Muscle strengthening agent and food composition

Abstract

To provide a method of preventing decrease in muscular strength by a method other than exercise.SOLUTION: As a result of examining lactic acid bacteria using various foods and the like as isolation sources, it was found that human-derived Bifidobacterium breve has a muscle-strengthening effect. That is, provided is a muscle strengthening agent containing cells of Bifidobacterium breve N106 strain as an active ingredient. Further, provided is a muscle strengthening agent which is in the form of a unit intake per meal and which contains Bifidobacterium breve N106 strain as an active ingredient in the unit.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] It is widely recognized that exercise is important for maintaining health. For example, there is data showing that people who are physically active or who exercise frequently have lower morbidity and mortality rates for all-cause mortality, ischemic heart disease, hypertension, diabetes, obesity, osteoporosis, colon cancer, and other conditions. It has also been reported that exercise can prevent sarcopenia, which is caused by a decrease in muscle strength, in elderly people, thereby reducing the risk of becoming bedridden or dying. Thus, exercise is essential for health.

[0002] However, in recent years, the amount of physical activity has clearly decreased due to the mechanization of housework and work, and the development of transportation. In particular, adults who tend to be sedentary have a significant decrease in muscle strength, coupled with the effects of aging. A decrease in muscle strength can also cause unexpected injuries in daily life. In addition, in elderly people, a decrease in the mass of skeletal muscles that support physical activity makes it difficult for the body to move as desired, increasing the risk of fractures due to falls, hospitalization, and becoming bedridden. Therefore, in order to live a healthy and cultured life, it is necessary to prevent a decrease in muscle mass and strength.

[0003] To prevent the decline of muscle mass and strength, it is desirable to engage in habitual exercise. However, those who have experienced muscle atrophy due to aging, those who are bedridden (not only the elderly) or cannot put strain on their locomotor organs (bones, joints) or circulatory system, and those who are busy every day are unable to exercise continuously even if they wish, making it difficult to prevent the decline of muscle strength.

[0004] There is also a method of actively ingesting concentrated fermented milk to prevent muscle loss (see Patent Document 1). However, people with small appetites, including not only the elderly, and those who tend to have irregular meal timing and frequency, are unable to actively ingest protein even if they wish, making it difficult to prevent muscle loss. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] JP 2020-014452 A Summary of the Invention [Problem to be solved by the invention]

[0006] The present invention has been made in consideration of the above problems. That is, an object of the present invention is to provide a method for preventing muscle weakness easily without exercise.

[0007] In order to solve the above problems, the inventors investigated lactic acid bacteria using various foods as isolation sources and discovered that human-derived Bifidobacterium breve has the effect of strengthening muscle strength, thereby completing the present invention. [Means for solving the problem]

[0008] The present invention includes the following features. (1) A muscle strengthening agent containing the Bifidobacterium breve N106 strain as an active ingredient. (2) A muscle strengthening agent which is in a unit intake form per meal, and in which the unit contains Bifidobacterium breve N106 strain as an active ingredient. (3) A muscle strengthening agent according to any one of claims 1 to 2, wherein the Bifidobacterium breve N106 strain cells are killed. (4) A food composition for strengthening muscles, comprising Bifidobacterium breve N106 strain cells as an active ingredient. (5) A food or drink containing the muscle strengthening agent according to any one of claims 1 to 3.

[0009] Here, the term "muscle strength" in the claims refers to the force generated when a muscle contracts. Also, "muscle strength strengthening" is a concept that includes not only strengthening muscle strength based on muscle hypertrophy, but also strengthening muscle strength based on the contraction of more muscle fibers due to the improvement of the nervous system that contracts muscle fibers.

[0010] In addition, the term "unit intake form" in the claims means that a predetermined amount is in the form of an individual packet, tablet, capsule, granule, etc., so that a fixed amount can be easily taken at one time. Effect of the Invention

[0011] According to the present invention, it is possible to provide a muscle strengthening agent and a food composition that can maintain or strengthen muscle mass and muscle strength by a method other than exercise, which is expected to extend healthy lifespan and ultimately reduce medical costs. [Brief description of the drawings]

