Agent for improving joint movement or alleviating joint pain, food / beverage or feed for improving joint movement or alleviating joint pain, and medicinal composition for improving joint movement or alleviating joint pain

Abstract

An agent for improving joint movement or alleviating joint pain which contains Bacillus subtilis bacterium and / or a Bacillus subtilis culture product.

Description

Agent for improving joint movement or alleviating joint pain, food, drink or feed for improving joint movement or alleviating joint pain, and pharmaceutical composition for improving joint movement or alleviating joint pain

[0001] The present invention relates to an agent for improving joint movement or relieving joint pain, a food, drink or feed for improving joint movement or relieving joint pain, and a pharmaceutical composition for improving joint movement or relieving joint pain.

[0002] In Japan, the proportion of elderly people is increasing year by year. Therefore, it is becoming increasingly important to improve the quality of life of the elderly. One of the symptoms associated with aging is osteoarthritis of the knee. This condition is caused by the wear of the cartilage that cushions the knee joint and a decrease in muscle strength, which leads to inflammation and deformation of the knee joint, resulting in pain. The estimated number of people aged 50 and over suffering from osteoarthritis of the knee is said to be as high as 24 million. Joint disease is also a major cause of the need for assistance and nursing care, and more proactive measures are needed.

[0003] To date, glucosamine, Acanthosoma extract, salmon nasal cartilage-derived proteoglycan, undenatured type II collagen, collagen peptides, and the like have been reported as ingredients that have the effect of promoting the repair of damaged cartilage in knee osteoarthritis.

[0004] It has been reported that Bacillus subtilis has various effects and can be used as an active ingredient in various agents, such as an agent for increasing intestinal butyric acid-producing bacteria (see, for example, Patent Document 1), an agent for suppressing the increase and / or decrease of bifidobacteria in the large intestine (see, for example, Patent Document 2), an agent for preventing norovirus infection (see, for example, Patent Document 3), an agent for improving functional gastrointestinal disorders (see, for example, Patent Document 4), an agent for improving bone metabolism or bone density (see, for example, Patent Document 5), an agent for preventing and treating inflammatory bowel disease (see, for example, Patent Document 6), an agent for improving lipid metabolism (see, for example, Patent Document 7), and an agent for improving renal function (see, for example, Patent Document 8).

[0005] JP 2013-147469 A, International Publication No. 2011 / 111783, JP 2016-121086 A, JP 2017-109973 A, JP 2017-226649 A, International Publication No. 2008 / 069102, International Publication No. 2008 / 023608, International Publication No. 2008 / 023607

[0006] As mentioned above, although ingredients useful for knee osteoarthritis have been reported, there remains a strong demand for new materials that are highly safe and can improve the movement of joints, including the knee, and alleviate joint pain, and there is currently a strong demand for such materials to be provided promptly.

[0007] The present invention aims to solve the above-mentioned problems of the prior art and to achieve the following object: That is, the present invention aims to provide a joint movement improving or joint pain relieving agent, a food, drink, or feed for improving joint movement or relieving joint pain, and a pharmaceutical composition for improving joint movement or relieving joint pain, which are highly safe and have an excellent effect of improving joint movement or relieving joint pain.

[0008] In order to solve the above problems, the present inventors conducted extensive research and found that oral administration of highly safe Bacillus subtilis cells and / or a culture thereof significantly improves joint movement and alleviates joint pain.

[0009] The present invention is based on the findings of the present inventors, and provides the following means for solving the above problems. That is, <1> An agent for improving joint movement or relieving joint pain, characterized by containing Bacillus subtilis cells and / or a culture. <2> The agent for improving joint movement or relieving joint pain according to <1>, wherein the culture is a soybean culture. <3> The agent for improving joint movement or relieving joint pain according to <1> or <2>, wherein the culture is a soybean culture. <4> The agent for improving joint movement or relieving joint pain according to any of <1> to <3>, wherein the Bacillus subtilis is Bacillus subtilis C-3102 strain (FERM BP-1096). <5> The agent for improving joint movement or relieving joint pain according to any of <1> to <4>, which is taken orally. <6> A dose of 2.0 x 10 per day ８ ~2.5 x 10 １１ <7> The agent for improving joint movement or relieving joint pain according to any one of <1> to <5>, which is formulated so that Bacillus subtilis CFU can be ingested. ９ ~2.0 x 10 １０ The agent for improving joint movement or alleviating joint pain according to any one of <1> to <6>, which is formulated so that CFU of Bacillus subtilis can be ingested. <8> A food, drink, or feed for improving joint movement or alleviating joint pain, which contains Bacillus subtilis cells and / or a culture. <9> A pharmaceutical composition for improving joint movement or alleviating joint pain, which contains Bacillus subtilis cells and / or a culture.

