JP7475155B2 - Composition for improving endurance - Google Patents

Description

NPMD NITE BP-02621 IPOD FERM BP-11175

This technology relates to compositions for promoting glycogen storage, compositions for improving endurance, compositions for supporting endurance, and compositions for preventing, improving, or suppressing fatigue.

In recent years, in today's society where people lead irregular lifestyles and are subject to a lot of stress, people of all ages and genders are encouraged to take part in moderate exercise and sports to prevent lifestyle-related diseases, promote health, relieve stress, and so on.
For example, in order to maintain or improve the quality of daily life activities, it is recommended that elderly people engage in moderate exercise, moderate sports, and maintain the amount of daily activity. Such physical activity is said to be effective for elderly people in preventing sarcopenia (age-related muscle weakness), relieving stress, and preventing forgetfulness. In general, the decline in muscle mass, muscle strength, and physical function that occurs with aging is called sarcopenia.
In general, in sports and exercise, importance is placed on physical strength, particularly endurance, fatigue resistance, fatigue recovery, and the like.

For example, Patent Document 1 discloses a physical activity promoter that contains Lactobacillus gasseri as an active ingredient, with the aim of providing lactic acid bacteria that promote bodily functions.

JP 2013-47192 A JP 2015-590103 A JP 2005-289861 A

The present inventor has focused on endurance, believing that endurance is important when continuously performing sports, moderate exercise, daily activities, etc. Furthermore, as a result of extensive research into endurance, the present inventor has focused on glycogen, which serves as an energy source for activity, rather than the conventional methods of promoting metabolic functions or increasing muscle mass (especially slow-twitch muscle mass).
Therefore, a main objective of the present technology is to provide technology relating to endurance.

As a conventional technique related to promoting glycogen storage, Patent Document 2 describes a muscle glycogen storage promoter that contains processed powder of a plant of the genus Panax notoginseng of the family Araliaceae, which can increase muscle glycogen storage safely and easily without exerting an exercise load and without causing injury. Patent Document 3 describes a composition used to promote glycogen storage, which contains milk whey protein as an active ingredient, with the aim of providing a composition for promoting glycogen storage.

As a result of intensive research, the present inventors discovered that Bifidobacterium longum and/or breve, particularly Bifidobacterium longum NITE BP-02621 and/or Bifidobacterium breve FERM BP-11175, have endurance-improving effects, endurance-supporting effects, glycogen storage-promoting effects, and the like, and thus completed the present invention. The technology is as follows.

The present technology provides a composition for promoting glycogen storage, which contains Bifidobacterium longum NITE BP-02621 and/or Bifidobacterium breve FERM BP-11175 as an active ingredient.
The present technology provides a composition for improving or supporting endurance, which contains Bifidobacterium longum NITE BP-02621 and/or Bifidobacterium breve FERM BP-11175 as an active ingredient.
The present technology provides a composition for preventing, improving or suppressing fatigue, which contains Bifidobacterium longum NITE BP-02621 and/or Bifidobacterium breve FERM BP-11175 as an active ingredient.
Preferably, said endurance is muscular endurance.
Preferably, the composition is a pharmaceutical composition.
Preferably, the composition is a food composition.

This technology can provide technology related to endurance. Note that the effects described here are not necessarily limited, and may be any of the effects described in this technology.

The following describes a preferred embodiment for implementing the present technology. Note that the embodiment described below is an example of a representative embodiment of the present technology, and is not intended to narrow the scope of the present technology. Note that percentages in this specification are expressed by mass unless otherwise specified. Furthermore, the upper and lower limits of each numerical range can be combined in any desired way.

The present technology is a composition containing Bifidobacterium longum and/or Bifidobacterium breve as an active ingredient, and more preferably a composition containing Bifidobacterium longum NITE BP-02621 and/or Bifidobacterium breve FERM BP-11175 as an active ingredient.
The composition according to the present technology is a composition used for promoting glycogen storage, improving endurance, assisting endurance, or preventing, improving, suppressing fatigue, etc. The composition can also be used for pharmaceuticals, foods, etc.

The Bifidobacterium bacteria used in the present technology are one or more species selected from the group consisting of, for example, Bifidobacterium longum subsp. longum; Bifidobacterium breve, etc.

Among the Bifidobacterium bacteria, Bifidobacterium longum (more preferably Bifidobacterium longum subsp. longum) and/or Bifidobacterium breve are more preferable in terms of the effects of the present technology, such as the glycogen storage promoting effect and endurance improving effect.

From the viewpoint of the effects of the present technology, such as promoting glycogen storage and supporting endurance, the Bifidobacterium bacteria used in the present technology include, for example, the following strains, and one or more species selected from the group consisting of these are preferred. In other words, the Bifidobacterium bacteria used in the present technology are more preferably Bifidobacterium longum NITE BP-02621 and/or Bifidobacterium breve FERM BP-11175.

As an example of Bifidobacterium subsp. longum, Bifidobacterium longum subsp. longum NITE BP-02621 (BB536) (deposited on January 26, 2018) is suitable.

Examples of Bifidobacterium breve include Bifidobacterium breve FERM BP-11175 (MCC1274; B3) (deposited on August 25, 2009) and Bifidobacterium breve NITE BP-02622 (deposited on January 26, 2018) (M-16V), and one or more species selected from the group consisting of these are preferred.

More specifically, the Bifidobacterium longum subsp. longum NITE BP-02621 (ATCC BAA-999; BB536) was internationally deposited under the Budapest Treaty on January 26, 2018 at the National Institute of Technology and Evaluation, Patent Microorganisms Depositary (NPMD) (Address: Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, 292-0818, Japan) under the accession number NITE BP-02621. Bifidobacterium longum NITE BP-02621 is the same bacterium as Bifidobacterium longum ATCC BAA-999, which has been deposited with the American Type Culture Collection (ATCC), a US preservation institution (10801 University Boulevard, Manassas, Virginia, 20110, USA) under the accession number ATCC BAA-999. This strain is also available as a commercial product (BB536) from Morinaga Milk Industry Co., Ltd.

