JP2020079309A - Muscle-enhancing agent - Google Patents

Abstract

To provide a novel muscle-enhancing agent.SOLUTION: The present invention discloses 1. a muscle differentiation induction promoter containing, as an active ingredient, a fermented extract obtained by fermenting rice bran and/or rice germ to obtain a fermented product and then extracting the fermented rice with a polar solvent; 2. a muscle differentiation induction promoter containing, as an active ingredient, a distribution obtained by dividing the fermented extract described in the above 1 with a polar solvent; 3. a muscle differentiation induction promoter containing, as an active ingredient, at least one compound of stigmasterol, ergosterol peroxide, β-sitosterol glucoside, glucosylceramide; 4. a muscle bulking agent containing, as an active ingredient, a fermented extract obtained by fermenting rice bran and/or rice germ to obtain a fermented product and then extracting the fermented rice with a polar solvent; 5. a muscle bulking agent containing, as an active ingredient, a distribution obtained by dividing the fermented extract described in the above 4 with a polar solvent; and 6. a muscle bulking agent containing stigmasterol as an active ingredient.SELECTED DRAWING: None

Description

The present invention relates to a muscle enhancing agent containing a novel ingredient as an active ingredient. INDUSTRIAL APPLICABILITY The present invention can be widely used for foods and drinks (health foods, foods for specified health uses, foods with functional claims), quasi drugs, pharmaceuticals, and the like.

Aspergillus has been used for a long time in Japan for the production of sake, miso and soy sauce, and is an essential bacterium in our daily lives. These fermented products are attracting attention not only as simple foods but also as functional foods, and there is a possibility that various physiologically active substances will be newly produced during fermentation.

On the other hand, sphingolipids are known to have functions such as skin moisturizing effect, water regulation, elasticity retention, surface protection (barrier effect), collagen protection, and antioxidant (stabilization of vitamins C and E). Widely used in the field. In addition, recently, as a function of sphingolipids, it has been clarified that the invasion of antigens from the outside to the body is suppressed and atopic dermatitis is suppressed, and the development as a pharmaceutical is being promoted.

As a method for producing a sphingolipid-containing composition from rice, for example, a method is known in which the gum obtained in the production process of rice bran oil is defatted with an organic solvent, and the defatted product is extracted with water or alcohol ( Patent Document 1).

On the other hand, it is known that ingestion of phytosterol lowers blood cholesterol. Phytosterols are plant sterols found in small amounts in vegetable oils such as corn, beans or other vegetable oils, which occur as free sterols, fatty acid esters and glycosides. Phytosterols are structurally similar to cholesterol, with the major differences occurring in the carbon backbone of their side chains. Many different phytosterol structures are found in nature. The most common are campesterol, beta-sitosterol and stigmasterol. In particular, stigmasterol has a remarkable cholesterol lowering effect in blood (Non-patent Document 1).

In addition, Japanese Patent Laid-Open No. 2005-73502 (Patent Document 2) describes a food or food material having an apoptosis-inducing ability, which comprises a mushroom extract obtained by extracting a mushroom. In addition, the same publication describes that this extract of mushrooms exhibits cancer suppressing and preventive effects, and that the food or food material having an apoptosis-inducing ability consists of ergosterol peroxide derived from mushrooms. ing.

On the other hand, the following problems were tried for the purpose of comprehensively preventing and treating diseases derived from lifestyle-related diseases. We have already discovered resveratrol which activates the longevity gene in polyphenols containing a large amount of quercetin and chlorogenic acid in addition to the hypoglycemic action of mulberry stem powder over that of mulberry leaf powder (Japanese Patent Application No. 2011-90489). Furthermore, evening primrose powder, which contains a large amount of β-sitosterol glycoside that has a strong hemostatic effect on bleeding caused by vascular leaky disease, is a bleeding caused by lifestyle-related diseases such as hyperglycemia, hypertension and hyperlipidemia. It is known to have a comprehensive preventive effect against sexually transmitted diseases.

JP, 2001-97983, A JP, 2005-73502, A Batta et al. 2006, Metabolism Clinical and Experimental 55: 292-29.

Under such a background, the present inventor has found that glucosylceramide, stigmasterol, ergosterol peroxide, and β-sitosterol glycoside obtained by fermenting rice bran and rice germ have a muscle-enhancing effect. The present invention has been completed.
That is, an object of the present invention is to provide a new muscle enhancing agent.