[0010] According to the present invention, the above-mentioned problems of the prior art can be solved, the above-mentioned objects can be achieved, and it is possible to provide a joint movement improving or joint pain relieving agent, a food, drink or feed for improving joint movement or relieving joint pain, and a pharmaceutical composition for improving joint movement or relieving joint pain, which are highly safe and have an excellent effect of improving joint movement or relieving joint pain.

[0011] FIG. 1 is a graph showing the test results of Test Example 1. In FIG. 1, "*" represents "p<0.05". FIG. 2 is a graph showing the change over time in the "JKOM score" in the test of Test Example 2. In FIG. 2, "*" represents "p<0.05". FIG. 3 is a graph showing the change over time in the "daily life status score" in the test of Test Example 2. In FIG. 2, "*" represents "p<0.05".

[0012] (Agent for Improving Joint Mobility or Relieving Joint Pain) The agent for improving joint mobility or relieving joint pain of the present invention contains at least Bacillus subtilis cells and / or a culture thereof as an active ingredient, and may further contain other ingredients as necessary.

[0013] <Bacterial Cells and / or Culture of Bacillus subtilis> Bacillus subtilis has long been deeply involved in human life, for example, in enzyme production and food production, and there is a wealth of information about its functionality; however, its usefulness for improving joint movement or alleviating joint pain has not been known. Bacillus subtilis is an obligately aerobic, Gram-positive bacterium that forms spores that are resistant to high temperatures and digestive enzymes. One embodiment of the agent for improving joint movement or alleviating joint pain of the present invention contains Bacillus subtilis in a spore state, and therefore, even when orally ingested, it can reach the intestine alive and exert its effects.

[0014] The mycological properties of Bacillus subtilis are described in, for example, Burgess' Manual of Bacteriology, Vol. 11 (1986), and specifically, it has the following characteristics: (1) Gram-positive (2) Forms oval spores (3) Bacillus (4) Motility: Yes (5) Aerobic (6) Catalase: Positive (7) Growth at 50°C: + (8) Growth at pH 5.7: + (9) Utilization of citrate: + (10) Presence or absence of acid production from sugars: arabinose, glucose, xylose, mannitol: + (11) VP reaction: + (12) Starch hydrolysis: + (13) Nitrate reduction: + (14) Indole production: - (15) Gelatin hydrolysis: + (16) Casein hydrolysis: + (17) Film formation in liquid medium: + (18) Milk coagulation: - (19) Milk peptonization: +

[0015] The Bacillus subtilis used in the present invention is not particularly limited and can be appropriately selected depending on the purpose, but a preferred example is Bacillus subtilis C-3102. Bacillus subtilis C-3102 was domestically deposited on December 25, 1985, at the Fermentation Research Institute, Agency of Industrial Science and Technology, Ministry of International Trade and Industry (1-1-3 Higashi, Yatabe-cho, Tsukuba-gun, Ibaraki Prefecture). Subsequently, on June 28, 1986, a request for transfer to international deposit under the Budapest Treaty was made, and the strain is currently continuously deposited at the Patent Organism Depositary, National Institute of Technology and Evaluation (Room 120, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture) under accession number FERM BP-1096. The depositor of the Bacillus subtilis C-3102 strain at the time of its deposition was Calpis Food Industry Co., Ltd. (2-20-3 Ebisu Nishi, Shibuya-ku, Tokyo), and the current depositor is Asahi Group Holdings, Ltd. (1-23-1 Azumabashi, Sumida-ku, Tokyo).

[0016] Bacillus subtilis C-3102 strain is known to have various useful effects, such as an intestinal regulating effect (WO 2011 / 111783), a lipid metabolism improving effect (WO 2008 / 23608), and a renal function improving effect (WO 2008 / 023607). Food products containing Bacillus subtilis C-3102 strain are already commercially available, and its safety when taken over a long period of time has been established.

[0017] Soybean cultures of Bacillus subtilis C-3102 strain have effects on birds and livestock, such as improving the intestinal flora, promoting weight gain, preventing infection, strengthening eggshells, improving meat quality, and reducing fecal odor, and are used as feed additives (JP-B 4-24022) and as an eggshell strength enhancer for birds (JP-A 10-108628). This strain is also known to have health benefits such as regulating the intestines and reducing intestinal putrefactive products (Journal of the Society of Intestinal Microbiology, Vol. 18, No. 2, pp. 93-99 (2004)).

