaginal It can be administered by intraoral insertion, ocular administration, otic administration, nasal administration, inhalation, spraying through the mouth or nose, dermal administration, transdermal administration, and the like.

The pharmaceutical composition of the present invention is determined according to the type of drug as an active ingredient together with various related factors such as the disease to be treated, the route of administration, 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 patient's age, sex, and weight, and is generally 0.001 to 150 mg per 1 kg of body weight, preferably 0.01 to 100 mg per day or every other day, or 1 It can be administered in 1 to 3 divided doses per day. However, since it may increase or decrease depending on the route of administration, severity of obesity, gender, weight, age, etc., the dosage is not limited to the scope of the present invention in any way.

In the present invention, "subject" means a subject in need of treatment of a disease, and more specifically, a human or non-human primate, mouse, rat, dog, cat, horse, cow, etc. It may be a mammal of, but is not limited thereto.

In the present invention, "administration" means providing a given composition of the present invention to a subject by any suitable method.

In the present invention, “prevention” refers to any action that suppresses or delays the onset of a desired disease, and “treatment” means that the desired disease and its resulting metabolic abnormality are improved or improved by administration of the pharmaceutical composition according to the present invention. All actions that are advantageously altered are meant, and "improvement" means any action that reduces a parameter related to a target disease, for example, the severity of a symptom, by administration of the composition according to the present invention.

In addition, the present invention provides a food composition for preventing or improving inflammatory diseases comprising Lactobacillus acidophilus-derived vesicles as an active ingredient.

The food composition may be a health functional food composition, but is not limited thereto.

When the vesicle of the present invention is used as a food additive, it can be added as it is or used together with other foods or food ingredients, and can be appropriately used according to a conventional method. The mixing amount of the active ingredient may be appropriately determined according to the purpose of use (prevention, health or therapeutic treatment). In general, the antifoam 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 when preparing food or beverage. However, in the case of long-term intake for the purpose of health and hygiene or health control, 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.

There is no particular limitation on the type of food. Examples of foods to which the above substances can be added include meat, sausages, bread, chocolates, candies, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice creams, various soups, beverages, tea, drinks, There are alcoholic beverages and vitamin complexes, and includes all health functional foods in a conventional sense.

The health beverage composition according to the present invention may contain various flavoring agents or natural carbohydrates as additional components, like conventional beverages. The aforementioned natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrins and cyclodextrins, and sugar alcohols such as xylitol, sorbitol and erythritol. As the sweetener, natural sweeteners such as thaumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame may be used. The proportion of the natural carbohydrate is generally about 0.01-0.20 g, or about 0.04-0.10 g per 100 mL of the composition of the present invention.

In addition to the above, the composition of the present invention contains various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, A carbonation agent used in carbonated beverages and the like may be contained. In addition, the composition of the present invention may contain fruit flesh for preparing natural fruit juice, fruit juice beverages and vegetable beverages. These components may be used independently or in combination. The ratio of these additives is not critical, but is generally selected in the range of 0.01-0.20 parts by weight per 100 parts by weight of the composition of the present invention.

In addition, the present invention relates to a cosmetic composition for preventing or improving inflammatory skin disease, comprising Lactobacillus acidophilus-derived vesicles as an active ingredient.

The formulation of the cosmetic composition according to the present invention is skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrient lotion, massage cream, nutrient cream, mist, moisture cream, hand cream, hand lotion, foundation, It may be in the form of essence, nutritional essence, pack, soap, cleansing foam, cleansing lotion, cleansing cream, cleansing oil, cleansing balm, body lotion, or body cleanser.

The cosmetic composition of the present invention may further include a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, high-molecular peptides, high-molecular polysaccharides, and sphingolipids.

Any water-soluble vitamin can be used as long as it can be incorporated into cosmetics, but examples include vitamin B1, vitamin B2, vitamin B6, pyridoxine, pyridoxine hydrochloride, vitamin B12, pantothenic acid, nicotinic acid, nicotinic acid amide, folic acid, vitamin C, and vitamin H. salts (thiamin hydrochloride, sodium ascorbate, etc.) or derivatives (sodium ascorbic acid-2-phosphate, magnesium salt of ascorbic acid-2-phosphate, etc.) included Water-soluble vitamins can be obtained by conventional methods such as a microbial transformation method, a purification method from a microbial culture, an enzymatic method, or a chemical synthesis method.

Any useful vitamin can be used as long as it can be formulated into cosmetics, but examples include vitamin A, carotene, vitamin D2, vitamin D3, vitamin E (d1-alpha tocopherol, d-alpha tocopherol, d-alpha tocopherol), etc. , their derivatives (ascorbine palmitate, ascorbine stearate, ascorbine dipalmitate, dl-alpha tocopherol acetate, dl-alpha tocopherol nicotinate, vitamin E, DL-pantothenyl alcohol, D-pantothenyl alcohol, pantothenylethyl ether, etc.) and the like are also included in the useful vitamins used in the present invention. The useful vitamins can be obtained by conventional methods such as microbial transformation, purification from microbial culture, enzymatic or chemical synthesis.