The Bifidobacterium breve FERM BP-11175 (MCC1274; B-3) was internationally deposited under the Budapest Treaty on August 25, 2009 at the National Institute of Advanced Industrial Science and Technology (currently the National Institute of Technology and Evaluation (NITE) Patent Organism Depositary (IPOD) (NITE-IPOD)) (Address: Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, 292-0818, Japan) under the accession number FERM BP-11175. This strain is also available as a commercial product (B-3) from Morinaga Milk Industry Co., Ltd.

The strains with the accession number "NITE" are those deposited at the National Institute of Technology and Evaluation, Patent Microorganisms Depositary (NPMD) (Address: Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, 292-0818, Japan), an international depositary institution based on the Budapest Treaty.
The strains with the accession number "FERM" are strains that have been deposited at the International Patent Organism Depositary of the National Institute of Advanced Industrial Science and Technology (currently the International Patent Organism Depositary of the National Institute of Technology and Evaluation (NITE) (NITE-IPOD) (Address: 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, 292-0818, Japan), an international depositary institution based on the Budapest Treaty.

The strains specified by the strain names exemplified in the present technology are not limited to the strains themselves that have been deposited or registered in a specified institution under the strain name (hereinafter, for convenience of explanation, also referred to as "deposited strains"), but also include strains that are substantially equivalent thereto (also referred to as "derived strains" or "derived strains"). That is, for example, the above Bifidobacterium longum subsp. longum NITE BP-02621 (ATCC BAA-999; BB536) has the accession number NITE BP-02621, and the above Bifidobacterium breve FERM BP-11175 (MCC1274; B-3) has the accession number FERM BP-11175, and are not limited to the strains themselves that have been deposited in a depository institution under the accession number NITE BP-02621, but also include strains that are substantially equivalent thereto.
Regarding the strain, "a strain substantially equivalent to the deposited strain" means a strain that belongs to the same species as the deposited strain and that can obtain the effects of the present invention, such as improved glycogen storage capacity and endurance. A strain substantially equivalent to the deposited strain may be, for example, a derived strain that uses the deposited strain as a parent strain. Examples of derived strains include strains bred from the deposited strain and strains that naturally arise from the deposited strain.

Substantially identical strains and derived strains include the following strains.
(1) Strains determined to be identical by the RAPD (Randomly Amplified Polymorphic DNA) and PFGE (Pulsed-field gel electrophoresis) methods (described in Probiotics in food/Health and nutritional properties and guidelines for evaluation 85, page 43)
(2) A strain that contains only genes derived from the deposited strain, does not contain any genes of foreign origin, and has a DNA identity of 95% or more (preferably 98% or more). (3) A strain bred from the deposited strain (including genetic engineering modifications, mutations, and natural mutations), or a strain that has the same characteristics.

The Bifidobacterium bacteria used in the present technology can be easily grown by culturing the bacteria. In addition, the Bifidobacterium bacteria used in the present technology can be obtained by appropriate production, or a commercially available product can be used.
The method for culturing the Bifidobacterium bacteria used in the present technology is not particularly limited as long as the Bifidobacterium bacteria can grow, and a method commonly used for culturing Bifidobacterium bacteria can be used with appropriate modifications as necessary. For example, the culture temperature may be 25 to 50° C., and preferably 30 to 40° C. The culture is preferably performed under anaerobic conditions, and for example, the culture can be performed while aerating anaerobic gas such as carbon dioxide gas.

The medium for culturing the Bifidobacterium genus bacteria is not particularly limited, and a medium commonly used for culturing Bifidobacterium genus bacteria can be used with appropriate modifications as necessary. As the carbon source of the medium, for example, sugars such as galactose, glucose, fructose, mannose, cellobiose, maltose, lactose, sucrose, trehalose, starch, starch hydrolysates, and blackstrap molasses can be used depending on the assimilation ability. As the nitrogen source of the medium, for example, ammonium salts such as ammonia, ammonium sulfate, ammonium chloride, and ammonium nitrate, and nitrates can be used. In addition, as the inorganic salt of the medium, for example, sodium chloride, potassium chloride, potassium phosphate, magnesium sulfate, calcium chloride, calcium nitrate, manganese chloride, ferrous sulfate, and the like can be used. In addition, organic components such as peptone, soybean flour, defatted soybean meal, meat extract, and yeast extract can be used in the medium. The medium may be prepared from a known medium, and as a prepared medium, for example, MRS medium can be suitably used.

The form of the Bifidobacterium bacteria used in the present technology is not particularly limited, and may be any of bacterial bodies, a bacterial culture, or a treated bacterial product.
The bacterial cells may be, for example, live bacterial cells, dead bacterial cells, or a mixture of live bacterial cells and dead bacterial cells, and the dead bacterial cells may be sterilized or have broken cell walls. These bacterial cells may be wet or dry. From the viewpoint of the effects of the present technology, such as the glycogen storage promoting effect and the endurance improving effect, these bacterial cells are preferably live bacterial cells and/or dead bacterial cells.

The bacterial culture may be used as it is, or may be diluted or concentrated, or bacterial cells recovered from the culture may be used. The bacteria to be recovered may be live or killed, or may be both live and killed. The bacterial culture may be a culture supernatant or a culture fraction.
Examples of processed bacterial products that can be used include bacterial cells or cultures that have been subjected to various treatments such as heat treatment, crushing treatment, and drying treatment (e.g., freeze-drying), as well as diluted products, dried products, and fractions of such treatments.

The Bifidobacterium longum and/or Bifidobacterium breve in the present technology, or a composition containing said bacteria, has the effect of promoting glycogen storage, the effect of improving endurance, the effect of supporting endurance, and the effect of preventing, improving, or suppressing fatigue.
As used herein, "promoting glycogen storage" means that when the composition of the present technology is ingested, the glycogen content (g) per gram of muscle tissue or liver tissue increases compared to when the composition is not ingested.
As used herein, "endurance" refers to the ability to continuously perform physical activities such as sports and daily life (standing, walking, running, climbing stairs, etc.).
As used herein, the term "effect of supporting endurance" refers to supplying an energy source to tissues to extend the durability (duration) of a person's activity (endurance) or to maintain improved endurance.
As used herein, the term "effect of supporting muscular endurance" refers to supplying an energy source to muscles to extend the duration of a person's muscular activity, and to supporting the improvement and sustainability of muscular endurance.
As used herein, "fatigue" refers to physical fatigue resulting from sports and physical activities in daily life (standing, walking, running, climbing stairs, etc.).