[0012] [Figure 1] Figure 1 shows a comparison of grip strength (actual measured values) before intake began, after 6 weeks of intake, and after 12 weeks of intake. [Diagram 2] Figure 2 shows a comparison of the number of seconds (actual measurements) required for standing on one leg with eyes open before, after 6 weeks of intake, and after 12 weeks of intake. [Diagram 3] FIG. 3 is a graph comparing the 6-minute walking distance (actual measurements) before the start of intake, after 6 weeks of intake, and after 12 weeks of intake. [Figure 4] Figure 4 shows the rate of change in score for the question "Do you find it difficult to climb stairs?" from a subjective symptom questionnaire before, 6 weeks after, and 12 weeks after intake. [Diagram 5] Figure 5 shows the rate of change in score for the question "Do you find it difficult to lift heavy objects?" from a subjective symptom questionnaire before, 6 weeks after, and 12 weeks after intake. [Figure 6] Figure 6 shows the rate of change in score for the question "Do you find it difficult to open hard lids?" from a subjective symptom questionnaire before, 6 weeks after, and 12 weeks after intake. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] In this specification, "muscle strengthening" can be rephrased as "muscle strength enhancement," "strengthening muscle output," "enhancing muscle output," etc. Therefore, the muscle strengthening agent of this embodiment can be rephrased as "muscle strength enhancing agent," "agent for enhancing muscle output," "agent for enhancing muscle output," etc.

[0014] The muscle strengthening agent of this embodiment is based on the discovery that a composition containing the bacterial cells of Bifidobacterium breve N106 strain (NITE BP-03231) as an active ingredient is effective in preventing a decrease in muscle mass and strength.

[0015] (Bifidobacterium breve strain N106) Bifidobacterium breve strain N106 has been deposited under the following conditions: (1) Name of depository institution: National Institute of Technology and Evaluation, Patent Microorganisms Depository Center (2) Contact: Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, 292-0818 (3) Accession number: NITE BP-03231 (4) Identification marking: N106 (5)Deposit date: June 18, 2020

[0016] A known method can be used as a method for preparing the Bifidobacterium breve N106 strain cells used in this embodiment. For example, in the pre-culture and main culture, a GAM medium (Nissui Pharmaceutical Co., Ltd.) is prepared by a predetermined method, and the Bifidobacterium breve N106 strain is inoculated and cultured at 37°C for 24 hours. The cells in the main culture are collected by centrifugation or membrane concentration, washed with water, and then heat sterilized. Sterilization may be performed before the cell collection process. A dry sterilized powder can be obtained by freeze-drying after sterilization and washing of the cells. The method for powdering the cells is not limited to freeze-drying, and may be spray-drying or drum-drying. In addition, a live cell powder can be obtained without sterilization.

[0017] In the present embodiment, the bacterial cells of the Bifidobacterium breve N106 strain may be live or killed. When live bacteria are used, dead bacteria may be mixed in. It is preferable to ingest 20 mg or more of the bacterial cells of the Bifidobacterium breve N106 strain per day. The daily amount may be ingested at once or in several divided doses, and the timing of ingestion does not matter. For example, if there is a tablet containing 5 mg of killed bacteria of the Bifidobacterium breve N106 strain per tablet as a unit intake form, 4 tablets may be ingested at once or 4 tablets may be ingested in several divided doses. In addition, if each tablet contains 20 mg of killed bacteria of the Bifidobacterium breve N106 strain, one tablet may be ingested per day. There is no upper limit to the daily intake amount of the bacterial cells of the Bifidobacterium breve N106 strain, but if an upper limit is to be set, it is considered that 10 g is sufficient. It is believed that there will be no difference in effectiveness if taken for a longer period of time. It is also recommended that Bifidobacterium breve N106 strain cells be taken continuously for 12 weeks or more.

[0018] The method of ingesting the Bifidobacterium breve N106 strain bacteria is not particularly limited, but is preferably oral ingestion. Specifically, it is preferable to ingest the Bifidobacterium breve N106 strain bacteria as a food composition, a drug or the like containing the Bifidobacterium breve N106 strain bacteria as a pharmaceutical composition.

[0019] When ingested as a food or drink, for example, the Bifidobacterium breve N106 strain can be used as a food composition and as an ingredient in fermented milk and lactic acid bacteria drinks, dairy products such as butter, egg products such as mayonnaise, and sweet breads such as butter cake. It can also be suitably used in processed foods such as instant noodles and cookies. Furthermore, it may be dissolved in water or juice and ingested as a drink. The Bifidobacterium breve N106 strain can be ingested as a food or drink by itself. In addition to general drinks and foods, it may be contained in foods for specified health uses, foods with functional claims, nutritional supplements, supplements, etc.