Any polymer peptide may be used as long as it can be incorporated into cosmetics, and examples thereof include collagen, hydrolyzed collagen, gelatin, elastin, hydrolyzed elastin, and keratin. Polymer peptides can be purified and obtained by conventional methods such as a purification method from a culture medium of microorganisms, an enzyme method, or a chemical synthesis method, or can be used after being purified from natural products such as pig or cow dermis or silkworm silk fibers.

The polymeric polysaccharide may be any material as long as it can be incorporated into cosmetics, and examples thereof include hydroxyethyl cellulose, xanthan gum, sodium hyaluronate, chondroitin sulfate or salts thereof (sodium salt, etc.). For example, chondroitin sulfate or a salt thereof can be used after being purified from mammals or fish.

Any sphingolipid can be used as long as it can be incorporated into cosmetics, and examples thereof include ceramide, phytosphingosine, and sphingoglycolipid. Sphingolipids can be purified by conventional methods or obtained by chemical synthesis from mammals, fish, shellfish, yeast, or plants.

In the cosmetic composition of the present invention, in addition to the above essential ingredients, other ingredients normally formulated in cosmetics may be blended as necessary.

Other ingredients that may be added include fats and oils, humectants, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, bactericides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, fragrances, A blood circulation accelerator, a cooling agent, an antiperspirant, purified water, etc. are mentioned.

Examples of the oil and fat component include ester oils, hydrocarbon oils, silicone oils, fluorine oils, animal fats and vegetable oils.

Examples of ester oils include glyceryl tri-2-ethylhexanoate, cetyl 2-ethylhexanoate, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isocetyl isostearate, and stearic acid. Butyl, ethyl linoleate, isopropyl linoleate, ethyl oleate, isocetyl myristate, isostearyl myristate, isostearyl palmitate, octyldodecyl myristate, isocetyl isostearate, diethyl sebacate, adipine Diisopropyl acid, isoalkyl neopentanoate, tri(capryl, capric acid) glyceryl, trimethylolpropane tri2-ethylhexanoate, trimethylolpropane triisostearate, pentaelislitol tetra2-ethylhexanoate , Cetyl caprylate, decyl laurate, hexyl laurate, decyl myristate, myristyl myristate, cetyl myristate, stearyl stearate, decyl oleate, cetyl ricinooleate, isostea laurate Lil, isotridecyl myristate, isocetyl palmitate, octyl stearate, isocetyl stearate, isodecyl oleate, octyldodecyl oleate, octyldodecyl linoleate, isopropyl isostearate, 2-ethylhexanoate Aryl, 2-ethyl stearyl hexanoate, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate, propylene glycol dicaprate, di(caprylic, capric acid) propylene glycol, propylene glycol dicaprylate, dicap Neopentyl glycol prinate, neopentyl glycol dioctanoate, glyceryl tricaprylate, glyceryl triundecylate, glyceryl triisopalmitate, glyceryl triisostearate, octyldodecyl neopentanoate, isostearyl octanoate , Octyl isononanoate, hexyldecyl neodecanoate, octyldodecyl neodecanoate, isocetyl isostearate, isostearyl isostearate, octyldecyl isostearate, polyglycerin oleate, polyglycerin isostearate, Triisocetyl citrate, triisoalkyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactate, cetyl lactate, octyldecyl lactate, triethyl citrate, acetyltriethyl citrate, acetyltributyl citrate, trioctyl citrate, dimalate Isostearyl, 2-ethylhexyl hydroxystearate, di2-ethylhexyl succinate, diisobutyl adipate, diisopropyl sebacate, dioctyl sebacate, Cholesteryl stearate, cholesteryl isostearate, cholesteryl hydroxystearate, cholesteryl oleate, dihydrocholesteryl oleate, phyteryl isostearate, physteryl oleate, 12-stealoyl and esters such as isocetyl hydroxystearate, stearyl 12-stealoylhydroxystearate, and isostearyl 12-stealoylhydroxystearate.

Examples of hydrocarbon-based fats and oils include hydrocarbon-based fats and oils such as squalene, liquid paraffin, alpha-olefin oligomer, isoparaffin, ceresin, paraffin, liquid isoparaffin, polybuden, microcrystalline wax, and vaseline.

Examples of silicone oils include polymethylsilicone, methylphenylsilicone, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, dimethylsiloxane/methylcetyloxysiloxane copolymer, dimethylsiloxane/methylstealoxysiloxane copolymer, alkyl Modified silicone oil, amino modified silicone oil, etc. are mentioned.

Examples of fluorine-based fats and oils include perfluoropolyether and the like.

