JP7401156B1 - Agents for preventing and/or treating inflammation in the uterus, fallopian tubes and ovaries - Google Patents

Abstract

The present invention provides a novel preventive and/or therapeutic agent for inflammation in the uterus, fallopian tubes, and ovaries. SOLUTION: An orally administered preventive and/or therapeutic agent for inflammation in the uterus, fallopian tubes, and ovaries, which contains Clostridium butyricum or a culture thereof as an active ingredient. [Selection diagram] None

Description

IPOD FERM BP-2789

The present invention relates to an agent for preventing and/or treating inflammation in the uterus, fallopian tubes, and ovaries.

Inflammation is known to occur in reproductive organs such as the uterus, fallopian tubes, and ovaries due to chlamydia infection, gonococcal infection, herpesvirus infection, bacterial vaginosis, and the like.

In the treatment of inflammation caused by bacterial infections such as chlamydial infections, gonococcal infections, and bacterial vaginosis, drugs such as ceftriaxone, azithromycin, and minocycline are used depending on the causative bacteria (non-patent literature). 1).

Edited and supervised by the Japan Society of Obstetrics and Gynecology, Japan Gynecology Association, Obstetrics and Gynecology Practice Guidelines Gynecology Outpatient Edition 2020, April 23, 2020

An object of the present invention is to provide a novel preventive and/or therapeutic agent for inflammation in the uterus, fallopian tubes, and ovaries.

The inventors have surprisingly found that orally administered Clostridium butyricum can exert anti-inflammatory effects in the uterus, fallopian tubes and ovaries. Based on this knowledge, the present invention was completed.

That is, according to one embodiment of the present invention, there is provided an orally administered preventive and/or therapeutic agent for inflammation in the uterus, fallopian tubes, and ovaries, which contains Clostridium butyricum or a culture thereof as an active ingredient. be done.

FIG. 1 shows a schematic diagram of an experiment using mice in an example. 2 is a graph showing the results of measuring the mass of the removed uterine cervix to ovary in each group of Examples. FIG. 2 is a graph showing the results of measuring the cytokine concentration in the excised cervical canal to ovary region in each group of Examples. FIG.

Hereinafter, an embodiment according to one form of the present invention will be described. The present invention is not limited only to the following embodiments.

In this specification, the range "X to Y" means "more than or equal to X and less than or equal to Y." Further, unless otherwise specified, operations and measurements of physical properties, etc. are performed under conditions of room temperature (20 to 25°C)/relative humidity of 40 to 50% RH.

<Prophylactic and/or therapeutic agents for inflammation in the uterus, fallopian tubes, and ovaries>
One form of the present invention is an orally administered preventive and/or therapeutic agent for inflammation in the uterus, fallopian tubes, and ovaries, which contains Clostridium butyricum or a culture thereof as an active ingredient.

Another form of the invention is Clostridium butyricum or a culture thereof for use as an orally administered prophylactic and/or therapeutic agent for inflammation in the uterus, fallopian tubes and ovaries.

Another form of the invention is the use of Clostridium butyricum or a culture thereof for the manufacture of an orally administered prophylactic and/or therapeutic agent for inflammation in the uterus, fallopian tubes and ovaries.

In this specification, "a prophylactic and/or therapeutic agent for inflammation in the uterus, fallopian tubes, and ovaries" and "an orally administered prophylactic and/or therapeutic agent for inflammation in the uterus, fallopian tubes, and ovaries" are simply referred to as "preventive and/or therapeutic agents for inflammation in the uterus, fallopian tubes, and ovaries." Also called "therapeutic agent".

Examples of inflammation in the uterus, fallopian tubes, and ovaries include endometritis, cervicitis, ovariitis, and salpingitis. In one embodiment, the inflammation in the uterus, fallopian tubes and ovaries according to the invention is preferably at least one selected from the group consisting of endometritis, cervicitis, ovariitis and salpingitis. Bacterial infections, viral infections, and the like are known causes of these inflammations. The preventive and/or therapeutic agent according to the present invention is more effective against inflammation caused by bacterial infection.

