JP2021123561A - Sleep promoting composition and food, pharmaceutical, and feed containing composition - Google Patents

Abstract

To provide a sleep promoting composition that can promote sleep introduction at night and has excellent stability and safety and to provide a food, a pharmaceutical, and feed containing the sleep promoting composition.SOLUTION: A sleep promoting composition contains, as an active ingredient, microorganisms belonging to any of Lactobacillus, Lactococcus, Leuconostoc, Pediococcus, Weissella, or Bifidobacterium or a culture thereof.SELECTED DRAWING: Figure 1

Description

The present invention relates to a sleep-promoting composition capable of promoting sleep and having excellent stability and safety. The present invention also relates to foods, pharmaceuticals, and feeds containing the sleep-promoting composition.

Sleep is an action necessary for animals to survive, and sleep states or sleep-like states exist not only in mammals but also in non-mammals such as fish, flies, and nematodes. In recent years, changes in lifestyle have disturbed the rhythm of life, and the prevalence of sleep disorders such as insomnia has increased. According to a survey by the Ministry of Health, Labor and Welfare, one in five people has some sort of sleep problem (Non-Patent Document 1). It has been clarified that sleep disorders are closely related not only to the decrease in labor productivity due to drowsiness, but also to mental illnesses such as dementia and depression, and lifestyle-related diseases such as obesity and diabetes. The economic loss due to this lack of sleep is estimated to be about 3.5 trillion yen (Non-Patent Document 2).

For people with sleep disorders such as insomnia, the use of pharmaceutical sleeping pills may be considered. As the hypnotic, barbituric acid hypnotics, benzodiazepine hypnotics, and non-benzodiazepine hypnotics are mainly used. However, barbituric acid hypnotics have strong side effects such as dependence, and sleep with benzodiazepine hypnotics induces sleep showing an abnormal frequency range called benzodiazepine hypnotics, so it should not lead to physiological and good quality sleep. It has been known.

For this reason, materials that can be taken on a daily basis instead of pharmaceuticals and that can induce physiological sleep are being actively developed, and various natural ingredients and food ingredients have been proposed. ing. For example, there are a sleep disorder improving agent using glycine, which is an amino acid (Patent Document 1), and a sleep promoting composition using theanine (Patent Document 2). In addition, even in lactic acid bacteria that have been used in fermented foods for a long time and are highly safe for living organisms, hypnotic activity by specific strains of non-pathogenic lactic acid bacteria Lactobacillus acidophilus, Lactobacillus herbeticus, and Streptococcus thermophilus ( Improvement of sleep quality by increasing REM sleep and decreasing non-REM sleep) (Patent Document 3), sleep quality improving agent by Lactobacillus casei (Patent Document 4), circadian rhythm improving agent by Lactobacillus brevis (Patent Document 5), etc. There is.

However, there are still not enough options to meet the diverse needs of consumers.

Drosophila melanogaster (hereinafter referred to as Drosophila melanogaster) is used as a model organism in various fields of biology. In 2000, the entire genome sequence was determined, and it is estimated that about 75% of genes related to human diseases are also present in Drosophila. In recent years, disease models such as Alzheimer's disease, Parkinson's disease, and metabolic syndrome have been developed. It is actually used for elucidation of the disease onset mechanism and screening of therapeutic agents (Non-Patent Documents 3 and 4).

Many studies using Drosophila have been conducted in the field of sleep research, and it is clear that Drosophila not only exhibit sleep-like behavior similar to humans, but also have many of their characteristics and molecular basis in common with mammalian sleep. (Non-Patent Documents 5 and 6). Specific examples include age-dependent changes in sleep patterns (Non-Patent Document 7), sleep constancy mechanism (sleep behavior that compensates for lost sleep (Non-Patent Document 8), and arousal action by serotonin (Non-Patent Document 7). 9), sleep-promoting action by glycine (Non-Patent Document 10), neurotransmitters such as dopamine, serotonin, and GABA, and sleep-wake control action of EGF (Non-Patent Documents 11, 12, and 13) are common to Drosophila and mammals. In addition, the Drosophila pars intercerebralis, which is developmentally and functionally similar to the hypothalamus of mammals, plays an important role in sleep and arousal-promoting nerves. Despite differences in the structure of the nervous system, such as the fact that circuits and sleep-promoting neural circuits are in communication with each other in Drosophila as well as mammals, sleep-related neural circuits are evolutionarily conserved. It is known (Non-Patent Documents 5 and 6).

