Compositions, foods, cosmetics, quasi-drugs, or pharmaceuticals containing lactic acid bacteria that have a blood pressure-lowering effect.

Abstract

To provide lactic acid bacterium or bifidobacteria having a hypotensive action effective to a human body.SOLUTION: A blood pressure lowering composition comprises, as an active ingredient, at least one of bacterial cells obtained from Bifidobacterium longum N714 strain (NITE BP-03004), secretions, culture solution or fermentation products, or bacterial cell components and bacterial cell processed products. In addition, a blood pressure lowering composition is composed of a unit intake form per meal, and contains, in the unit, at least one of bacterial cells obtained from Bifidobacterium longum N714 strain, secretions, culture solution or fermentation products, or bacterial cell components and bacterial cell processed products as an effective ingredient.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to a composition, food or drink, cosmetic, quasi-drug or drug having a blood pressure-lowering effect and containing lactic acid bacteria.

Background Art

[0002] Currently, it is estimated that 1.28 billion people worldwide have hypertension. Hypertension increases the risk of stroke, heart disease, kidney disease and other diseases, and is a major cause of premature death. According to the National Health and Nutrition Survey, the prevalence of hypertension increases with age, and it is said that it exceeds 50% in men in their 50s and women in their 60s and above. Furthermore, with the global aging in the future, the prevalence of hypertension is expected to increase further. Therefore, as one of the global goals for non-communicable diseases, it is stated that the prevalence of hypertension should be reduced by 33% between 2010 and 2030.

[0003] Hypertension is known to progress without any symptoms. Furthermore, it has been reported that even if the blood pressure value does not correspond to hypertension, the risk of developing hypertension or cerebrovascular and cardiovascular diseases increases as the blood pressure value increases. Here, there are essential hypertension and secondary hypertension. Essential hypertension is considered to be caused by constitution, excessive salt intake, obesity, excessive drinking, lack of exercise, stress, smoking, etc. Secondary hypertension is considered to be caused by hormonal secretion disorders, kidney diseases, side effects of drugs, etc.

[0004] Whether it is essential hypertension or secondary hypertension, once hypertension develops, treatment with antihypertensive drugs becomes necessary. Therefore, it is necessary to take measures before developing hypertension. In particular, since essential hypertension is considered to be caused by lifestyle habits, it is considered important to maintain normal blood pressure by improving lifestyle habits including diet and exercise.

[0005] As a method for improving blood pressure, a blood pressure-lowering composition containing an aqueous extract of Bifidobacterium bacteria as an active ingredient has been reported (see Patent Document 1). According to Patent Document 1, when an aqueous extract of Bifidobacterium bacteria was administered to spontaneously hypertensive rats, age-related increases in blood pressure were suppressed.

[0006] On the other hand, it is known that the effects of bacterial cells can vary depending on the strain (see paragraph 0006 of Patent Document 2). Furthermore, even if an effect is observed in animal experiments, it is not guaranteed that the same effect will occur in humans. [Prior art documents] [Patent Documents]

[0007] [Patent Document 1] Patent No. 2687229 [Patent Document 2] Japanese Patent Publication No. 2019-172661 [Overview of the project] [Problems that the invention aims to solve]

[0008] This invention was made in view of the above-mentioned problems. Specifically, a screening was conducted to find any lactic acid bacteria or bifidobacteria that have an effective blood pressure-lowering effect on the human body. As a result, it was newly discovered that ingesting Bifidobacterium strain N714, which has the ability to suppress IL-17 production, is effective in lowering blood pressure, and this led to the completion of the present invention. [Means for solving the problem]

[0009] The present invention encompasses the following features. (1) A blood pressure lowering composition containing at least one of the following as an active ingredient: bacterial cells, secretions, culture medium or fermentation products obtained from Bifidobacterium longum strain N714 (NITE BP-03004), or bacterial cell components or bacterial cell processed products. (2) Food and beverages containing at least one of the following as an active ingredient for lowering blood pressure: bacterial cells, secretions, culture solution or fermentation products obtained from Bifidobacterium longum strain N714 (NITE BP-03004), bacterial cell components, or bacterial cell processed products. (3) Cosmetics, quasi-drugs, or pharmaceuticals containing at least one of the following as an active ingredient for lowering blood pressure: bacterial cells, secretions, culture medium or fermentation products obtained from Bifidobacterium longum strain N714 (NITE BP-03004), bacterial cell components, or bacterial cell processed products.