Clostridium butyricum is a spore-forming, anaerobic, Gram-positive bacterium that repeats division and growth (vegetative cells) when nutrients are balanced, but when the balance is disrupted, spores are produced within the bacterial body. It is. Many bacteria, not only anaerobic bacteria, easily die when left in a dry state when they have the form of vegetative cells. However, since spores are dormant cells, they have strong resistance to various external environments such as dryness, heat, and chemicals, making them convenient for storage.

Furthermore, as described above, Clostridium butylicum is spore-forming, and when in the spore state, it is resistant to various external environments. For this reason, when Clostridium butylicum is orally administered to humans or animals in the form of spores, even if it comes into contact with digestive fluids such as stomach acid, intestinal fluid, and bile acids, Clostridium butylicum is not completely killed and is removed from the lower part of the small intestine. It is now possible to reach and proliferate at fermentation sites up to the large intestine.

In addition, Clostridium butylicum is widely available commercially as a live bacterial agent, feed additive, and food, and it is highly safe with no side effects observed even when administered to humans, livestock, and other mammals over long periods of time. ing.

Among Clostridium butyricum, Clostridium butyricum Miyairi, Clostridium butyricum NIP1020 (Clostridium butyricum NIP1020), Clostridium butyricum NIP1021 (Clostridium butyricum NIP1021), Clostridium butyricum NIP1021 Clostridium butyricum (FERM P-11868), Clostridium butyricum (FERM P-11868), Clostridium butyricum (FERM P-11869), and Clostridium butyricum (FERM P-11870), Clostridium butyricum ATCC859 (Clostridium butyricum ATCC859), Clostridium butyricum NBRC3315 (Clostridium ridium butyricum NBRC3315), Clostridium Clostridium butyricum ATCC 860 or Clostridium butyricum ATCC 19398 is preferred. More preferably Clostridium butyricum MIYAIRI 588 (FERM BP-2789), Clostridium butyricum MIYAIRI 585 (FERM BP-06815), Clostridium butyricum MIYAIRI 595 (FERM BP-) 06816) and Clostridium butyricum One or more species selected from the group consisting of Clostridium butyricum MIYAIRI 588 (FERM BP-06817), and more preferably Clostridium butyricum MIYAIRI 588 (FERM BP-2789). Clostridium butyricum Miyairi strain 588 was approved as of May 1, 1981 by the Institute of Microbial Technology, Agency of Industrial Science and Technology, Ministry of International Trade and Industry (currently Patent Organism Depositary, National Institute of Technology and Evaluation, Independent Administrative Agency) (292-0818 It was deposited at Kazusa Kamatari 2-5-8, Kisarazu City, Chiba Prefecture, Japan as FERM BP-2789, and on March 6, 1990, it was transferred to an international depositary institution under the Budapest Treaty and has the accession number FERM BP-2789. It has been deposited. Furthermore, FERM BP-2789 will continue to be deposited until March 5, 2050.

Clostridium butyricum Miyairi is commercially available as a viable bacterial agent from Miyarisan Pharmaceutical Co., Ltd., and has no side effects even when administered to humans or animals over a long period of time, so it is particularly suitable for use in the present invention. In addition, as the active ingredient Clostridium butylicum, only one type may be used alone, or two or more types may be used in combination.

In the present invention, a culture of Clostridium butyricum means a culture solution in which Clostridium butyricum is cultured, a residue containing bacteria obtained by centrifuging the culture solution, and a dried product of the residue.