From these facts, Drosophila is considered to be suitable as a model organism for sleep research and also useful for searching for substances involved in sleep control (Non-Patent Documents 5 and 6).

Japanese Unexamined Patent Publication No. 2013-121961 Japanese Unexamined Patent Publication No. WO01 / 07432 Japanese Patent No. 45277922 Japanese Patent No. 6279714 Japanese Patent No. 5943342

2017 National Health and Nutrition Survey, Ministry of Health, Labor and Welfare <https://www.mhlw.go.jp/content/000451755.pdf> Makoto Uchiyama, Japan Psychiatric Hospital Association Magazine 31 (11), 1163-1169, 2012-11, 2012 Pandey, U.B. & Nichols, C.D. Human Disease Models in Drosophila melanogaster and the Role of the Fly in Therapeutic Drug Discovery. Drug Deliv. 63, 411-436 (2011) Ugur, B., Chen, K. & Bellen, H. J. Drosophila tools and assays for the study of human diseases. Dis. Model. Mech. 9, 235-244 (2016). Zimmerman, J.E., Naidoo, N., Raizen, D.M. & Pack, A.I. Conservation of sleep: insights from non-mammalian model systems. Trends Neurosci. 31, 371-376 (2008). Ly, S., Pack, A.I. & Naidoo, N. The neurobiological basis of sleep: Insights from Drosophila. Neurosci. Biobehav. Rev. 87, 67-86 (2018). Koh, K., Evans, JM, Hendricks, JC & Sehgal, A. A Drosophila model for age-associated changes in sleep: wake cycles. Proc. Natl. Acad. Sci. USA 103 (37), 13843-13847 (2006) ) Huber, R. et al. Sleep homeostasis in Drosophila melanogaster. Sleep 27, 628-639 (2004) Wu, M.N. et al. The effects of caffeine on sleep in Drosophila require PKA activity, but not the adenosine receptor. J Neurosci 29 (35), 11029-11037 (2009) Kazuhiko Kume, Search for a New Sleep Control Mechanism Using Drosophila, Uehara Memorial Foundation Research Report, 30 (2016) Ueno, T. et al. Identification of a dopamine pathway that regulates sleep and arousal in Drosophila. Nat. Neurosci. 15, 1516-1523 (2012) Sehgal, A. & Mignot, E. Genetics of sleep and sleep disorders. Cell. 146, 194-207 (2011). Foltenyi, K et al. Neurohormonal and neuromodulatory control of sleep in Drosophila. Cold Spring Harb Symp Quant Biol. 72, 565-571 (2007)

An object of the present invention is to provide a sleep-promoting composition capable of promoting sleep and having excellent stability and safety. Another object of the present invention is to provide foods, pharmaceuticals, and feeds containing the sleep-promoting composition.

As a result of diligent studies to solve the above problems, the present inventors have found that a plurality of microorganisms promote sleep.
That is, the present invention includes the following aspects.
(1) A sleep-promoting composition containing one or more microorganisms belonging to any one of the genus Lactobacillus, Lactococcus, and Bifidobacterium.
(2) The sleep-promoting composition according to (1), wherein the sleep-promoting composition is a sleep-inducing-promoting composition.
(3) Lactobacillus sp. The sleep-promoting composition according to any one or more of (1) and (2) selected from.
(4) The genus Lactococcus is one or more selected from the Lactococcus lactis subspecies lactis (Lactococcus lactis subsp. Lactis) species and the Lactococcus rafquinolactis species (1) or (2). ). The sleep-promoting composition.
(5) The genus Bifidobacterium is Bifidobacterium longum species, Bifidobacterium longum subspecies Longum (Bifidobacterium longum subsp. Longum) species, Bifidobacterium longum subspecies. Bifidobacterium longum subspecies, Bifidobacterium longum subspecies, Bifidobacterium longum subspecies, Bifidobacterium longum subspecies, Bifidobacterium longum subspecies, Bifidobacterium longum subspecies, Bifidobacterium longum subspecies, Bifidobacterium longum subspecies The sleep-promoting composition according to (1) or (2), which is one or more selected from the film (Bifidobacterium longum) species.
(6) A sleep-promoting food, a sleep-promoting drug, or a sleep-promoting feed containing the sleep-promoting composition according to any one of (1) to (5).