The technical features of the present invention for solving the above problems are as follows.
1. A muscle differentiation induction promoter comprising a fermented extract obtained by fermenting rice bran and/or rice germ to obtain a fermented product, and then extracting the fermented rice product with a polar solvent.
2. Above 1. An agent for promoting muscle differentiation induction, which comprises, as an active ingredient, a partition product obtained by partitioning the fermentation extract described in 1. in a polar solvent.
3. A muscle differentiation induction promoter containing as an active ingredient at least one compound selected from stigmasterol, ergosterol peroxide, β-sitosterol glucoside, and glucosylceramide.
4. A muscle bulking agent comprising a fermented extract obtained by fermenting rice bran and/or rice germ to obtain a fermented product, and then extracting the fermented rice product with a polar solvent.
5. Above 4. A muscle bulking agent comprising a partition product obtained by partitioning the fermentation extract described above in a polar solvent as an active ingredient.
6. A muscle bulking agent containing stigmasterol as an active ingredient.
7. A muscle mass gene expression promoter comprising a fermented extract obtained by fermenting rice bran and/or rice germ to obtain a fermented product, and then extracting the fermented rice product with a polar solvent.
8. Above 7. A muscle mass gene expression promoter comprising the partition product obtained by partitioning the above-mentioned fermentation extract as described in 1. in a polar solvent as an active ingredient.
9. A muscle mass gene expression promoter comprising at least one of stigmasterol and glucosylceramide as an active ingredient.
10. A muscle contraction improving agent comprising a fermented extract obtained by fermenting rice bran and/or rice germ to obtain a fermented product, and then extracting the fermented rice product with a polar solvent.
11. The above 10. An agent for improving muscle contraction force, which comprises as an active ingredient a partition product obtained by partitioning the fermentation extract described in 1. in a polar solvent.
12. A muscular contractile strength improving agent containing glucosylceramide as an active ingredient.
13. Above 1. ~12. A muscle strengthening agent containing the above agent as an active ingredient.

It is a schematic diagram which shows the method of isolating Compound 1-5 from a rice germ fermentation extract. It is a graph which shows the result of the comparison test with the rice origin protein in differentiation induction promotion. It is a graph which shows the evaluation result of the muscle differentiation induction promotion action by a precocious differentiation marker (myogenin). It is a graph which shows the evaluation result of the muscle differentiation induction promotion action by a maturation differentiation marker (MyHP). It is a graph which shows the result of the comparison test with a fermented extract of unfermented rice germ and a rice-derived protein in the muscle cell growth promoting action. It is a graph which shows the evaluation result of a muscle cell proliferation promotion effect. It is a graph which shows the evaluation result of muscle growth gene IGF-1 expression promotion activity. It is a graph which shows a muscle contraction promotion effect evaluation result.

The present invention will be described in detail below.
The present invention is characterized by fermenting rice bran and/or rice germ to obtain a fermented product, and then using the fermented extract obtained by extracting the fermented rice product with a polar solvent as an active ingredient.

For the "rice bran and/or rice germ", raw bran produced in the rice polishing process of brown rice may be used. Brown rice consists of pericarp, seed coat, endosperm and germ, and the outer layer of endosperm tissue has a thin layer called aleurone layer.
As rice polishing progresses, not only the pericarp and seed coat but also the aleurone layer are eliminated as rice bran (raw bran). If necessary, a mixture of pericarp, seed coat, rice germ or aleurone can be used, or one obtained by separating either pericarp, seed coat, rice germ or aleurone can be used.

The rice bran and/or rice germ is preferably defatted. This is because it is possible to obtain a composition containing a higher concentration of the active ingredient. This degreasing method is not particularly limited,
At this time, preferable degreasing solvents include n-hexane and acetone. In particular, it is preferable to use n-hexane as the degreasing solvent. This is because the extracted oil component can be used as an edible oil and the fermented rice germ extract can be easily used as a food material.
These degreasing solvents may be used alone or in combination of two or more.
The microorganism used in the "fermentation" is not particularly limited, but koji mold, yeast, citric acid bacterium, lactic acid bacterium, acetic acid bacterium and the like can be used. Of these, koji mold is preferable.
The above-mentioned "Koji mold" is not particularly limited, but for example, one or more selected from sake mold, shochu, awamori, miso or soy sauce can be used.
For example, Aspergillus kawachii, Aspergillus awamori, Aspergillus oryzae, Aspergillus sojae, Aspergillus saitoius, Aspergillus saitoi, Aspergillus saitoi One or more selected. The fermentation temperature is not particularly limited, but is preferably 10 to 60°C, more preferably 20 to 40°C, and particularly preferably 25 to 35°C. This is because the temperature within this range is the optimum temperature for the fermentation by the koji mold to proceed.
The fermentation time is not particularly limited, and it is preferably 1 to 7 days, more preferably 2 to 4 days.

