JP7808367B2 - composition - Google Patents

Description

The present invention relates to a composition containing digestible oligosaccharides and at least one selected from processed plant products, lactic acid bacteria, and polysaccharides.

Compositions containing processed plant products or polysaccharides have been known as health foods, etc. (Patent Document 1).

Japanese Patent Application Laid-Open No. 2003-267880

With regard to compositions used in health foods and the like, consumer needs have become more diverse in recent years, and there is an ever-increasing demand for further improved effects, particularly in terms of health and beauty.
However, conventional compositions have not been able to fully meet this demand.

The inventors therefore conducted extensive research into compositions containing at least one selected from processed plant products, lactic acid bacteria, and polysaccharides, to find compositions that would further enhance the health and beauty benefits. Surprisingly, they discovered that combining digestible oligosaccharides with at least one selected from processed plant products, lactic acid bacteria, and polysaccharides inhibits fat accumulation in adipocytes and activates myoblasts, making them useful for dieting and preventing or reducing locomotive syndrome, leading to the completion of the present invention.

The present invention is based on the above findings and provides a composition containing digestible oligosaccharides and at least one selected from processed plant products, lactic acid bacteria, and polysaccharides.

The present invention provides a composition that has excellent effects of inhibiting fat accumulation in adipocytes and activating myoblasts. The present invention also provides a diet composition and/or a composition for preventing locomotive syndrome that inhibits fat accumulation in adipocytes and activates myoblasts, thereby inhibiting the accumulation of body fat such as subcutaneous fat and visceral fat, maintaining or improving energy metabolic activity such as lipid breakdown and combustion, reducing or inhibiting weight gain, slimming the body, inhibiting rebound, building muscle or preventing muscle loss, and improving the efficiency of muscle damage recovery.

The composition of the present invention will be described below based on its preferred embodiments. Hereinafter, the term "composition of the present invention" applies to any of the following: "oral composition," "oral agent," "food and drink composition," "food and drink agent," and "food and drink" that can be safely ingested; and also applies to any of the following: "diet composition," "muscle-building composition," "composition for preventing muscle loss," "composition for preventing and/or ameliorating locomotor syndrome," "diet agent," "muscle-building agent," "muscle-loss prevention agent," "agent for preventing and/or ameliorating locomotor syndrome," "diet food," "muscle-building food," "food for preventing muscle loss," and "food for preventing and/or ameliorating locomotor syndrome," which allow the effects of the present invention to be obtained safely.

Digestive Oligosaccharides: In the present invention, digestible oligosaccharides are oligosaccharides of three or more sugars that are decomposed by human digestive enzymes, absorbed in the stomach or small intestine, and converted into energy. The number of constituent sugars of the digestible oligosaccharides usable in the present invention is preferably three to ten, more preferably three to eight, even more preferably three to six, and particularly preferably three to four from the viewpoint of effectiveness. Examples of digestible oligosaccharides include maltooligosaccharides, isomaltooligosaccharides, and soybean oligosaccharides. Maltooligosaccharides are particularly preferred from the viewpoints of availability, enhancing the effects of inhibiting fat accumulation in adipocytes and activating myoblasts, and ease of handling during production. Specific examples of maltooligosaccharides include maltotriose, maltotetraose, maltopentaose, and maltohexaose. Maltotriose is particularly preferred from the viewpoint of enhancing the effects of the present invention.

In order to further enhance the aforementioned effect of inhibiting fat accumulation in adipocytes and activating myoblasts, the content of digestible oligosaccharides in the solid content of the composition of the present invention is preferably 0.0001% by mass or more, more preferably 0.001% by mass or more, even more preferably 0.01% by mass or more, and particularly preferably 0.05% by mass or more from the standpoint of effectiveness. Furthermore, the composition of the present invention may consist solely of digestible oligosaccharides, processed plant products, lactic acid bacteria, and polysaccharides. However, from the standpoint of enabling high functionality and multi-functionality through combination with other components, the upper limit of the content of digestible oligosaccharides in the composition is preferably 30% by mass or less, more preferably 20% by mass or less, and even more preferably 10% by mass or less.

Processed Plant Products In the present invention, the processed plant products are obtained by subjecting an orally ingestible plant body to one or more of the following processes: drying, crushing, extraction, filtration, squeezing, slurrying, fermentation, heating, etc. Examples of plant body parts include leaves, roots, rhizomes, flowers, stems, fruits, seeds, etc., and the processed plant products may contain one or more parts.

When obtaining processed plant products from plants, it is preferable that the plants are immediately harvested or processed after harvest. If time is required before processing, it is preferable to store the plants using storage methods commonly used by those skilled in the art, such as low-temperature storage, to prevent deterioration of the plants.