The present invention provides a sleep-promoting composition capable of promoting sleep, and the term "promoting sleep (promoting sleep)" as used in the present application means promoting the introduction of sleep (the amount of sleep in the early night). It means that it can bring about an increase). That is, the present invention provides, in one aspect, a sleep-inducing promoting composition.

It is a figure which compared the relative value (sleep amount increase rate) of the median of the sleep amount of each test group with respect to the median of the sleep amount of a control group by the difference of the genus of a microorganism. It is a figure which shows the sleep onset latency of the group which administered each microorganism.

The sleep promoting composition of the present invention will be described. The active ingredient of the present invention is a microorganism belonging to the genus Lactobacillus, Lactococcus, Leukonostock, Pediococcus, Weissella, and Bifidobacterium, as long as it has an effect of promoting sleep induction. Both can be used.

The species is not particularly limited as long as it is Lactobacillus, Lactococcus, Leukonostock, Pediococcus, Weissera, or Bifidobacterium, but a preferred example is Lactobacillus acidophilus. , Lactobacillus amylovorus, Lactobacillus delbreckii subspecies Bulgaricus (Lactobacillus delbruecchii subsp. Bulgaricus) species, Lactobacillus fermentum (Lactobacillus) Lactobacillus helveticus, Lactobacillus jhonsiii, Lactobacillus mucosae, Lactobacillus mucosae, Lactobacillus orilus Bacillus paracasei subspecies Tolerance (Lactobacillus paracassei subsp. Tolerans) species, Lactobacillus plantarum species, Lactobacillus reuteri (Lactobacillus lactolas Lactobacillus sakei species, Lactobacillus salivalius subspecies Salicinus species, Lactobacillus subvarius subsp. Salicinius species, Lactobacillus baginalis species Lactobacillus raffinolactis species, Leuconost oc citreum, Leuconostoc mecenteroides subspecies Leuconostoc mecenteroides subsp. Pediococcus pentosaceus, Pediococcus stilesii, Weissella cibaria, Weissella Weissella Weissella Weissella Weissella Weissella Species, Bifidobacterium adolescentis Species, Bifidobacterium faecale Species, Bifidobacterium subspecies Longum (Bifidobacterium longum) Species Longum subspecies Infantis (Bifidobacterium longum subsp. Infantis) species, Bifidobacterium pseudocatenulatum species, Bifidobacterium pseudobisobisobisoboidum Examples include the Bifidobacterium thermofilm species.

These microorganisms may be used alone or in combination of two or more kinds of microorganisms as appropriate.

For culturing microorganisms, conditions suitable for each microorganism may be adopted. For example, as the medium, various media such as a milk medium or a medium containing a milk component, a semi-synthetic medium not containing the milk medium, and a chemically defined synthetic medium can be used. Examples of such a medium include reduced skim milk and MRS medium.

The cells separated from the obtained culture by means of collecting bacteria such as centrifugation can be used as they are as the active ingredient of the present invention. Live cells are preferable, but some treatment may be applied to the cells, and cells that have been concentrated, dried, freeze-dried, etc. may be used, or cells may be killed by heat drying, etc., and the treatment method thereof. Is not particularly limited.
Not only purely isolated cells, but also cultures, suspensions, other cell-containing substances, and cytoplasm and cell wall fractions obtained by treating cells using enzymes or physical means can be used. can.
The form of the culture is not only a culture using a medium generally used for culturing lactic acid bacteria such as a synthetic medium MRS medium (manufactured by DIFCO) and a reduced defatted milk medium, but also cheese, fermented milk, and milk. Products Dairy products such as lactic acid bacteria beverages can be exemplified, but are not particularly limited.

The sleep-promoting composition of the present invention can be added to foods, pharmaceuticals, feeds and the like. At that time, the sleep-promoting composition of the present invention may be used as it is, but since it can be used together with other raw materials usually contained in foods, pharmaceuticals, and feeds, powders, granules, tablets, etc. It can also be used for capsules, drinks and the like. It can also be added to foods and drinks such as yogurt, milk drinks and wafers, and feed.