Animal or vegetable oils include avocado oil, almond oil, olive oil, sesame oil, rice bran oil, birdflower oil, soybean oil, corn oil, rapeseed oil, algae oil, palm kernel oil, palm oil, castor oil, sunflower oil, grape seed oil , cottonseed oil, palm oil, kukui nut oil, wheat germ oil, rice germ oil, shea butter, moonshine colostrum, marker damian nut oil, meadowsweet oil, egg yolk oil, beef tallow, horse oil, mink oil, orange raffia oil, jojoba oil , animal or plant fats and oils such as candelilla wax, carnaba wax, liquid lanolin, and hydrogenated castor oil.

Examples of the moisturizer include water-soluble low-molecular moisturizers, fat-soluble molecular moisturizers, water-soluble polymers, and oil-soluble polymers.

Examples of water-soluble low-molecular moisturizers include serine, glutamine, sorbitol, mannitol, pyrrolidone-sodium carboxylate, glycerin, propylene glycol, 1,3-butylene glycol, ethylene glycol, polyethylene glycol B (polymerization degree n = 2 or more), polypropylene Glycol (polymerization degree n = 2 or more), polyglycerol B (polymerization degree n = 2 or more), lactic acid, lactate, etc. are mentioned.

Cholesterol, cholesterol ester, etc. are mentioned as a fat-soluble low-molecular moisturizer.

Examples of water-soluble polymers include carboxyvinyl polymer, polyaspartate, tragacanth, xanthan gum, methyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, carboxymethyl cellulose, water-soluble chitin, chitosan, dextrin, and the like. can

Examples of the fat-soluble polymer include polyvinylpyrrolidone/eicosene copolymer, polyvinylpyrrolidone/hexadecene copolymer, nitrocellulose, dextrin fatty acid ester, and high molecular silicone.

Examples of the emollient agent include long-chain acyl glutamic acid cholesteryl ester, hydroxystearic acid cholesteryl, 12-hydroxystearic acid, stearic acid, rosin acid, lanolin fatty acid cholesteryl ester, and the like.

Examples of the surfactant include nonionic surfactants, anionic surfactants, cationic surfactants, and amphoteric surfactants.

Nonionic surfactants include self-emulsifying glycerin monostearate, propylene glycol fatty acid esters, glycerin fatty acid esters, polyglycerin fatty acid esters, sorbitan fatty acid esters, POE (polyoxyethylene) sorbitan fatty acid esters, POE sorbitan fatty acid esters, POE Glycerin fatty acid ester, POE alkyl ether, POE fatty acid ester, POE hydrogenated castor oil, POE castor oil, POE·POP (polyoxyethylene·polyoxypropylene) copolymer, POE·POP alkyl ether, polyether modified silicone, lauric acid alkanolamides, alkylamine oxides, hydrogenated soybean phospholipids, and the like.

Anionic surfactants include fatty acid soaps, alpha-acyl sulfonates, alkyl sulfonates, alkyl allyl sulfonates, alkyl naphthalene sulfonates, alkyl sulfates, POE alkyl ether sulfates, alkyl amide sulfates, alkyl phosphates, POE alkyl phosphates, and alkyl amide phosphates. , alkyloylalkyl taurine salts, N-acylamino acid salts, POE alkyl ether carboxylate salts, alkyl sulfosuccinate salts, sodium alkyl sulfoacetate, acylated hydrolyzed collagen peptide salts, perfluoroalkyl phosphate esters, and the like. .

As the cationic surfactant, alkyltrimethylammonium chloride, stearyltrimethylammonium chloride, stearyltrimethylammonium bromide, cetostearyltrimethylammonium chloride, distearyldimethylammonium chloride, stearyldimethylbenzylammonium chloride, behenyltrimethylammonium bromide, chloride Benzalkonium, stearic acid diethylaminoethylamide, stearic acid dimethylaminopropylamide, lanolin derivative quaternary ammonium salt, etc. are mentioned.

Amphoteric surfactants include carboxybetaine type, amidebetaine type, sulfobetaine type, hydroxysulfobetaine type, amide sulfobetaine type, phosphobetaine type, aminocarboxylate type, imidazoline derivative type, amideamine type, etc. An amphoteric surfactant etc. are mentioned.

As organic and inorganic pigments, silicic acid, silicic anhydride, magnesium silicate, talc, sericite, mica, kaolin, bengala, clay, bentonite, titanium-coated mica, bismuth oxychloride, zirconium oxide, magnesium oxide, zinc oxide, titanium oxide, aluminum oxide inorganic pigments such as calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, iron oxide, ultramarine blue, chromium oxide, chromium hydroxide, calamine and complexes thereof; Polyamide, polyester, polypropylene, polystyrene, polyurethane, vinyl resin, urea resin, phenolic resin, fluororesin, silicon resin, acrylic resin, melamine resin, epoxy resin, polycarbonate resin, divinylbenzene/styrene copolymer, organic pigments such as silk powder, cellulose, CI pigment yellow and CI pigment orange, and composite pigments of these inorganic pigments and organic pigments.

