JP7209207B2 - muscle activator - Google Patents

Description

The present invention provides one or more plant extracts selected from the group consisting of bergamot, plantain coriander, wormwood wormwood, wildflower, soursop, giraffe, alligator pear, safflower, corn, tuna, buckwheat, udo, and gourd. The present invention relates to muscle activators containing substances.

Muscles are composed of myofibers that are myoblasts differentiated into myotubes and myotubes fused together. By activating these myoblasts and promoting cell proliferation, it is expected to be effective in muscle building or prevention of muscle wasting. Furthermore, similar effects can be expected by activating (phosphorylating) p70S6K and rpS6, which are downstream factors of the mTOR signal that promote muscle protein synthesis. Thus, activation of myoblasts or activation of p70S6K and rpS6 is associated with muscle diseases such as myopathy, sarcopenia associated with aging, locomotive syndrome (locomotive syndrome), long-term cast immobilization, and It is thought to be useful for prevention and/or treatment of disuse atrophy due to lack of exercise, inhibition of decrease in muscle protein synthesis during or after exercise, and the like. In addition, there are muscles under the skin that support the entire skin, but it is thought that when muscle function declines due to aging, the ability to support the skin weakens, leading to a decrease in skin elasticity, sagging, and wrinkle formation. Therefore, activation of myoblasts or activation of p70S6K and rpS6 is also expected to improve skin.
With the extension of the average life expectancy and the progress of the declining birthrate and aging population, it has an excellent myoblast activating effect, p70S6K activating effect, and rpS6 activating effect, and has a highly safe function in order to live long while maintaining health. There is a strong demand for the development of sex foods, cosmetic ingredients, and the like.

Special table 2012-529469 JP 2010-195696 JP 2008-156294

A main object of the present invention is to provide a novel muscle activator.

As a result of investigations to solve the above problems, the present inventors selected from a plant group consisting of bergamot, plantain root, wormwood, vadasou, soursop, giraffe, alligator pear, safflower, corn, tuna, buckwheat, udo and gourd It was found that one or two or more plant extracts having an excellent myoblast activating effect, and furthermore, bergamot, psyllium coriander, wormwood Niita, wildflower, and gourd were found to have a p70S6K activating effect. completed the invention.

The present invention includes the following.
(1) One or two or more plant extracts selected from the plant group consisting of bergamot, plantain coriander, wormwood wormwood, wildflower, soursop, giraffe, alligator pear, safflower, corn, tuna, buckwheat, udo and gourd as an active ingredient, a muscle activator.
(2) The muscle activator according to (1) above, wherein the muscle activator is a muscle-enhancing agent or an agent for preventing muscle wasting.
(3) The muscle activator according to (1) above, wherein the muscle activator is a skin improving agent.

The muscle activator according to the present invention is expected to be effective as a muscle-strengthening agent, a muscle-attenuation-preventing agent, and a skin-improving agent by activating myoblasts or promoting muscle protein synthesis. The muscle activator according to the present invention is highly safe because it is derived from edible plant components.