Specific examples of processed plant products include, but are not limited to, dried powders obtained by drying and grinding plants (hereinafter also referred to as "dried and ground powder"), shredded plant material and its dried product, squeezed plant juice and its dried powder, and plant extracts and its dried powder. In the present invention, a final powder form is preferred from the standpoint of ease of processing, storage, transportation, etc., and versatility of use. In this specification, the term "powder" generally includes any of dried and ground powders, dried powders of shredded material, dried powders of squeezed juice, dried powders of extracts, and dried powders of fermented products. Powder naturally includes granular and granular forms, etc.

Conventionally known methods can be used to dry and pulverize plant matter into powder. One such method is a combination of drying and pulverization of the plant matter. Either the drying or pulverization process can be performed first, but it is preferable to perform the drying process first. This method of drying and pulverization may further include one or more treatments selected from blanching, cooling, sterilization, etc., as needed. The pulverization process may be performed two or more times, and it is preferable to perform a coarse pulverization process followed by a fine pulverization process to pulverize the plant matter even more finely.

The drying process is not particularly limited, but preferably involves drying the barley stems and leaves so that their moisture content is 10% by mass or less, and particularly 5% by mass or less. This drying process can be carried out by any method known to those skilled in the art, such as hot air drying, high-pressure steam drying, electromagnetic wave drying, or freeze drying.

The grinding process is not particularly limited, but examples include grinding the plant body using grinding equipment or tools such as a crusher, mill, blender, or stone mill, using any method commonly used by those skilled in the art. The ground plant body is sieved as needed, and it is preferable to use plant body powder that passes through a 30-250 mesh sieve, for example. By using a particle size that passes through a 250 mesh or smaller, the plant body powder is easier to handle during further processing, and by using a particle size that passes through a 30 mesh or larger, it is easier to uniformly mix the plant body powder with other materials.

Specific methods for drying and pulverizing the plant include, for example, cutting the plant body, drying it so that the moisture content is 10% by mass or less, and then pulverizing it. Other methods include, for example, cutting the plant body, rolling it, drying it, and pulverizing it; or drying the plant body, coarsely pulverizing it, heating it at 110°C or higher, and then finely pulverizing it.

The method for shredding the plant body is not particularly limited, but methods commonly used by those skilled in the art for shredding plant bodies, such as slicing, crushing, or chopping, can be used. One example of shredding is slurrying. Slurrying can be achieved by putting the plant body through a mixer, juicer, blender, mass colloider, or the like to create a thick gruel-like substance (a suspension of liquid and solids). When shredded seeds are heated, water can be added to the liquid and boiled down, and the liquid can then be used after removing the coarse solids by sieving, filtration, or other means.

The method for extracting juice from the plant body is not particularly limited, but examples include squeezing the plant body or its fragments, and centrifuging or filtering the fragments of the plant body. Specific examples of juicing methods include extracting juice using mechanical crushing means such as a mixer or juicer, and then removing coarse solids by means of sieving or filtration, as needed, to obtain a squeezed juice. The squeezed juice may be concentrated, or dried and powdered, as needed. When powdering, an excipient may be used as needed.

There are no particular limitations on the method for obtaining a plant extract. Examples include adding an extraction solvent commonly used by those skilled in the art, such as ethanol, water, methanol, ethyl acetate, or acetone, to the plant, or its finely ground or dried form, and then stirring and/or heating as needed to extract. One or more extraction solvents can be selected and used as needed. When heating, the temperature is not particularly limited as long as it is between room temperature and the boiling point of the solvent. It is preferable to use an extract from which coarse solids have been removed by sieving, filtration, or other means. For example, the filtrate (extract) obtained by adding water to finely ground soybeans (ground material), boiling it down, and then filtering it is known as soy milk. The extract may be concentrated or dried and powdered as needed. When powdering, an excipient can be used as needed.

When the plant processed product is in powder form, it is preferable to keep the moisture content at 20% by mass or less, and particularly 10% by mass or less, from the standpoint of stability and preventing quality deterioration. For example, a moisture content of 1% by mass or more is preferable from the standpoint of ease of manufacturing the powdered plant processed product.

When the plant processed product is in powder form, it is preferable that the powder pass through a 30 to 250 mesh sieve, from the viewpoints of ease of mixing with other ingredients and ease of oral administration. From the same viewpoint, it is more preferable that 90% by mass or more of the powdered plant processed product pass through a 200 mesh sieve.

The processed plant products used in the present invention may be commercially available products. Examples of commercially available products include those described in the Examples below.