Examples of the organic powder include metal soaps such as calcium stearate; Alkyl phosphate metal salts, such as zinc sodium cetylrate, zinc laurylate, and calcium laurylate; polyvalent metal salts of acyl amino acids such as N-lauroyl-beta-alanine calcium, N-lauroyl-beta-alanine zinc, and N-lauroyl glycine calcium; polyvalent metal salts of amide sulfonic acids such as N-lauroyl-taurine calcium and N-palmitoyl-taurine calcium; N-epsilon-lauroyl-L-lysine, N-epsilon-palmitoylizine, N-alpha-paritoylolnithine, N-alpha-lauroylarginine, N-alpha-hardened beef fatty acid acylarginine, etc. -acyl basic amino acids; N-acyl polypeptides such as N-lauroyl glycyl glycine; alpha-amino fatty acids such as alpha-aminocaprylic acid and alpha-aminolauric acid; polyethylene, polypropylene, nylon, polymethyl methacrylate, polystyrene, divinylbenzene/styrene copolymer, tetrafluoroethylene, and the like.

Examples of UV absorbers include para-aminobenzoic acid, ethyl para-aminobenzoate, amyl para-aminobenzoate, octyl para-aminobenzoate, ethylene glycol salicylate, phenyl salicylate, octyl salicylate, benzyl salicylate, butylphenyl salicylate, homomenthyl salicylate, and benzyl cinnamate. , paramethoxycinnamic acid-2-ethoxyethyl, paramethoxycinnamic acid octyl, diparamethoxycinnamic acid mono-2-ethylhexane glyceryl, paramethoxycinnamic acid isopropyl, diisopropyl-diisopropylcinnamic acid ester mixture, uro Cannic acid, ethyl urocanate, hydroxymethoxybenzophenone, hydroxymethoxybenzophenonesulfonic acid and its salts, dihydroxymethoxybenzophenone, dihydroxymethoxybenzophenone sodium disulfonate, dihydroxybenzophenone , tetrahydroxybenzophenone, 4-tert-butyl-4'-methoxydibenzoylmethane, 2,4,6-trianilino-p-(carbo-2'-ethylhexyl-1'-oxy)-1 ,3,5-triazine, 2-(2-hydroxy-5-methylphenyl)benzotriazole, etc. are mentioned.

Bactericides include hinokitiol, triclosan, trichlorohydroxydiphenyl ether, chlorhexidine gluconate, phenoxyethanol, resorcin, isopropylmethylphenol, azulene, salicylic acid, zincophylthione, benzalkonium chloride, photosensitizer So No. 301, mononitroguacol sodium, undecyrenic acid, etc. are mentioned.

Examples of antioxidants include butylhydroxyanisole, propyl gallic acid, and elisorbic acid.

Examples of the pH adjuster include citric acid, sodium citrate, malic acid, sodium malate, fumaric acid, sodium fumalate, succinic acid, sodium succinate, sodium hydroxide, and sodium monohydrogenphosphate.

Examples of alcohol include higher alcohols such as cetyl alcohol.

In addition, the blending components that may be added are not limited to these, and any of the above ingredients can be blended within a range not impairing the objects and effects of the present invention, but 0.01 to 5% by weight percent or 0.01 to 3% based on the total weight. Can be formulated in % weight percentage.

When the formulation of the present invention is a lotion, paste, cream or gel, animal fiber, vegetable fiber, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide as a carrier component can be used

When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and in particular, in the case of a spray, additional chlorofluorohydrocarbon, propane / May contain a propellant such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solvating agent or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butyl glycol oil, fatty acid esters of glycerol, polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, an ethoxylated isostearyl alcohol, a suspending agent such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline Cellulose, aluminum metahydroxide, bentonite, agar or tracanth and the like may be used.

When the formulation of the present invention is surfactant-containing cleansing, as carrier components, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolin derivatives, or ethoxylated glycerol fatty acid esters may be used.

In addition, the present invention may be provided in the form of a composition for inhalation containing Lactobacillus acidophilus-derived vesicles as an active ingredient.

In the case of an inhalational dosage form, it may be formulated according to methods known in the art, and a suitable propellant such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas may be used. It can be conveniently delivered in the form of an aerosol spray from a pressurized pack or nebulizer. In the case of pressurized aerosols, dosage units may be determined by providing a valve that delivers a metered amount. For example, gelatin capsules and cartridges for use in inhalers or insufflators may be formulated to contain a powder mixture of the compound and a suitable powder base such as lactose or starch.

In addition, the present invention relates to a drug delivery composition for treating diseases, comprising Lactobacillus acidophilus-derived vesicles as an active ingredient.

The term "drug delivery" as used herein refers to any means or actions of loading and delivering a drug to a composition according to the present invention in order to deliver the drug to a specific organ, tissue, cell or organelle.

In the present invention, the drug delivery composition may deliver a drug to one or more organs selected from the group consisting of the stomach, small intestine, large intestine, lung, liver, and brain, but is not limited thereto.

In the present invention, the disease may be one or more selected from the group consisting of gastric disease, intestinal disease, lung disease, liver disease, and central nervous system disease, but is not limited thereto.