In general, when engaging in activities, including exercise, physical strength, especially endurance, resistance to fatigue, and ability to recover from fatigue, are important. Endurance limits and fatigue occur when glycogen in tissues is consumed and falls below a certain level. For example, when glycogen in tissues is depleted, it can become difficult to maintain activity.
It is generally known that there is a positive correlation between the amount of glycogen stored in tissues and endurance, so it is believed that increasing the amount of glycogen stored in tissues will help improve or support endurance, resistance to fatigue, and fatigue recovery.

It is also known that glycogen stored in muscles is converted to glucose and consumed, while glycogen stored in the liver is converted to glucose and supplied to tissues throughout the body, including muscles, via the blood (Reference 1: Laurence Cole et al., Human Physiology, Biochemistry and Basic Medicine, 2015). As exemplified in the Examples below, this technology uses Bifidobacterium longum and/or Bifidobacterium breve, and is effective in promoting glycogen storage not only in muscle tissue but also in liver tissue. This means that this technology can efficiently and stably supply glucose, an energy source necessary for tissues, to prevent it from being depleted, thereby supporting (assisting) endurance, thereby improving and sustaining endurance.

For this reason, this technology can be used by people of all ages and genders, and is also beneficial for people who tend to become physically inactive, and is particularly beneficial for athletes and those who actively participate in exercise and sports.
Therefore, Bifidobacterium longum and/or Bifidobacterium breve in the present technology, or a composition containing said bacteria, has the effect of improving endurance, the effect of sustaining endurance, the effect of supporting endurance, and the effect of preventing, improving, and suppressing fatigue, etc.

This technology can maintain a person's endurance in daily activities without exerting a strong exercise load, which makes it easier to avoid the risk of injury that comes with exerting a strong exercise load, making it particularly useful for the elderly and those undergoing rehabilitation.
Bifidobacterium longum and/or Bifidobacterium breve in the present technology have the effect of improving muscle endurance, the effect of sustaining muscle endurance, the effect of supporting muscle endurance, and the effect of preventing, improving, and suppressing muscle fatigue, etc. In this way, the present technology is more effective against muscle endurance and muscle fatigue.

In addition, the present technology can increase glycogen storage in both muscle tissue and liver tissue even after intense exercise by ingesting Bifidobacterium longum and/or Bifidobacterium breve. Therefore, Bifidobacterium longum and/or Bifidobacterium breve in the present technology can be applied to people who wish to recover from fatigue (for example, people who are recovering from illness, or people who are active or have just finished working).
Therefore, Bifidobacterium longum and/or Bifidobacterium breve in the present technology have the effect of promoting recovery from fatigue in muscles and the liver, assisting in fatigue recovery, and the like.

The Bifidobacterium longum and/or Bifidobacterium breve of the present technology can be used as the bacteria themselves, or can be used in combination with a physiologically or pharma- ceutical acceptable conventional carrier or diluent.
In addition, the Bifidobacterium longum and/or Bifidobacterium breve of the present technology can be used in a variety of applications and compositions, such as medicines, foods, and feeds.

The present technology can also provide Bifidobacterium longum and/or Bifidobacterium breve or use thereof for the purposes of achieving the above-mentioned effects of the present technology, such as promoting glycogen storage, improving endurance, assisting endurance, preventing, improving, suppressing fatigue, etc. Furthermore, the Bifidobacterium longum and/or Bifidobacterium breve of the present technology can be used as an active ingredient in a method for promoting glycogen storage, a method for improving endurance, a method for assisting endurance, or a method for preventing, improving, or suppressing symptoms such as fatigue.
Furthermore, the Bifidobacterium longum and/or Bifidobacterium breve of the present technology can be used for producing various preparations or compositions having the above-mentioned effects or intended uses.
Furthermore, the Bifidobacterium longum and/or Bifidobacterium breve or a composition containing said bacteria of the present technology can be used in a method for promoting glycogen storage, a method for improving endurance, a method for supporting endurance, or for preventing, improving, suppressing, or treating symptoms such as fatigue.

The Bifidobacterium longum and/or Bifidobacterium breve of the present technology may be used on humans or non-human animals (preferably primates), preferably humans, pets (dogs, cats, etc.), and livestock (cows, sheep, pigs, etc.), and more preferably humans.

Additionally, the technology may be for therapeutic or non-therapeutic uses.
"Non-therapeutic purposes" is a concept that does not include medical procedures, i.e., treatment of the human body through therapy. Examples include health promotion and prevention of lifestyle-related diseases (e.g., borderline pre-diseases).
"Prevention" refers to preventing or delaying the onset of a disease or condition, or reducing the risk of the disease or condition to which it is applied.
"Amelioration" refers to improvement of a disease, symptom or condition, prevention or slowing of deterioration, or reversal, prevention or slowing of progression.
"Suppression" refers to improving, preventing or slowing the worsening, or reversing, preventing or slowing the progression of a disease, symptom or condition.

In the present technology, in addition to Bifidobacterium longum and/or Bifidobacterium breve, optional components may be used in combination as necessary. As optional components, components acceptable for use in pharmaceuticals, foods, feeds, etc. may be used as appropriate. Examples of optional components include sugars, sugar alcohols, polysaccharides, pH adjusters, fatty acid esters, flavorings, fragrances, excipients, etc.

The composition of the present technology can be produced by using Bifidobacterium longum and/or Bifidobacterium breve. The composition of the present technology can be obtained by mixing the Bifidobacterium bacteria with various raw materials of the composition to a predetermined content as described below. The content or amount of each raw material used in the composition of the present technology may be the same or similar amount with reference to the amount of each raw material normally used in the target composition.
The composition of the present technology can be produced using known production methods.