FIG. 3 shows myoblast-activating effects of a coriander psyllium extract and a bergamot extract. The vertical axis represents the cell activation activity (%) when the control group is set to 100. The horizontal axis represents the treatment concentration (ppm) of coriander and bergamot extracts. FIG. 4 is a diagram showing the myoblast activating action of a soursop extract and a giraffe extract. The vertical axis represents the cell activation activity (%) when the control group is set to 100. The horizontal axis represents the treatment concentration (ppm) of the soursop extract and eucharis extract. It is a figure which shows the myoblast activating effect of the alligator pear extract. The vertical axis represents the cell activation activity (%) when the control group is set to 100. The horizontal axis represents the treatment concentration (ppm) of alligator pear extract. FIG. 3 shows the myoblast-activating action of a safflower extract. The vertical axis represents the cell activation activity (%) when the control group is set to 100. The horizontal axis represents the treatment concentration (ppm) of the safflower extract. FIG. 4 is a diagram showing the myoblast activating effect of the Nitaka mugwort extract. The vertical axis represents the cell activation activity (%) when the control group is set to 100. The horizontal axis represents the treatment concentration (ppm) of the wormwood extract. It is a figure which shows the myoblast activation effect|action of a maize extract. The vertical axis represents the cell activation activity (%) when the control group is set to 100. The horizontal axis represents the treatment concentration (ppm) of corn extract. FIG. 2 is a diagram showing myoblast activating action of a gourd extract, a wild boar extract, a petiole extract, and a buckwheat extract. The vertical axis represents the cell activation activity (%) when the control group is set to 100. The horizontal axis represents the treatment concentration (ppm) of the gourd extract, the terracotta extract, the petroleum extract, and the buckwheat extract. FIG. 4 is a diagram showing the myoblast-activating effect of the udo extract. The vertical axis represents the cell activation activity (%) when the control group is set to 100. The horizontal axis represents the treatment concentration (ppm) of udo extract. FIG. 4 shows the p70S6K activation action of bergamot extract. The vertical axis represents the relative value of the amount of p70S6K phosphorylation when the control group is set to 1. The horizontal axis represents the treatment concentration (ppm) of bergamot extract. FIG. 3 shows the p70S6K activation effect of Psyllium coriander extract. The vertical axis represents the relative value of the amount of p70S6K phosphorylation when the control group is set to 1. The horizontal axis represents the treatment concentration (ppm) of coriander extract. FIG. 4 shows the p70S6K activating action of a mugwort extract. The vertical axis represents the relative value of the amount of p70S6K phosphorylation when the control group is set to 1. The horizontal axis represents the treatment concentration (ppm) of the wormwood extract. FIG. 3 shows the p70S6K activation action of the crocodile extract. The vertical axis represents the relative value of the amount of p70S6K phosphorylation when the control group is set to 1. The horizontal axis represents the treatment concentration (ppm) of the polliana extract. FIG. 3 shows the p70S6K activating action of the extract of P. gourd. The vertical axis represents the relative value of the amount of p70S6K phosphorylation when the control group is set to 1. The horizontal axis represents the treatment concentration (ppm) of P. gourd extract.