[0010] Here, "secretion" as described in the claims refers to substances secreted from bacterial cells into the culture medium. "Bacterial cell treatment product" refers to bacterial cell lysates obtained by ultrasonic treatment of bacterial cells, or bacterial cell lysates obtained by cell wall-degrading enzyme treatment, freeze-dried bacterial cells, or immobilized bacterial cells. "Culture medium" refers to the culture medium or culture supernatant containing secretions from bacterial cells and bacterial cells. Centrifugation or enzymatic treatment with proteases or lipases may also be performed as needed. "Fermentation product" refers to purified compounds or peptides produced during the fermentation process by the lactic acid bacteria. Specifically, examples include equol and GABA. [Effects of the Invention]

[0011] According to the present invention, it is possible to provide an effective blood pressure-lowering effect to the human body, particularly to individuals with high-normal blood pressure aged 55 years or older. [Brief explanation of the drawing]

[0012] [Figure 1] Figure 1 shows the change in blood pressure at the time of evaluation for individuals aged 55 and older with high-normal blood pressure, where (a) is the systolic blood pressure at the time of evaluation and (b) is the change from the start of intake. [Figure 2] Figure 2 shows the change in blood pressure at the time of evaluation for individuals with high-normal blood pressure who have low salt intake, where (a) is the systolic blood pressure at the time of evaluation and (b) is the change from the start of intake. [Modes for carrying out the invention]

[0013] In this specification, "lowering blood pressure" can be rephrased as "suppressing the rise in blood pressure," "improving blood pressure," "maintaining blood pressure values," etc. Therefore, the blood pressure lowering composition of this embodiment can also be rephrased as "blood pressure rise suppressing composition," "blood pressure improving composition," etc.

[0014] The blood pressure lowering composition of this embodiment is based on the discovery that a composition containing the bacterial cells of Bifidobacterium longum N714 strain (NITE BP-03004) as an active ingredient is effective in lowering blood pressure in individuals with high-normal blood pressure.

[0015] (Bifidobacterium longum N714 strain) This is Bifidobacterium longum. The symbol N714 in this invention is a number uniquely assigned to the strain by Nissin Foods Holdings Co., Ltd. The strain of this invention was first isolated by the inventor from infant feces.

[0016] The Bifidobacterium longum N714 strain of the present invention is deposited under the following conditions. (1) Name of depositary institution: National Institute of Technology and Evaluation, Patent Microorganism Depository Center (2) Contact information: Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture 292-0818 (3) Accession number: NITE BP-03004 (4) Identification mark: N714 (5) Deposit date: July 10, 2019

[0017] As a method for preparing the cells of Bifidobacterium longum N714 strain used in this embodiment, known methods can be used. For example, for preculture and main culture, GAM medium (Nissui) is prepared by a predetermined method, and after inoculating the Bifidobacterium longum N714 strain, it is cultured at 37°C for 24 hours. The cells in the main culture solution are collected by centrifugation or membrane concentration, and after washing the cells with water, heat sterilization treatment is performed. Note that sterilization treatment may be performed before the cell collection treatment. After sterilization and cell washing, a dry killed cell powder can be obtained by freeze-drying treatment. The method of powdering the cells is not limited to freeze-drying, and it may also be performed by spray drying or drum drying. Also, if sterilization treatment is not performed, a live cell powder can also be obtained.

