JP7438709B2 - Bifidobacterium improves cognitive function - Google Patents

Description

NPMD NITE BP-02214

TECHNICAL FIELD The present invention relates to Bifidobacterium having an effect of improving cognitive function. More specifically, the present invention relates to bifidobacteria that improve cognitive function by improving visual memory.

In recent years, the increase in the number of dementia patients has become a problem. It is estimated that there are 50 million people worldwide with dementia, and it is estimated that this number will reach 152 million by 2050. It is also predicted that approximately 10% of the population will suffer from dementia by 2025.

Aging is known to be the greatest risk factor for dementia. Using Japan as an example, the prevalence of dementia increases with age: 3% at age 65, 14% at age 75, 40% at age 85, and 80% at age 95. (See Non-Patent Document 1).

Dementia generally refers to a condition in which cognitive functions such as memory, recognition, and judgment deteriorate, which interferes with daily life. Alzheimer's disease (AD) is well known as a typical dementia, but the cause of AD is still unknown and no fundamental treatment has been established.

Takashi Asada: Prevalence of dementia in urban areas and response to daily life impairment due to dementia Ministry of Health, Labor and Welfare Chemical Research Grant Comprehensive Research Project on Dementia Countermeasures FY2011-FY2012 Comprehensive Research Report 2013 ) March

By the way, early measures are important to prevent dementia. Specifically, it is important to maintain current cognitive function or improve cognitive function before cognitive function deteriorates. Various foods have been proposed as foods that improve cognitive function. The present inventors investigated whether there are any new foodstuffs that can improve cognitive function, other than the foodstuffs that have been reported so far. Then, they newly discovered that ingesting Bifidobacteria strain N61, which has the ability to promote serotonin secretion, improves visual memory, leading to the completion of the present invention.

In order to solve the above problems, the present invention is a cognitive function improving agent containing the cells of Bifidobacterium N61 strain (NITE BP-02214) as an active ingredient.

In the above configuration, it is preferable that the bacterial cells of the N61 strain are dead bacteria.

In order to solve the above problem, the present invention is a cognitive function improving agent that is in the form of a unit intake per meal, and the unit contains N61 strain as an active ingredient.

The Bifidobacterium N61 strain of the present invention improves cognitive function by improving visual memory.

FIG. 1 is an explanatory diagram for explaining the result notation method for each test item in Cognitrax. FIG. 2 is an explanatory diagram for explaining the relationship between each test item and functional area of Cognitrax.

The present invention will be explained in detail below.
1. Bifidobacterium longum N61 strain (NITE BP-02214)
The Bifidobacterium of the present invention is Bifidobacterium longum. Among bifidobacteria belonging to Bifidobacterium longum, this strain is particularly Bifidobacterium longum N61 strain (NITE BP-02214). The symbol N61 referred to in the present invention is a number uniquely assigned to a bacterial strain by Nissin Foods Holdings, Inc., and this Bifidobacterium longum N61 strain was first isolated by the present inventor.

Bifidobacterium longum N61 strain of the present invention has been deposited under the following conditions.
(1) Name of depositary institution: National Institute of Technology and Innovation, Patent Microorganisms Depositary Center (2) Contact information: Room 122, 2-5-8 Kazusa Kamatari, Kisarazu, Chiba 292-0818 (3) Accession number: NITE BP -02214
(4) Display for identification: N61
(5) Date of original deposit: March 3, 2016 (6) Date of transfer to deposit under the Budapest Treaty: January 5, 2017

A known method can be used to prepare the Bifidobacterium longum N61 strain used in this embodiment. For example, for pre-culture and main culture, GAM medium (Nissui) is prepared according to a predetermined method, and Bifidobacterium longum N61 strain is inoculated at 37°C for 24 hours. The bacterial cells in the main culture solution are collected by centrifugation or membrane concentration, and the bacterial cells are washed with water and then heat sterilized. In addition, sterilization treatment may be performed before the bacterial collection treatment. After sterilization and washing of the bacterial cells, a dry sterilized powder can be obtained by freeze-drying. The method for pulverizing the bacterial cells is not limited to freeze-drying, but may also be performed by spray drying, drum drying, or the like. In addition, viable bacterial powder can be obtained without sterilization.