When the composition for promoting sleep of the present invention is blended in foods, pharmaceuticals, feeds and the like, the blending ratio of microorganisms is not particularly limited, and it may be appropriately adjusted according to the ease of production and the preferable daily dose. It is determined individually in consideration of the symptoms, age, etc. of the recipient, but in the case of adults, usually 10 to 200 g, 20 to 200 g, 50 to 200 g, 100 to 200 g, 150 to 200 g of microbial cultures, etc. 20 to 150 g, 50 to 150 g, 100 to 150 g, or 50 to 100 g, or the cells themselves, 0.1 to 5,000 mg, 0.2 to 5,000 mg, 0.5 to 5,000 mg, 1 to 5, 000 mg, 2 to 5,000 mg, 5 to 5,000 mg, 10 to 5,000 mg, 20 to 5,000 mg, 50 to 5,000 mg, 100 to 5,000 mg, 200 to 5,000 mg, 500 to 5,000 mg, 1,000 to 5,000 mg, 2,000 to 5,000 mg, 0.2 to 2,000 mg, 0.5 to 2,000 mg, 1 to 2,000 mg, 2 to 2,000 mg, 5 to 2,000 mg, 10-2,000 mg, 20-2,000 mg, 50-2,000 mg, 100-2,000 mg, 200-2,000 mg, 500-2,000 mg, 1,000-2,000 mg, 0.5-1 000 mg, 1-1,000 mg, 2-1,000 mg, 5-1,000 mg, 10-1,000 mg, 20-1,000 mg, 50-1,000 mg, 100-1,000 mg, 200-1,000 mg, 500-1,000 mg, 1-500 mg, 2-500 mg, 5-500 mg, 10-500 mg, 20-500 mg, 50-500 mg, 100-500 mg, 200-500 mg, 2-200 mg, 5-200 mg, 10-200 mg, The blending amount and the like may be adjusted so that 20 to 200 mg, 50 to 200 mg, 100 to 200 mg, 5 to 100 mg, 10 to 100 mg, 20 to 100 mg, 50 to 100 mg, 10 to 50 mg, or 20 to 50 mg can be ingested. By ingesting in this way, a desired effect can be exhibited.

(Evaluation method of sleep promoting action of sleep promoting composition)
The sleep-promoting action of the sleep-promoting composition of the present invention can be confirmed by the animal test on Drosophila described in Examples.

Examples and test examples are shown below, and the present invention will be described in detail, but these are merely examples, and the present invention is not limited thereto.

(Test Example 1) Sleep-promoting action of microorganisms Sleep using Drosophila for various microorganisms belonging to the genus Lactobacillus, Lactococcus, Leuconostocaceae, Pediococcus, Streptococcus, Weissella, and Bifidobacterium. The promoting effect was evaluated.
1. 1. Test method (1) Donated bacteria 21 species of Lactobacillus, 2 species of Lactococcus, 4 species of Leuconostock, 3 species of Pediococcus, 4 species of Streptococcus, Weissera shown in Table 1. Three species of the genus and nine species of the genus Bifidobacterium were used as donors, one strain each.
(2) Preparation of donor cells After culturing each of the donor bacteria in (1) above under the conditions shown in Table 1 (OD600 = 0.7 or more), centrifugation (7,000 × g, 4 ° C., 20 min). ) Separated the cells. The cells were washed twice with physiological saline and then suspended in 1/10 of the culture volume in a 12.5% trehalose solution. Further, this suspension was mixed in equal amounts with a 2% agar and a 10% sucrose solution to prepare a feed containing the donor cells.
(3) Test procedure A wild-type Drosophila CS ^2202u strain was bred at a temperature of 25 ° C. under a light-dark cycle every 12 hours, and an unmated female individual 5 days after emergence was used for evaluation. Drosophila is placed in a glass tube having a diameter of 5 mm together with the feed (test group) of (2) above or a feed composed of 1% agar and 5% sucrose without lactic acid bacteria, and Drosophila is measured. Behavior was measured for 3 days in an activity monitoring system (manufactured by Trikinetics). In Drosophila, a period of immobility (a period of inactivity) of 5 minutes or more is defined as "sleep". Therefore, the test group and the control group were compared for the amount of sleep in the first 3 hours (ZT12 to 15 in approximate day time) at night on the 3rd day of measurement. Each group was evaluated using about 96 Drosophila, and after performing the Wilcoxon rank sum test, it was corrected by the Holm method, and it was judged that there was a significant difference at p <0.05.