The plant body in the processed plant product is preferably a processed green leaf product. In the present invention, green leaves refer to the leaves and/or stems of orally ingestible green plants that contain chlorophyll. Examples of green leaves that can be used in the present invention include those of grasses, Asteraceae, Umbelliferae, Mulberry, Houttuynia cordata, Lamiaceae, Liliaceae, Tilia, Convolvulaceae, and Theaceae. These green leaves are preferred because of their high fat accumulation inhibitory activity in adipocytes and myoblast-activating activity, as well as the ability to ingest plant-derived nutrients. Examples of grasses include wheat, durum wheat, rye, triticale, barley, oats, Job's tears, corn, rice, barnyard millet, foxtail millet, and Japanese bamboo grass. Examples of Asteraceae include mugwort. Examples of Umbelliferae include Angelica keiskei, parsley, celery, and longevity grass (also known as Peucedanum japonicum). Examples of plants in the Moraceae family include mulberry and the like. Examples of plants in the Houttuynia family include Houttuynia cordata and the like. Examples of plants in the Lamiaceae family include perilla and the like. Examples of plants in the Brassicaceae family include komatsuna, kale, cabbage, broccoli and the like. Examples of plants in the Liliaceae family include asparagus and the like. Examples of plants in the Tilia family include mulukhiyah and the like. Examples of plants in the Convolvulaceae family include sweet potato and the like. Examples of plants in the Theaceae family include tea and the like. Among these, barley, kale, chomeiso, kumazasa, tea, angelica tree, mulberry, sweet potato, mugwort, and cabbage are particularly preferred in terms of effectiveness.

When the green leaves used in the present invention are barley, it is preferable to use young barley leaves. Young barley leaves are leaves and/or stems harvested before maturity, i.e., from the time when tillers begin to emerge until the time when heading begins. When the green leaves are sweet potato, it is preferable to use leaves including leaves and/or stems that are exposed above ground during sweet potato cultivation, and young sweet potato leaves are particularly preferable.

When the plant body is green leaves, the green leaves are preferably in the form of dried and crushed powder. As described above, dried and crushed powder is a dry powder obtained by drying and crushing plant body that has not been subjected to juice squeezing or extraction processes.

The processed plant products of the present invention also include fermented plants due to their high activity in inhibiting fat accumulation in adipocytes and activating myoblasts. In addition to the various plants listed above as examples of processed green leaves, other vegetables, mushrooms, grains, seeds, beans, fruits, seaweed, wild plants, etc. can be used for fermented plant products. Specific examples of other vegetables, mushrooms, grains, seeds, beans, fruits, seaweed, and wild plants include burdock, carrots, lotus root, cabbage, pumpkin, radish, tomato, bell pepper, shiitake mushroom, barley, millet, brown rice, Job's lettuce, corn, sesame, soybeans, kidney beans, pineapple, bananas, apples, lemons, melons, prunes, kiwi, kelp, wakame seaweed, gynostemma pentaphyllum, horsetail, muira puama, and cat's claw. These may be used alone or in combination of two or more. When the processed plant product is a fermented product, the plant body is preferably a fermented product of 5 or more, more preferably 10 or more, more preferably 20 or more, and particularly preferably 40 or more types of plant body selected from vegetables such as cabbage, mushrooms, grains, seeds, beans, fruits, seaweed, and wild plants. As the fermented processed plant product, for example, commercially available fermented products called "fermented plant extract" or "fermented wild plant extract" may be used.

When the processed plant product is a fermented product, the fermented product may be in any of the following forms: liquid, slurry, paste, or powder. Fermented plant products include those obtained by fermenting the plant, or its crushed product, juice, extract, or shredded product. When the product is in liquid, slurry, or paste form after fermentation, it can be dried using any method known to those skilled in the art, such as hot air drying, high-pressure steam drying, electromagnetic wave drying, or freeze-drying. In this case, an excipient may be added, and the product may be dried and then crushed to form a dry powder.

Fermented plant matter can be obtained, for example, by adding lactic acid bacteria, yeast, koji mold, natto bacteria, acetic acid bacteria, etc. to the plant matter or its crushed material, shredded material, juice, or extract. These may be used alone or in combination of two or more species, and when two or more species are used, they may be added simultaneously or different species may be added in stages. It is preferable that the bacteria used for fermentation be at least one species selected from lactic acid bacteria, yeast, and acetic acid bacteria, as this allows for the ingestion of useful components such as physiologically active substances produced by these species.

Lactic acid bacteria used for fermentation refer to bacteria that produce lactic acid through metabolism, and specific examples include bacteria described as lactic acid bacteria that can be used in the present invention.
Examples of yeasts used for fermentation include sake yeast, wine yeast, brewer's yeast, baker's yeast, etc. For example, yeasts belonging to the genus Saccharomyces, Schizosaccharomyces, etc. are used, and preferred examples include Saccharomyces cerevisiae, Saccharomyces pastorianus, Schizosaccharomyces pombe, etc. In particular, Saccharomyces cerevisiae and isolated strains thereof are preferred in terms of producing useful substances such as amino acids and vitamins.
Examples of koji molds used for fermentation include black koji mold, white koji mold, yellow koji mold, red koji mold, etc., and commercially available products can be suitably used. Specific examples include microorganisms belonging to the genus Aspergillus, such as Aspergillus awamori (black koji mold), Aspergillus saitoi (black koji mold), Aspergillus nakazawai (black koji mold), Aspergillus usamii (black koji mold), Aspergillus luchensis (black koji mold), Aspergillus niger (black koji mold), Aspergillus kawachii (white koji mold), and Aspergillus oryzae (yellow koji mold).
Examples of acetic acid bacteria include microorganisms belonging to the genus Acetobacter, such as Acetobacter aceti, Acetobacter pasteurianus, and Acetobacter hansenii.
Examples of natto bacteria include bacteria of the genus Bacillus, such as Bacillus natto, Bacillus subtilis, and Bacillus circulans.