The manufacturing method of the composition of the present technology is described below, but the manufacturing method of the present technology is not limited to this. This makes it possible to provide a new composition, and examples of uses of the composition include promoting glycogen storage, improving endurance, assisting endurance, and recovering from fatigue.

In addition, it is preferable to use the manufacturing method of the present technology to manufacture a food composition, because the Bifidobacterium longum and/or Bifidobacterium breve of the present technology is ingestible and highly safe, making it easy to handle during the manufacturing stage, and fermentation technology (preferably fermented milk technology) is easily applicable, and raw materials such as milk components or plant-based nutrient substances can be used as desired. In addition, the manufacturing method of the present technology can also provide health-conscious food compositions, and among these, food compositions for promoting glycogen storage, improving endurance, supporting endurance, and recovering from fatigue are preferable.

In addition, the composition of the present technology may be fermented milk, and when producing fermented milk of the present technology, the production process of the production method of the present technology may include a fermentation step using Bifidobacterium longum and/or Bifidobacterium breve, or may include a step of mixing a fermented product produced on a separate line.
Furthermore, the composition obtained by the manufacturing method of the present technology contains Bifidobacterium longum and/or Bifidobacterium breve of the present technology, and therefore can be used as a food ingredient to impart the effects of the present technology, and can be used in foods to which the effects of the present technology have been imparted or in the manufacturing method thereof.

Depending on the form of the composition of the present technology (e.g., tablet, drink, capsule, pill, etc.), the manufacturing method of the present technology may include a step for forming the form, whereby the composition containing Bifidobacterium longum and/or Bifidobacterium breve of the present technology can be made into various forms. In this formation step, the Bifidobacterium bacteria can be mixed with excipients and the like to obtain a mixture, and examples of such steps include compressing the mixture in the case of a tablet; filling the mixture into a container in the case of a drink; and filling the mixture into a capsule in the case of a capsule.

The method for producing the composition of the present technology will be described as a first embodiment and a second embodiment, but the uses of the composition are not limited to the above-mentioned compositions for promoting glycogen storage, improving endurance, supporting endurance, and recovering from fatigue.

The present technology provides a method for producing the food composition according to a first embodiment, the method including at least one of the following steps (a) and (b):
(a) mixing the Bifidobacterium longum and/or Bifidobacterium breve of the present technology and a prebiotic; or (b) mixing the Bifidobacterium longum and/or Bifidobacterium breve of the present technology and a milk component;
can be provided.
Furthermore, in the mixing step (a), it is preferable to further mix a milk component.
The step (A) of obtaining a culture in the second embodiment can also be carried out in the first embodiment in order to obtain the Bifidobacterium bacteria used in the first embodiment. The step (B) of obtaining bacterial powder in the second embodiment can also be carried out appropriately in the first embodiment.

The manufacturing method according to the first embodiment of the present technology can produce compositions for promoting glycogen storage and compositions for improving endurance, etc., with high production efficiency in terms of cost and workability, and with which the effects of the present technology can be satisfactorily exhibited.

In addition, the present technology provides a second embodiment of a method for producing a food composition, the method including the following steps (A) and (B), and the food composition is a composition related to the present technology, such as the above-mentioned composition for promoting glycogen storage or composition for improving endurance:
(A) culturing Bifidobacterium longum and/or Bifidobacterium breve of the present technology in a medium to obtain a culture;
(B) drying the culture to obtain a bacterial powder;
The drying is preferably freeze-drying.
The steps (A) and (B) can be carried out with reference to the method for culturing the Bifidobacterium bacteria used in the present technology. For example, milk components may be mixed into the culture medium for the culture, and the medium may be appropriately fermented to produce fermented milk. Furthermore, lactic acid bacteria and/or other Bifidobacterium bacteria may be appropriately used for fermentation.
In addition, in the manufacturing method of the second embodiment, configurations that overlap with those of the first embodiment can be omitted as appropriate.

The manufacturing method according to the second embodiment of the present technology can produce compositions related to the present technology, such as compositions for promoting glycogen storage and compositions for improving endurance, with high production efficiency and capable of satisfactorily demonstrating the effects of the present technology. The manufacturing method according to the second embodiment can produce food compositions, pharmaceutical compositions, and feed compositions, and among these, it is preferable to produce food compositions such as fermented milk and bacterial powder from the viewpoint of production efficiency.

The form of the culture containing Bifidobacterium longum and/or Bifidobacterium breve is not particularly limited and may be either liquid or solid, but a dry form (e.g., bacterial powder or dried culture product) is preferred from the viewpoint of ease of handling during production and storage. In addition, examples of the culture include the bacterial cells themselves from which the culture solution has been separated, and a culture containing the bacterial cells, as described above.

The drying method is not particularly limited, but examples thereof include freeze-drying, spray-drying, retort sterilization, freeze-drying, UHT sterilization, pressure sterilization, high-pressure steam sterilization, dry heat sterilization, circulating steam disinfection, electromagnetic wave sterilization, electron beam sterilization, high frequency sterilization, radiation sterilization, ultraviolet sterilization, ethylene oxide gas sterilization, hydrogen peroxide gas plasma sterilization, and chemical sterilization (alcohol sterilization, formalin fixation, and electrolyzed water treatment).

The cells may be disrupted, and the disrupted material may be either live or dead cells that have been disrupted, or may have been subjected to heating, freeze-drying, or the like after disruption.
Of these, from the viewpoint of increasing the viable cell rate, it is preferable to subject the culture to freeze-drying, and from the viewpoint of increasing production efficiency, it is preferable to subject the culture to spray-drying.