As the "polar solvent", polar solvents such as water, methanol, ethanol, isopropyl alcohol, 1,3-butylene glycol, ethylene glycol, propylene glycol, glycerin, ethyl acetate and the like can be used. You may mix 2 or more types of these solvents.
Desirably, when water ethanol or water-containing ethanol which is a mixture thereof is used as an extraction solvent, the active ingredient is efficiently extracted.

  When hydrous ethanol is used as the extraction solvent, the concentration of ethanol is not particularly limited, but the ethanol concentration is preferably 10 to 90% (wt/wt), and more preferably 20 to 80% (wt/wt). The reason why the ethanol concentration is 90% (wt/wt) or less is that if the ethanol concentration is too high, the oil component of the fermentation extract easily dissolves in the hydrous ethanol.

  The extraction temperature is 20 to 80°C, preferably about 40 to 50°C. This is because if the extraction temperature is too low, the active ingredient becomes difficult to extract, and if the extraction temperature is too high, the active ingredient is decomposed and the physiological activity (health functionality) is reduced.

  As the extraction method, any method such as stirring extraction, continuous extraction, immersion extraction, countercurrent extraction, supercritical extraction can be adopted, and any apparatus can be used at room temperature or under reflux heating.

Further, in the present invention, the above-mentioned fermentation extract may be used as it is, but it is also possible to use a product obtained by dividing the fermentation extract with a polar solvent for liquid separation thereafter.
The method of “distribution” is not particularly limited, but liquid-liquid distribution is preferable. The solvent used in the liquid-liquid distribution is not particularly limited, and for example, water, methanol, ethanol, isopropyl alcohol, 1,3-butylene glycol, ethylene glycol, propylene glycol, glycerin, hexane, ethyl acetate, chloroform and the like can be used. Of these, hexane, methanol and chloroform are preferably used.

By the above method, compounds such as stigmasterol, ergosterol peroxide, β-sitosterol glucoside, and glucosylceramide can be obtained, but are not limited thereto. In addition, these may use what isolate|separated only one type of compound, these mixtures, or a crude thing may be used.

Further, the present invention is characterized in that any one of stigmasterol, ergosterol peroxide, β-sitosterol glucoside, and glucosylceramide is used as an active ingredient.
The method for obtaining the above compound is not particularly limited, but it can be obtained by purifying and isolating the above-mentioned fermented product as described above.

The present invention can be applied to various uses. Examples of applications include pharmaceuticals, reagents, and compositions for oral ingestion (for example, health foods, foods with functional claims, supplements, foods for specific purposes, supplements for pets, etc.).

The pharmaceutical composition containing the present invention may contain an excipient, a carrier or an additive. Excipients, carriers, and additives are not particularly limited as long as they are commonly used and pharmaceutically acceptable, and their types and compositions can be appropriately changed.

Examples of the excipient include sodium chloride, sodium citrate and the like, and examples of the carrier include sterilized water, physiological saline, various buffer solutions and the like. Examples of the additives include thickeners, buffers, preservatives, preservatives and the like.

The dosage form of the pharmaceutical composition is not particularly limited and may be appropriately selected as necessary. For example, oral preparations such as tablets, capsules, granules, fine granules and powders; injections , Suppositories,
Parenteral agents such as a coating agent may be mentioned.

Oral preparations such as tablets, capsules, granules, fine granules, powders, etc. are manufactured according to a conventional method using starch, lactose, sucrose, trehalose, mannitol, carboxymethylcellulose, corn starch, inorganic salts, etc., The compounding amount of the compound of the present invention in these preparations is not particularly limited and can be appropriately set. Binders, disintegrants, surfactants, lubricants, fluidity promoters, corrigents, coloring agents, flavors and the like can be appropriately used in this type of preparation.

In the case of a parenteral preparation, the dose is adjusted according to the age, body weight, degree of disease, etc. of the patient and administered by, for example, intravenous injection, intravenous drip injection, subcutaneous injection, intramuscular injection or the like. This parenteral preparation is manufactured according to a conventional method, and distilled water for injection, physiological saline or the like can be generally used as a diluent. Further, if necessary, a bactericide, a preservative, a stabilizer and the like may be added. Further, from the viewpoint of stability, this parenteral preparation may be filled in a vial or the like and then frozen, the water content thereof may be removed by an ordinary freeze-drying treatment, and the liquid preparation may be re-prepared from the freeze-dried product immediately before use. Further, if necessary, a tonicity agent, a stabilizer, a preservative, a soothing agent, etc. may be added. Examples of other parenteral agents include liquid preparations for external use, coating agents such as ointments, suppositories for rectal administration, and the like, which are also manufactured according to a conventional method.