[0018] The Bacillus subtilis C-3102 strain is characterized in that a fragment of approximately 700 bps is amplified when PCR is performed using the PCR primers of Sequences 1 and 2 below. There is no other Bacillus subtilis strain in which amplification occurs with both of these PCR primers. The approximately 700 bp fragment amplified using the genome of the Bacillus subtilis C-3102 strain as a template is characterized by not having any homology to amylase sequences, and in this respect, the C-3102 strain is clearly distinguished from other Bacillus subtilis strains. Sequence 1 (forward primer): 5'-GCCCCGCACATACGAAAAGACTGGCTGAAA-3' (SEQ ID NO: 1) Sequence 2 (reverse primer): 5'-GGATCCCACGTTGTGATTAAAGCAGCGAT-3' (SEQ ID NO: 2)

[0019] Furthermore, the Bacillus subtilis C-3102 strain has the following properties: (1) it does not contain plasmid DNA; (2) its genomic DNA was prepared, digested with the restriction enzyme NotI or SfiI, and separated by agarose gel electrophoresis; the digestion pattern was as shown in previous reports (FIG. 1 in WO 2008 / 023607 and FIG. 1 in WO 2008 / 023608); and (3) it has no resistance to ampicillin, chloramphenicol, ciprofloxacin, erythromycin, gentamicin, kanamycin, linezolid, quinupristin / dalfopristin, rifampin, streptomycin, tetracycline, trimethoprim, or vancomycin (minimum inhibitory concentrations of each were 0.03 to 4 μg / ml).

[0020] Bacillus subtilis can be cultured using a liquid or solid medium containing carbon sources, nitrogen sources, inorganic substances, and the like commonly used in microbial culture. Carbon sources may be any carbon source assimilable by Bacillus subtilis, such as glucose, fructose, sucrose, starch, and molasses. Nitrogen sources include, for example, peptone, casein hydrolysate, meat extract, and ammonium sulfate. Furthermore, salts of phosphate, potassium, magnesium, calcium, sodium, iron, manganese, and the like, vitamins, amino acids, surfactants, and the like may also be added as needed. In addition to these synthetic media, natural substances such as soybean meal may also be used for culture. Aerobic conditions are preferred, and aerated and agitated liquid culture using a jar fermenter is preferred. The culture temperature is 20 to 50°C, particularly 30 to 45°C. The culture time is 12 hours to 7 days. The initial culture pH is 5 to 9, particularly preferably 6 to 8.

[0021] The culture thus obtained contains Bacillus subtilis cells, a medium, and a fermentation product. The culture may be used as an agent for improving joint movement or relieving joint pain, or the culture may be concentrated. Alternatively, excipients may be added to the concentrated culture and the concentrated culture may be used as a formulation such as a dry powder, granules, or tablets. Alternatively, cells isolated from the culture may be used (e.g., spores), or the culture may be used after removing the cells (e.g., culture supernatant), or a culture containing the cells may be used. In one embodiment, Bacillus subtilis is cultured using edible natural products such as soybean meal, boiled soybeans, boiled azuki beans, cooked rice, barley rice, wheat bran, boiled corn, or other grains, and the culture is incorporated directly into foods, beverages, feed, etc., without separating the cells from the culture. In another embodiment, the culture supernatant of Bacillus subtilis is isolated and incorporated into foods, pharmaceuticals, etc. In yet another embodiment, Bacillus subtilis cells (spores) are incorporated into foods, medicines, etc.

[0022] Among the above cultures, soybean cultures cultured using soybean-derived substances are preferred. Examples of soybean-derived substances include soybean oil cake and boiled soybeans, with soybean oil cake being particularly preferred.

[0023] The Bacillus subtilis may be any of spores, a mixture of spores and germinants, or germinants, but the agent for improving joint movement or relieving joint pain preferably contains Bacillus subtilis spores.

[0024] <Other Components> The other components in the agent for improving joint mobility or relieving joint pain are not particularly limited and can be selected appropriately depending on the purpose as long as they do not impair the effects of the present invention, and examples thereof include components that can be used when formulating the agent as described below. The other components may be used alone or in combination of two or more. The content of the other components in the agent for improving joint mobility or relieving joint pain is not particularly limited and can be selected appropriately depending on the purpose.

[0025] The agent for improving joint movement or relieving joint pain of the present invention safely and simply improves joint movement or relieves joint pain. The agent for improving joint movement or relieving joint pain of the present invention is preferably taken orally. The agent for improving joint movement or relieving joint pain of the present invention can particularly improve knee joint movement or relieve knee joint pain, and therefore has the function of supporting knee joint movement and relieving pain and discomfort in the knee joint.