[0020] When taken as a medicine, it can be used together with additives and other pharma- ceutical carriers. Examples of the form of the medicine include tablets, capsules, granules, etc. For preparation, known methods can be used.

[0021] Additives used in pharmaceuticals include lubricants such as calcium stearate, sweeteners such as sucrose and lactose, flavoring agents, stabilizers, buffers, solubilizing agents, antioxidants, preservatives, and the like. EXAMPLES

[0022] The present invention will now be described in more detail with reference to examples, but the present invention is not limited to the following examples.

[0023] An open-label, single-arm study was conducted on healthy Japanese men and women aged 45 years or older and 65 years or younger. An open-label study is a study method in which the doctor, subjects, and staff know which treatment group the subjects have been assigned to. A single-arm study is a study method in which only the intake group is included, without a placebo control group.

[0024] [Manufacturing of test food tablets] Tablets were manufactured by mixing powdered Bifidobacterium breve N106 strain cells and excipients, which included reduced maltose syrup, cellulose, calcium stearate, and fine silicon dioxide.

[0025] The test food tablets were formulated with 20 mg of killed Bifidobacterium breve N106 powder per tablet.

[0026] [Test implementation] Healthy Japanese men and women aged 45 years or older and 65 years or younger were recruited, and 37 subjects who gave their voluntary consent after explaining the contents of the study were selected. Screening (SCR) was conducted on the 37 study volunteers to determine their eligibility for the study. In the screening, the study volunteers were interviewed by a doctor, had their body measurements taken, their vital signs (blood pressure, pulse), blood tests taken, and exercise ability tests taken. Based on the results of the screening, 10 subjects who were not sick and were deemed eligible for the study were selected.

[0027] The test schedule is shown in Table 1. Next, the main test consisted of the subjects ingesting the test food for 12 weeks.

[0028] [Table 1]

[0029] The test food was administered as one tablet per day with water or lukewarm water. The time of administration was after breakfast to prevent forgetting to take the food, but if subjects forgot to take the food after breakfast, they were to take it on the same day and not carry it over to the next day. Subjects were instructed to observe the following restrictions during the study period.

[0030] <Matters to be observed during the exam period> 1. During the study period, you will maintain your normal lifestyle by limiting excessive exercise, dieting, overeating, and excessive drinking that significantly deviate from your daily routine. 2. During the study period, try not to change your lifestyle or environment (e.g., eating, drinking, exercise, sleep, work, etc.) from before participating in the study. 3. During the study period, participants who are used to exercising will not exercise more strenuously than usual, and will not change the amount or quality of their exercise. Also, participants who are not used to exercising will not start a new exercise routine. 4. During the study period, do not take any medicines, supplements, health foods, etc. that may affect muscles. If it becomes necessary to take any new medicines, please contact the consultation desk as far in advance as possible. If you do take any new medicines, please record the details of your intake in the lifestyle questionnaire. 5. During the study period, participants will record in the lifestyle questionnaire any medications, supplements, health foods, etc. that they have been taking or taking without restrictions, and will not change their habits. In addition, participants who do not have any habits will not start taking any new medications. If it becomes necessary to take new medications for unavoidable reasons, participants will contact the consultation desk in advance, if possible. 6. During the study period, you may continue to consume foods rich in lactic acid bacteria, but do not significantly change the amount or frequency of your intake. If there are any changes, record them in the lifestyle questionnaire. 7. Do not participate in any other clinical trials (studies) during the study period. 8. Do not donate whole blood or blood components during the study period. 9. If you undergo any vaccinations, health checkups, or medical examinations during the study period, you must report this to the consultation desk in advance and record it on the lifestyle questionnaire. 10. Do not disclose to anyone else any information about the exam or about the exam content. 11. Overseas travel and business trips are prohibited during the exam period. 12. Smokers will not change their smoking habits during the study period.

[0031] <Things to observe before the test> 1. Avoid strenuous exercise for 3 days prior to the test. 2. Do not drink alcohol the day before the test. 3. Try to sleep as consistently as possible for the same amount of time and quality the night before the test. 4. Maintain a fasting state from 6 hours before the start of the test until the end of the test. In addition, avoid fatty foods and large amounts of sugary drinks as your last meal before fasting. 5. Be sure to urinate before the test begins and refrain from drinking any fluids until body composition measurement is completed.