A culture of Clostridium butylicum can be obtained by a known method for culturing microorganisms, for example, the method disclosed in JP-A-08-252088. One embodiment thereof is shown below: Clostridium butyricum at 1.0 (w/v)% peptone, 1.0 (w/v)% yeast extract, 1.0 (w/v)% cornstarch and 0.2%. By inoculating 10 ⁵ to 10 ⁶ cells/mL into a medium containing (w/v)% precipitated calcium carbonate and culturing for 48 hours at 37°C, a "Culture of Clostridium butyricum" was prepared. obtain. Next, the obtained culture solution is centrifuged (2,000-6,000g x 10-30 minutes) to separate "residue containing bacteria obtained by centrifuging the culture solution", and this residue is , 0 to 80°C, preferably 10 to 20°C, for 1 to 24 hours, preferably 5 to 18 hours, or drying at 0 to 80°C, preferably 10 to 20°C, 0.05 to 500 Torr (7Pa 66.7 kPa), preferably 1 to 100 Torr (133 Pa to 13.3 kPa) for 1 to 24 hours, preferably 2 to 15 hours, to obtain a "dried residue". To obtain a dried product, spray drying, freeze drying, etc. may be used.

The medium used for culturing Clostridium butylicum according to the present invention varies depending on the type of bacterial strain used, but it contains a carbon source that can be assimilated by Clostridium butylicum, an appropriate amount of nitrogen source, inorganic salts, vitamins, etc. As long as the medium contains other nutrients, either a synthetic medium or a natural medium may be used.

For example, the carbon source used in the culture medium according to the present invention is not particularly limited as long as it is a carbon source that can be assimilated by the strain used. The carbon source is not necessarily limited to sugars, but in consideration of the growth of bacterial cells, sugars or substances containing sugars that can be utilized by the bacteria used are preferably used. Specific examples of carbon sources that can be used include cellobiose, glucose, fructose, galactose, lactose, maltose, mannose, melibiose, raffinose, salicin, starch, sucrose, trehalose, xylose, dextrin, and molasses. etc. Among these carbon sources, starch, glucose, fructose, sucrose and molasses are preferably used. One or more of the above carbon sources may be selected and used in consideration of Clostridium butyricum to be used. At this time, the concentration of the carbon source added varies depending on the Clostridium butylicum used, the type of carbon source, and the composition of the medium other than the carbon source, but is usually 0.5 to 5 (w/v)%. Preferably it is 2 to 4 (w/v)%.

Nitrogen sources and vitamins include, for example, meat extract, peptone, yeast extract, soybean and wheat hydrolysates such as flavoring liquid, soybean powder, milk casein, casamino acids, various amino acids, corn steep liquor, and other Examples include organic nitrogen compounds such as hydrolysates of animals, plants, and microorganisms, and ammonium salts such as ammonium sulfate. Among these nitrogen sources, peptone, yeast extract, meat extract, corn steep liquor and flavor liquid are preferably used. One or more of the above nitrogen sources and vitamins may be selected and used in order to improve the growth of Clostridium butyricum. At this time, the concentration of the nitrogen source added will vary depending on the strain used, the type of nitrogen source, and the composition of the medium other than the nitrogen source, but when using peptone containing a large amount of nitrogen source, it is usually The amount is 0.5 to 4 (w/v)%, preferably 1 to 3 (w/v)%, and is usually 0. 5 to 5 (w/v)%, preferably 1 to 4 (w/v)%, and furthermore, when using yeast extract or meat extract containing a lot of vitamins, it is usually 0.5 to 4 (w/v)%. (w/v)%, preferably 1 to 3 (w/v)%.

Additionally, inorganic salts include phosphates, hydrochlorides, sulfates, butyrates, propionates and acetic acid, such as magnesium, manganese, calcium, sodium, potassium, molybdenum, strontium, boron, copper, iron, tin and zinc. One or more selected salts can be used. In addition, antifoaming agents, vegetable oils, surfactants, blood and blood components, drugs such as antibiotics, physiologically active substances such as plant or animal hormones, etc. may be appropriately added to the medium, if necessary.

The conditions for culturing in the present invention vary depending on the physiological properties of Clostridium butylicum used in the present invention, such as the growth range (pH, temperature, etc.), but since Clostridium butylicum is obligately anaerobic, It is necessary to culture under anaerobic conditions, such as by lowering the redox potential by aerating nitrogen or carbon dioxide gas, or by adding a reducing agent to the medium. The culture conditions at that time are appropriately selected depending on the growth range of the strain used, the composition of the medium, and the culture method, and are not particularly limited as long as the conditions allow the growth of the present strain. Specifically, the culture temperature is usually 20 to 42°C, preferably 35 to 40°C.