[0026] The agent for improving joint movement or relieving joint pain of the present invention may be appropriately formulated together with pharmacologically acceptable carriers and additives and administered in the form of a liquid, powder, granules, capsules, tablets, etc., or may be added to food, drink, or feed.

[0027] When the agent for improving joint movement or relieving joint pain of the present invention is formulated as a tablet, examples of pharmacologically acceptable carriers to be used include excipients, lubricants, binders, disintegrants, preservatives, antioxidants, coloring agents, and flavoring agents, but are not limited to these, and other commonly used carriers can also be used as appropriate.

[0028] Examples of excipients include lactose, sucrose, D-mannitol, D-sorbitol, starch, pregelatinized starch, dextrin, crystalline cellulose, low-substituted hydroxypropyl cellulose, sodium carboxymethylcellulose, gum arabic, pullulan, light anhydrous silicic acid, synthetic aluminum silicate, and magnesium aluminometasilicate.

[0029] Examples of lubricants include sugar esters such as sucrose fatty acid esters and glycerin fatty acid esters, calcium stearate, magnesium stearate, stearic acid, stearyl alcohol, hydrogenated oils such as powdered vegetable oils and fats, waxes such as white beeswax, talc, silicic acid, and silicon.

[0030] Examples of binders include pregelatinized starch, sucrose, gelatin, gum arabic, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, crystalline cellulose, sucrose, D-mannitol, trehalose, dextrin, pullulan, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, and polyvinylpyrrolidone.

[0031] Examples of disintegrants include lactose, sucrose, starch, carboxymethylcellulose, carboxymethylcellulose calcium, croscarmellose sodium, carboxymethylstarch sodium, light anhydrous silicic acid, and low-substituted hydroxypropylcellulose.

[0032] Examples of preservatives include parahydroxybenzoates, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid, and sorbic acid, while examples of antioxidants include sulfites and ascorbic acid salts.

[0033] Examples of coloring agents include water-soluble food tar dyes, water-insoluble lake dyes, and natural dyes such as β-carotene, chlorophyll, and red iron oxide.

[0034] Examples of sweeteners include sodium saccharin, dipotassium glycyrrhizinate, aspartame, and stevia.

[0035] In addition to the above, the composition may contain, for example, vitamins, minerals, amino acids, tea extracts such as catechin and sweet tea, glucosamine, chondroitin, plant extracts such as saw palmetto and apple, polyphenols contained in grapes and apples, lactic acid bacteria, bifidobacteria, butyric acid bacteria, natto bacteria, saccharifying bacteria, yeast, starch, dextrin, gum arabic, sugar alcohols such as mannitol and xylitol, cellulose derivatives such as crystalline cellulose and hydroxypropyl cellulose, gelatin, glycerin, fermented milk, lactoferrin, dietary fiber, flavors, etc.

[0036] In particular, tablets containing the agent for improving joint movement or relieving joint pain of the present invention preferably contain monosaccharides such as glucose and fructose, disaccharides such as sucrose and lactose, oligosaccharides such as isomalt and cyclodextrin, sugars such as granulated sugar, maltose, and lactose, and starch, and it is particularly preferable to contain maltose and starch.

[0037] The tablets may be sugar-coated, coated, or plain. Sugar-coated and coated tablets generally have lower disintegration properties than plain tablets, but have the advantage of making the flavor and odor of the active ingredient less noticeable. There are no particular restrictions on the shape, structure, or size of the tablets, and they can be selected appropriately depending on the purpose.

[0038] The amount of intake of the agent for improving joint movement or relieving joint pain of the present invention is not particularly limited and can be appropriately selected depending on the purpose. ８ ~2.5 x 10 １１ It is preferable that the formulation is such that 2.0 x 10 CFU of Bacillus subtilis can be ingested. ９ ~2.0 x 10 １０ More preferably, the formulation is such that CFU of Bacillus subtilis can be ingested.

[0039] The number of times the agent for improving joint mobility or relieving joint pain of the present invention is taken is not particularly limited and can be appropriately selected depending on the purpose, but is preferably taken once to five times a day, more preferably once to four times a day, and particularly preferably taken in divided doses of once to three times a day or once a day. The agent for improving joint mobility or relieving joint pain of the present invention is desirably formulated as a tablet suitable for administration as described above.