As the bacterial cells of Bifidobacterium longum N61 strain in this embodiment, live bacteria or dead bacteria may be used. Moreover, when using live bacteria, dead bacteria may be mixed. Furthermore, it is preferable to ingest 20 mg or more of Bifidobacterium longum N61 strain per day. Regarding intake, the daily amount may be taken at once or divided into several doses, and the timing of intake does not matter. For example, if there is a tablet containing 10 mg of killed bacteria of Bifidobacterium longum N61 per tablet as a unit dosage form, you can take two tablets at once or divide the two tablets into several doses. You may. Furthermore, if each tablet contains 20 mg of killed bacteria of Bifidobacterium longum N61 strain, it is sufficient to take one tablet per day. There is no upper limit to the daily intake of Bifidobacterium longum strain N61, but if we were to set an upper limit, we believe that 1 g would be sufficient. This is because it is thought that there is no difference in efficacy even if more than that is taken. Furthermore, it is preferable that the bacterial cells of Bifidobacterium longum N61 be continuously ingested for 12 weeks or more.

The method of ingesting Bifidobacterium longum N61 strain is not particularly limited, but oral ingestion is preferred. Specifically, it is preferable to ingest Bifidobacterium longum N61 strain cells as a food or drink containing them as a food composition, a medicine containing them as a pharmaceutical composition, or the like.

When ingested as food or drink, for example, Bifidobacterium longum N61 strain can be used as a food composition in fermented milk and lactic acid bacteria drinks, dairy products such as butter, processed egg products such as mayonnaise, sweet breads such as butter cake, etc. It can also be used as a material. 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 drink. In addition, Bifidobacterium breve strain N61 alone may be ingested as is as a food or drink. In addition to general drinks and foods, it may also be included in foods for specified health uses, foods with functional claims, nutritional supplements, supplements, and the like.

When ingested as a medicine, it can be used together with excipients and other pharmaceutically acceptable carriers. Examples of the shape of the pharmaceutical include tablets, capsules, and granules. Note that a known method can be used for formulation.

Hereinafter, the present invention will be specifically explained based on Examples. In this example, an ingestion test was conducted on 56 healthy adults to examine the effects of the present invention.

<Creation of test food and control food (placebo food)>
The test foods and placebo foods were as shown in Table 1.

試験食品の組成（1日摂取量当たり）

Composition of test food (per daily intake)

<Test>
In this example, 60 people were selected through screening and divided into two groups of 30 people each using a random number table for allocation. Subjects divided into two groups were given either a test food or a placebo food for 12 weeks. At the end of the intake, there were 56 subjects (29 subjects in the intake group (group A) and 27 subjects in the non-consumption group (group P), excluding those who violated the rules or declined the test). The general background of the subjects at the time of screening is shown in Table 2.

被験者背景因子
数値は、性別は該当被験者数、それ以外は平均値 ± 標準偏差で示す。
P群との群間比較（性別：χ2検定、性別以外の項目：2標本t検定）

Subject background factor values are expressed as the number of applicable subjects for gender, and as mean ± standard deviation for other factors.
Intergroup comparison with group P (gender: χ2 test, items other than gender: two-sample t-test)

Subjects were asked to take one packet (two tablets) a day with water or lukewarm water without chewing. However, the time of intake was not specified. Also, even if I forgot to take a dose, I made sure not to take too much the next day. During the study, subjects were given diaries and had to keep records every day.

<Cognitive function evaluation test (Cognitrax)>
Cognitrax basic (manufactured by Health Solution Co., Ltd.) was implemented. Cognitrax test items and their contents are shown in Table 3. The order of inspection was the order in which each inspection item was listed in Table 3.

Cognitraxの検査項目

Cognitrax inspection items

Further, the results of each test are expressed as items listed in the right column of FIG. To give an example, the results of a verbal memory test are expressed as the number of correct hits (immediate), the number of correct passes (immediate), the number of correct hits (delayed), and the number of correct passes (delayed).

Furthermore, as shown in FIG. 2, the above inspection tests are related to each functional area. Here, the results of each inspection test are used to calculate scores for each functional area. In FIG. 2, "0" indicates that it is directly used to calculate the score of the functional area. "△" indicates that it is used indirectly.