Furthermore, in the present invention, when culturing Clostridium butyricum, growth is promoted by neutralizing the acid produced during the culture with an alkali, so it is preferable to add calcium carbonate to the medium in advance. At this time, the amount of calcium carbonate added is usually 0.1 to 4 (w/v)%, preferably 0.2 to 2.5 (w/v)%. Alternatively, it is also preferable to carry out the neutralization step while keeping the pH of the medium within the set pH range with an aqueous alkaline solution such as sodium hydroxide, sodium bicarbonate, sodium carbonate, potassium hydroxide, and potassium carbonate. In addition, when using an alkaline aqueous solution, "set pH" means the pH of the medium that is preset during the culture period, and "set pH range" means the pH that is allowed during the culture period. It is a pH range, and is generally expressed as a set pH±tolerance. According to the present invention, the set pH is usually set within a range of 5.0 to 7.5, preferably 5.5 to 6.5, and the range of the set pH is set within a range of ±0.5, preferably set pH. The pH is ±0.2.

In the present invention, the pH of the medium during culturing is around neutral, preferably 6.5 to 7.5, at the time of inoculation of the bacteria. In addition, when using an alkaline aqueous solution, it is preferable to maintain the pH within a set pH range while stirring gently to prevent oxygen from being mixed in. By controlling the pH during inoculation and multiplication of bacteria in this way, the density of bacteria can be dramatically increased.

In the culture according to the present invention, the initial culture concentration of Clostridium butylicum is not particularly limited as long as Clostridium butylicum can grow, and is generally the same as that used for culturing Clostridium butylicum. Specifically, it is usually 10 ⁴ to 10 ⁷ cells/mL, preferably 10 ⁵ to 10 ⁶ cells/mL.

The cultures of Clostridium butyricum thus obtained, especially those of Clostridium butyricum MIYAIRI 588 (FERM BP-2789), are useful for preventing inflammation in the uterus, fallopian tubes and ovaries and/or Can exert therapeutic effects.

Clostridium butyricum contained in the prophylactic and/or therapeutic agent according to the present invention may be a living bacterium or a dead bacterium. Clostridium butyricum is preferably a living bacterium (including spores) from the viewpoint of being able to more effectively exhibit the effects of the present invention.

As shown in the Examples below, by orally administering Clostridium butyricum or a culture thereof according to the present invention, it is possible to suppress the increase in mass of the uterus, fallopian tubes, and ovaries caused by inflammation. It can also increase anti-inflammatory cytokines and decrease inflammatory cytokines in the uterus, fallopian tubes, and ovaries where inflammation occurs.

The prophylactic and/or therapeutic agent according to the present invention contains Clostridium butylicum or a culture thereof in an amount (ie, an effective amount) sufficient to exert the desired effect. The prophylactic and/or therapeutic agent may be Clostridium butylicum or a culture thereof itself (consisting of Clostridium butylicum or a culture thereof), or may be in the form of a composition containing Clostridium butylicum or a culture thereof. For example, the prophylactic and/or therapeutic agent may be prepared as a formulation according to a conventional method using additives that are acceptable for formulation. Acceptable additives for formulation include, for example, excipients, stabilizers, preservatives, wetting agents, emulsifiers, lubricants, sweeteners, colorants, flavors, buffers, antioxidants, pH Conditioners, binders, thickeners, dispersants, suspending agents, disintegrants, bacteriostatic agents, surfactants and the like can be mentioned.

The dosage form is not particularly limited as long as it can be administered orally, and can be determined as appropriate. Dosage forms include, for example, tablets, powders, fine granules, granules, capsules, pills, sustained release agents, solutions, suspensions, emulsions, and the like.

The prophylactic and/or therapeutic agent according to the present invention can contain a pharmacologically acceptable carrier. Examples of pharmacologically acceptable carriers include, but are not limited to, excipients such as lactose and starch; binders such as dextrin and cellulose; and solvents such as water and organic solvents.