Hereinafter, a preferred embodiment is presented to aid understanding of the present invention. However, the following examples are provided to more easily understand the present invention, and the content of the present invention is not limited by the following examples.

[Example]

Example 1: Isolation of vesicles derived from Lactobacillus acidophilus

In order to isolate Lactobacillus acidophilus -derived vesicles (extracellular vesicles; EV), Lactobacillus acidophilus was inoculated into MRS (de Man-Rogosa and Sharpe) medium, and at 37 ° C. at 200 rpm. It was cultured until the absorbance (OD _{600 nm} ) was 1.0 to 1.5. Subsequently, it was re-inoculated and cultured in LB (Luria Bertani) medium. Then, the culture solution containing the cells was recovered and centrifuged at 10,000 g at 4° C. for 20 minutes to obtain a supernatant from which the cells were removed. The obtained supernatant was filtered again using a 0.22 μm filter. The filtered supernatant was concentrated to a volume of 50 mL or less using a 100 kDa Pellicon 2 Cassette filter membrane (Merck Millipore) and a MasterFlex pump system (Cole-Parmer). Then, the concentrated supernatant was filtered again using a 0.22 μm filter to separate Lactobacillus acidophilus-derived vesicles. To quantify the vesicles contained in the supernatant, the amount of protein contained in the vesicles was measured using the Pierce BCA Protein Assay kit (Thermo Fisher Scientific).

Example 2: Pharmacokinetics of Lactobacillus acidophilus-derived vesicles

In order to confirm the pharmacokinetic characteristics of absorption, distribution, and excretion by orally administering Lactobacillus acidophilus-derived vesicles, an experiment was conducted as follows. Lactobacillus acidophilus-derived vesicles labeled with phosphor Cy7 were orally administered to mice, and 0, 1, 3, 6, 24, and 48 hours later, images of the entire mouse and various organs were removed to evaluate their distribution to organs. Optical imaging equipment (Davinch-Invivo Fluoro Chemi (western); Davinch-K) was used to observe the fluorescence signal.

As shown in Figure 1, it was confirmed that the vesicles were absorbed systemically at 1 hour after administration of the vesicles, which continued until 24 hours after administration, and most of the vesicles were excreted after 48 hours. In addition, when the distribution pattern to major organs was evaluated after oral administration of the vesicles, it was found that the vesicles were distributed to various organs 1 hour after oral administration.

After oral administration of the vesicles, absorption, distribution, and excretion into the stomach, small intestine, and large intestine were evaluated over time.

As shown in Figure 2, at 1 hour after administration, it was mainly absorbed into the stomach and small intestine and distributed to tissues, and most of it was excreted 48 hours after administration.

In addition, after orally administering the vesicles, absorption, distribution, and excretion into the lungs and liver were evaluated over time.

As shown in Figure 3, at 1 hour after administration, it was absorbed into the lungs and distributed to lung tissue, and most of them were excreted 48 hours after administration. On the other hand, absorption into the liver occurred 3 hours after vesicle administration, and most of them were excreted 48 hours after administration.

In addition, after orally administering the vesicles, absorption, distribution, and excretion into the central nervous system were evaluated over time.

As shown in Figure 4, at 3 hours after administration, it was mainly absorbed into brain tissue, distributed most in brain tissue at 24 hours after administration, and most of it was excreted after 48 hours of administration.

In addition, after oral administration of the vesicles, cardiac and renal absorption, distribution, and excretion patterns were evaluated over time.

As shown in Figure 5, the vesicles were not absorbed and distributed to cardiac and renal tissues.

Through the above results, when Lactobacillus acidophilus-derived vesicles are orally administered, the vesicles are mainly absorbed through the stomach and small intestine, move to blood vessels through the lymphatic vessels in the gastrointestinal tissue, and then first into the pulmonary artery (pulmonary artery). ), it was distributed to the lung tissue, and then to the central nervous system and liver via the systemic artery.

Example 3: Comparison of proinflammatory effects of E. coli-derived vesicles and Lactobacillus acidophilus-derived vesicles