Details of each functional area are as follows.
(1) Overall memory ability: Indicates the ability to memorize, and is evaluated by the total score of verbal memory ability and visual memory ability. An increase in the number indicates improvement.
(2) Verbal memory ability: Indicates the ability to memorize words and is evaluated using a verbal memory test. An increase in the number indicates improvement.
(3) Visual memory ability: Indicates the ability to memorize figures and is evaluated using a visual memory test. An increase in the number indicates improvement.
(4) Cognitive function speed: This test measures the ability to quickly process visual information. Evaluation is based on the sum of the right-hand average and left-hand average of the finger-tapping test, and the total number of correct responses on the SDC test. An increase in the number indicates improvement.
(5) Reaction time: This test measures the ability to quickly respond to instructions. It is evaluated by the combined reaction time of the Stroop test and the average value of the Stroop reaction time. A decrease in the number indicates improvement.
(6) General attentiveness: This test measures the ability to maintain attention and respond accurately. Evaluation is based on the total score of the number of incorrect Stroop responses in the Stroop test, the number of incorrect responses in the attention shift test, the number of missed correct answers, and the number of incorrect responses in the sustained processing test. A decrease in the number indicates improvement.
(7) Cognitive flexibility: This test measures the ability to respond to and process changes in instructions. Evaluation is based on the total score obtained by subtracting the number of incorrect responses in the Attention Shift Test and the number of Stroop incorrect responses in the Stroop Test from the number of correct responses in the Attention Shift Test. An increase in the number indicates improvement.
(8) Processing speed: This test measures the ability to quickly process information. Evaluation is based on the total score obtained by subtracting the number of incorrect answers on the SDC test from the number of correct answers on the SDC test. An increase in the number indicates improvement.
(9) Executive function: This test measures the ability to understand rules and concepts and make decisions. Evaluation is based on the total score obtained by subtracting the number of incorrect answers on the Attention Shift Test from the number of correct answers (responses) on the Attention Shift Test. An increase in the number indicates improvement.
(10) Simple attention: This is a test that measures the ability to pay attention for a long period of time. Evaluation is based on the total score obtained by subtracting the number of incorrect responses in the sustained processing test from the number of correct responses in the sustained processing test. An increase in the number indicates improvement.
(11) Movement speed: This is a test that measures the ability to repeat fine movements quickly. Evaluation is based on the total score of the right-hand average and left-hand average of the finger tapping test. An increase in the number indicates improvement.

Cognitrax test results are shown in Tables 4 to 10.

Cognitrax言語記憶テストの測定値
いずれの数値も、平均値 ± 標準偏差で示す。
数値の増加が改善を示す。
P群：n＝27、A群：n＝29
摂取前との群内比較 * p ＜ 0.05、** p ＜ 0.01（1標本t検定）
P群との群間比較 † p ＜ 0.1、# p ＜ 0.05、## p ＜ 0.01（2標本t検定）

Cognitrax Verbal Memory Test Measurements All values are expressed as mean ± standard deviation.
An increase in the number indicates improvement.
Group P: n=27, Group A: n=29
Intra-group comparison with before intake * p < 0.05, ** p < 0.01 (one sample t-test)
Between-group comparison with P group † p < 0.1, # p < 0.05, ## p < 0.01 (two-sample t-test)

Cognitrax視覚記憶テストの測定値
いずれの数値も、平均値 ± 標準偏差で示す。
P群：n＝27、A群：n＝29
数値の増加が改善を示す。
摂取前との群内比較 * p ＜ 0.05、** p ＜ 0.01（1標本t検定）
P群との群間比較 † p ＜ 0.1、# p ＜ 0.05、## p ＜ 0.01（2標本t検定）

Cognitrax Visual Memory Test Measurements All values are expressed as mean ± standard deviation.
Group P: n=27, Group A: n=29
An increase in the number indicates improvement.
Intra-group comparison with before intake * p < 0.05, ** p < 0.01 (one sample t-test)
Between-group comparison with P group † p < 0.1, # p < 0.05, ## p < 0.01 (two-sample t-test)

From the results in Table 5, superiority was observed in the number of correct hits (immediate), and a significant difference was observed in the number of correct hits (delayed). As shown in Figure 2, the visual memory test is related to the visual memory domain, and the visual memory test is directly used to calculate the score of the visual memory domain. These results suggest that it has an effect on visual memory.

Cognitrax指たたきテストの測定値
いずれの数値も、平均値 ± 標準偏差で示す。
P群：n＝27、A群：n＝29
数値の増加が改善を示す。
摂取前との群内比較 * p ＜ 0.05、** p ＜ 0.01（1標本t検定）
P群との群間比較 † p ＜ 0.1、# p ＜ 0.05、## p ＜ 0.01（2標本t検定）

Cognitrax Finger Tapping Test Measurements All values are expressed as mean ± standard deviation.
Group P: n=27, Group A: n=29
An increase in the number indicates improvement.
Intra-group comparison with before intake * p < 0.05, ** p < 0.01 (one sample t-test)
Between-group comparison with P group † p < 0.1, # p < 0.05, ## p < 0.01 (two-sample t-test)

From the results in Table 6, a superiority was observed in the left hand average, and a significant difference was observed in the right hand average. As shown in Figure 2, the finger tapping test is related to the cognitive function speed domain and the motor speed domain. Additionally, the finger tapping test is directly used to calculate scores in the cognitive function speed domain and the motor speed domain. These results suggest that it is effective in the cognitive function speed area and the motor speed area.