The prophylactic and/or therapeutic agent according to the present invention can contain other auxiliary ingredients as necessary. Other auxiliary ingredients include antibiotics, vitamins (e.g. vitamin C, vitamin E), amino acids, peptides, minerals (e.g. zinc, iron, copper, manganese, etc.), nucleic acids, polysaccharides, fatty acids. , crude drugs, etc.

The blending ratio of active ingredients in the prophylactic and/or therapeutic agent according to the present invention is not particularly limited. The blending ratio may be 0.01% by weight to 100% by weight based on the entire preventive and/or therapeutic agent.

The dosage of the prophylactic and/or therapeutic agent according to the present invention may be changed as appropriate depending on the symptoms to be treated, pathological condition, age, etc., and is, for example, 0.1 to 1000 mg/kg body weight as the active ingredient.

The preventive and/or therapeutic agent according to the present invention can be administered to mammals, preferably mammals in which inflammation in the uterus, fallopian tubes, and ovaries has occurred or is likely to occur. Here, mammals include primates such as humans, monkeys, gorillas, chimpanzees, orangutans, and non-human animals such as mice, rats, hamsters, guinea pigs, rabbits, dogs, cats, pigs, cows, horses, sheep, camels, and goats. Includes both humans and mammals. Among these, humans are preferred.

(Food and drink composition)
One form of the present invention is a food or drink composition for preventing and/or treating inflammation in the uterus, fallopian tubes, and ovaries, which includes the above-mentioned preventive and/or therapeutic agent and is ingested orally.

The food and drink products according to the present invention should contain the preventive and/or therapeutic agent in an appropriate amount so as to contain the active ingredient of the preventive and/or therapeutic agent according to the present invention, that is, an effective amount of Clostridium butyricum or a culture thereof. is preferred. In this form, "effective amount" means that the active ingredient is contained in an amount that can exhibit the effect of the active ingredient when the individual food or drink is ingested in the amount normally consumed.

The food and drink compositions according to the present invention are prepared as food and drink products by adding conventional additive ingredients such as stabilizers to preventive and/or therapeutic agents, various proteins, sugars, fats, trace elements, vitamins, etc. It may be prepared by further blending them, in liquid, semi-liquid or solid form, in paste form, or in general food and drink products with preventive and/or therapeutic agents added. .

In the present invention, the "food and beverage composition" is not particularly limited as long as it is something other than medicine and can be orally ingested by mammals, and its form is also liquid (solution, suspension). , emulsion, etc.), semi-liquid, powder, or solid molded product. Therefore, the food or drink may be in the form of a drink, for example, or may be in the form of a tablet of a nutritional supplement such as a supplement.

Specifically, the food and drink compositions include, for example, instant noodles, retort foods, canned foods, microwave foods, instant soups, miso soups, freeze-dried foods, and other instant foods; soft drinks, fruit juice drinks, vegetable drinks, and soy milk drinks. Beverages such as coffee drinks, tea drinks, powdered drinks, concentrated drinks, nutritional drinks, and alcoholic drinks; Flour products such as bread, pasta, noodles, cake mixes, fried chicken powder, and bread crumbs; Candy, caramel, chewing gum, chocolate, Sweets such as cookies, biscuits, cakes, pies, snacks, crackers, Japanese sweets, and desserts; sauces, processed tomato seasonings, flavor seasonings, cooking mixes, sauces, dressings, dipping sauces, ingredients for curry and stews Seasonings such as processed oils and fats, butter, margarine, and mayonnaise; Dairy products such as milk drinks, yogurts, lactic acid bacteria drinks, ice creams, and creams; Processed seafood products such as fish, ham, sausage, and seafood paste products Processed livestock products such as meat hams and sausages; Processed agricultural products such as canned agricultural products, jams and marmalades, pickled foods, boiled beans, and cereals; Frozen foods; and nutritional foods.

In the present invention, "food and beverage compositions" include those classified as health foods, functional foods, foods for specified health uses, nutritional supplements, foods with disease risk reduction labeling, or foods for the sick. is also included. Furthermore, when used for mammals other than humans, the term "food/beverage composition" can be used to include feed.