[0040] For example, when formulated as tablets, the amount of Bacillus subtilis per tablet is not particularly limited and can be appropriately selected depending on the purpose. ７ ~2.5 x 10 １１ CFU is preferred, 2.0 x 10 ８ ~9.0 x 10 １０ CFU is more preferably 4.0 x 10 ８ ~4.0 x 10 １０ More preferably, 4.0 x 10 CFU ８ ~2.0 x 10 １０ CFUs are particularly preferred.

[0041] The number of tablets to be taken at one time is not particularly limited and can be appropriately selected depending on the purpose, and examples include 1 to 5 tablets.

[0042] The target of the agent for improving joint movement or relieving joint pain of the present invention is not particularly limited, and can be selected appropriately depending on the purpose, so long as it is anyone who needs to take the agent, and examples include people who have joint pain or discomfort, and people who need support for joint movement due to aging.

[0043] Bacillus subtilis, the active ingredient of the agent for improving joint mobility or relieving joint pain of the present invention, is effective in small amounts and for a short period of time, has excellent storage stability and acid resistance, and easily reaches the intestines and proliferates there, so that a sustained improvement in joint mobility or relief of joint pain can be expected. Meanwhile, Bacillus subtilis of the present invention can be safely ingested over a long period of time. Therefore, the agent for improving joint mobility or relieving joint pain of the present invention is preferably used continuously for 4 weeks or more, preferably 8 weeks or more, and more preferably 12 weeks or more.

[0044] The agent for improving joint movement or relieving joint pain of the present invention can improve joint movement or relieve joint pain. The active ingredient, Bacillus subtilis, has long been used in foods and the like and can be safely taken orally over a long period of time. Therefore, the agent for improving joint movement or relieving joint pain of the present invention is useful not only as a pharmaceutical product, but also as a quasi-drug, food or drink, or feed intended to improve joint movement or relieve joint pain.

[0045] (Food, drink, or feed for improving joint movement or relieving joint pain) The food, drink, or feed of the present invention for improving joint movement or relieving joint pain contains at least Bacillus subtilis cells and / or a culture as an active ingredient, and may further contain other ingredients as necessary.

[0046] The Bacillus subtilis cells and / or culture are the same as the Bacillus subtilis cells and / or culture described in the above section of the present invention (Agent for improving joint movement or relieving joint pain).

[0047] The other ingredients in the food, drink, or feed for improving joint movement or alleviating joint pain are not particularly limited and can be selected appropriately depending on the type of food, drink, or feed. The content of the other ingredients in the food, drink, or feed for improving joint movement or alleviating joint pain is also not particularly limited and can be selected appropriately depending on the type of food, drink, or feed.

[0048] The type of food or beverage is not particularly limited and can be appropriately selected depending on the purpose, and examples include beverages, confectionery tablets, pastes, bread, processed fish products, dairy products, jellies, candies, retort pouch foods, cookies, castella cakes, biscuits, chocolates, etc. Alternatively, the agent for improving joint mobility or relieving joint pain of the present invention may be added to these various food materials. The food or beverage of the present invention can be provided as a health drink, health food, or functional food.

[0049] The amount of Bacillus subtilis cells and / or cultures contained in a food or drink is determined appropriately depending on the type and purpose of the food or drink. Taking into account the normal intake of food or drink, the amount of Bacillus subtilis cells and / or cultures contained in a food or drink is determined appropriately depending on the type and purpose of the food or drink. ８ ~2.5 x 10 １１It is preferable that the composition is such that CFU of Bacillus subtilis can be ingested, and 2.0 x 10 ９ ~2.0 x 10 １０ It is more preferable that the composition be formulated so that CFU of Bacillus subtilis can be ingested.

[0050] The number of times of ingestion per day can also be appropriately selected depending on the type of food or drink and the purpose. For example, in the case of health foods and functional foods, it is preferable to ingest them once to five times a day, more preferably once to four times a day, and particularly preferably once to three times a day or once a day. Continuous ingestion of the food or drink of the present invention can improve joint movement and relieve joint pain.

[0051] The target of the feed is not particularly limited and can be appropriately selected depending on the purpose, and examples include birds such as chickens, mammals such as cows, pigs, sheep, and goats, fish, etc. The feed may be produced by adding the agent for improving joint movement or relieving joint pain of the present invention to an existing feed.

[0052] The amount of Bacillus subtilis cells and / or cultures contained in the feed is determined appropriately depending on the target and purpose of the feed. For example, in the case of cattle, for example, 1 x 10 per day ６ It is preferable to mix it so that the CFU / kg or more is 5 x 10 ６ ~6 x 10 ８ It is more preferable to compound the amount of CFU / kg per day. In the case of birds, for example, 1 x 10 ３ ~1 x 10 １１ It is preferable to mix it so that it becomes CFU / g, and 1 × 10 ３ ~1 x 10 ７ It is more preferable to mix it so that the CFU / g is 1 x 10 ４ ~1 x 10 ６ It is particularly preferable to compound the amount so as to be CFU / g.