[0018] In this embodiment, the cells of Bifidobacterium longum N714 strain may use live cells or dead cells. Also, when using live cells, dead cells may be mixed. Furthermore, cell debris obtained by ultrasonic treatment of the cells, or cell lysate obtained by cell wall degrading enzyme treatment may also be used. It is preferable to ingest 20 mg or more of the cells of Bifidobacterium longum N714 strain per day. Regarding ingestion, the amount per day may be ingested at once or divided into several times, and the timing of ingestion is not limited. For example, if there is a tablet containing 5 mg of dead cells of Bifidobacterium longum N714 strain per tablet as a unit ingestion form, 4 tablets may be ingested at once or 4 tablets may be ingested in several divided times. Also, if each tablet contains 20 mg of dead cells of Bifidobacterium longum N714 strain, 1 tablet may be ingested per day. Note that there is no upper limit to the daily ingestion amount of the cells of Bifidobacterium longum N714 strain, but if an upper limit is deliberately set, it is considered sufficient to ingest 200 mg. This is because it is considered that there is no difference in the effect even if more than that is ingested. Also, it is preferable to continuously ingest the cells of Bifidobacterium longum N714 strain for 12 weeks or more.

[0019] As a method for ingesting Bifidobacterium longum N714 strain cells, there are no particular restrictions, but oral ingestion is preferred. Specifically, it is preferably ingested as a food or drink containing Bifidobacterium longum N714 strain cells as a food composition, a pharmaceutical containing the same as a pharmaceutical composition, a cosmetic containing the same as a cosmetic composition, etc.

[0020] When ingested as a food or drink, for example, Bifidobacterium longum N714 strain cells can also be used as a material for fermented milk, lactic acid bacteria beverages, dairy products such as butter, processed egg products such as mayonnaise, and confectionery breads such as butter cakes as a food composition. It can also be suitably used for processed foods such as instant noodles and cookies. Furthermore, it may be dissolved in water, juice, etc. and ingested as a beverage. In addition, Bifidobacterium longum N714 strain cells alone may be ingested as a food or drink as they are. In addition to general beverages and foods, it may also be contained in foods for specified health uses, foods with functional claims, dietary supplements, supplements, etc.

[0021] When ingested as a pharmaceutical, it can be used together with additives and other pharmaceutically acceptable carriers. Examples of the form of the pharmaceutical include tablets, capsules, granules, etc. In addition, known methods can be used for formulation.

[0022] Examples of additives used in pharmaceuticals include lubricants such as calcium stearate, sweeteners such as sucrose and lactose, as well as flavoring agents, stabilizers, buffers, solubilizing agents, antioxidants, preservatives, etc.

[0023] When used as a cosmetic, examples include forms such as ointments, creams, lotions, etc. In the case of an ointment, it is possible to prepare by mixing the lactic acid bacteria powder or its suspension with an oily base material. In the case of a cream, it can be produced by mixing it with an oil-in-water (O / W type) or water-in-oil (W / O type) liquid substance. Furthermore, in the case of a lotion, there are emulsion lotions and suspension lotions. The emulsion type is an emulsion, and the suspension type is a lotion with alcohols or water as the main ingredient.

[0024] Examples of additives used in cosmetics include sugars, proteins, amino acids, lipids, water, vitamins, and minerals. [Examples]

[0025] Next, the present invention will be described in more detail with reference to examples, but the present invention is not limited to the following examples.

[0026] [Manufacturing of test foods] The Bifidobacterium longum N714 strain cells were washed with water, sterilized by heat treatment, and then freeze-dried to produce a powder for use. The N714 capsules and placebo capsules were made of hydroxypropyl methylcellulose (HPMC) hard capsules of the same shape and color. The N714 capsules (example food) contained 20 mg of the N714 strain cells, and the placebo capsules (placebo food) contained 20 mg of maltodextrin. The other ingredients were prepared in the same amounts for both capsules.