Cognitrax SDCテストの測定値
いずれの数値も、平均値 ± 標準偏差で示す。
※で示した項目は数値の低下が改善を示し、その他は数値の増加が改善を示す。
P群：n＝27、A群：n＝29
摂取前との群内比較 * p ＜ 0.05、** p ＜ 0.01（1標本t検定）
P群との群間比較 † p ＜ 0.1、# p ＜ 0.05、## p ＜ 0.01（2標本t検定）

Cognitrax SDC test measurements All values are expressed as mean ± standard deviation.
For items marked with *, a decrease in the value indicates improvement, and for other items, an increase in the value indicates improvement.
Group P: n=27, Group A: n=29
Intra-group comparison with before intake * p < 0.05, ** p < 0.01 (one sample t-test)
Between-group comparison with P group † p < 0.1, # p < 0.05, ## p < 0.01 (two-sample t-test)

Cognitraxストループテストの測定値
いずれの数値も、平均値 ± 標準偏差で示す。
数値の低下が改善を示す。
P群：n＝27、A群：n＝28
摂取前との群内比較 * p ＜ 0.05、** p ＜ 0.01（1標本t検定）
P群との群間比較 † p ＜ 0.1、# p ＜ 0.05、## p ＜ 0.01（2標本t検定）

Cognitrax Stroop test measurements All values are expressed as mean ± standard deviation.
A decrease in the number indicates improvement.
Group P: n=27, Group A: n=28
Intra-group comparison with before intake * p < 0.05, ** p < 0.01 (one sample t-test)
Between-group comparison with P group † p < 0.1, # p < 0.05, ## p < 0.01 (two-sample t-test)

Cognitrax注意シフトテストの測定値
いずれの数値も、平均値 ± 標準偏差で示す。
※で示した項目は数値の低下が改善を示し、その他は数値の増加が改善を示す。
P群：n＝27、A群：n＝29
摂取前との群内比較 * p ＜ 0.05、** p ＜ 0.01（1標本t検定）
P群との群間比較 † p ＜ 0.1、# p ＜ 0.05、## p ＜ 0.01（2標本t検定）

Cognitrax Attention Shift Test Measurements All values are expressed as mean ± standard deviation.
For items marked with *, a decrease in the value indicates improvement, and for other items, an increase in the value indicates improvement.
Group P: n=27, Group A: n=29
Intra-group comparison with before intake * p < 0.05, ** p < 0.01 (one sample t-test)
Between-group comparison with P group † p < 0.1, # p < 0.05, ## p < 0.01 (two-sample t-test)

Cognitrax持続処理テストの測定値
いずれの数値も、平均値 ± 標準偏差で示す。
※で示した項目は数値の低下が改善を示し、その他は数値の増加が改善を示す。
P群：n＝27、A群：n＝29
摂取前との群内比較 * p ＜ 0.05、** p ＜ 0.01（1標本t検定）
P群との群間比較 † p ＜ 0.1、# p ＜ 0.05、## p ＜ 0.01（2標本t検定）

Cognitrax sustained processing test measurements All values are expressed as mean ± standard deviation.
For items marked with *, a decrease in the value indicates improvement, and for other items, an increase in the value indicates improvement.
Group P: n=27, Group A: n=29
Intra-group comparison with before intake * p < 0.05, ** p < 0.01 (one sample t-test)
Between-group comparison with P group † p < 0.1, # p < 0.05, ## p < 0.01 (two-sample t-test)

Furthermore, the scores for each functional area were as shown in Table 11.

いずれの数値も、平均値 ± 標準偏差で示す。
数値の増加が改善を示す。
P群：n＝27、A群：n＝29
摂取前との群内比較 * p ＜ 0.05、** p ＜ 0.01（1標本t検定）
P群との群間比較 † p ＜ 0.1、# p ＜ 0.05、## p ＜ 0.01（2標本t検定）

All values are expressed as mean value ± standard deviation.
An increase in the number indicates improvement.
Group P: n=27, Group A: n=29
Intra-group comparison with before intake * p < 0.05, ** p < 0.01 (one sample t-test)
Between-group comparison with P group † p < 0.1, # p < 0.05, ## p < 0.01 (two-sample t-test)

From the results in Table 11, a significant tendency was observed in the visual memory area in the intergroup comparison.