In order to evaluate the extent to which Lactobacillus acidophilus-derived vesicles induce inflammation in inflammatory cells, Lactobacillus acidophilus-derived vesicles were added to Raw 264.7, a mouse macrophage cell line, at concentrations of 0.1, 1, and 10 μg/mL. After treatment and reaction, the amount of secretion of TNF-α and IL-6, which are representative inflammatory mediators that cause inflammatory diseases, was measured. In order to evaluate the degree of causing an inflammatory response, E. coli-derived vesicles (E coli EV) were treated with a positive control pathogenic factor and compared. More specifically, Raw 264.7 cells were dispensed in 1X10 ⁵ each in a 48-well plate, treated with Lactobacillus acidophilus-derived vesicles or E. coli-derived vesicles diluted in DMEM serum-free medium, and cultured for 12 hours. Thereafter, the cell culture medium was placed in a 1.5 mL tube and centrifuged at 3,000 g for 5 minutes to obtain only the supernatant, which was stored at -80 ° C and the amount of TNF-α and IL-6 was measured by ELISA (R&D Systems). In the ELISA method, Capture antibody (Abcam) was diluted in phosphate buffer solution, dispensed in 50 μL each in a 96-well plate, and reacted at 4° C. for 16 hours. After washing three times with 100 μL of phosphate buffer solution (PBST) to which 0.05% Tween-20 was added, 100 μL of phosphate buffer solution (RD) to which 1% BSA (bovine serum albumin) was added was added per well. It was added and blocked for 1 hour at room temperature. In addition, 50 μL of the sample and standard to be tested were added according to the concentration, reacted at room temperature for 2 hours, and washed three times with PBST. Then, the detection antibody was diluted in RD, added in 50 μL per well, reacted at room temperature for 2 hours, washed three times with PBST to remove unbound antibody, and Streptavidin-HRP diluted 1/40 in RD (R&D system) was added by 50 μL per well and reacted at room temperature for 20 minutes. Finally, after washing three times with 100 μL of PBST, 50 μL of TMB substrate (SurModics) was added per well, and after 5 to 20 minutes, when the color developed, 50 μL of 1 M sulfuric acid solution was added per well to quench the reaction. After termination, absorbance was measured at 450 nm using a SpectraMax M3 microplate reader (Molecular Devices).

As a result, as shown in FIGS. 6a and 6b, when treated with 1 μg/mL of E. coli-derived vesicles, secretion of TNF-α and IL-6 was induced in inflammatory cells, but the same amount of Lactobacillus acidophilus When the derived vesicles were treated with inflammatory cells, secretion of TNF-α and IL-6 was not induced. In addition, even when Lactobacillus acidophilus-derived vesicles were treated with inflammatory cells at a concentration 10 times that of E. coli-derived vesicles, TNF-α and IL-6 secretion was hardly induced.

Through the above results, it can be seen that the vesicles derived from Lactobacillus acidophilus are significantly less effective in inducing inflammation than the vesicles derived from Escherichia coli, which is a pathogenic factor.

Example 4: Anti-inflammatory effect of Lactobacillus acidophilus-derived vesicles on pathogenic factor-induced inflammatory responses

It is known that vesicles secreted by pathogenic bacteria are important pathogenic factors causing various inflammatory diseases. Thus, the therapeutic effect of Lactobacillus acidophilus-derived vesicles on the pathogenesis of inflammatory diseases caused by pathogenic factors was confirmed. Raw 264.7, a mouse macrophage cell line, was pretreated with Lactobacillus acidophilus-derived vesicles at concentrations of 0.1, 1, and 10 μg/mL and cultured, and then E. coli-derived vesicles at 1 μg/mL concentration were treated with inflammatory cells Secretion amounts of TNF-α and IL-6 were measured. More specifically, Raw 264.7 cells were seeded in a 48-well plate, treated with Lactobacillus acidophilus-derived vesicles diluted in DMEM serum-free medium, and cultured for 12 hours. Then, after treatment with E. coli-derived vesicles at a concentration of 1 μg/mL, the cells were further cultured for 12 hours. The secretion amounts of TNF-α and IL-6 were measured in the same manner as in Example 3.

As a result, as shown in FIGS. 7A and 7B , it was confirmed that secretion of TNF-α and IL-6 by E. coli-derived vesicles was inhibited in a dose-dependent manner in the samples pretreated with Lactobacillus acidophilus-derived vesicles.

Through the above results, it was confirmed that the Lactobacillus acidophilus-derived vesicles effectively inhibited inflammation induced by pathogenic factors causing inflammatory diseases.

Through the above results, it was confirmed that the Lactobacillus acidophilus-derived vesicles of the present invention effectively suppress the occurrence of inflammatory diseases mediated by TNF-α or IL-6 by being absorbed and distributed to major organs, the present invention Lactobacillus acidophilus-derived vesicles of Lactobacillus acidophilus are expected to be used for the improvement, prevention, or treatment of inflammatory diseases.

The above description of the present invention is for illustrative purposes, and those skilled in the art can understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, the embodiments described above should be understood as illustrative in all respects and not limiting.