[0027] [Conducting the test] A randomized, double-blind, placebo-controlled, parallel-group comparative trial was conducted with Japanese men and women aged 20 or older who reported having elevated blood pressure. 345 participants were selected based on their responses to an application questionnaire. Inclusion and exclusion criteria are described in the Appendix. 115 participants received a full explanation of the study content and methods from a physician, and provided written informed consent.

[0028] The person responsible for assigning the test food was assigned subjects to two groups (57 subjects in the sample food group and 58 subjects in the placebo group) to ensure no bias in age, sex, or BMI. The person responsible for assigning the test food was kept strictly confidential until the key was opened, and blinding was maintained for all parties except the person responsible for assigning the test food.

[0029] Two participants withdrew before the start of the study, so the study was conducted with the remaining 113 participants. The study was conducted from August to December 2020. Participants took one capsule of the test food once a day after breakfast for 12 weeks. During the intake period, participants recorded their capsule intake using a web questionnaire. During the study period, participants were instructed to comply with prohibitions and precautions, and otherwise maintained their normal lifestyle.

[0030] [Efficacy Evaluation] The subjects' blood pressure was measured 4 weeks before ingestion of the test food, on the first day of ingestion, and at 4, 8, and 12 weeks after ingestion. Blood pressure was measured multiple times after the subjects rested in a seated position for 10-15 minutes upon arrival at the clinic. The average of three measurements with an error margin of 5 mmHg or less was used for both systolic blood pressure (SBP) and diastolic blood pressure (DBP). Salt intake was estimated using the Tanaka formula, based on the amount of sodium and creatinine in urine before ingestion and at 12 weeks after ingestion.

[0031] [Safety Evaluation] Subject weight measurement and blood tests were performed 4 weeks before ingestion of the test food, on the day of ingestion, and at 4, 8, and 12 weeks after ingestion. Blood tests measured red blood cell count, white blood cell count, hemoglobin level, hematocrit value, platelet count, aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), gamma-glutamyltransferase (γ-GT), lactate dehydrogenase (LD), uric acid, glucose, creatinine, sodium, chloride, potassium, total cholesterol (TC), high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol, triglycerides (TG), total bilirubin, total protein, and urea nitrogen.

[0032] [Statistical analysis] Data were calculated as mean ± standard error. An unpaired t-test (two-tailed test) was used to compare the food group with the placebo group, while a paired t-test was used to compare the start date of intake with the period after intake. The statistical significance level was set at 5%, and statistical analysis software R was used for calculating statistics and performing tests.

[0033] [Test Results] During the study period, four participants in the placebo group dropped out due to illness. Therefore, at the end of the study, there were 53 participants in the placebo group and 56 in the sample food group, for a total of 109 participants. Safety evaluation was performed on these 109 participants. Efficacy evaluation was performed on 106 participants, after excluding three who met the exclusion criteria during the study period. Furthermore, the blood pressure of the 106 participants included in the efficacy evaluation on the start date of intake was classified according to the criteria of the Japanese Society of Hypertension's "Hypertension Treatment Guidelines 2014." The results are shown in Table 1.

[0034] [Table 1]

[0035] Furthermore, 89 individuals with high-normal blood pressure were included in the analysis. Table 2 shows the basic attributes of the analysis participants before the start of intake. No significant differences were observed between the placebo group and the food sample group in age, height, weight, BMI, and blood pressure.

[0036] [Table 2]

[0037] [Results of effectiveness evaluation] In a study of 89 individuals with high-normal blood pressure, divided into a placebo group and a group receiving the test food, changes in blood pressure before and after intake were compared at 4, 8, and 12 weeks. The results showed that the reduction in blood pressure in the test food group was greater than in the placebo group, both for systolic and diastolic blood pressure. However, no statistically significant difference was observed between the groups (Table 3).

