KR20200047097A - A pharmaceutical composition for treating arthritis comprising Lactobacillus which induces secretion of IL-10 from dendritic cells - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for treating arthritis using lactobacillus which promotes IL-10 secretion of dendritic cells as an active component. By using the composition of the present invention, it is possible to effectively treat autoimmune diseases or arthritis. An objective of the present invention is to provide the composition for treating autoimmune diseases or arthritis, or relieving or alleviating symptoms which contains selected lactobacillus or a culture thereof as an active component.

Description

A pharmaceutical composition for treating arthritis comprising Lactobacillus which induces secretion of IL-10 from dendritic cells}

The present invention relates to a pharmaceutical composition for treating arthritis using lactic acid bacteria that promote IL-10 secretion of dendritic cells as an active ingredient.

Arthritis is a typical cartilage-related disease, and is classified into rheumatoid arthritis and degenerative arthritis (osteogenic arthritis), and is often found in cases where the knee cartilage is damaged due to a traffic accident or excessive exercise. Currently, there are about 1 million arthritis patients in Korea, and women are more than twice as many as men, especially among menopausal women (Belmonte-S. MA et al; Arthritis Rheum, 30, pp 1214- 1225, 1987). Cartilage tissue makes up the joints and absorbs the power of joint movement. When such cartilage tissue is damaged, arthritis with swelling, heat, and pain is caused, and arthritis occurs regardless of race and is divided into more than 100 types depending on the cause. The most common forms are osteoarthritis, a degenerative joint disease mainly caused by aging, and other autoimmune diseases, rheumatoid arthritis, psoriatic arthritis, and infection. And septic arthritis.

Rheumatoid arthritis is the most common inflammatory arthritis in adults, the cause of which is not fully known, but is understood as an inflammatory reaction of the synovial membrane in the joint due to the autoimmune mechanism. Antirheumatic drugs and biological agents used to treat rheumatoid arthritis have the effect of improving arthritis through inflammation control, but are still insufficient to induce remission, the ultimate therapeutic goal. In many patients, the therapeutic effect is incomplete, and the side effects associated with the therapeutic agent have become an obstacle to treatment, and when a biological agent is used for a long time, an antibody against the drug is produced, and thus the therapeutic effect may be reduced.

In order to treat such arthritis, studies such as autografting, allografting, and artificial bone grafting have been conducted, but complications such as infection, hematoma, and the like of the bone-collecting site have been performed. Transplantation), the possibility of disease transmission from donors (allogeneic bone graft), and underlying bone formation (artificial bone graft).

Therefore, there is an urgent need for a new new treatment or treatment that overcomes the shortcomings of the existing treatment.

An object of the present invention is to provide a composition for treating autoimmune diseases or arthritis or alleviating or improving symptoms using selected lactic acid bacteria or a culture thereof as an active ingredient.

Hereinafter, various embodiments described herein are described with reference to the drawings. In the following description, for the full understanding of the present invention, various specific details, such as specific forms, compositions and processes, have been described. However, certain embodiments may be practiced without one or more of these specific details, or in combination with other known methods and forms. In other instances, well-known processes and manufacturing techniques are not described in specific details in order not to unnecessarily obscure the present invention. Reference throughout this specification to “one embodiment” or “an embodiment” means that a particular feature, form, composition or characteristic described in connection with the embodiment is included in one or more embodiments of the invention. Thus, the context of “in one embodiment” or “an embodiment” expressed in various places throughout this specification does not necessarily represent the same embodiment of the invention. Additionally, special features, shapes, compositions, or properties can be combined in any suitable way in one or more embodiments. Unless otherwise specified in the present invention, all scientific and technical terms used herein have the same meaning as commonly understood by those skilled in the art to which the present invention pertains.

In one embodiment of the invention, Bifidobacterium latis lactis ) provides a pharmaceutical composition for the treatment of arthritis comprising lactic acid bacteria or a culture medium thereof as an active ingredient, and the arthritis is osteoarthritis, degenerative arthritis, exfoliative osteochondritis, joint ligament damage, meniscal cartilage damage, arthritis due to infection, psoriasis Arthritis, ankylosing spondylitis, rheumatoid arthritis and childhood rheumatoid arthritis provides at least one pharmaceutical composition selected from the group consisting of powders, granules, capsules, tablets, aqueous suspensions, external preparations, suppositories or injection solutions. , To provide a pharmaceutical composition, the lactic acid bacteria or a culture medium thereof provides a pharmaceutical composition in which dendritic cells promote IL-10 secretion.