In addition, the composition of the present technology may contain known or future components having probiotic effects or components that support the probiotic effects, as long as they do not impair the effects of the present technology. Here, probiotics generally refer to bacteria that have beneficial effects in the intestines. In addition, substances that serve as selective nutrient sources for bacteria that have beneficial effects in the intestines and promote their growth are generally called prebiotics.
When a prebiotic is used or contained to promote the growth of Bifidobacterium longum and/or Bifidobacterium breve in the present technology, the amount of prebiotic is preferably 1 to 1,000,000 parts by mass, and more preferably 10 to 10,000 parts by mass, per 100 parts by mass of the bacteria.

Examples of the prebiotics include various proteins such as whey protein, casein protein, soy protein, or pea protein (pea protein), or mixtures or hydrolysates thereof; amino acids such as leucine, valine, isoleucine, or glutamine; vitamins such as vitamin B6 or vitamin C; creatine; citric acid; fish oil; or oligosaccharides such as isomaltooligosaccharides, galactooligosaccharides, xylooligosaccharides, soybean oligosaccharides, fructooligosaccharides, lactulose, and human milk oligosaccharides (HMO). One or more types selected from the group consisting of these may be used.
Furthermore, the composition of the present technology can be produced by combining the prebiotic component with the bacteria of the present technology or a culture thereof.

In addition, the "human milk oligosaccharides" that can be used with this technology include 2'-fucosyllactose, 3-fucosyllactose, 2',3-difucosyllactose, lacto-N-triose II, lacto-N-tetraose, lacto-N-neotetraose, lacto-N-fucopentaose I, lacto-N-neofucopentaose, lacto-N-fucopentaose II, lacto-N-fucopentaose III, lacto-N-fucopentaose V, lacto-N-neofucopentaose V, lacto-N Neutral human milk oligosaccharides such as difucohexaose I, lacto-N-difucohexaose II, 6'-galactosyl lactose, 3'-galactosyl lactose, lacto-N-hexaose, and lacto-N-neohexaose, and acidic human milk oligosaccharides such as 3'-sialyllactose, 6'-sialyllactose, 3-fucosyl-3'-sialyllactose, and disialyl-lacto-N-tetraose can be used, and one or more types selected from the group consisting of these may be used.

The intake (administration) amount of the composition of the present technology is appropriately selected depending on the age, sex, condition, and other conditions of the subject to be ingested (administered).
Regardless of the amount or period of ingestion (administration), the composition of the present technology can be ingested (administered) once or multiple times a day.
The content or dosage of Bifidobacterium longum and/or Bifidobacterium breve cells (live or killed) in the present technology is not particularly limited, but is preferably 1×10 ³ to 1×10 ¹² (CFU/g or CFU/mL), more preferably 1×10 ⁵ to 1×10 ¹¹ (CFU/g or CFU/mL), and even more preferably 1×10 ⁷ to 1×10 ¹⁰ (CFU/g or CFU/mL) in the composition. Note that CFU stands for colony forming unit. In this specification, when the cells are killed, "CFU" is replaced with "cells".
The content or dosage of the bacteria may be the content of only Bifidobacterium longum bacteria, the content of only Bifidobacterium breve bacteria, or the total amount of both. These may be within the range of the content when the composition is usually distributed as an oral composition.
The bacterial cell count (CFU) can be measured by spreading and culturing an appropriately diluted bacterial suspension on a suitable agar medium, such as BCP-added plate count agar medium (manufactured by Eiken Chemical Co., Ltd.), and counting the number of colonies that appear.

The daily dosage of the Bifidobacterium bacteria (live or killed) of the present technology is not particularly limited, but is preferably 1 x ¹⁰ to 1 x ¹⁰ CFU/day, more preferably 1 x ¹⁰ to 1 x ¹⁰ CFU/day, and even more preferably 1 x ¹⁰ to 1 x ¹⁰ CFU/day.
The dosage of the bacteria may be the dosage of only Bifidobacterium longum, or the dosage of only Bifidobacterium breve, or the total dosage of both.
The dosage of the Bifidobacterium bacteria of the present technology is, for example, preferably in the range of 0.5 to 5 g/day for an adult, more preferably in the range of 1 to 4 g/day, and even more preferably 2 to 3 g/day.

The administration timing and interval of the present technology are not particularly limited, and may be once a day or may be divided into multiple doses, but once a day is preferable because it is simple and effective.
Since the Bifidobacterium longum and/or Bifidobacterium breve of the present technology are highly safe, they can be administered or ingested for the above-mentioned symptoms or diseases such as decreased glycogen storage, decreased endurance, fatigue, etc. Furthermore, from the viewpoint of prevention, it is preferable to administer or ingest the present technology before symptoms or diseases caused by decreased glycogen storage, decreased endurance, fatigue, etc. are observed. From the viewpoint of easily obtaining the preventive effect and the subsequent improving effect, it is preferable to administer or ingest the present technology one week (more preferably two weeks) before performing exercise.
Furthermore, in the case of the present technology, after symptoms such as decreased glycogen storage, decreased stamina, and fatigue have been observed, it is preferable from the standpoint of effectiveness to administer or ingest the product for a long period of time, more preferably for two weeks or more, and even more preferably for four weeks or more, from the standpoint of making it easier to obtain improvement effects.

Pharmaceutical Compositions
When Bifidobacterium longum and/or Bifidobacterium breve are used in pharmaceutical compositions or medical applications, the pharmaceuticals may be administered orally or parenterally, but oral administration and administration acting on mucous membranes are preferred. Parenteral administration includes, for example, injection (blood, skin, muscle, etc.), rectal administration, inhalation, etc. Oral dosage forms include, for example, tablets, capsules, lozenges, syrups, granules, powders, ointments, etc.
The dosage and administration of the pharmaceutical composition of the present technology is as described above.

When formulating the product, in addition to Bifidobacterium longum and/or Bifidobacterium breve, ingredients such as excipients, pH adjusters, colorants, and flavorings that are typically used in formulations can be used. Furthermore, ingredients that have known or future disease-preventing or therapeutic effects can also be used in combination depending on the purpose.

Furthermore, depending on the method of administration, the compound can be formulated into a suitable dosage form as desired. For example, in the case of oral administration, the compound can be formulated into solid preparations such as powders, granules, tablets, and capsules; or liquid preparations such as solutions, syrups, suspensions, and emulsions. In addition, in the case of parenteral administration, the compound can be formulated into suppositories, sprays, ointments, patches, injections, and the like.