Lactic acid bacteria of the present invention refer to bacteria that produce lactic acid through metabolism. The lactic acid bacteria that can be used in the present invention may be live or dead, and in the case of dead bacteria, the cells may be disrupted. Lactic acid bacteria powder can be obtained, for example, by removing unnecessary components such as the medium from cultured lactic acid bacteria or a lactic acid bacteria culture used in fermentation, and then powdering the lactic acid bacteria cells using a known method. The type of lactic acid bacteria is not particularly limited as long as it produces lactic acid as a metabolic product, and examples include those that have traditionally been taken orally by animals such as humans, such as those of the genera Bifidobacterium, Lactobacillus, Enterococcus, Leuconostoc, Pediococcus, Staphylococcus, Tetragenococcus, and Bacillus.
Examples of the genus Bifidobacterium include Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium lactis, Bifidobacterium longum, Bifidobacterium adolescentis, and Bifidobacterium mongoliense.
Lactobacillus genus includes Lactobacillus brevis, Lactobacillus gasseri, Lactobacillus acidophilus, Lactobacillus buchneri, Lactobacillus bulgaricus, Lactobacillus delburvecki, Lactobacillus casei, Lactobacillus crispatus, Lactobacillus curvatus, Lactobacillus halivaticus, Lactobacillus Examples include Lactobacillus pentosus, Lactobacillus plantarum, Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus salivarius, Lactobacillus sporogenes, Lactobacillus sakei, Lactobacillus fructivorans, Lactobacillus hilgardii, Lactobacillus reuteri, and Lactobacillus fermentum.
Enterococcus includes Enterococcus faecalis (sometimes called Streptococcus faecalis), Enterococcus faesium (sometimes called Streptococcus faesium), Streptococcus thermophilus, and Lactococcus lactis (sometimes called Streptococcus lactis).
The genus Leuconostoc includes Leuconostoc mesenteroides and Leuconostoc oenos.
The genus Pediococcus includes Pediococcus acidilactici and Pediococcus pentosaceus.
Examples of the genus Staphylococcus include Staphylococcus carnosus and Staphylococcus xylosus.
The genus Tetragenococcus includes Tetragenococcus halophilus.
Examples of the genus Bacillus include Bacillus coagulans and Bacillus mesentericus.
Among these, Bacillus coagulans, Enterococcus faecalis, and Bifidobacterium bifidum are preferred because of their high effects of inhibiting fat accumulation in adipocytes and activating myoblasts. These may be used alone or in combination of two or more.

Polysaccharides of the present invention are those that are not digested or are extremely difficult to digest (resistant to digestion) by human digestive enzymes. Examples of polysaccharides that can be used in the present invention include raffinose, fructooligosaccharides, galactooligosaccharides, soybean oligosaccharides, xylooligosaccharides, polydextrose, resistant dextrin, psyllium, pectin, arabinoglucan, xylan, galactomannan, glucomannan, galactan, tamarind gum, xanthan gum, etc., and polydextrose, resistant dextrin, tamarind gum, xanthan gum, etc. are preferred. Examples of water-soluble dietary fibers include psyllium, pectin, arabinoglucan, xylan, galactomannan, glucomannan, galactan, tamarind gum, and xanthan gum. Examples of water-soluble dietary fibers include polydextrose, indigestible dextrin, tamarind gum, and xanthan gum, and more preferably polydextrose, indigestible dextrin, xanthan gum, and tamarind gum. Xanthan gum and tamarind gum are particularly preferred because of their high effect of inhibiting fat accumulation in adipocytes and myoblast activation.

The contents of the processed plant products, lactic acid bacteria, and polysaccharides in the present invention are not particularly limited. However, in order to further enhance the effect of inhibiting fat accumulation in adipocytes and activating myoblasts, the total dry mass of the solid content of the composition of the present invention is preferably 10% by mass or more, more preferably 20% by mass or more, even more preferably 30% by mass or more, and particularly preferably 40% by mass or more. Furthermore, the composition of the present invention may consist solely of digestible oligosaccharides, processed plant products, lactic acid bacteria, and polysaccharides. However, in order to enable the composition to be combined with other components to achieve high functionality and multi-functionality, the upper limit of the content of the processed plant products, lactic acid bacteria, and polysaccharides in the composition is preferably 90% by mass or less, more preferably 80% by mass or less, and even more preferably 70% by mass or less. The total amount of processed plant products, lactic acid bacteria, and polysaccharides in the composition referred to here refers to the amount of each of the processed plant products, lactic acid bacteria, or polysaccharides when only one type is contained, and refers to the total amount when two or more types are contained.