[0053] The feed may contain vitamins, minerals, amino acids, flavorings, acidulants, baking soda, emulsifiers, stabilizers, etc. depending on the subject. Continuously feeding the feed of the present invention to livestock etc. can improve joint movement and alleviate joint pain.

[0054] As a method for producing the food, drink, or feed, any known method can be appropriately selected, except that the bacterial cells and / or culture of Bacillus subtilis are blended.

[0055] (Pharmaceutical composition for improving joint movement or relieving joint pain) The pharmaceutical composition of the present invention for improving joint movement or relieving joint pain contains at least Bacillus subtilis cells and / or a culture as an active ingredient, and may further contain other ingredients as necessary.

[0056] The pharmaceutical composition of the present invention for improving joint movement or relieving joint pain can be used, for example, to prevent or treat discomfort in joint movement, joint pain, and the like.

[0057] The Bacillus subtilis cells and / or culture are the same as the Bacillus subtilis cells and / or culture described in the above section of the present invention (Agent for improving joint movement or relieving joint pain).

[0058] The other ingredients in the pharmaceutical composition for improving joint movement or relieving joint pain are not particularly limited and can be selected appropriately depending on the type of pharmaceutical composition. The content of the other ingredients in the pharmaceutical composition for improving joint movement or relieving joint pain is also not particularly limited and can be selected appropriately depending on the type of pharmaceutical composition.

[0059] The route of administration of the pharmaceutical composition of the present invention for improving joint movement or relieving joint pain is not particularly limited and can be appropriately selected depending on the purpose. However, as described above, Bacillus subtilis cells or a culture thereof are effective in a small amount and for a short period of time, their safety when administered orally is guaranteed, they have excellent storage stability and acid resistance, and they easily reach the intestines and grow there. Therefore, oral administration is simple and preferable, and can be expected to have a sustained effect of improving joint movement and relieving joint pain.

[0060] When administered orally, the pharmaceutical composition of the present invention for improving joint mobility or relieving joint pain is appropriately formulated with pharmacologically acceptable carriers and additives and administered in the form of a liquid, powder, granules, capsules, tablets, etc. Examples of pharmacologically acceptable carriers used include, but are not limited to, excipients, lubricants, binders, disintegrants, preservatives, antioxidants, colorants, and flavoring agents, and other commonly used carriers can also be used as appropriate. Specific examples of the carriers are the same as those described above in the section of the present invention (Agent for Improving Joint Mobility or Relieving Joint Pain).

[0061] The dosage of the pharmaceutical composition of the present invention for improving joint movement or alleviating joint pain is not particularly limited and can be appropriately selected depending on the purpose. ８ ~2.5 x 10 １１ The preferred dose of Bacillus subtilis is 2.0 x 10 CFU. ９ ~2.0 x 10 １０ A dose that allows administration of CFU of Bacillus subtilis is more preferable.

[0062] The number of times the pharmaceutical composition of the present invention for improving joint movement or alleviating joint pain is taken is not particularly limited and can be appropriately selected depending on the purpose, but is preferably taken once to five times a day, more preferably once to four times a day, and particularly preferably taken in divided doses of once to three times a day, or once a day.

[0063] For example, when formulated as tablets, the amount of Bacillus subtilis per tablet is not particularly limited and can be appropriately selected depending on the purpose. ７ ~2.5 x 10 １１ CFU is preferred, 2.0 x 10 ８ ~9.0 x 10 １０ CFU is more preferably 4.0 x 10 ８ ~4.0 x 10 １０ More preferably, 4.0 x 10 CFU ８ ~2.0 x 10 １０ CFUs are particularly preferred.

[0064] The number of tablets to be taken at one time is not particularly limited and can be appropriately selected depending on the purpose, and examples include 1 to 5 tablets.

[0065] The duration of administration of the pharmaceutical composition of the present invention for improving joint movement or relieving joint pain is not particularly limited and can be appropriately selected depending on the purpose, but it is recommended to use it continuously for 4 weeks or more, preferably 8 weeks or more, and more preferably 12 weeks or more.

[0066] The agent for improving joint movement or relieving joint pain, food, drink or feed for improving joint movement or relieving joint pain, and pharmaceutical composition for improving joint movement or relieving joint pain of the present invention may be used in combination with other pharmaceuticals, food, drink or feed for improving joint movement or relieving joint pain.