In addition, formulation can be carried out by a known method as appropriate depending on the dosage form. When formulating, only Bifidobacterium bacteria or each isolated/purified fraction may be formulated, or a formulation may be prepared by appropriately mixing a formulation carrier, etc.

As the pharmaceutical carrier, various organic or inorganic carriers can be used depending on the dosage form. In the case of a solid formulation, examples of the carrier include excipients, binders, disintegrants, lubricants, stabilizers, flavoring agents, etc.

Examples of excipients include sugar derivatives such as lactose, sucrose, glucose, mannitol, and sorbitol; starch derivatives such as corn starch, potato starch, α-starch, dextrin, and carboxymethyl starch; cellulose derivatives such as crystalline cellulose, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, carboxymethyl cellulose, and calcium carboxymethyl cellulose; gum arabic; dextran; pullulan; silicate derivatives such as light anhydrous silicic acid, synthetic aluminum silicate, and magnesium aluminometasilicate; phosphate derivatives such as calcium phosphate; carbonate derivatives such as calcium carbonate; and sulfate derivatives such as calcium sulfate.

Examples of binders include the above excipients, gelatin, polyvinylpyrrolidone, macrogol, etc.

Disintegrants include, for example, the above-mentioned excipients as well as chemically modified starch or cellulose derivatives such as croscarmellose sodium, sodium carboxymethyl starch, and cross-linked polyvinylpyrrolidone.

Examples of lubricants include talc; stearic acid; metal stearates such as calcium stearate and magnesium stearate; colloidal silica; waxes such as pea gum and geranium stearate; boric acid; glycol; carboxylic acids such as fumaric acid and adipic acid; sodium carboxylates such as sodium benzoate; sulfates such as sodium sulfate; leucine; lauryl sulfates such as sodium lauryl sulfate and magnesium lauryl sulfate; silicic acids such as silicic anhydride and silicic acid hydrate; and starch derivatives.

Examples of stabilizers include paraoxybenzoic acid esters such as methylparaben and propylparaben; alcohols such as chlorobutanol, benzyl alcohol, and phenylethyl alcohol; benzalkonium chloride; acetic anhydride; and sorbic acid.

Examples of flavoring agents include sweeteners, acidulants, and fragrances.
In the case of a liquid preparation for oral administration, examples of the carrier used include a solvent such as water, a flavoring agent, and the like.

[Food composition]
The present technology can provide a food composition containing Bifidobacterium longum and/or Bifidobacterium breve as an active ingredient. The food composition of the present technology also includes beverages. The food composition of the present technology can be used to prevent or improve the above-mentioned symptoms such as decreased glycogen storage, decreased endurance, and fatigue. For example, it can be used as a health food or functional food with the concept of improving endurance and anti-fatigue.

Furthermore, the food composition according to the present technology can be infant formula. Infant formula refers to infant formula for infants aged 0-12 months, follow-up milk for infants aged 6-9 months and older and young children (up to 3 years old), low birth weight infant formula for newborns weighing less than 2500g at birth (low birth weight infants), various therapeutic milks used to treat children with pathological conditions such as milk allergy and lactose intolerance, etc.

When Bifidobacterium longum and/or Bifidobacterium breve is used in foods for humans or animals, it can be prepared by adding it to known foods, or it can be mixed into food ingredients to produce new foods.
The method of use, amount used, and content in the food composition of the present technology are as described above.

The foods may be in any form, such as liquid, paste, solid, or powder, and may include tablet confectionery, liquid food, and feed (including for pets), as well as, for example, flour products, instant foods, processed agricultural products, processed marine products, processed livestock products, milk and dairy products, oils and fats, basic seasonings, compound seasonings and foods, frozen foods, confectionery, beverages, and other commercially available products.

Examples of flour products include bread, macaroni, spaghetti, noodles, cake mixes, fried chicken flour, and breadcrumbs.
Examples of instant foods include instant noodles, cup noodles, retort/prepared foods, canned foods, microwave foods, instant soups/stews, instant miso soup/cleansing liquids, canned soups, freeze-dried foods, and other instant foods.
Examples of processed agricultural products include canned agricultural products, canned fruit, jams and marmalades, pickles, boiled beans, dried agricultural products, and cereals (processed grain products).
Examples of processed seafood products include canned seafood, fish ham and sausage, seafood paste products, seafood delicacies, and tsukudani.
Examples of processed livestock products include canned livestock products and pastes, livestock ham and sausages, etc.
Examples of milk and dairy products include processed milk, milk drinks, yogurt, lactic acid bacteria drinks, cheese, ice cream, infant formula, cream, and other dairy products.
Examples of fats and oils include butter, margarines, vegetable oils, and the like.
Examples of basic seasonings include soy sauce, miso, sauces, tomato-processed seasonings, mirin, vinegars, etc., and examples of the compound seasonings and foods include cooking mixes, curry bases, sauces, dressings, noodle soups, spices, and other compound seasonings.
Examples of frozen foods include raw frozen foods, semi-cooked frozen foods, and cooked frozen foods.
Examples of sweets include caramel, candy, chewing gum, chocolate, cookies, biscuits, cakes, pies, snacks, crackers, Japanese sweets, rice sweets, bean sweets, dessert sweets, and other sweets.

Examples of beverages include carbonated beverages, natural fruit juices, fruit juice drinks, soft drinks containing fruit juice, fruit pulp drinks, fruit drinks containing fruit particles, vegetable beverages, soy milk, soy milk beverages, coffee beverages, tea beverages, powdered beverages, concentrated beverages, sports drinks, nutritional beverages, alcoholic beverages, and other beverages of your choice.
Other commercially available foods include, for example, baby food, furikake, and ochazuke nori.

In addition, the food defined by the present technology may be provided and sold as a food with a health use label.
The act of "displaying" includes all acts aimed at informing consumers of the above-mentioned uses, and any expression that can recall or infer the above-mentioned uses falls under the act of "displaying" of this technology, regardless of the purpose of the display, the content of the display, the object or medium on which it is displayed, etc.