Claims (17)

A pharmaceutical composition for preventing or treating inflammatory diseases, comprising vesicles derived from Lactobacillus acidophilus as an active ingredient.
According to claim 1,
The inflammatory disease is at least one inflammatory skin selected from the group consisting of eczema, atopic dermatitis, psoriasis, acne, alopecia, contact dermatitis, seborrheic dermatitis, and mastocytosis disease;
one or more inflammatory oral diseases selected from the group consisting of caries, gingivitis, and periodontitis;
Ulcerative colitis, Crohn's disease, intestinal Behcet's disease, indeterminate colitis, chronic gastritis, chronic peptic ulcer, celiac disease disease), and at least one inflammatory gastrointestinal disease selected from the group consisting of inflammatory bowel disease;
At least one inflammatory disease selected from the group consisting of non-alcoholic steatohepatitis (NASH), alcoholic steatohepatitis, liver cirrhosis, cholangitis, cholecystitis, and pancreatitis hepato-biliary-pancreatic disease;
one or more inflammatory nasal diseases selected from the group consisting of rhinitis, sinusitis, and sinusitis;
bronchitis, asthma, chronic obstructive pulmonary disease (COPD), emphysema, cystic fibrosis, pneumonia, chronic interstitial pneumonitis, bronchopulmonary dysplasia, and idiopathic pulmonary fibrosis ( at least one inflammatory lung disease selected from the group consisting of idiopathic pulmoniary fibrosis;
Hyperinsulinemia, dyslipidemia, arrhythmias, atherosclerosis, angina, metabolic syndrome, myocardial infartion, cardiomyopathy, stroke, ischemic heart disease, one or more inflammatory cardiovascular diseases selected from the group consisting of heart failure, thrombosis, and atherosclerosis;
Atony, muscular atrophy, muscular dystrophy, myasthenia, cachexia, Gout, sarcopenia, osteoporosis, Paget's disease, one or more inflammatory musculoskeletal disorders selected from the group consisting of rheumatoid arthritis and osteoarthritis;
Mild cognitive impairment, dementia, Alzheimer's disease, Parkinson's disease, Huntington's disease, Amyotrophic lateral sclerosis (ALS), Batten disease ), Kearns-Sayre syndrome (KSS), chronic progressive external ophthalmoplegia (CPEO), mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes (MELAS), MERRF syndrome ( myoclonic epilepsy with ragged-red fibers (MERRF), neurogenic weakness with ataxia and retinitis pigmentosa (NARP), Leigh syndrome (LS), mitochondrial recessive ataxia syndrome (MIRAS syndrome), anxiety disorder, post-traumatic stress disorder Post-traumatic stress disorder (PTSD), panic disorder, depression, autism spectrum disorders, attention deficit/hyperactivity disorder (ADHD), schizophrenia ), Dementia with Lewy Bodies (DLB), Multi-Infarct Dementia (MID), frontotemporal lobar degeneration (FTLD), Pick's disease, and cortical degeneration (Co rticobasal degeneration (CBD), progressive supranuclear palsy (PSP), age-related macular degeneration (AMD), dysosmia, deafness, and diabetic retinopathy ) At least one inflammatory central nervous system disease selected from the group consisting of; and
Proliferative vitreoretinopathy, macular degeneration, retinopathy pigmentosa, diabetic retinopathy, choroidal neovascularization, neovascular glaucoma, ischemic optic nerve disorder, retinopathy of prematurity, retinopathy of prematurity, epidemic keratoconjunctivitis, neovascular iris disease, postphakic fibroplasia, atopic A pharmaceutical composition, characterized in that selected from the group consisting of at least one chronic inflammatory eye disease selected from the group consisting of keratitis, limbal keratitis, phase patch psoriatic keratitis, plyctenic keratoconjunctivitis, scleritis and diabetic macular edema.
According to claim 1,
The inflammatory disease is characterized in that mediated by TNF-α or IL-6, the pharmaceutical composition.
According to claim 1,
The pharmaceutical composition, characterized in that the vesicles have an average diameter of 10 to 300 nm.
According to claim 1,
The pharmaceutical composition, characterized in that the vesicles are secreted naturally or artificially from Lactobacillus acidophilus.
According to claim 1,
The vesicles are characterized in that separated from the culture medium of Lactobacillus acidophilus or food cultured by adding Lactobacillus acidophilus, a pharmaceutical composition.
A food composition for preventing or improving inflammatory diseases, comprising vesicles derived from Lactobacillus acidophilus as an active ingredient.
According to claim 7,
The inflammatory disease is at least one inflammatory skin selected from the group consisting of eczema, atopic dermatitis, psoriasis, acne, alopecia, contact dermatitis, seborrheic dermatitis, and mastocytosis disease;
one or more inflammatory oral diseases selected from the group consisting of caries, gingivitis, and periodontitis;
Ulcerative colitis, Crohn's disease, intestinal Behcet's disease, indeterminate colitis, chronic gastritis, chronic peptic ulcer, celiac disease disease), and at least one inflammatory gastrointestinal disease selected from the group consisting of inflammatory bowel disease;
At least one inflammatory disease selected from the group consisting of non-alcoholic steatohepatitis (NASH), alcoholic steatohepatitis, liver cirrhosis, cholangitis, cholecystitis, and pancreatitis hepato-biliary-pancreatic disease;