[0067] The present invention also relates to a method for improving joint movement or alleviating joint pain, which comprises administering the above-mentioned Bacillus subtilis cells and / or culture to a subject.

[0068] The present invention will be specifically explained below by way of test examples, but the present invention is not limited to these test examples.

[0069] In the following, as an example of Bacillus subtilis, Bacillus subtilis C-3102 strain (deposited on December 25, 1985 at the Fermentation Research Institute, Agency of Industrial Science and Technology, Ministry of International Trade and Industry (currently the Patent Organism Depositary Center, National Institute of Technology and Evaluation, Room 120, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture) under accession number FERM BP-1096) was used.

[0070] (Test Example 1: Human Test on Healthy Middle-aged and Elderly Volunteers) <Tablet Preparation> 5 kg of water was added to 10 kg of commercially available soybean meal granules, and the mixture was sterilized at 121°C for 60 minutes. A pre-cultured culture of Bacillus subtilis C-3102 strain (accession number FERM BP-1096) was inoculated and cultured at approximately 37°C for 60.5 hours to produce a soybean culture of Bacillus subtilis C-3102 strain. The resulting culture was dried and pulverized (hereinafter, sometimes referred to as "fermented soybean powder") and blended with the other ingredients shown in the "Composition of Sugar-Coated Tablets" below to produce sugar-coated tablets. The "Nutritional Information of Sugar-Coated Tablets" is also shown below.

[0071] - Composition of sugar-coated tablets - Sugars: 70% by weight Fermented soybean powder: 9% by weight Starch: 7% by weight Lubricant: 0.9% by weight Others: 13.1% by weight Total: 100% by weight * Fermented soybean powder is made by fermenting 100% of Bacillus subtilis C-3102 strain. １０ The Bacillus subtilis C-3102 strain is contained in the form of spores.

[0072] - Nutritional information for sugar-coated tablets (nutritional analysis values ​​per 3 tablets (1.14g)) - Energy: 4.02kcal Protein: 0.08g Fat: 0.01-0.03g Carbohydrates: 0.88g Sodium equivalent: 0-0.00001g

[0073] <Test Method and Results> A double-blind, randomized, placebo-controlled, parallel-group comparative study was conducted to investigate the effect of Bacillus subtilis strain C-3102 on joint pain. The subjects in this study were 100 healthy middle-aged to elderly people (ages: 50-79 years). In this study, healthy subjects excluded those with or a history of serious diseases such as diabetes, brain disease, circulatory system disease, liver disease, kidney disease, or heart disease. The subjects were randomly divided into a Bacillus subtilis strain C-3102 intake group and a placebo intake group.

[0074] The test food group taking Bacillus subtilis C-3102 strain was given 4.6 x 10 per 3 tablets. ９The subjects were given sugar-coated tablets containing CFU of Bacillus subtilis C-3102 strain, and the placebo group was given placebo tablets containing dextrin instead of fermented soybean powder, three tablets per day for four weeks.

[0075] Before and after the start of the study, the subjects were asked to fill out a questionnaire about joint pain using the following five-point scale. Figure 1 shows the average change in the scores of the questionnaire results before and after the start of the study (after the study - before the study). 1 point: Not bothering me at all. 2 points: A little bothersome. 3 points: A little bothersome. 4 points: A lot of bothersome. 5 points: So bothersome I can't bear it.

[0076] As shown in FIG. 1, the group receiving Bacillus subtilis C-3102 strain (mean change: −0.2) showed a significant improvement in score compared to the placebo group (mean change: 0.2).

[0077] (Test Example 2: Human test on healthy adults) <Preparation of tablets> Fermented soybean powder was produced in the same manner as in Test Example 1. Coated tablets were produced by blending the fermented soybean powder with other ingredients shown in the "Composition of coated tablets" below. The "Nutritional information of coated tablets" is also shown below.

[0078] - Composition of coated tablets - Sugars: 71.3% by mass Fermented soybean powder: 16.2% by mass Starch: 3.3% by mass Lubricant: 1.9% by mass Others: 7.3% by mass Total: 100% by mass * Fermented soybean powder is made by fermenting 100% of Bacillus subtilis C-3102 strain. １０ The Bacillus subtilis C-3102 strain is contained in the form of spores.