In the food and drink composition according to the present invention, in addition to the above-mentioned active ingredients, components having other functions may be further added. For example, the active ingredient of the present invention may be added to foods ingested in daily life, health foods, functional foods, and supplements (for example, foods containing one or more minerals such as calcium and magnesium, and vitamins such as vitamin K). By blending, in addition to the effects of the present invention, it is possible to provide food and drink products that have functions based on other ingredients.

The proportion of the prophylactic and/or therapeutic agent in the food or drink is not particularly limited, but the amount of the preventive and/or therapeutic agent is, for example, 0.001 to 50% by mass based on the dry weight of the food or drink.

<Method for preventing and/or treating inflammation in the uterus, fallopian tubes, and ovaries>
One form of the present invention is a method for preventing and/or treating inflammation in the uterus, fallopian tubes, and ovaries, which comprises orally administering an effective amount of the above-mentioned prophylactic and/or therapeutic agent to a subject in need thereof. be.

In this embodiment, the "subject" and "inflammation in the uterus, fallopian tubes, and ovaries" are the same as those explained in the above-mentioned preventive and/or therapeutic agent, so their explanation will be omitted.

In this form, "effective amount" refers to the active ingredient of the prophylactic and/or therapeutic agent that is at least required to exert the desired effect of preventing and/or treating inflammation in the uterus, fallopian tubes, and ovaries (i.e. , Clostridium butyricum or its culture).

<An agent for promoting the production of anti-inflammatory cytokines and/or suppressing the production of inflammatory cytokines in the uterus, fallopian tubes, and ovaries>
One form of the present invention includes Clostridium butyricum or a culture thereof as an active ingredient, and is orally administered to promote the production of anti-inflammatory cytokines and/or to stimulate the production of inflammatory cytokines in the uterus, fallopian tubes, and ovaries. It is an agent for suppressing production.

In this specification, "an agent for promoting the production of anti-inflammatory cytokines and/or suppressing the production of inflammatory cytokines in the uterus, fallopian tubes, and ovaries" is simply referred to as "an agent for promoting and/or suppressing the production of cytokines." Also called.

As shown in the Examples below, by orally administering Clostridium butylicum or a culture thereof according to the present invention, anti-inflammatory cytokines can be increased and inflammatory cytokines can be decreased. It is thought that an increase in anti-inflammatory cytokines and a decrease in inflammatory cytokines exerts an anti-inflammatory effect and suppresses the increase in mass caused by inflammation in the uterus, fallopian tubes, and ovaries.

Examples of anti-inflammatory cytokines include IL-10, TGF-β, and IL-4. In one embodiment, the anti-inflammatory cytokine is at least one selected from the group consisting of IL-10, TGF-β and IL-4.

Examples of inflammatory cytokines include TNF-α, IFN-γ, IL-17A, and IL-6. In one embodiment, the inflammatory cytokine is at least one selected from the group consisting of TNF-α, IFN-γ, IL-17A and IL-6.

In this embodiment, "Clostridium butylicum or a culture thereof" is the same as that explained in the above-mentioned preventive and/or therapeutic agent, so the explanation will be omitted.

The agent for promoting and/or suppressing cytokine production according to the present invention contains Clostridium butylicum or a culture thereof in an amount (ie, an effective amount) sufficient to exert the desired effect. The agent for promoting and/or suppressing cytokine production may be Clostridium butylicum or a culture thereof itself (consisting of Clostridium butylicum or a culture thereof), or may be in the form of a composition containing Clostridium butylicum or a culture thereof. There may be. For example, an agent for promoting and/or suppressing cytokine production may be prepared as a formulation according to a conventional method using additives that are acceptable for formulation. In this form, "additives and dosage forms acceptable for formulation" are the same as the above-mentioned prophylactic and/or therapeutic agents, and therefore their explanations will be omitted.