In one embodiment of the invention, Bifidobacterium latis lactis ) or a culture solution thereof to provide a food composition for alleviating or improving arthritis, which is an active ingredient, wherein the arthritis is osteoarthritis, degenerative arthritis, exfoliative osteochondritis, joint ligament damage, meniscal cartilage damage, arthritis caused by infection, tendon Provided is a food composition of at least one selected from the group consisting of adenomatous arthritis, ankylosing spondylitis, rheumatoid arthritis and childhood rheumatoid arthritis.

In one embodiment of the present invention, Lactobacillus paracasei ( Lactobacillus paracasei ), Lactobacillus cactus ( Lactobacillus curvatus ) and Lactobacillus plantarum ( Lactobacillus plantarum ) selected from the group consisting of at least one lactic acid bacteria or a culture medium thereof as an active ingredient It provides a pharmaceutical composition for the treatment of arthritis, including osteoarthritis, degenerative arthritis, exfoliative osteochondritis, joint ligament damage, meniscal cartilage damage, arthritis caused by infection, psoriatic arthritis, ankylosing spondylitis, rheumatoid arthritis and childhood Provides a pharmaceutical composition, at least one selected from the group consisting of rheumatoid arthritis, the pharmaceutical composition provides a pharmaceutical composition, prepared as a powder, granule, capsule, tablet, aqueous suspension, external preparation, suppository or injection solution, lactic acid bacteria or In its culture, dendritic cells secrete IL-10 It provides a pharmaceutical composition to promote.

In one embodiment of the present invention, Lactobacillus paracasei ( Lactobacillus paracasei ), Lactobacillus cactus ( Lactobacillus curvatus ) and Lactobacillus plantarum ( Lactobacillus plantarum ) selected from the group consisting of lactic acid bacteria or a culture medium thereof as an active ingredient Provides a food composition for alleviating or improving arthritis, including arthritis, degenerative arthritis, exfoliative osteochondritis, joint ligament damage, meniscal cartilage damage, arthritis caused by infection, psoriatic arthritis, ankylosing spondylitis, rheumatoid arthritis And at least one selected from the group consisting of childhood rheumatoid arthritis, provides a food composition.

In one embodiment of the invention, Bifidobacterium latis autoimmunity comprising at least one lactic acid bacteria selected from the group consisting of lactis ), Lactobacillus paracasei , Lactobacillus curvatus , and Lactobacillus plantarum or a culture medium thereof as an active ingredient Providing a pharmaceutical composition for the treatment of diseases, the autoimmune diseases include rheumatoid arthritis, inflammatory vertebral arthritis, adult Stills' disease, macrophage activation syndrome, systemic sclerosis, polymyositis, dermatomyositis, vasculitis, mixed connective tissue disease, Sjogren's syndrome, Ulcerative colitis, Crohn's disease, allergic asthma, allergic rhinitis, atopic dermatitis, margin, fistula, conjunctivitis, psoriasis, vulgar rash, Parkinson's disease, amyotrophic lateral sclerosis, myasthenia gravis, multiple sclerosis, Alzheimer's disease, stroke, Arteriosclerosis, vascular restenosis, type I diabetes, type II diabetes, diabetic retinopathy At least one from the group consisting of chronic thyroiditis provides pharmaceutical compositions.

In the present invention, "IL-10" forms a dimer with one of the water-soluble cytokines, and interacts with a heterodimeric receptor composed of IL-10Rα and IL-10β. It plays a protective role in autoimmune diseases such as rheumatoid arthritis and chronic inflammatory diseases such as Crohn's disease and psoriasis. In particular, IL-10, which is mainly secreted from Breg cells, Treg cells, M2 macrophages, etc., which suppress inflammation, inhibits activation of neutrophils and penetration into inflammatory sites, converts macrophages into M2 phenotype, and TNF- Rheumatism symptoms can be alleviated by inhibiting the expression of inflammatory cytokines, which play a key role in rheumatism symptoms such as α (Chen, Z., Bozec, A., Ramming, A. & Schett, GJNRR Anti-inflammatory and immune-regulatory cytokines in rheumatoid arthritis. 1 (2018)).