[0079] -Nutritional information for coated tablets (nutritional analysis values ​​per 3 tablets (0.615g))- Energy: 2.41kcal Protein: 0.07g Fat: 0.01-0.03g Carbohydrates: 0.49g Sodium equivalent: 0-0.00001g

[0080] <Test Method and Results> A double-blind, randomized, placebo-controlled, parallel-group comparative study was conducted to investigate the effect of Bacillus subtilis strain C-3102 on joint pain. Eighty-eight healthy adults (ages: 20 to 69 years) were used as subjects in this study. In this study, healthy subjects were defined as those diagnosed with a Kellgren-Lawrence classification (KL grade) of 0 or 1 in a knee X-ray examination. The subjects were randomly divided into a Bacillus subtilis C-3102 intake group and a placebo intake group so that the JKOM score and KL grade based on the Japanese Knee Osteoarthritis Measure (JKOM) were uniform.

[0081] The test food group taking Bacillus subtilis C-3102 strain was given 3.4 x 10 per 3 tablets. ９ The subjects were given three coated tablets containing CFU of Bacillus subtilis C-3102 strain per day, and the placebo group was given three placebo tablets containing dextrin instead of fermented soybean powder per day for 12 weeks.

[0082] The subjects were evaluated using JKOM before the start of the study (week 0), and four weeks after the start (week 4), eight weeks after the start (week 8), and 12 weeks after the start (week 12).

[0083] The JKOM consists of 25 questions about knee pain and related movements, in addition to the degree of knee pain perceived. The condition of knee pain can be understood from the "JKOM Score" calculated from the subscores made up of 25 questions such as "knee pain and stiffness," "daily life status," "usual activities," and "health status." The higher the score obtained, the greater the degree of knee pain.

[0084] Figure 2 shows the changes in the "JKOM score" (average value) for each group, and Figure 3 shows the changes in the "daily life status score" (average value).

[0085] As shown in Figures 2 and 3, the overall score, "JKOM score" (Figure 2), and the subscale, "daily life status score" (Figure 3), were significantly improved in the Bacillus subtilis C-3102 strain intake group compared to the placebo intake group.

[0086] (Formulation Example 1: Capsules) A ​​soybean culture of Bacillus subtilis strain C-3102 and edible oil were mixed and filled into a soft capsule shell consisting of the following ingredients by a conventional method to obtain soft capsules weighing 380 mg each. <Contents> - Soybean culture of Bacillus subtilis strain C-3102... 100 mg - Edible oil... 150 mg <Shell> - Gelatin... 100 mg - Glycerin... 30 mg

[0087] (Formulation Example 2: Powder) The following ingredients were blended and granulated according to a conventional method to produce 5 g stick granules: Bacillus subtilis C-3102 strain soybean culture... 5% by mass Sodium carboxymethylcellulose... appropriate amount Dextrin... appropriate amount

[0088] (Formulation Example 3: Liquid) The following ingredients were blended and 10 kg of water was added according to a conventional method to prepare a liquid: Bacillus subtilis C-3102 strain soybean culture 100 g, liquid sugar 4000 g, DL-sodium tartrate 1 g, citric acid 50 g, vitamin C 50 g, vitamin E 150 g, cyclodextrin 25 g, potassium chloride 5 g, magnesium sulfate 2 g

[0089] This application claims priority based on Japanese Patent Application No. 2023-180349, filed on October 19, 2023, the entire contents of which are incorporated herein by reference.

[0090] FERM BP-1096

Claims

1. An agent for improving joint movement or relieving joint pain, comprising Bacillus subtilis cells and / or a culture thereof.

2. The agent for improving joint movement or relieving joint pain according to claim 1, wherein the culture is a soybean culture.

3. The agent for improving joint movement or relieving joint pain according to claim 1 or 2, which contains Bacillus subtilis spores.

4. The agent for improving joint movement or relieving joint pain according to claim 1 or 2, wherein the Bacillus subtilis is Bacillus subtilis C-3102 strain (FERM BP-1096).

5. A method for improving joint movement or relieving joint pain according to claim 1 or 2, which is taken orally.

6. 2.0 x 10 per day ８ ~2.5 x 10 １１ 3. The agent for improving joint movement or relieving joint pain according to claim 1 or 2, which is formulated so that CFU of Bacillus subtilis can be ingested.

7. 2.0 x 10 per day ９ ~2.0 x 10 １０ 3. The agent for improving joint movement or relieving joint pain according to claim 1 or 2, which is formulated so that CFU of Bacillus subtilis can be ingested.

8. A food, drink, or feed for improving joint movement or relieving joint pain, characterized by containing Bacillus subtilis cells and / or a culture product.

9. A pharmaceutical composition for improving joint movement or relieving joint pain, comprising Bacillus subtilis cells and / or a culture.

Images