The agent for promoting and/or suppressing cytokine production according to the present invention can contain a pharmacologically acceptable carrier. Furthermore, the agent for promoting and/or suppressing cytokine production according to the present invention can contain other auxiliary components as necessary. In this form, the "pharmacologically acceptable carrier" and "other auxiliary ingredients" are the same as those of the above-mentioned prophylactic and/or therapeutic agent, and therefore their explanations will be omitted.

The blending ratio of active ingredients in the agent for promoting and/or suppressing cytokine production according to the present invention is not particularly limited. The blending ratio may be 0.01% by mass to 100% by mass based on the entire agent for promoting and/or suppressing cytokine production.

The dosage of the agent for promoting and/or suppressing cytokine production according to the present invention may be changed as appropriate depending on the symptoms to be treated, pathological condition, age, etc. It is.

The agent for promoting and/or suppressing cytokine production according to the present invention can be administered to mammals. The mammal is preferably a mammal that needs to promote the production of anti-inflammatory cytokines and/or suppress the production of inflammatory cytokines in the uterus, fallopian tubes, and ovaries (e.g., when inflammation is occurring or is likely to occur). mammals). In this embodiment, "mammal" is the same as the above-mentioned prophylactic and/or therapeutic agent, so the explanation will be omitted.

<Embodiment>
Embodiments of the present invention are illustrated below.
[1] An orally administered preventive and/or therapeutic agent for inflammation in the uterus, fallopian tubes, and ovaries, containing Clostridium butyricum or a culture thereof as an active ingredient.
[2] The prophylactic and/or therapeutic agent according to [1], wherein the Clostridium butyricum is Clostridium butyricum MIYAIRI 588 (FERM BP-2789).
[3] In [1] or [2], the inflammation in the uterus, fallopian tubes and ovaries is at least one selected from the group consisting of endometritis, cervicitis, oophoritis and salpingitis. Prophylactic and/or therapeutic agents as described.
[4] A food or drink for the prevention and/or treatment of inflammation in the uterus, fallopian tubes and ovaries, which contains the preventive and/or therapeutic agent according to any one of [1] to [3] as an active ingredient and is ingested orally. Composition.
[5] Treatment of inflammation in the uterus, fallopian tubes, and ovaries, including orally administering an effective amount of the prophylactic and/or therapeutic agent according to any one of [1] to [3] to a subject in need thereof. Prevention and/or treatment methods.
[6] Clostridium butyricum or a culture thereof for use as a prophylactic and/or orally administered therapeutic agent for inflammation in the uterus, fallopian tubes, and ovaries.
[7] Use of Clostridium butyricum or a culture thereof for the production of an orally administered prophylactic and/or therapeutic agent for inflammation in the uterus, fallopian tubes, and ovaries.
[8] Contains Clostridium butyricum or a culture thereof as an active ingredient and is administered orally for promoting the production of anti-inflammatory cytokines and/or suppressing the production of inflammatory cytokines in the uterus, fallopian tubes and ovaries. agent.
[9] The agent according to [8], wherein the anti-inflammatory cytokine is at least one selected from the group consisting of IL-10, TGF-β, and IL-4.
[10] The agent according to [8] or [9], wherein the inflammatory cytokine is at least one selected from the group consisting of TNF-α, IFN-γ, IL-17A, and IL-6.

The present invention will be explained below using specific examples, but the present invention is not limited to these examples. In addition, unless otherwise specified, the work was performed at room temperature (25° C.).

Animal experiments using mice were conducted with approval from the Animal Experiment Committee, an ethics committee established by Aichi Medical University (approval number: 2022-50).

A schematic diagram of the experiment in mice is shown in Figure 1.

Balb/c mice (8-9 weeks old, female, wild type, purchased from Charles River Japan Co., Ltd.) were subjected to a 7-day quarantine and acclimatization period, and all mice had no abnormalities in health status and weight loss was observed. After confirming that there was no Mice were allowed free access to food and water in a breeding environment with 12 hours of light, temperature of 20-26°C, and humidity of 30-70%.

Forty mice were divided into Ope- group (n=10), Ope+ group (n=10), Ope+LPS group (n=10) and Ope+LPS CBM group (n=10).