It is preferable that the "labeling" be done in a way that allows consumers to directly recognize the above-mentioned uses. Specifically, this includes the acts of transferring, delivering, displaying for transfer or delivery, or importing food products or product packaging on which the above-mentioned uses are written, displaying or distributing advertisements, price lists, or transaction documents related to the products and writing the above-mentioned uses in information containing the above-mentioned uses and providing them by electromagnetic means (such as the Internet), etc.

On the other hand, it is preferable that the content of the labeling be approved by the government or the like (for example, a labeling approved based on various systems established by the government and made in a manner based on such approval). It is also preferable that such content of the labeling be affixed to promotional materials at the point of sale such as packaging, containers, catalogs, pamphlets, and POP, as well as other documents, etc.

In addition, "labeling" also includes labeling as health food, functional food, enteral nutritional food, special purpose food, health functional food, food for specified health uses, nutritional functional food, functional food, functional labeling food, medical drug, etc. Among these, in particular, labeling approved by the Consumer Affairs Agency, such as labeling approved under the Food for Specified Health Uses system or similar systems, etc. Examples of the latter include labeling as a food for specified health uses, labeling as a conditional food for specified health uses, labeling that indicates an effect on the structure or function of the body, labeling that reduces the risk of disease, etc. More specifically, typical examples are labeling as a food for specified health uses (especially labeling of health uses) and similar labeling as stipulated in the Enforcement Regulations of the Health Promotion Act (Ministry of Health, Labour and Welfare Ordinance No. 86 of April 30, 2003) and labeling similar thereto.

Examples of such claims include "For those who want to improve their endurance," "Supports improving endurance," "For those who want to have a body that does not tire easily," "For fatigue recovery," "For those who are concerned about physical fatigue," "Promotes fatigue recovery," etc.

[Feed composition]
When Bifidobacterium longum and/or Bifidobacterium breve are used in feed, they can be added to known feed to prepare the feed, or they can be mixed into the raw materials of the feed to produce a new feed.
The method of use, amount used, and content in the feed composition of the present technology are as described above.

Examples of the raw materials for the feed include grains such as corn, wheat, barley, and rye; bran, wheat bran, rice bran, and defatted rice bran; manufacturing residues such as corn gluten meal and corn jam meal; animal feeds such as skim milk powder, whey, fish meal, and bone meal; yeasts such as brewer's yeast; mineral feeds such as calcium phosphate and calcium carbonate; oils and fats; amino acids; sugars, etc. Examples of the form of the feed include pet feed (pet food, etc.), livestock feed, and fish feed.

In this way, this technology can be used in a wide range of fields, including food, food compositions, functional foods, pharmaceuticals, and feed.

The present technology can also employ the following configuration.
[1] A composition for promoting glycogen storage, comprising, as an active ingredient, Bifidobacterium longum and/or Bifidobacterium breve, a bacterium of the genus Bifidobacterium, preferably promoting muscle glycogen storage and/or liver glycogen storage.
[2] A composition for improving or supporting endurance, comprising, as an active ingredient, Bifidobacterium longum and/or Bifidobacterium breve, among bacteria of the genus Bifidobacterium, wherein the endurance is preferably muscular endurance.
[3] A composition for preventing, improving, or suppressing fatigue, comprising, as an active ingredient, Bifidobacterium longum and/or Bifidobacterium breve, among bacteria of the genus Bifidobacterium. The fatigue is preferably muscular fatigue.
[4] A composition for maintaining endurance, a composition for promoting recovery from muscle fatigue, and a composition for assisting recovery from fatigue, each of which contains as an active ingredient Bifidobacterium longum and/or Bifidobacterium breve, among bacteria of the genus Bifidobacterium.
[5] It is preferable that any of the compositions according to [1] to [4] above is a pharmaceutical composition or a food composition. It is also preferable that the composition is a food, a beverage, or a supplement.
[6] It is preferable that any of the compositions according to [1] to [5] above is an oral composition or an ingestible composition.

[7] A method for promoting glycogen storage, a method for improving or supporting endurance, a method for preventing, improving or suppressing fatigue, a method for sustaining endurance, a method for promoting recovery from muscle and liver fatigue, or a method for supporting recovery from fatigue, which comprises administering Bifidobacterium longum and/or Bifidobacterium breve, or a composition containing the same, among the Bifidobacterium bacteria.

[7] Bifidobacterium bacteria or use thereof for promoting glycogen storage, improving or supporting endurance, preventing, improving or suppressing fatigue, maintaining endurance, promoting recovery from fatigue in muscles and the liver, or supporting recovery from fatigue. The use may be for non-therapeutic purposes.
[8] Bifidobacterium longum and/or Bifidobacterium breve, among bacteria of the genus Bifidobacterium, or use thereof, for producing a composition such as a composition for promoting glycogen storage, a composition for improving or supporting endurance, a composition for preventing, improving or suppressing fatigue, a composition for maintaining endurance, a composition for promoting recovery from fatigue in muscles and the liver, a composition for supporting recovery from fatigue, etc. The composition is preferably for use in medicines or foods, and may also be for use as a supplement.

[9] A method for producing a food composition, comprising the following steps (A) to (B):
(A) culturing Bifidobacterium longum and/or Bifidobacterium breve, which are bacteria of the genus Bifidobacterium, in a medium to obtain a fermentation product;
(B) A step of subjecting the fermented product to freeze-drying or heat-drying to obtain bacterial powder.
The food composition is preferably one or more selected from the group consisting of compositions for promoting glycogen storage; compositions for improving or assisting endurance; compositions for preventing, improving or suppressing fatigue; compositions for maintaining endurance; compositions for promoting recovery from fatigue in muscles and liver; and compositions for assisting recovery from fatigue.