When the present invention is used for health foods (including functionally labeled foods, foods for specified health uses, health drinks and supplements), the compound of the present invention may be added to health foods as a raw material for various health foods, or dextrin may be added as necessary. A health food can be produced by processing into pellets, tablets, granules or the like together with excipients such as lactose, starch and the like, flavors, pigments or the like, or by coating with gelatin or the like and molding into capsules.

The blending ratio of the muscle enhancing agent to be incorporated into the health food is not particularly limited as long as the effect expected from the muscle enhancing agent can be obtained, but usually, the intake amount is about 0.0001 to 2000 mg. is there.

These health foods can be blended with various components depending on their types, for example, glucose, fructose, sucrose, maltose, sorbitol, stevioside, corn syrup, lactose, citric acid, tartaric acid, malic acid, Succinic acid, lactic acid, L-ascorbic acid, dl-α-tocopherol, sodium erythorbate, glycerin, propylene glycol, glycerin fatty acid ester, polyglycerin fatty acid ester, sucrose fatty acid ester, sorbitan fatty acid ester, propylene glycol fatty acid ester, gum arabic Food materials such as carrageenan, casein, gelatin, pectin, agar, vitamin Bs, nicotinic acid amide, calcium pantothenate, amino acids, calcium salts, pigments, flavors and preservatives can be used. Furthermore, muscle strengthening agents having a health maintenance function include other antioxidants and health food materials such as antioxidants (reduced ascorbic acid (vitamin C), vitamin E, reduced glutatin, tocotrienol). , Vitamin A derivative, lycopene, lutein, astaxanthin, zeaxanthin, fucoxanthin, uric acid, ubiquinone, coenzyme Q10, folic acid, garlic extract, allicin, sesamin, lignans, catechin, isoflavone, chalcone, tannins, flavonoids, coumarin, isocoumarin , Blueberry extract), health food ingredients (V. (vitamin) A, V.B1, V.B2, V.B6, V.B12, VC, VD, VE, VP, choline, niacin, pantothenic acid, calcium folate) , EPA, oligosaccharides, dietary fiber, squalene, soy lecithin, taurine, donariella, protein, octacosanol, DHA, egg yolk lecithin, linoleic acid, lactoferrin, magnesium, zinc, chromium, selenium, potassium, heme iron, oyster meat extract, chitosan , Chitin oligosaccharide, collagen, chondroitin, turmeric, liquorice, kokushi, keihi, hawthorn, ginger, ganoderma lucidum, clam extract, terrapin, chamomile, chamomile, dandelion, hibiscus, honey, boren, royal jelly, lime, lavender, rosehip, Rosemary, bifidobacteria, faecalis fungus, lacris, wheat germ oil, sesame oil, perilla oil, soybean oil, medium chain fatty acid, agarix, ginkgo biloba extract, turmeric, chondroitin, brown rice germ extract, litchi, onion, DPA, beetroot tea, caterpillar , Garlic, bee pup, papaya, puer, propolis, megusuri tree, yam mushroom, saw palmetto, hyaluronic acid, collagen, gaba, harp seal oil, shark cartilage, glucosamine, lecithin, phosphatidylserine, field carrot, mulberry leaf, soybean Extract, Echinacea, Eleuthero, barley extract, olive leaf, olive fruit, Gymnema, Banaba, Salacia, Garcinia, chitosan, St. John's wort, jujube, carrot, passion flower, broccoli, placenta, coix, grape seed, peanut seed coat, bilberry , Black cohosh, thistle, laurel, sage, rafuma, black vinegar, bitter melon, maca, safflower, flax, oolong tea, flower spines, caffeine, capsaicin, xylooligosaccharide, glucosamine, buckwheat, citrus, dietary fiber, Protein, prune, spirulina, barley leaf, nucleic acid, yeast, shiitake mushroom, plum meat, amino acid, deep sea shark extract, noni, oyster meat, champignon, plantain, acerola, pineapple, banana, peach, apricot, melon, strawberry, raspberry, Orange, Fucoidan, Phellinus linteus, Cranberry, Chondroitin Sulfate, Zinc, Iron, Ceramide, Silk Peptide, Glycine, Niacin, Chest Tree, L-Cysteine, Red Wine Leaf, Millet, Horsetail, Biotin, Centella Asiatica, Hascup, Pycnogenol, Butterbur, rhubarb, cloves, catechin, puer, citric acid, brewer's yeast, merrilot, black zinger, ginger, gadgets, nattokinase, benizi, tocotrienols, lactoferrin, cinnamon, Chinese barley buckwheat, cocoa, yuzu seed extract, perilla seed extract, litchi Seed extract, evening primrose extract, black rice extract, α-lipoic acid, green coffee bean extract, Satsuma mandarin orange extract, triterpenoid, kiwi seed extract, red ginger extract, astaxanthin, walnut extract, resveratrol, red rice extract, white fungus polysaccharide, Strawberry seed extract, strawberry seed extract, leek seed extract, lingonberry extract, cherry blossom extract, Pleurotus cornucopia extract, maqui berry extract, rice polyamine, wheat polyamine, black ginger extract, jungsai extract, swallow nest extract, purple tea extract. , Chrysanthemum flower extract, seaberry extract) and the like can also be added.