“Autoimmune disease” in the present invention is a non-malignant disease or disorder that occurs and is directed against an individual's own tissues. One of the most important characteristics of all normal individuals is that they do not react detrimentally to the antigens that make up themselves, but they can recognize and remove them from non-self antigens. The body's non-response to an autoantigen is called immunologic unresponsiveness or tolerance. However, if there is an abnormality in inducing or maintaining self-tolerance, an immune response occurs against the self-antigen, and as a result, a phenomenon that attacks one's own tissue occurs. The disease caused by the above process is called an autoimmune disease. The autoimmune disease is an inflammatory disease in which antibodies are produced against its organ tissues or its components, and may refer to a disease that causes chronic systemic inflammation in a number of tissues and organs. The autoimmune disease is, but is not limited to, rheumatoid arthritis, inflammatory vertebral arthritis, adult Stills' disease, macrophage activation syndrome, systemic sclerosis, polymyositis, dermatomyositis, vasculitis, mixed connective tissue disease, Sjogren's syndrome, ulcerative colitis , Crohn's disease, allergic asthma, allergic rhinitis, atopic dermatitis, phimosis, fistula, conjunctivitis, psoriasis, vulgar erythema, Parkinson's disease, amyotrophic lateral sclerosis, myasthenia gravis, multiple sclerosis, Alzheimer's disease, stroke, atherosclerosis, One or more may be selected from the group consisting of vascular restenosis, type I diabetes, type II diabetes, diabetic retinopathy, and chronic thyroiditis.

The bones support the soft tissues and weight of the human body and surround the internal organs to protect the internal organs from external shocks. In addition, it is an important part of the human body that not only structurally supports muscles or organs, but also stores substances such as calcium and other essential minerals in the body, such as phosphorus and magnesium. Joints exist between the bones that make up the human body. Joints are immobilized joints with little or no mobility between two bones or cartilage that are in contact with each other, such as the roots of the skull or teeth, and movable joints or joints with many connective tissues and large mobility between the bones, such as the bones or jaws of the arms and legs of animals. And semi-joints. Generally, a joint refers to a movable joint, and the movable joint is divided into a ligament bond and a synovial joint, in which both bones are joined only by ligaments. The synovial membrane is a joint where the joints are stacked with connective tissue pockets (joint capsules), and the inside of the joint sac secretes synovial fluid, which is a lubricating oil.

In the present invention, the "arthritis" may mean chronic inflammation of the joints in tissues around the joints, such as tendons, ligaments, and muscles, as well as other organs of the body, preferably osteoarthritis, degenerative arthritis, exfoliative osteochondritis , Joint ligament damage, meniscal cartilage damage, arthritis due to infection, psoriatic arthritis, ankylosing spondylitis, rheumatoid arthritis, and childhood rheumatoid arthritis.

In the present invention, "prevention" may include, without limitation, any action to block or suppress or delay arthritis symptoms using the pharmaceutical composition of the present invention.

In the present invention, "treatment" may include, without limitation, any act of improving or benefiting arthritis symptoms using the pharmaceutical composition of the present invention.

In the present invention, the pharmaceutical composition may be characterized in that it is in the form of capsules, tablets, granules, injections, ointments, powders or beverages, and the pharmaceutical composition may be characterized for humans.

The pharmaceutical composition of the present invention is not limited to these, but may be used in the form of oral dosage forms such as powders, granules, capsules, tablets, aqueous suspensions, external preparations, suppositories, and sterile injectable solutions, respectively, according to a conventional method. You can. The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers may include binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, pigments, fragrances, etc. when administered orally, and buffers, preservatives, and analgesics for injections. Agents, solubilizers, isotonic agents, stabilizers, etc. can be used in combination, and for topical administration, bases, excipients, lubricants, preservatives, etc. can be used. The formulation of the pharmaceutical composition of the present invention can be variously prepared by mixing with a pharmaceutically acceptable carrier as described above. For example, when administered orally, tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc. may be prepared, and in the case of injection, unit dosage ampoules or multiple doses may be prepared. have. Others can be formulated as solutions, suspensions, tablets, capsules, and sustained release preparations.

On the other hand, examples of carriers, excipients and diluents suitable for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, malditol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil and the like can be used. In addition, fillers, anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers, preservatives, etc. may be further included.

The route of administration of the pharmaceutical composition according to the present invention is not limited to these, but oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical , Sublingual or rectal. Oral or parenteral administration is preferred.