In terms of inhibiting lipid accumulation in adipocytes and activating myoblasts, the mass ratio of digestible oligosaccharides to the total amount of processed plant products, lactic acid bacteria, and polysaccharides per 1 part by mass of digestible oligosaccharides is preferably 0.01 parts by mass or more, more preferably 0.1 parts by mass to 10,000 parts by mass, even more preferably 1 part by mass to 1,000 parts by mass, and particularly preferably 5 parts by mass to 600 parts by mass.

In particular, from the viewpoint of further enhancing the fat accumulation prevention effect and myoblast activation activity, the mass ratio of digestible oligosaccharides to processed green leaf products is preferably 1 part by mass or more of each processed product by dry mass to 1 part by mass of digestible oligosaccharides, more preferably 25 parts by mass or more and 10,000 parts by mass or less, even more preferably 50 parts by mass or more and 1,000 parts by mass or less, and particularly preferably 100 parts by mass or more and 500 parts by mass or less from the viewpoint of effectiveness.
For the same reasons, the mass ratio of digestible oligosaccharides to plant fermentation products is preferably 0.01 mass parts or more of plant fermentation products per 1 mass part of digestible oligosaccharides, more preferably 0.01 mass parts or more and 100 mass parts or less, even more preferably 0.1 mass parts or more and 50 mass parts or less, and particularly preferably 1 mass part or more and 10 mass parts or less from the standpoint of effectiveness.
For the same reasons, the mass ratio of digestive oligosaccharides to lactic acid bacteria is preferably 0.0001 to 100 parts by mass, more preferably 0.001 to 50 parts by mass, and particularly preferably 0.01 to 20 parts by mass, in terms of effectiveness, of lactic acid bacteria per 1 part by mass of digestive oligosaccharides.
From the same viewpoint, the mass ratio of digestible oligosaccharides to polysaccharides is preferably 0.1 mass parts or more of polysaccharides on a dry mass basis to 1 mass part of digestible oligosaccharides, more preferably 1 mass part or more and 5,000 mass parts or less, even more preferably 10 mass parts or more and 1,000 mass parts or less, and particularly preferably 20 mass parts or more and 500 mass parts or less from the viewpoint of effectiveness.

The composition of the present invention may contain other ingredients in addition to digestible oligosaccharides, processed plant products, lactic acid bacteria, and polysaccharides. Examples of such other ingredients include vitamins, proteins, minerals, dairy products, citric acid, acidulants, coloring agents, and glazing agents, as well as manufacturing agents such as talc and calcium stearate. Other ingredients include various excipients, binders, lubricants, stabilizers, diluents, bulking agents, emulsifiers, coloring agents, flavorings, food additives, and seasonings. The content of such other ingredients can be appropriately selected depending on the form of the composition. In this embodiment, the amount of ingredients other than digestible oligosaccharides, processed plant products, lactic acid bacteria, and polysaccharides contained in the composition is preferably 50% by mass or less of the solid content, more preferably 30% by mass or less, and particularly preferably 10% by mass or less from the standpoint of effectiveness.

The composition of the present invention may be in any form, such as a solid, semi-solid, or fluid. For example, solid forms include powder, granules, granules, tablets, capsules, sticks, plates, blocks, solids, rounds, candies, gummies, wafers, biscuits, cookies, cakes, and chewable forms. Semi-solid forms include pastes and jellies. Fluid forms include syrups, liquids, and jellies.

The composition of the present invention is preferably in powder, granule, or granular form and can be taken orally by mixing with water, as this prevents spoilage and allows for long-term storage. If the composition of the present invention is in solid form, it can be taken orally by mixing with water to form a liquid and drinking the liquid, as described above. However, depending on the preferences of the user, the solid form may also be taken orally. It may also be added to milk, soy milk, fruit juice drinks, whey drinks, soft drinks, yogurt, pancake mix, etc., in addition to water. It may also be used as a supplement, health food, nutrient-functional food, functional food, food for specified health uses, and pharmaceutical drug.

The composition of the present invention is an oral composition, and when the composition contains processed green leaves as the processed plant product, it is preferable to use the composition of the present invention as an edible or drinkable composition for green juice. An edible or drinkable composition for green juice is a beverage containing processed green leaves. Examples of edible or drinkable compositions for green juice include this beverage and solids that are dispersed or dissolved in liquid to obtain this beverage. In addition to commonly known green juice products, edible or drinkable compositions for green juice include smoothies, jellies, etc.

The form of the composition of the present invention is not limited to the beverages listed as examples of edible compositions for green juice and the solid form that is dispersed or dissolved in a liquid to obtain this beverage, and any form can be adopted.