When the present invention is used as a health food, the muscle enhancing agent can be used in the form of various general foods and drinks. Here, "it can be used as a dosage form of various general foods and drinks." means that general foods and drinks can be selected as a dosage form for the purpose of exerting the effect as the muscle enhancing agent. Is intended to be eaten only by those who desire the effect expected from the muscle strengthening agent, and can be widely eaten by everyone including those who do not expect the effect as the muscle strengthening agent. is not.
Further, the compounding amount for exerting the effect as the muscle enhancing agent is not particularly limited, but it is preferable that the total content of the active ingredient is 1 to 20 wt% with respect to the food or drink. In addition, although it is not particularly limited as a general food or drink to be mixed, for example, edible oil (salad oil), confectionery (gum, candy, caramel, chocolate, cookie, snack, jelly, gummy, tablet confectionery, etc.), noodles (soba, soba, Udon, ramen, etc.), dairy products (milk, ice cream, yogurt, etc.), seasonings (miso, soy sauce, etc.), soups, drinks (juice, coffee, tea, tea, carbonated drinks, sports drinks, etc.), etc. Be done.

  These general foods and drinks can be blended with various components depending on their types, for example, glucose, fructose, sucrose, maltose, sorbitol, stevioside, corn syrup, lactose, citric acid, tartaric acid, malic acid, Succinic acid, lactic acid, L-ascorbic acid, dl-α-tocopherol, sodium erythorbate, glycerin, propylene glycol, glycerin fatty acid ester, polyglycerin fatty acid ester, sucrose fatty acid ester, sorbitan fatty acid ester, propylene glycol fatty acid ester, gum arabic Food materials such as carrageenan, casein, gelatin, pectin, agar, vitamin Bs, nicotinic acid amide, calcium pantothenate, amino acids, calcium salts, pigments, flavors and preservatives can be used. In addition, the muscle enhancing agent having a health maintenance function includes other antioxidants and health food materials such as antioxidants (reduced ascorbic acid (vitamin C), vitamin E, reduced glutatin, Tocotrienols, vitamin A derivatives, lycopene, lutein, astaxanthin, zeaxanthin, fucoxanthin, uric acid, ubiquinone, coenzyme Q10, folic acid, garlic extract, allicin, sesamin, lignans, catechins, isoflavones, chalcones, tannins, flavonoids, coumarin, Isocoumarins, blueberry extract), health food materials (V. (vitamins) A, V.B1, V.B2, V.B6, V.B12, VC, VD, VE, VP, choline, niacin, pantothenic acid, folic acid Calcium, EPA, oligosaccharide, dietary fiber, squalene, soy lecithin, taurine, donariella, protein, octacosanol, egg yolk lecithin, linoleic acid, lactoferrin, magnesium, chromium, selenium, potassium, heme iron, oyster meat extract, chitosan, chitin oligo Sugar, collagen, chondroitin, turmeric, licorice, kokushi, cinnamon beetle, hawthorn, ginger, ganoderma lucidum, hydrangea extract, kanzo, kokushi, keihi, hawthorn, ginger, ganoderma lucidum, psyllium, chamomile, chamomile, dandelion, hibiscus, honey, boren , Royal jelly, lime, lavender, rosehip, rosemary, sage, bifidobacteria, faecalis fungus, lacris, wheat germ oil, sesame oil, perilla oil, soybean oil, medium chain fatty acid, agarix, ginkgo biloba extract, turmeric, chondroitin, brown rice Germ extract, litchi, onion, DHA, EPA, DPA, beet tea, caterpillar, garlic, bee pup, papaya, puer, propolis, megusuri tree, yab mushroom, royal jelly, saw palmetto, hyaluronic acid, collagen, gaba, harp seal oil , Shark cartilage, glucosamine, lecithin, phosphatidylserine, field carrot, mulberry leaf, soybean extract, echinacea, eleuthero, barley extract, olive leaf, olive fruit, gymnema, banaba, salacia, garcinia, chitosan, St. John's wort, jujube , Carrot, passionflower, broccoli, placenta, pearl barley, grape seeds, peanut seed coat, bilberry, black cohosh, maria thistle, laurel, sage, rosemary, rafuma, black vinegar, bitter melon, maca, safflower, flax, u -Long tea, flower spines, caffeine, capsaicin, xylooligosaccharide, glucosamine, buckwheat, citrus, dietary fiber, protein, prune, spirulina, barley leaf, nucleic acid, yeast, shiitake mushroom, plum meat, amino acid, deep sea shark extract, noni, Oyster meat, soft-shelled turtle, champignon, plantain, acerola, pineapple, banana, peach, apricot, melon, strawberry, raspberry, orange, fucoidan, messimacob, cranberry, chondroitin sulfate, zinc, iron, ceramide, silk peptide, glycine, niacin, chest Tree, L-cysteine, red wine leaf, millet, horsetail, biotin, centella asiatica, haskap, pycnogenol, butterbur, rhubarb, clove, rosemary, catechin, puer, citric acid, brewer's yeast, meliloth, black zinger, ginger , Gautu, Nattokinase, Benikouji, Tocotrienol, Lactoferrin, Cinnamon, Buckwheat buckwheat, Cocoa, Yuzu seed extract, Perilla seed extract, Litchi seed extract, Evening primrose extract, Black rice extract, α-lipoic acid, Gaba, Green coffee bean extract, Japanese butterbur extract , Kiwi seed extract, Wenzhou tangerine extract, red ginger extract, astaxanthin, walnut extract, leek seed extract, red rice extract, citrus extract, white fungus, polysaccharides, fucoxanthin, lingonberry extract, cherry blossom extract, maquiberry extract, sacred cherry extract. , Rice polyamine, wheat polyamine, black ginger extract, Junsai extract, swallow's nest extract, purple tea extract, chrysanthemum flower extract, seaberry extract) and the like can also be blended.