[0038] [Table 3]

[0039] Next, stratified analysis was performed based on age and estimated salt intake. In age-based stratified analysis, to examine the effects on older age groups, where blood pressure is generally more likely to be elevated, a threshold of 55 years old was used, representing the upper third of the age distribution of the 89 subjects analyzed. When comparing the changes in blood pressure between the placebo group (n = 14) and the sample food group (n = 14) aged 55 and older, the changes in systolic blood pressure at 8 and 12 weeks were +6.4 mmHg and +4.4 mmHg, respectively, in the placebo group, while they were -5.2 mmHg and -4.0 mmHg in the sample food group, indicating a significant decrease in the sample food group (Figure 1(a); P = 0.0013 at 8 weeks and P = 0.047 at 12 weeks). On the other hand, the reduction in diastolic blood pressure in the sample food group at 8 and 12 weeks of intake was greater than in the placebo group, but no significant difference was observed between the groups (Figure 1(b). Error bars in the figure represent the standard deviation, and '*' represents P<0.05).

[0040] Next, a stratified analysis of salt intake was performed. The analysis was conducted using the target daily salt intake of 8.0g set by Health Japan 21 (Ministry of Health, Labour and Welfare) as the threshold value. As a result, in the group where the estimated salt intake at 12 weeks after intake was less than 8.0g, the decrease in systolic blood pressure at 12 weeks was -0.2 mmHg in the placebo group (n = 13) and -8.5 mmHg in the example food group (n = 15), with the example food group showing a significantly larger decrease (Figure 2(a); P = 0.022). Furthermore, the decrease in diastolic blood pressure at 12 weeks was +1.8 mmHg in the placebo group and -3.6 mmHg in the example food group, with the example food group showing a significantly larger decrease (Figure 2(b); P = 0.033). Note that error bars in the figures represent the standard deviation, and '*' represents P<0.05.

[0041] To investigate changes in salt intake among the subjects during the study period, we compared salt intake at the start of the test food intake with that at 12 weeks. As a result, no change was observed in the salt intake of the subjects before and after the intake of the test food (start of intake = 9.23 ± 0.2 g, 12 weeks = 9.22 ± 0.2 g; paired t-test: P=0.97). This suggests that the salt intake trend of the subjects did not change during the study period, and that the magnitude of the blood pressure reduction observed at 12 weeks of test food intake in the example food group was not due to a change in salt intake.

[0042] [Results of safety evaluation] The BMI and weight results are shown in Table 4.

[0043] [Table 4]

[0044] As is clear from Table 4, no significant differences were observed in changes in BMI or body weight between the placebo group and the group that received the sample food.

[0045] Next, the results of hematological and biochemical tests are shown in Tables 5 and 6. In Tables 5 and 6, '#' indicates P<0.05 (vs 0 W), and in Table 6, '*' indicates P<0.05 (example food group vs placebo group).

[0046] [Table 5]

[0047] [Table 6]

[0048] A significant difference in total bilirubin was observed between the placebo group and the sample food group at 12 weeks of intake (Table 6). However, no significant differences were observed between the groups in other items (Tables 5 and 6). When comparing the difference between the start date of intake and 12 weeks of intake between the placebo group and the sample food group, significant differences were observed in several items (Tables 5 and 6).

[0049] As explained above, the present invention has been shown to have the effect of lowering elevated blood pressure in elderly adults and adults who are classified as having high-normal blood pressure despite not having excessive salt intake in their daily diet.

Claims

1. A blood pressure lowering composition containing at least one of the following as an active ingredient: bacterial cells, bacterial cell components, or bacterial cell treatment products obtained from Bifidobacterium longum strain N714 (NITE BP-03004).

2. A blood pressure-lowering food or beverage containing at least one of the following as an active ingredient for lowering blood pressure: bacterial cells, bacterial cell components, or bacterial cell processed products obtained from Bifidobacterium longum N714 strain (NITE BP-03004).

3. A quasi-drug or pharmaceutical product for lowering blood pressure containing at least one of the following as an active ingredient for lowering blood pressure: bacterial cells, bacterial components, or bacterial treatment products obtained from Bifidobacterium longum N714 strain (NITE BP-03004).

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