2. Test Results From Table 2, Lactobacillus acidophilus species, Lactobacillus amylovourus species, Lactobacillus delbreckii subspecies Lactobacillus delbrucyllus vulsyllus vulsyllus vulsyllus delbulus delbulus fermentum species, Lactobacillus gasseri species, Lactobacillus helveticus species, Lactobacillus joussonii species, Lactobacillus johnsonii species, Lactobacillus jhonsonii species oris) species, Lactobacillus parabuchneri species, Lactobacillus paracasei subspecies Tolerance (Lactobacillus paracassei subsp. Tolarans) species, Lactobacillus plantarlus talus Lactobacillus rhamnosus species, Lactobacillus sakei species, Lactobacillus salivarius subspecies Lactobacillus salivarius salivarius subs Subspecies Lactobacillus lactis subsp. , Leukonostock Pseudobacterium pseudomesenteroides, Pediococcus acidilactici, Pediococcus pentosaceus, Pediococcus pestisicossis Weissella cybaria, Weissella confusa, Weissella paramesenteroides, Bifidobacterium adresentis, Bifidobacterium longum bacterium adolicis , Bifidobacterium longum subspecies Longum (Bifidobacterium longum subsp. Longum) species, Bifidobacterium longum subspecies Infantis (Bifidobacterium longum subspec. ) Species, Bifidobacterium pseudolongum subspecies, Bifidobacterium pseudolonegum subsp. Globosum species, Bifidobacterium thermofilm (Bifidobacterium thermofilm) species compared with the control group at night Was found to be significantly increased. Similarly, sleep onset latency was also significantly reduced in most of the species tested (see also Figure 2).

The relative value of the median sleep amount of each test group to the median sleep amount of the control group was defined as the sleep increase rate, and the results of comparison by different genera are shown in FIG. Since Non-Patent Document 3 states that Streptococcus thermophilus has hypnotic activity, a comparison of the sleep increase rates of each genus with respect to the sleep increase rate of the genus Streptococcus shows that the genus Bifidobacterium tends to be large. The genera Lactobacillus, Leuconostoc, and Pediococcus were found to be significantly larger. That is, it was shown that the genus Bifidobacterium, the genus Lactobacillus, the genus Leuconostoc, and the genus Pediococcus have a particularly remarkable sleep promoting effect.

(Example 1) Production of sleep-promoting agent (granule) 5 weights of Lactobacillus plantarum MSN16 strain was added to an edible synthetic medium (0.5% yeast extract, 0.1% trypticase peptone addition). % Inoculated, cultured at 38 ° C. for 15 hours, and then the cells were collected by centrifugation. The recovered cells were freeze-dried to obtain a freeze-dried powder of the early cells. 1 g of this freeze-dried powder was mixed with 5 g of lactose and molded into granules to obtain the sleep promoter of the present invention.

(Example 2) Manufacture of sleep-promoting agent (powder) In accordance with the provisions of the 13th revised Japanese Pharmacopoeia Manual of Pharmaceutical Formula "Powder", lactose (10 g of freeze-dried powder of MSN16 strain obtained in Example 1 above) is added to lactose (Example 2). 400 g of Japanese Pharmacopoeia and 600 g of potato starch (Japanese Pharmacopoeia) were added and mixed uniformly to produce the sleep-promoting agent of the present invention.

(Example 3) Production of nutritional composition for promoting sleep 5 weights of Lactobacillus plantarum MSN16 strain was added to an edible synthetic medium (0.5% yeast extract, 0.1% trypticase soybean peptone addition). % Inoculated, cultured at 38 ° C. for 15 hours, and then the cells were collected by centrifugation, and the collected cells were lyophilized. To 40 g of vitamin C or an equal amount mixture of vitamin C and citric acid, 100 g of granulated sugar, and 60 g of an equal amount mixture of cornstarch and lactose, 40 g of the freeze-dried powder of this freeze-dried powder was added and mixed. The mixture was packed in a bag to produce the sleep-promoting nutritional composition of the present invention.