  As a specific manufacturing method, the muscle enhancing agent is spray-dried or freeze-dried together with powdered cellulose, and the powder, granules, tableting or solution is used to easily prepare food and drink (health foods, foods with functional claims, etc. Can be included). In addition, the muscle enhancing agent can be dissolved in, for example, fats and oils, ethanol, glycerin, or a mixture thereof to form a liquid and then added to a beverage or a solid food. If necessary, it may be mixed with a binder such as gum arabic or dextrin to give a powder or granules, which may be added to a beverage or a solid food.

Example: Isolation of rice germ fermented extract and active ingredient (1) Preparation of rice germ fermented extract 1.4 kg of defatted rice germ and an equal amount of distilled water were mixed and autoclaved, and 560 mg of koji mold and incense 42 g was added and mixed, and the mixture was fermented at 35° C. for 3 days to obtain a koji mold fermented rice germ. The fermented rice germ was extracted with 3 kg of methanol with heating three times, and the extract was concentrated under reduced pressure to obtain 156.06 g of a methanol extract (fermented rice germ extract).
(2) Isolation of active ingredient 83 g of this methanol extract was sequentially separated with hexane, ethyl acetate, and chloroform:methanol = 6:1 to give 17.67 g of hexane-soluble portion, 5.73 g of ethyl acetate-soluble portion, and chrometa. 3.73 g of a soluble part was obtained. Each soluble part was fractionated according to the fractionation scheme of FIG. 1 to isolate 5 components of Compound 1-5. Structural analysis by NMR was performed on the five isolated components. Of these, structural analysis was performed on Compounds 1, 2, 4, and 5.
Here, the chemical formulas of Compounds 1, 2, 4, and 5 are shown below.

Test Example 1: Muscle Differentiation Induction Promoting Action (1) Test on Rice Germ Fermented Extract The rice germ fermented extract was added to mouse myoblast C2C12 at a concentration of 1-100 μg/mL, and at the same time, differentiation was induced. It was After culturing for 1 week, the cells were collected and quantitatively analyzed for the mRNA expression levels of myogenin which is a muscle precocious differentiation marker and myosin, heavy polypeptide (MyHP). At that time, GAPDH was used as an endogenous control for correction. In addition, as a comparative example, the same test was conducted on rice extract having no fermentation step and oryza protein (manufactured by Oryza Yuka Co., Ltd.) which is a rice-derived protein having no fermentation step. The result is shown in FIG.
(2) Test on isolated component In the same manner as the above method, the component (Compound. 1-5) isolated from the fermented rice germ extract was tested at a concentration of 1 and 10 µg/mL. The results are shown in Fig. 3 (myogenin) and Fig. 4 (MyHP).