As described in the Examples below, the green leaf powder of the present invention contains digestible oligosaccharides and at least one selected from the group consisting of processed plant products, lactic acid bacteria, and polysaccharides, thereby exhibiting excellent effects of inhibiting fat accumulation in adipocytes and/or activating myoblasts. Specifically, ingesting a composition containing digestible oligosaccharides and at least one selected from the group consisting of processed plant products, lactic acid bacteria, and polysaccharides can prevent lipid accumulation in adipocytes and activate myoblasts in skeletal muscle, etc. Therefore, ingesting the composition of the present invention can prevent body fat accumulation, reduce body fat, maintain or promote energy metabolism in muscles, promote muscle growth, prevent muscle wasting and loss, be used for dieting, and prevent or improve locomotive syndrome, etc.
That is, the composition of the present invention can be excellent for preventing body fat accumulation, reducing body fat, activating myoblasts, promoting muscle cell proliferation, building muscle, preventing muscle loss, preventing muscle wasting, promoting metabolism, preventing obesity, preventing obesity, eliminating obesity, reducing or suppressing weight gain, slimming the body, preventing rebound, dieting, and preventing and/or ameliorating locomotive syndrome, etc. Here, building muscle means increasing muscle mass, and preventing muscle loss means preventing muscle mass loss.

The present invention will be explained in more detail below using examples, but the scope of the present invention is not limited to these examples.

[Examples 1 to 20, Comparative Examples 1 to 24]
The test samples described in Tables 1 and 2 were as follows:
Maltotriose: A commercially available powdered product was used.
Trehalose: A commercially available powdered product was used.
Young barley leaves: Dried and pulverized powder of young barley leaves was used, manufactured by Toyo Shinyaku Co., Ltd. The dried and pulverized powder had a content of 90% by mass or more passing through a 200 mesh sieve and a moisture content of 1% by mass or more and 7% by mass or less.
Kale: Dried and crushed powder of kale leaves manufactured by Toyo Shinyaku Co., Ltd. The dried and crushed powder had 90% or more by mass of particles passing through a 200 mesh sieve and a moisture content of 1% to 7% by mass.
Chomeisou: Dried and crushed powder of Chomeisou leaves manufactured by Toyo Shinyaku Co., Ltd. The dried and crushed powder had 90% or more by mass of particles passing through a 200 mesh sieve and a moisture content of 1% to 7% by mass.
Kumazasa: Commercially available dried and crushed Kumazasa leaves were used. The dried and crushed powder had 90% or more by mass of particles passing through a 200 mesh and a moisture content of 1% to 7% by mass.
Matcha: Dried and pulverized powder of Tencha manufactured by Toyo Shinyaku Co., Ltd. The dried and pulverized powder had a content of 90% by mass or more passing through a 200 mesh sieve and a moisture content of 1% by mass or more and 7% by mass or less.
Angelica keiskei: commercially available dried and pulverized powder of Angelica keiskei leaves was used. The dried and pulverized powder had 90% or more by mass of particles passing through a 200 mesh sieve and a moisture content of 1% to 7% by mass.
Mulberry: Dried and crushed powder of mulberry leaves manufactured by Toyo Shinyaku Co., Ltd. The dried and crushed powder had 90% or more by mass of particles passing through a 200 mesh sieve and a moisture content of 1% to 7% by mass.
Young sweet potato leaves: Dried and crushed powder of sweet potato leaves was used, manufactured by Toyo Shinyaku Co., Ltd. The dried and crushed powder had 90% or more by mass of material passing through a 200 mesh sieve and a moisture content of 1% to 7% by mass.
Mugwort: Dried and crushed powder of mugwort leaves manufactured by Toyo Shinyaku Co., Ltd. The dried and crushed powder had 90% or more by mass of particles passing through a 200 mesh sieve and a moisture content of 1% to 7% by mass.
- Spore-forming lactic acid bacteria: Commercially available dried powder of spores of the scientific name Bacillus coagulans was used.
- Lactic acid bacteria (killed bacteria): Commercially available dried powder of killed bacteria of the scientific name Enterococcus faecalis was used.
- Lactic acid bacteria (mixture): A commercially available mixture of dry powder of live bacteria with the scientific names Enterococcus faecium and Bifidobacterium bifidum was used.
Polydextrose: Commercially available polydextrose (powder) with a dietary fiber content of 90% by mass or more was used.
- Indigestible dextrin: Commercially available indigestible dextrin (powder) with a dietary fiber content of 85 to 95% by mass was used.
- Xanthan gum: Commercially available xanthan gum (powder) derived from Xanthomonas Campestris was used.
- Tamarind gum: Commercially available tamarind gum (dried powder) derived from tamarind seeds was used.
Fermented cabbage extract: Toyo Shinyaku Co., Ltd., product name "Fermented cabbage extract" (dried and pulverized powder obtained by fermenting raw cabbage with lactic acid bacteria, then drying and pulverizing) was used.
- Multi-plant fermented extract: A commercially available multi-plant fermented extract was used (raw materials including more than 80 types of vegetables, fruits, grains, and wild plants such as burdock, carrot, lotus root, cabbage, pumpkin, radish, tomato, bell pepper, shiitake mushroom, barley, millet, brown rice, pearl barley, corn, sesame, soybean, kidney bean, pineapple, banana, apple, lemon, melon, prune, kiwi, kelp, wakame seaweed, gynostemma pentaphyllum, horsetail, muira puama, and cat's claw, which are fermented with lactic acid bacteria, yeast, and acetic acid bacteria, and then the fermented product is dried and turned into a powder).