[0032] <Things to observe on the day of the test> 1. On the day of the examination, please come to the hospital wearing comfortable clothing and shoes. 2. On the day of the examination, refrain from smoking until all examinations are completed. 3. If for any reason you are unable to attend the hospital on the day of the test, please contact the test consultation desk immediately.

[0033] [Exam Evaluation] The effects of taking the test food were evaluated based on the following items.

[0034] [Body composition] After urination, body composition items (body weight, total body water, protein, body fat, muscle mass, skeletal muscle mass, BMI, body fat percentage, upper limb muscle mass, trunk muscle mass, lower limb muscle mass, and basal metabolic rate) were measured using InBody270 (InBody Japan Co., Ltd.).

[0035] [Lower limb strength] Lower limb muscle strength was measured using Locomoscan (registered trademark) II (Alcare Corporation).

[0036] [Grip strength] Grip strength was measured using a Grip-D (digital grip strength meter) TKK5401 (Takei Machinery Industry Co., Ltd.) according to the method described in the New Physical Fitness Test Implementation Guidelines (Japan Sports Agency).

[0037] [Open-eyes one-legged standing test] The open-eyed one-legged standing test was conducted according to the method described in the New Physical Fitness Test Implementation Guidelines (Japan Sports Agency).

[0038] [6-minute walking test] A 6-minute walking test was conducted according to the method described in the New Physical Fitness Test Implementation Guidelines (Japan Sports Agency).

[0039] [6m walking test] A 6m walking test was performed according to the method described in the 2017 edition of the Sarcopenia Treatment Guidelines (Japanese Society of Sarcopenia and Frailty).

[0040] [2-step test] The two-step test, which is included in the Locomotion Test (Japanese Orthopaedic Association), was conducted. The two-step score was calculated using the following formula. 2 step value = 2 strides (cm) ÷ height (cm)

[0041] [10-time chair rise test] The chair-stand test was performed 10 times according to the method described in the Exercise Guidelines for Health Promotion 2006 (Ministry of Health, Labour and Welfare).

[0042] [Chalder Fatigue Questionnaire] The Chalder Fatigue Questionnaire was administered.

[0043] [VAS Survey] A VAS questionnaire was administered to assess related subjective symptoms.

[0044] [Subjective Symptom Questionnaire] A questionnaire was administered regarding subjective muscle symptoms and lower back pain symptoms.

[0045] [Lifestyle Survey] The subjects were asked to record their daily living conditions. The lifestyle questionnaire items were recorded according to Table 2.

[0046] [Table 2]

[0047] Statistical analysis was performed on the following populations: [Per Protocol Set (PPS)] The population excluding cases that met the following analysis exclusion criteria was determined as the per protocol population (PPS). (1) Cases in which the intake rate of the test food was less than 80% (2) Patients in whom non-compliance was identified during the study period (3) Cases in which actions that significantly undermine the reliability of clinical test results during the study period are evident (4) Cases in which it became clear after the start of intake that they met the exclusion criteria (5) Cases in which significant deviations from the study protocol were observed (6) Cases for which there were clear reasons to exclude them based on other information such as clinical test values ​​and daily diary entries.

[0048] First, the 10 subjects enrolled in this study underwent item tests before the start of intake (see Table 1). After the tests were completed, they began taking the test food. The intake period was from the day of the pre-intake test to the day before the 12-week intake test. For each test, subjects were required to maintain a fasting state from 6 hours before the start of the test until the end of the test. For the 6-week intake test, subjects were required to come to the hospital without taking the test food in order to maintain a fasting state, and were required to take the test food on the same day after the test and measurements were completed.

[0049] [result] Next, the results of the statistical analysis of each test item will be explained. First, one person was excluded from the statistical analysis because their intake rate of the test food during the test period was 61%. Therefore, statistical analysis was performed on the nine people in the PPS. The results of the body composition measurements and physical ability tests were statistically analyzed using the Dunnett method, with the values ​​before ingestion of the test food as the baseline, and the values ​​6 and 12 weeks after ingestion. P<0.01 is marked with **, and P<0.05 is marked with *. Furthermore, P<0.1 is marked with †.

[0050] Regarding the physical ability tests, the results of the test items in which significant differences (P<0.05) or significant trends (P<0.1) were observed are shown in Table 3 and Figures 1 to 3.