In the present invention, "parenteral" includes subcutaneous, intradermal, intravenous, intramuscular, intra-articular, intra-synovial, intrasternal, epidural, intralesional and intracranial injection or infusion techniques. The pharmaceutical composition of the present invention can also be administered in the form of suppositories for rectal administration.

The pharmaceutical composition of the present invention depends on a number of factors, including the activity, age, weight, general health, sex, diet, administration time, route of administration, release rate, drug combination and severity of the particular disease to be prevented or treated, of the specific compound used. It may vary, and the dosage of the pharmaceutical composition varies depending on the patient's condition, body weight, disease severity, dosage form, administration route and duration, but can be appropriately selected by those skilled in the art, and 0.0001 to 50 mg / day It can be administered in kg or 0.001 to 50 mg / kg. The administration may be administered once a day, or may be divided into several times. The above dosage does not limit the scope of the present invention in any way. The pharmaceutical composition according to the present invention may be formulated as pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions.

In the present invention, "improvement" refers to all actions in which symptoms of arthritis are improved or beneficially changed by administration of the food composition of the present invention.

The food composition of the present invention may be prepared in the form of various foods, for example, beverages, gum, tea, vitamin complexes, powders, granules, tablets, capsules, cookies, rice cakes, breads, and the like. Since the food composition of the present invention is composed of plant extracts having little toxicity and side effects, it can be safely used even for a long period of time for prevention purposes.

Here, when the food composition is prepared in the form of a beverage, there are no particular limitations other than containing the food composition in an indicated ratio, and it may contain various flavoring agents or natural carbohydrates, etc., as additional components, like a conventional beverage. That is, natural carbohydrates include monosaccharides such as glucose, disaccharides such as fructose, sucrose, and common sugars such as polysaccharides, dextrins, cyclodextrins, and sugar alcohols such as xylitol, sorbitol, and erythritol. can do. Examples of the flavoring agent include natural flavoring agents (taumatine, stevia extract (for example, rebaudioside A, glycyrrhizine, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.).

Other food compositions of the present invention include various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners , pH adjusting agents, stabilizers, preservatives, glycerin, alcohol, carbonic acid used in carbonated beverages, and the like.

Using the composition of the present invention, autoimmune diseases or arthritis can be effectively treated.

Figure 1 shows the secretion of IL-10 when co-hanging the target lactic acid bacteria and dendritic cells.

Hereinafter, the present invention will be described in more detail through examples. These examples are only intended to illustrate the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .

Example  One

With dendritic cells  Lactobacillus Coculture

After obtaining white blood cells from the spleen of mice (C57BL / 6), dendritic cells were separated using CD11c microbeads (Miltenyi Biotech) and LS column (Miltenyi Biotech). CD11c microbeads were used at 50 μl per 1 × ^{10 8} white blood cells. Dendritic cells and lactic acid bacteria in the genus Bifidobacterium and Lactobacillus in a culture ratio of 200 μl (RPMI1640, 10% FBS, 5 mM HEPES, 1 mM sodium pyruvate, 1 mM non-essential amino acid, 2 mM L-glutamine, 55 µM 2-mercaptoethanol, 100 U / ml penicillin, 100 µg / ml streptomycin, 150 µg / ml gentamycin, 10 ng / ml GM-CSF) was dispensed into a 96-well plate.

Example  2

IL-10 secretion from dendritic cells Promotion  Screening of lactic acid bacteria through confirmation

After dendritic cells and lactic acid bacteria of Example 1 were co-cultured at 37 ° C. for 16 hours, a culture supernatant was obtained, and the amount of IL-10 was measured using an ELISA kit (eBiosciences). As a control group, Lactobacillus helveticus (HY7801), which is known to promote IL-10 secretion and the group without lactic acid bacteria, was used as a positive control ( Lactobacillus helveticus suppresses experimental rheumatoid arthritis by reducing inflammatory T cell responses.Journal of Functional Foods (2015) 13 : 350) As a result of experiments on a large number of lactic acid bacteria, the following four lactic acid bacteria having excellent ability to induce IL-10 secretion from dendritic cells were selected.

The types of lactic acid bacteria used in the culture were as follows:

Bifidobacterium lactis (B1))

Lactobacillus paracasei (L6)

Lactobacillus curvatus (L8))

Lactobacillus plantarum (L15)

In particular, the lactic acid bacteria induces IL-10 secretion in the existing literature and is a positive control Lactobacillus, which has been found to be effective in rheumatoid arthritis It was confirmed that it showed superior IL-10 secretion inducing efficacy than helveticus (HY7801) (see FIG. 1).