The test samples were subjected to a fat accumulation test according to the following procedures (a) to (L).
[Fat accumulation test]
(a) Mouse fibroblast cells 3T3-L1 were cultured in a 75 ^cm2 flask in DMEM medium containing 10% (v/v) FBS at 37°C in a 5% _CO2 incubator.
(b) Fibroblast 3TL-L1 cells were suspended by trypsinization and seeded at a cell density of 2 x ¹⁰ cells/well from a 75 ^cm2 flask into each well of a 96-well plate. They were pre-cultured in DMEM medium containing 10% (v/v) FBS for 2 days in a 37°C, 5% CO2 _incubator . The medium was then replaced with differentiation-inducing medium containing the test sample (control: differentiation-inducing medium only), and the cells were cultured for 3 days to induce differentiation. The differentiation-inducing medium was 10% (v/v) FBS-containing DMEM medium containing 0.5 mM isobutylmethylxanthine, 0.5 μM dexamethasone, and 10 μg/mL insulin. The test samples were prepared in the differentiation-inducing medium to achieve the desired concentration (300 μg/mL). Tables 1 and 2 show the mass percentage of each test sample relative to the total test sample.
(c) The differentiation-inducing medium was replaced with a differentiation-maintenance medium containing the test sample (control: differentiation-maintenance medium only), and the cells were cultured for 4 days. The differentiation-maintenance medium was a 10% (v/v) FBS-containing DMEM medium containing 10 μg/mL insulin. The test samples were prepared in the differentiation-maintenance medium so that the total amount reached a predetermined concentration (300 μg/mL). Tables 1 and 2 show the mass percentage of each test sample in the total amount of test sample.
(d) After culturing in (c), the culture medium supernatant was removed from the adipocytes. Next, 10% (v/v) formalin-containing PBS was added to the cells in an equal volume to the culture medium, and the cells were left to stand at room temperature for 10 minutes in the dark. The formalin solution was then removed from the cells, and the cells were washed once with PBS.


(e) 100 μL/well of 10% (v/v) formalin-containing PBS was added to the cells, and the cells were fixed by leaving the plate at room temperature for 10 minutes in the dark.
(f) The formalin solution was removed, and the sections were washed twice with PBS.
(g) 50 μL/well of a 60% by volume solution of 3 mg/mL Oil Red in isopropanol (staining solution) was added to the adipocytes and blank wells, and the wells were left at room temperature for 30 minutes in the dark to stain lipids (lipid droplets). The staining solution was then removed, and 150 μL/well of a 60% by volume aqueous solution of isopropanol was added, and the cells were washed twice.
(h) 100% isopropanol (volume) was added to the cells and blank wells at 100 μL/well and shaken for approximately 10 minutes to extract the staining solution. The absorbance of the extracted isopropanol solution was measured at 520 nm and 650 nm.
(k) After the isopropanol solution was completely removed by air drying, the protein amount of each cell was calculated using a Pierce ^™ BCA Protein Assay kit (Thermo Fisher Scientific).
(L) The amount of fat accumulation per protein was calculated using the formula below, and the relative values to the control were calculated. The results are shown in Tables 1 and 2. A smaller relative value indicates greater suppression of fat accumulation in adipocytes.

Relative fat accumulation (%) = [[(Abs520 sample - Abs520 blank) - (Abs650 sample - Abs650 blank)]/(Protein sample)] / [(Abs520 control - Abs520 blank) - (Abs650 control - Abs650 blank)]/(Protein control)]] × 100 (%)
Abs520 sample, Abs650 sample: absorbance of each example or comparative example at 520 nm and 650 nm
Abs520 control, Abs650 control: Absorbance of control at 520nm and 650nm
Abs520 blank, Abs650 blank: Blank absorbance at 520nm and 650nm
Protein sample: Amount of protein in the cells in each example or comparative example
Protein control: The amount of protein in the cells in the control

It is generally known that differentiation of fibroblasts induces the expression of PPARγ, which stimulates fat synthesis and causes fat accumulation within the cells, resulting in large, round adipocytes.
As shown in Table 1, the compositions of each Example, which combined digestive oligosaccharides with at least one selected from processed plant products, lactic acid bacteria, and polysaccharides, had low relative absorbance values (turbidity), indicating that little fat had accumulated in adipocytes. In contrast, the compositions containing only digestive oligosaccharides (Comparative Example 1), or only processed plant products, lactic acid bacteria, or polysaccharides (Comparative Examples 2 to 18), and the compositions combining trehalose and processed plant products (Comparative Examples 19 and 20) had high relative absorbance values (turbidity), indicating that much fat had accumulated in adipocytes.
From the above, it was shown that by combining digestible oligosaccharides with at least one selected from plant processed products, lactic acid bacteria, and polysaccharides, fat accumulation in adipocytes can be effectively suppressed and obesity can be prevented.