one or more inflammatory nasal diseases selected from the group consisting of rhinitis, sinusitis, and sinusitis;
bronchitis, asthma, chronic obstructive pulmonary disease (COPD), emphysema, cystic fibrosis, pneumonia, chronic interstitial pneumonitis, bronchopulmonary dysplasia, and idiopathic pulmonary fibrosis ( at least one inflammatory lung disease selected from the group consisting of idiopathic pulmoniary fibrosis;
Hyperinsulinemia, dyslipidemia, arrhythmias, atherosclerosis, angina, metabolic syndrome, myocardial infartion, cardiomyopathy, stroke, ischemic heart disease, one or more inflammatory cardiovascular diseases selected from the group consisting of heart failure, thrombosis, and atherosclerosis;
Atony, muscular atrophy, muscular dystrophy, myasthenia, cachexia, Gout, sarcopenia, osteoporosis, Paget's disease, one or more inflammatory musculoskeletal disorders selected from the group consisting of rheumatoid arthritis and osteoarthritis;
Mild cognitive impairment, dementia, Alzheimer's disease, Parkinson's disease, Huntington's disease, Amyotrophic lateral sclerosis (ALS), Batten disease ), Kearns-Sayre syndrome (KSS), chronic progressive external ophthalmoplegia (CPEO), mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes (MELAS), MERRF syndrome ( myoclonic epilepsy with ragged-red fibers (MERRF), neurogenic weakness with ataxia and retinitis pigmentosa (NARP), Leigh syndrome (LS), mitochondrial recessive ataxia syndrome (MIRAS syndrome), anxiety disorder, post-traumatic stress disorder Post-traumatic stress disorder (PTSD), panic disorder, depression, autism spectrum disorders, attention deficit/hyperactivity disorder (ADHD), schizophrenia ), Dementia with Lewy Bodies (DLB), Multi-Infarct Dementia (MID), frontotemporal lobar degeneration (FTLD), Pick's disease, and cortical degeneration (Co rticobasal degeneration (CBD), progressive supranuclear palsy (PSP), age-related macular degeneration (AMD), dysosmia, deafness, and diabetic retinopathy ) At least one inflammatory central nervous system disease selected from the group consisting of; and
Proliferative vitreoretinopathy, macular degeneration, retinopathy pigmentosa, diabetic retinopathy, choroidal neovascularization, neovascular glaucoma, ischemic optic nerve disorder, retinopathy of prematurity, retinopathy of prematurity, epidemic keratoconjunctivitis, neovascular iris disease, postphakic fibroplasia, atopic A food composition, characterized in that selected from the group consisting of at least one chronic inflammatory eye disease selected from the group consisting of keratitis, limbal keratitis, phase patch psoriatic keratitis, plyctenic keratoconjunctivitis, scleritis and diabetic macular edema.
According to claim 7,
The inflammatory disease is characterized in that the disease mediated by TNF-α or IL-6, food composition.
According to claim 7,
Characterized in that the vesicles are secreted naturally or artificially from Lactobacillus acidophilus, food composition.
An inhalant composition for preventing or treating inflammatory diseases or cancer, comprising vesicles derived from Lactobacillus acidophilus as an active ingredient.
A cosmetic composition for preventing or improving inflammatory skin disease, comprising vesicles derived from Lactobacillus acidophilus as an active ingredient.
According to claim 12,
The inflammatory skin disease is at least one selected from the group consisting of eczema, atopic dermatitis, psoriasis, acne, alopecia, contact dermatitis, seborrheic dermatitis, and mastocytosis Characterized in, cosmetic composition.
According to claim 12,
The inflammatory skin disease is characterized in that mediated by TNF-α or IL-6, cosmetic composition.
A composition for drug delivery for the treatment of diseases, comprising vesicles derived from Lactobacillus acidophilus as an active ingredient.
According to claim 15,
The disease is characterized in that at least one selected from the group consisting of gastric disease, intestinal disease, lung disease, liver disease, and central nervous system disease, disease treatment drug delivery composition.
According to claim 16,
The central nervous system disease is mild cognitive impairment, dementia, Alzheimer's disease, Parkinson's disease, Huntington's disease, Lou Gehrig's disease (Amyotrophic lateral sclerosis (ALS)), Batten disease, Kearns-Sayre syndrome (KSS), chronic progressive external ophthalmoplegia (CPEO), mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes (MELAS) ), MERRF syndrome (myoclonic epilepsy with ragged-red fibers, MERRF), NARP syndrome (neurogenic weakness with ataxia and retinitis pigmentosa, NARP), Leigh syndrome (LS), MIRAS syndrome (mitochondrial recessive ataxia syndrome), anxiety disorder , Post-traumatic Stress Disorder (PTSD), panic disorder, depression, autism spectrum disorders, attention deficit/hyperactivity disorder (ADHD) , schizophrenia, Dementia with Lewy Bodies (DLB), Multi-Infarct Dementia (MID), frontotemporal lobar degeneration (FTLD), Pick's disease e), Corticobasal degeneration (CBD), progressive supranuclear palsy (PSP), age-related macular degeneration (AMD), dysosmia, deafness, And diabetic retinopathy (diabetic retinopathy) characterized in that at least one disease selected from the group consisting of, disease treatment drug delivery composition.

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