Since the specific parts of the present invention have been described in detail above, it is clear that for those skilled in the art, these specific techniques are only preferred embodiments, and the scope of the present invention is not limited thereto. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims (14)

Bifidobacterium lactis ) A pharmaceutical composition for the treatment of arthritis comprising lactic acid bacteria or a culture medium thereof as an active ingredient. According to claim 1,
The arthritis is at least one selected from the group consisting of osteoarthritis, degenerative arthritis, exfoliative osteochondritis, joint ligament damage, meniscal cartilage damage, arthritis due to infection, psoriatic arthritis, ankylosing spondylitis, rheumatoid arthritis and childhood rheumatoid arthritis, pharmaceutical Composition. According to claim 1,
The pharmaceutical composition is prepared as a powder, granule, capsule, tablet, aqueous suspension, external preparation, suppository or injection solution, pharmaceutical composition. According to claim 1,
Lactic acid bacteria or a culture medium thereof is a pharmaceutical composition in which dendritic cells promote IL-10 secretion. Bifidobacterium lactis ) or a food composition for alleviating or improving arthritis, comprising a culture medium thereof as an active ingredient. The method of claim 5,
The arthritis is at least one selected from the group consisting of osteoarthritis, degenerative arthritis, exfoliative osteochondritis, joint ligament damage, meniscal cartilage damage, arthritis due to infection, psoriatic arthritis, ankylosing spondylitis, rheumatoid arthritis and childhood rheumatoid arthritis, food Composition. Lactobacillus paracasei ( Lactobacillus paracasei ), Lactobacillus cactus ( Lactobacillus curvatus ) and Lactobacillus plantarum ( Lactobacillus plantarum ) selected from the group consisting of a pharmaceutical composition for the treatment of arthritis containing at least one lactic acid bacteria or a culture medium thereof as an active ingredient . The method of claim 7,
The arthritis is at least one selected from the group consisting of osteoarthritis, degenerative arthritis, exfoliative osteochondritis, joint ligament damage, meniscal cartilage damage, arthritis due to infection, psoriatic arthritis, ankylosing spondylitis, rheumatoid arthritis and childhood rheumatoid arthritis, pharmaceutical Composition. The method of claim 7,
The pharmaceutical composition is prepared as a powder, granule, capsule, tablet, aqueous suspension, external preparation, suppository or injection solution, pharmaceutical composition. The method of claim 7,
Lactic acid bacteria or a culture medium thereof is a pharmaceutical composition in which dendritic cells promote IL-10 secretion. Lactobacillus paracasei ( Lactobacillus paracasei ), Lactobacillus cactus ( Lactobacillus curvatus ) and Lactobacillus plantarum ( Lactobacillus plantarum ) selected from the group consisting of lactic acid bacteria or a culture medium thereof for the relief or improvement of arthritis comprising as an active ingredient Food composition. The method of claim 11,
The arthritis is at least one selected from the group consisting of osteoarthritis, degenerative arthritis, exfoliative osteochondritis, joint ligament damage, meniscal cartilage damage, arthritis due to infection, psoriatic arthritis, ankylosing spondylitis, rheumatoid arthritis and childhood rheumatoid arthritis, food Composition. Bifidobacterium autoimmunity comprising at least one lactic acid bacteria selected from the group consisting of lactis ), Lactobacillus paracasei , Lactobacillus curvatus , and Lactobacillus plantarum or a culture medium thereof. Pharmaceutical composition for the treatment of diseases. The method of claim 13,
The autoimmune diseases include rheumatoid arthritis, inflammatory vertebral arthritis, adult Stills disease, macrophage activation syndrome, systemic sclerosis, polymyositis, dermatomyositis, vasculitis, mixed connective tissue disease, Sjogren's syndrome, ulcerative colitis, Crohn's disease, allergic Asthma, allergic rhinitis, atopic dermatitis, margin, fistula, conjunctivitis, psoriasis, vulgar pylori, Parkinson's disease, amyotrophic lateral sclerosis, myasthenia gravis, multiple sclerosis, Alzheimer's disease, stroke, atherosclerosis, vascular restenosis, type I One or more pharmaceutical compositions from the group consisting of diabetes, type II diabetes, diabetic retinopathy and chronic thyroiditis.

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