In the Ope+ group, Ope+LPS group, and Ope+LPS CBM group, under anesthesia, incisions were made in the left dorsal region on the start day of the experiment (Day 0), and on the right back on the 5th day after the start of the experiment (Day 5), and the uterine horns and ovaries of the mice were incised. After the site was exposed, the tissue was returned to the body and the endothelium and epidermis were sutured. In the Ope+ group, 0.1 mL of phosphate buffered saline (PBS) was administered into the uterine horn using a syringe at the time of exposure of the uterine horn on the experiment start day (Day 0) and the 5th day after the experiment start (Day 5). , Ope+LPS group, and Ope+LPS CBM group, 0.1 mL of lipopolysaccharide (LPS, 2.5 mg/mL) was administered into the uterine horn using a syringe.

In the Ope+LPS CBM group, 3.4 x 10 8 cfu of Clostridium butyricum MIYAIRI 588 (FERM BP-2789) culture (hereinafter also simply referred to as "CBM588") suspended in physiological saline was ^used . /mouse body weight (30 g)/day (500 mg/kg/day) was orally administered every day using a sonde from the start of the experiment (Day 0) to the 9th day after the start of the experiment. The amount of bacteria in CBM588 was 2.2×10 ¹⁰ cfu/g.

In the Ope- group, there was no incision, administration of PBS and LPS, and oral administration of CBM588.

On the 10th day after the start of the experiment (Day 10), the mice were euthanized. After ligating the cervix with a thread, the region from the cervix to the ovaries (uterine tissue: uterus, fallopian tubes, and ovaries) was removed. The mass of the excised site was measured. The results are shown in Figure 2.

Next, the excised area was treated with an extract solution (RIPA buffer: 50 mmol/L Tris-HCl buffer (pH 7.6), 150 mmol/L NaCl, 1% Nonidet P40 Substitute, 0.5% Sodium Deoxycholate, Protease Inhibito). r Cocktail (1× )) and then centrifuged, and the cytokine protein concentration of the supernatant was measured by ELISA. The results are shown in Figure 3.

The cytokines measured and the ELISA kits used are shown in Table 1.

As shown in FIG. 2, it can be seen that the increase in uterine mass caused by inflammation was significantly suppressed by oral administration of CBM588 in the Ope+LPS CBM group compared to the Ope+LPS group.

Furthermore, as shown in Figure 3, regarding cytokines in the excised site, administration of CBM588 decreased the concentration of anti-inflammatory cytokines IL-10, TGF-β, and IL-4 in the Ope+LPS CBM group. It can be seen that the concentrations of the inflammatory cytokines TNF-α, IFN-γ, IL-17A and IL-6 decreased.

Claims (7)

It contains Clostridium butyricum MIYAIRI 588 (FERM BP-2789) or a culture thereof as an active ingredient and is orally administered , RI 588, FERM BP-2789) , a prophylactic and/or therapeutic agent for inflammation in the uterus, fallopian tubes and ovaries, which is a live bacterium . The prevention and/or treatment according to claim 1 , wherein the inflammation in the uterus, fallopian tubes, and ovaries is at least one selected from the group consisting of endometritis, cervicitis, oophoritis, and salpingitis. agent. A food or drink composition for preventing and/or treating inflammation in the uterus, fallopian tubes, and ovaries, which is orally ingested and contains the preventive and/or therapeutic agent according to claim 1 or 2 as an active ingredient. It contains Clostridium butyricum MIYAIRI 588 (FERM BP-2789) or a culture thereof as an active ingredient and is orally administered , RI 588, FERM BP-2789) , a viable bacterial agent for suppressing the production of inflammatory cytokines in the uterus, fallopian tubes and ovaries. The agent according to claim 4 , wherein the inflammatory cytokine is at least one selected from the group consisting of TNF-α, IFN-γ, IL-17A, and IL-6. The agent according to claim 4 or 5 , which is used for promoting the production of anti-inflammatory cytokines in the uterus, fallopian tubes, and ovaries. The agent according to claim 4 or 5 , which is used for the prevention and/or treatment of inflammation in the uterus, fallopian tubes, and ovaries.