[0051] [Table 3]

[0052] As shown in Table 3 and Figure 1, grip strength increased significantly after 12 weeks of intake. In addition, as shown in Figure 2, there was a tendency for the time to stand on one leg with eyes open to be extended after 12 weeks of intake. Furthermore, as shown in Figure 3, the distance of 6-minute walking significantly increased after 12 weeks of intake.

[0053] Next, the subjective symptoms were statistically analyzed using the values ​​before intake of the test food as a baseline and the values ​​after 6 and 12 weeks of intake. The VAS questionnaire scores were analyzed using the Dunnett method, and the results of the Chalder Fatigue Questionnaire and the subjective symptoms questionnaire were analyzed using the Wilcoxon signed-rank test. The amount of change and rate of change were also calculated and analyzed. Change: Each visit - Value before intake started Rate of change: Amount of change ÷ Value before intake started × 100 P<0.05 is marked with *, and P<0.1 is marked with †.

[0054] Regarding subjective symptoms, the results of the test items in which significant differences or significant trends were observed are shown in Table 4 and Figures 4 to 6.

[0055] [Table 4]

[0056] As shown in Table 4 and Figure 4, the score change rate for the question "Do you feel it is difficult to climb stairs?" showed significant improvement after 6 weeks and 12 weeks of intake. Also, as shown in Figure 5, the score change rate for the question "Do you feel it is difficult to carry heavy objects? *About 5 kg. A 5 kg bag of rice, 2-3 2 L plastic bottles, a work bag filled with a computer and documents, etc." showed significant improvement after 6 weeks and 12 weeks of intake. Furthermore, as shown in Figure 6, the score change rate for the question "Do you feel it is difficult to open a hard lid?" showed significant improvement after 6 weeks and 12 weeks of intake. Note that this questionnaire was created independently, and the answers were converted into scores by selecting from five levels: "No feeling: 0, Not feeling much: 1, Feeling a little: 2, Feeling: 3, Feeling a lot: 4".

[0057] As mentioned above, in this example, healthy Japanese men and women aged 45 years or older and 65 years or younger were given 20 mg of Bifidobacterium N106 strain powder for 12 weeks, and a test was carried out to examine the effect on muscles. As a result, significant improvements were confirmed in exercise test items such as grip strength and 6-minute walking.

[0058] According to Richard WB's Grip Strength: An Indispensable Biomarker For Older Adults (Clin Interv Aging. 2019; 14: 1681-1691.), there is a correlation between grip strength and overall motor ability. In addition, because it is easy to measure, it is also used to diagnose sarcopenia, and is also designated as a new physical fitness test conducted by the Ministry of Education, Culture, Sports, Science and Technology. From the above, it can be said that grip strength is an important health indicator of motor ability. It was suggested that the improvement in grip strength due to the intake of the N106 strain according to the present invention may reflect an improvement in the muscle condition of the entire body.

[0059] Next, the 6-minute walk test is generally performed as a method for testing walking ability. It is known that walking ability decreases with age. Walking is not just a movement movement, but also a part of physical activity. A decrease in the average number of steps taken per day can lead to obesity, lifestyle-related diseases, and frailty. In the above examples, it was suggested that the improvement in walking ability by taking the N106 strain according to the present invention may lead to the prevention of the above diseases.

[0060] Although the data is not shown, in this example, no significant increase in muscle mass was observed in the primary endpoint of body composition, but at the same time, no decrease was observed. This suggests that the intake of this test food may suppress the decrease in muscle mass due to lack of exercise and aging. [Industrial Applicability]

[0061] According to the present invention, it is possible to provide a muscle strengthening agent and a food composition that can maintain or strengthen muscle mass and muscle strength by a method other than exercise, which is expected to extend healthy lifespan and ultimately reduce medical costs.

Claims

1. A muscle strengthening agent containing the bacteria of Bifidobacterium breve N106 strain as an active ingredient.

2. A muscle strengthening agent which is in a unit intake form per meal and contains Bifidobacterium breve N106 strain as an active ingredient in said unit.

3. The muscle strengthening agent according to any one of claims 1 to 2, wherein the Bifidobacterium breve N106 strain cells are killed.

4. A food composition for strengthening muscles, comprising Bifidobacterium breve N106 strain cells as an active ingredient.

Images