Effect of Example in Test Example 1 As shown in FIG. 2, the rice germ fermented extract significantly promoted myocyte proliferation. The positive control IGF-1 also proliferated significantly. In addition, unfermented rice germ extract has an action of suppressing cell growth, and rice-derived protein (Oryza protein), which is a non-fermented rice extract, has a muscle growth promoting effect as compared with the rice germ fermented extract. It was weak. From the above, it has been confirmed that fermenting rice germ has an unprecedented muscle-enhancing effect, which is stronger than ingesting protein.
Further, as shown in FIG. 3, the expression of myogenin, which is a muscle precocious differentiation marker, was promoted by the addition of Compound 1-5 (1 and 10 μg/mL). In particular, the addition of Compound 1 (1 μg/mL) and Compound 5 (1 and 10 μg/mL) significantly promoted the expression. On the other hand, as shown in FIG. 4, in MyHP which is a marker for differentiation of muscle maturation, Compound 1 (1 and 10 μg/mL), Compound 3 (10 μg/mL), Compound 4 (1 μg/mL), Compound 5 (1 And 10 μg/mL) promoted the expression. In particular, the addition of Compound 1 (1 μg/mL) and Compound 5 (10 μg/mL) significantly promoted the expression. From the above, it was confirmed that Compound 1-5 has an activity of promoting the induction of muscle differentiation in the precocious stage and the mature stage. From the above, it was confirmed that the fermented rice germ extract and Compound 1-5 function as a muscle differentiation induction promoter.

Test Example 2: Muscle cell proliferation promoting action (muscle mass increasing action)
(1) Evaluation by rice fermented rice germ extract To the mouse myoblast C2C12, rice fermented rice germ extract, as a comparative example, rice-derived protein (Oriza protein) was added at 1-100 μg/mL, and at the same time, differentiation induction I went. After culturing for 1 week, MTT assay was performed to evaluate the effect on cell proliferation. In addition, as a comparative example, the same test was conducted on rice extract having no fermentation step and oryza protein (manufactured by Oryza Yuka Co., Ltd.) which is a rice-derived protein having no fermentation step. The result is shown in FIG.
(2) Evaluation by isolated components Rice germ fermented extract and component (Compound 1) isolated from rice germ fermented extract were added to mouse myoblast C2C12 at a concentration of 0.1-30 μg/mL, and at the same time differentiation induction I went. After culturing for 1 week, MTT assay was performed to evaluate the effect on cell proliferation. The result is shown in FIG.

Effect of Example in Test Example 2 As shown in FIG. 5, the rice germ fermented extract significantly promoted myocyte proliferation. In addition, the positive control IGF-1 also proliferated significantly. In addition, the unfermented rice germ extract showed an action of suppressing cell growth, and the protein derived from rice had a weaker muscle growth promoting effect than the fermented rice germ extract. From the above, it was confirmed that fermenting rice germ had an unprecedented muscle-enhancing effect, which was stronger than ingesting protein.
Moreover, as shown in FIG. 6, cell proliferation was significantly increased by adding Compound 1 at a concentration of 3 to 30 μg/mL. The positive control rice fermented extract (10 μg/mL) also increased significantly. From the above, it was suggested that the rice germ fermented extract and its component Compound 1 have the activity of promoting the proliferation of muscle cells. From the above, it was confirmed that the fermented rice germ extract and its component Compound 1 are useful as a muscle mass increasing agent.

Test Example 3: Muscle hypertrophy promoting factor IGF-1 expression promoting activity To the mouse myoblast C2C12, the components (Compound 1-5) isolated from the rice germ fermentation extract were added at a concentration of 1 and 10 μg/mL, and At the same time, differentiation was induced. After culturing for 1 week, the cells were collected and quantitatively analyzed for the mRNA expression level of the muscle growth factor IGF-1. At that time, GAPDH was used as an endogenous control for correction. The result is shown in FIG. 7.

Effects of Examples in Test Example 3 As shown in FIG. 7, the expression of IGF-1, which is a muscle mass increasing factor, was promoted by the addition of Compound 1 (1 μg/mL) and Compound 5 (1 μg/mL). From the above, Compound 1 and 5 were confirmed to be useful as a muscle mass increasing gene expression promoter.