The mean age of this study (n=56) was 58.2±5.3 years. According to standard data for CNS vital signs (corresponding to the basic Cognitrax package) reported by Gualtieri and Johnson, the average overall memory domain score for 20- to 29-year-olds was 99.7. Overall, the subjects in this study were considered to be elderly adults with poor memory function, as their overall memory domain score was <99.7. In addition, in the normative data, the overall memory domain score for those aged 50 to 59 was 97.4, so subjects with scores <97.4 to 1SD (89.9) are considered to have considerably lower memory function due to aging. Estimated. For these reasons, a stratified analysis was performed using a baseline total memory domain score >=90. Among subjects with a baseline total memory domain score of 90 or higher, the total memory domain score ranged from 90 to 98, suggesting that these subjects were healthy but had slightly poor memory ( low normal memory function group, n=34). On the other hand, in subjects whose baseline total memory domain score was less than 90, the average value of the total memory domain score was 83.9, which was lower than the standard data for patients with mild cognitive impairment (86.9). However, the scores on the Mini Mental State Test (MMSE-J), a dementia screening test often used to diagnose dementia, were within the normal range (below normal memory function group, n = 22).

Table 12 shows 34 subjects with an overall memory score of 90 to 98 points (17 subjects in the P group, 17 subjects in the A group) and 22 subjects with an overall memory score of less than 90 points (10 subjects in the P group, 12 subjects in the A group). The subjects' background factors were shown.

数値は、性別は該当被験者数、それ以外は平均値 ± 標準偏差で示す。
P群との群間比較（性別：χ2検定、性別以外の項目：2標本t検定）

Figures for gender are expressed as the number of applicable subjects, and other values are expressed as mean ± standard deviation.
Intergroup comparison with group P (gender: χ2 test, items other than gender: two-sample t-test)

Next, we reanalyzed the scores of each functional area for the 34 subjects. The results are shown in Table 13.

いずれの数値も、平均値 ± 標準偏差で示す。数値の増加が改善を示す。
P群：n＝17、A群：n＝17
摂取前との群内比較 * p ＜ 0.05、** p ＜ 0.01（1標本t検定）
P群との群間比較 † p ＜ 0.1、# p ＜ 0.05、## p ＜ 0.01（2標本t検定）

All values are expressed as mean value ± standard deviation. An increase in the number indicates improvement.
Group P: n=17, Group A: n=17
Intra-group comparison with before intake * p < 0.05, ** p < 0.01 (one sample t-test)
Between-group comparison with P group † p < 0.1, # p < 0.05, ## p < 0.01 (two-sample t-test)

As shown in Table 13, significant differences were observed in overall memory and visual memory before and after ingestion. Additionally, a significant difference in visual memory ability was observed between the groups.

Next, we reanalyzed the Stroop test results. The results are shown in Table 14.

いずれの数値も、平均値 ± 標準偏差で示す。数値の低下が改善を示す。
P群：n＝17、A群：n＝17
摂取前との群内比較 * p ＜ 0.05、** p ＜ 0.01（1標本t検定）
P群との群間比較 † p ＜ 0.1、# p ＜ 0.05、## p ＜ 0.01（2標本t検定）

All values are expressed as mean value ± standard deviation. A decrease in the number indicates improvement.
Group P: n=17, Group A: n=17
Intra-group comparison with before intake * p < 0.05, ** p < 0.01 (one sample t-test)
Between-group comparison with P group † p < 0.1, # p < 0.05, ## p < 0.01 (two-sample t-test)

As shown in Table 14, significant differences were observed in simple reaction time, complex reaction time (correct answer), and Stroop reaction time (correct answer) before and after ingestion. Furthermore, in the comparison between groups, a significant difference was observed in composite reaction time (correct answer). As shown in Figure 2, the Stroop test is related to the reaction time domain, general attention domain, and cognitive flexibility domain. In addition, the Stroop test is directly used to calculate scores in the reaction time domain, general attention domain, and cognitive flexibility domain. These results suggest that it is effective in reaction time, general attention, and cognitive flexibility.

The above results suggested that ingestion of N61 bifidobacteria improves cognitive function, especially visual-based cognitive function.

Claims (3)

A cognitive function improving agent that improves visual memory , containing Bifidobacterium strain N61 strain (NITE BP-02214) as an active ingredient . The agent for improving cognitive function by improving visual memory according to claim 1 , wherein the bacterial cells of the N61 strain are dead bacteria. A cognitive function improving agent for improving visual memory, which is in the form of a unit intake per meal, and each unit contains strain N61 as an active ingredient.

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