(Example 4) Production of sleep-promoting beverage After dissolving 1 g of the MSN16 strain obtained in Example 3 in 699 g of deionized water and heating to 40 ° C., an ultra disperser (ULTRA-TURRAX T-25; The mixture was stirred and mixed at 9,500 rpm for 20 minutes at IKA Japan Co., Ltd.). After adding 100 g of maltitol, 2 g of acidulant, 20 g of reduced starch syrup, 2 g of flavor and 176 g of deionized water, the mixture is filled in a 100 ml glass bottle, sterilized at 95 ° C. for 15 seconds, sealed, and 10 beverages of the present invention (100 ml). Included) was manufactured.

(Example 5) Production of feed for promoting sleep Soybean meal 12 kg, defatted milk powder 14 kg, soybean oil 4 kg, corn oil 2 kg, palm oil 23.2 kg, corn starch 14 kg, wheat flour 9 kg, bran 2 kg, vitamin mixture 5 kg, cellulose 2. 8 kg and 2 kg of a mineral mixture were blended and sterilized at 120 ° C. for 4 minutes, and 10 kg of the MSN16 strain obtained in Example 3 was blended to produce the feed of the present invention.

(Example 6) Production of fermented milk Lactobacillus plantarum MSN16 strain was inoculated into an edible synthetic medium (0.5% yeast extract, 0.1% trypticase soybean addition) in an amount of 5% by weight. After culturing at 37 ° C. for 17 hours, the cells were collected by centrifugation and lyophilized. 5 g of this freeze-dried mycelium, 1700 g of skim milk powder, 300 g of glucose, and 7695 g of deionized water were mixed and kept at 95 ° C. for 2 hours for heat sterilization. This was cooled to 37 ° C., 300 g of a lactic acid bacterium starter (Lb. Casei) was inoculated, stirred and mixed, and then fermented to pH 4.0 in an incubator maintained at 37 ° C. After reaching pH 4.0, the mixture was cooled to 10 ° C. or lower to produce 10 kg of the fermented milk of the present invention.

The present invention includes microorganisms belonging to the genus Lactobacillus, Lactococcus, Leukonostock, Pediococcus, Weissella, Bifidobacterium, and compositions containing the microorganisms, as compositions that promote sleep. It provides foods, pharmaceuticals, and feeds containing compositions. Sleep can be promoted by ingesting the composition of the present invention or the like.

Claims (6)

A sleep-promoting composition comprising one or more microorganisms belonging to any one of the genus Lactobacillus, Lactococcus, and Bifidobacterium. The sleep-promoting composition according to claim 1, wherein the sleep-promoting composition is a sleep-inducing-promoting composition. The genus Lactobacillus is a subspecies of Lactobacillus paracasei subsp. Tolerans, a species of Lactobacillus rhamnosus, a subspecies of Lactobacillus salivalis The sleep-promoting composition according to claim 1 or 2, which is one or more of the above. The first or second claim, wherein the genus Lactococcus is one or more selected from the Lactococcus lactis subspecies lactis (Lactococcus lactis subsp. Lactis) species and the Lactococcus rafquinolactis species. Sleep-promoting composition. The genus Bifidobacterium is Bifidobacterium longum faecale species, Bifidobacterium longum subspecies Longum (Bifidobacterium longum subsp. Longum) species, Bifidobacterium longum subspecies Infantis. Bifidobacterium longum subspecies, Bifidobacterium longum subspecies, Bifidobacterium longum subspecies, Bifidobacterium longum subspecies, Bifidobacterium longum subspecies, Bifidobacterium longum subspecies, Bifidobacterium longum subspecies, Bifidobacterium longum subspecies, Bifidobacterium longum subspecies. The sleep-promoting composition according to claim 1 or 2, which is one or more selected from the thermophilum species. A sleep-promoting food, a sleep-promoting drug, or a sleep-promoting feed containing the sleep-promoting composition according to any one of claims 1 to 5.

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