Test Example 4: Muscle contraction promoting action For mouse myoblast C2C12 embedded in collagen gel, rice germ fermentation extract (10 μg/ml) and Compound 5 (10 μg/ml), insulin-like growth factor 1 (IGF-1, 1 ng/ml) was added, and at the same time, differentiation was induced. After inducing differentiation for 2 weeks, the size of the collagen gel was measured and the effect on the contractile force was evaluated. The result is shown in FIG.

Effect of Example in Test Example 4 A collagen gel contraction test was performed as an index of muscle contraction. As a result, as shown in FIG. 8, the contraction of collagen gel was significantly recognized by the addition of the rice germ fermentation extract and Compound 5 as in the positive control IGF-1. From the above, it was confirmed that Compound 5 is useful as a muscle contraction strength improving agent.

Effects of Examples From the above, it was confirmed that the fermented rice germ extract and Compound 1-5 obtained thereby are useful as a muscle strengthening agent.

The formulation examples of the muscle enhancing agent of the present invention are shown below, but the following formulation examples do not limit the present invention.
Formulation 1: Chewing gum (food with functional claims or food for specified health uses)
Sugar 53.0wt%
Gum base 20.0
Glucose 10.0
Starch syrup 16.0
Fragrance 0.5
Muscle enhancer 0.5
100.0 wt%

Formulation 2: Gummy (food with functional claims or food for specified health uses)
Reduced starch syrup 40.0wt%
Granulated sugar 20.0
Glucose 20.0
Gelatin 4.7
Water 9.68
Kiwi juice 4.0
Kiwi flavor 0.6
Pigment 0.02
Muscle enhancer 1.0
100.0 wt%

Formulation 3: Candy (food with functional claims or food for specified health uses)
Sugar 50.0wt%
Starch syrup 33.0
Water 14.4
Organic acid 2.0
Fragrance 0.2
Muscle enhancer 0.4
100.0 wt%

Formulation 4: Yogurt (hard/soft) (food with functional claims or food for specified health uses)
Milk 41.5wt%
Skim milk powder 5.8
Sugar 8.0
Agar 0.15
Gelatin 0.1
Lactic acid bacteria 0.005
Muscle enhancer 0.4
Fragrance
Water residue
100.0 wt%

Formulation 5: Soft drink (food with functional claims or food for specified health use)
Fructose glucose liquid sugar 30.0 wt%
Emulsifier 0.5
Muscle enhancer 0.05
Fragrance suitable amount
Purified water residue
100.0 wt%

Formulation 6: Soft capsules (foods with functional claims or foods for specified health uses)
Rice germ oil 87.0 wt%
Emulsifier 12.0
Muscle enhancer 1.0
100.0 wt%

Formulation 7: Tablets (foods with functional claims or foods for specified health uses)
Lactose 54.0wt%
Crystalline cellulose 30.0
Starch degradation product 10.0
Glycerin fatty acid ester 5.0
Muscle enhancer 1.0
100.0 wt%

Formulation 8: Oral granule (medicine)
Muscle enhancer 1.0 wt%
Lactose 30.0
Corn starch 60.0
Crystalline cellulose 8.0
Polyvinyl pyrrolidone 1.0
100.0 wt%

Formulation 9: Tablets
Sugar 76.4 wt%
Glucose 19.0
Sucrose fatty acid ester 0.2
Muscle enhancer 0.5
Purified water 3.9
100.0 wt%

Formulation 15: Cat food
Corn 34.0wt%
Flour 35.0
Meat meal 15.0
Beef tallow 8.9
Salt 1.0
Skipjack extract 4.0
Muscle enhancer 1.0
Taurine 0.1
Vitamins 0.5
Minerals 0.5
100.0 wt%

Formulation 16: dog food
Corn 30.0 wt%
Meat (chicken) 15.0
Defatted soybean 10.0
Flour 25.0
Rice bran 5.0
Muscle enhancer 5.0
Animal fats and oils 8.9
Oligosaccharide 0.1
Vitamin 0.5
Mineral 0.5
100.0 wt%

As described above, the present invention can provide a novel muscle enhancing agent.

Claims (5)

A muscular differentiation induction promoter containing at least one compound selected from stigmasterol, ergosterol peroxide, β-sitosterol glucoside, and glucosylceramide as an active ingredient. A muscle bulking agent containing stigmasterol as an active ingredient. A muscular mass increase gene expression promoter comprising at least one of stigmasterol and glucosylceramide as an active ingredient. A muscular contractile strength improving agent containing glucosylceramide as an active ingredient. A muscle strengthening agent comprising the agent according to any one of claims 1 to 4 as an active ingredient.