[Examples 21 to 39, Comparative Examples 25 to 36]
Each of the above test sample powders was subjected to a myoblast activation test according to the following procedures (1) to (7).
[Myoblast activation test]
(1) Mouse skeletal muscle-derived myoblasts (product name C2C12, manufactured by RIKEN BioResource Center) were cultured in a 75 ^cm2 flask containing 10 vol% FBS-containing DMEM medium at 37°C in a 5% _CO2 incubator.
(2) After the culture in (1), cells were suspended by trypsinization and recovered from the 75 ^cm2 flask. The cells were then counted. Then, the cells were seeded in a collagen-coated 96-well plate at a cell density of 2,000 cells/well, along with the medium, and pre-cultured for 24 hours in a 37°C, 5% CO2 _incubator .
(3) Separately from (1) and (2), the powders marked with "●" in Table 3 or Table 4 were dispersed or dissolved in 10 vol% FBS-containing DMEM medium to the concentrations shown in the left column of Table 3 or Table 4, respectively, to prepare solutions. The resulting solutions were filter-sterilized using a 0.2 μm filter (Advantec) to prepare sample solutions. However, for spore-forming lactic acid bacteria, lactic acid bacteria (killed bacteria), and bifidobacteria, dispersions were used as sample solutions. As a control, 10 vol% FBS-containing DMEM medium itself was used as the sample solution.
(4) After removing the medium from each well, 200 μL of the sample solution prepared in (3) was added to each well, and the plates were cultured at 37°C in a 5% CO ₂ incubator for 24 hours.
(5) After the incubation in (4), the medium was removed, and each well was washed once with 200 μL/well of PBS. Then, 150 μL/well of Cell Counting Kit-8 solution (Dojindo Laboratories) diluted 30-fold with serum-free DMEM was added.
(6) After adding the solution from (5), the plate was placed in a 37°C, 5% _CO2 incubator to allow for adequate color development, and the absorbance at 450 nm of each well was measured. Based on the obtained data, the percentage of cells relative to the control (% of control) was calculated using the following formula, and this was taken as myoblast-activating activity.
% of control=(Data sample - Data blank)/(Data control - Data blank)×100
Data sample: absorbance of each example or comparative example
Data control: control absorbance
Data blank: Absorbance when no cells are present

(7) Evaluation The calculation results of the percentage of cell counts for each Example and Comparative Example, with the control set at 100%, are summarized in Tables 3 and 4. In Tables 3 and 4, a black circle indicates that the sample solution of the Example or Comparative Example contained the test sample in the left column at the concentration shown in the left column.

As shown in Table 3, the compositions of each Example, which combined digestible oligosaccharides with at least one selected from a processed plant product, lactic acid bacteria, and polysaccharides, exhibited a myoblast-activating effect. In contrast, as shown in Table 4, the compositions containing only digestible oligosaccharides (Comparative Examples 25 and 26), the compositions containing trehalose and a processed plant product (Comparative Example 27), and the compositions containing only a processed plant product, lactic acid bacteria, or polysaccharides, or only polysaccharides (Comparative Examples 28 to 36), did not exhibit a myoblast-activating effect.
From the above, it was shown that by combining digestible oligosaccharides with at least one selected from lactic acid bacteria and polysaccharides, myoblasts are effectively activated and energy metabolism in muscle tissue is enhanced.

Example 40 (Production of a composition for green juice)
Raw materials were prepared according to the blending ratios described in the following formulation examples, and a composition for green juice exhibiting the effects of the present invention was manufactured.

Example 41 (Production of a composition for green juice)
Raw materials were prepared according to the blending ratios described in the following formulation examples, and a composition for green juice exhibiting the effects of the present invention was manufactured.

Example 42 (Production of a composition for green juice)
Raw materials were prepared according to the blending ratios described in the following formulation examples, and a composition for green juice exhibiting the effects of the present invention was manufactured.

Example 43 (Production of a composition for green juice)
Raw materials were prepared according to the blending ratios described in the following formulation examples, and a composition for green juice exhibiting the effects of the present invention was manufactured.

Example 44 (Production of a composition for green juice)
Raw materials were prepared according to the blending ratios described in the following formulation examples, and a composition for green juice exhibiting the effects of the present invention was manufactured.

Example 45 (Production of tablets)
After preparing the raw materials according to the compounding ratios described in the following prescription examples, tablets exhibiting the effects of the present invention were produced using a tableting machine.

Claims (1)

A solid green juice composition containing 0.01 to 5% by mass of maltotriose, which is dispersed or dissolved in a liquid and consumed, and further containing one or more selected from (1) to (3) described below.
(1) Dried and crushed powder of one or more green leaves selected from the group consisting of young barley leaves, tencha tea, kumazasa leaves, angelica leaves, young sweet potato leaves, chomeiso grass leaves, and mugwort leaves; (2) lactic acid bacteria; (3) one or more polysaccharides selected from the group consisting of indigestible dextrin, polydextrose, xanthan gum, and tamarind gum.