[10] Among the Bifidobacterium bacteria according to any one of [1] to [9] above, Bifidobacterium longum and/or Bifidobacterium breve are preferably in the form of live cells and/or killed cells.
[11] In any one of [1] to [10] above, the Bifidobacterium longum and/or Bifidobacterium breve is preferably one or more species selected from the group consisting of Bifidobacterium longum subsp. longum and Bifidobacterium breve.
[12] The Bifidobacterium bacterium according to any one of [1] to [11] above is preferably Bifidobacterium longum subsp. longum NITE BP-02621 (BB536) and/or Bifidobacterium breve FERM BP-11175 (B-3).

The present technology will be described in more detail below based on examples and test examples. Note that the examples and test examples described below are representative examples and test examples of the present technology, and should not be construed as narrowing the scope of the present technology.

[Sample preparation]
Lyophilized powders of live cells of Bifidobacterium longum NITE BP-02621 (Accession No. NITE BP-02621 (BB536): hereinafter sometimes simply referred to as "BB536") and Bifidobacterium breve FERM BP-11175 (Accession No. FERM BP-11175 (MCC1274): hereinafter sometimes simply referred to as "B-3") were diluted with physiological saline to a concentration of 1 x 10^9 CFU/mL. These strains are available from NITE-IPOD.
Each diluted solution of the bacteria was used as it was as a solution containing live bacteria without any treatment, and each diluted solution of the bacteria was heat-treated at 90° C. for 30 minutes and used as a solution containing killed bacteria.
Comparative sample (control): Physiological saline (0.9 w/v % sodium chloride aqueous solution)
Test sample 1: A solution containing live cells of B. longum NITE BP-02621 (BB536) Test sample 2: A solution containing dead cells of B. longum NITE BP-02621 (BB536) Test sample 3: A solution containing live cells of B. breve FERM BP-11175 (B-3) Test sample 4: A solution containing dead cells of B. breve FERM BP-11175 (B-3)

[Animal testing]
Seven-week-old C57BL/6J mice (Charles River Japan: 45 healthy mice) were used as experimental animals. Each mouse was allowed to freely consume Labo MR Stock Feed (Nosan Kogyo Co., Ltd.) and tap water. After one week of acclimation, the mice were divided into five groups (9 mice per group; comparison sample group, test sample groups 1 to 4), and the comparison sample or test samples 1 to 4 were orally administered using an oral sonde. The dose of test samples 1 to 4 was 1 mL per mouse, 1 x 10^9 CFU/day/mouse, and the number of administrations and period were once a day for four weeks. The dose of the comparison sample was 1 mL per mouse, and the number of administrations and period were once a day for four weeks, similar to the test sample group.

Three weeks after the start of administration of the test sample, all experimental animals were subjected to a treadmill maximum running distance test. The running speed of the mice was changed according to the program shown in Table 1, and the maximum running distance was determined as the distance the mice could run until they touched the rear electrode three times within five seconds.

One hour after the final administration of the sample, the muscles (plantaris muscle) and liver of each mouse were removed under sevoflurane anesthesia. Each tissue was cut into a block (approximately 20 mg) and each was weighed. The weight of the tissue was measured according to the method described in Reference 2 (S Lo, et. al., J Appl Physiol, 28 (2), 234-236. Determination of Glycogen in Small Tissue Samples). The amount of glycogen per gram of tissue was calculated by correcting the measured weight.

The maximum running distance for each group is shown in Table 2. An increase in the maximum running distance was confirmed in all groups, including the live and killed BB536 groups and the live and killed B-3 groups, compared to the comparison sample group.

The amount of glycogen in the muscle and liver for each group is shown in Table 3. Increases in the amount of glycogen in the muscle and liver were confirmed in all groups administered live and killed BB536 bacteria, and live and killed B-3 bacteria, compared to the control group.

From the results of this experiment, it was confirmed that the administration of Bifidobacterium longum NITE BP-02621 (live and killed bacteria) and Bifidobacterium breve FERM BP-11175 (live and killed bacteria) increased the glycogen content in the muscles and liver, resulting in improved endurance.
From this, it has become clear that Bifidobacterium bacteria (more preferably, B. longum and/or B. breve) have excellent endurance improving effects, excellent muscle glycogen storage promoting effects, and excellent liver glycogen storage promoting effects.

Claims (7)

The present invention relates to a method for producing a medicament for the treatment of a disease comprising administering to the patient an effective amount of Bifidobacterium longum NITE BP-02621 or a strain substantially equivalent to Bifidobacterium longum NITE BP-02621 ,
The substantially equivalent strain has a DNA identity of 98% or more with the deposited strain, and is a strain bred from the deposited strain or a strain naturally derived from the deposited strain.
A composition for promoting glycogen storage. The present invention relates to a method for producing a medicament for the treatment of a disease comprising administering to the patient an effective amount of Bifidobacterium longum NITE BP-02621 or a strain substantially equivalent to Bifidobacterium longum NITE BP-02621 ,
The substantially equivalent strain has a DNA identity of 98% or more with the deposited strain, and is a strain bred from the deposited strain or a strain naturally derived from the deposited strain.
A composition for improving or assisting endurance. The composition of claim 2, wherein the endurance is muscle endurance. The present invention relates to a method for producing a medicament for the treatment of a disease comprising administering to the patient an effective amount of Bifidobacterium longum NITE BP-02621 or a strain substantially equivalent to Bifidobacterium longum NITE BP-02621 ,
The substantially equivalent strain has a DNA identity of 98% or more with the deposited strain, and is a strain bred from the deposited strain or a strain naturally derived from the deposited strain.
A composition for preventing, improving or suppressing physical fatigue. Furthermore, the present invention contains as an active ingredient Bifidobacterium breve FERM BP-11175 or a strain substantially equivalent to Bifidobacterium breve FERM BP-11175 ,
The composition according to claims 1 to 4, wherein the substantially equivalent strain has a DNA identity of 98% or more with the deposited strain and is a strain bred from the deposited strain or a strain naturally occurring from the deposited strain. The composition according to any one of claims 1 to 5 , wherein the composition is a pharmaceutical composition. The composition according to any one of claims 1 to 5 , wherein the composition is a food composition.