Plant extract Bergamot (Citrus bergamia) is a plant belonging to the genus Citrus of the family Rutaceae. Bergamot extracts according to the present invention can be extracts from various parts of bergamot, such as leaves, stems, roots, fruits, pericarp, seeds, seed coats, aerial parts, underground parts, or whole plants. A pericarp extract is preferred.
Eryngium foetidum is a plant belonging to the genus Eryngium foetidum of the family Apiaceae. For the coriander extract according to the present invention, extracts of various parts of coriander, such as leaves, stems, roots, fruits, pericarp, seeds, seed coats, aboveground parts, underground parts, or whole plants can be used. Whole plant extracts are preferred.
Artemisia campestris is a plant belonging to the genus Artemisia of the Asteraceae family. For the wormwood extract according to the present invention, it is possible to use an extract of each part of wormwood, such as leaves, stems, branches, roots, fruits, pericarp, seeds, seed coats, aboveground parts, underground parts, or whole plants. can. Preferred are twig and leaf extracts.
Pseudostellaria heterophylla is a plant belonging to the family Caryophyllaceae. For the extract of the crocodile plant according to the present invention, an extract of each part of the crocodile plant, such as leaves, stems, roots, fruits, pericarp, seeds, seed coats, aerial parts, underground parts, or the whole plant can be used. Preferably, it is a root extract.
Annona muricata is a plant belonging to the genus Annuaceae of the family Annuaceae. The extract of soursop sorrel according to the present invention can be an extract of each part of soursop, such as leaves, stems, roots, fruits, pericarp, seeds, seed coats, aerial parts, underground parts, or whole plants. Fruit extracts are preferred.
Kirinketu (Daemonorops draco) is a plant belonging to the genus Coleoptera of the Palm family. The extract of Eucheuma muricatum according to the present invention can be an extract of each part of Eucheuma muricatum, such as leaves, stems, roots, fruits, pericarp, seeds, seed coats, aerial parts, underground parts, or whole plants. Fruit extracts are preferred.
Alligator pear (Persea americana) is an extract of a plant belonging to the genus Alligator in the family Lauraceae. For the alligator pear extract according to the present invention, an extract from each part of alligator pear, such as leaves, stems, roots, fruits, pericarp, seeds, seed coats, aerial parts, underground parts, or whole plant can be used. Fruit extracts are preferred.
Safflower (Carthamus tinctorius) is an extract of a plant belonging to the genus Carthamus of the Asteraceae family. For the safflower extract according to the present invention, extracts of various parts of safflower, such as flowers, leaves, stems, roots, fruits, pericarp, seeds, seed coats, aboveground parts, underground parts, or whole plants can be used. Flower extracts are preferred.
Maize (Zea mays) is a plant belonging to the genus Maize of the Poaceae family. The corn extract according to the present invention can be an extract of each part of corn, such as pistil (whisker), stem, root, fruit, pericarp, seed, seed coat, above ground part, below ground part, or whole plant. . A pistil extract is preferred.
Tsuruna (Tetragonia tetragonioides) is a plant belonging to the genus Tetragonia of the family Tetragonaceae. The extract of tetula according to the present invention can be an extract of each part of tetula, such as leaves, stems, roots, fruits, pericarp, seeds, seed coats, aerial parts, underground parts, or the whole plant. Whole plant extracts are preferred.
Buckwheat (Fagopyrum esculentum) is a plant belonging to the genus Fagopyrum of the Polygonaceae family. As the buckwheat extract according to the present invention, extracts from buckwheat parts such as leaves, stems, roots, fruits, pericarp, seeds, seed coats, aboveground parts, belowground parts, or whole plants can be used. Preferably, it is a seed extract.
Aralia cordata is a plant belonging to the Araliaceae family Aralia. For the udo extract according to the present invention, extracts of various parts of udo, such as leaves, flowers, stems, roots, fruits, pericarp, seeds, seed coats, above-ground parts, underground parts, or whole plants can be used. Preferred are stem and leaf extracts.
Trichosanthes kirilowii var. japonica is a plant belonging to the family Cucurbitaceae, genus Trichosanthes. For the extract of the gourd of the present invention, an extract of each part of the gourd, such as leaves, stems, roots, fruits, pericarp, seeds, seed coats, aerial parts, underground parts, or the whole plant can be used. Preferably, it is a seed extract.

The method for producing the plant extract according to the present invention is not particularly limited, and it can be produced by a commonly used method. For example, it can be produced by cutting each part of the plant as it is or cutting it into an appropriate size, drying it if necessary, and extracting it with juice or a solvent.
The extraction solvent is not particularly limited, but examples include water, lower alcohols such as methanol, ethanol, propanol, isopropanol and n-butanol, polyhydric alcohols such as 1,3-butylene glycol, propylene glycol and glycerin, ethyl ether, Examples include ethers such as propyl ether, esters such as ethyl acetate, and ketones such as acetone and ethyl methyl ketone. Also, these solvents can be used singly or in combination of two or more. Among the extraction solvents, water, methanol, ethanol, and hydrous ethanol are particularly preferred.

The amount of the solvent used for extraction varies depending on the extraction conditions and the like. It is preferably within the range of 1:5 to 1:10. A suitable extraction time is in the range of 1 hour to 15 days. The extraction temperature is suitably in the range of 5-100°C, preferably in the range of 60-100°C.
The extraction method is not particularly limited, and any method such as batch extraction or continuous extraction using a column can be applied.

The obtained extract can be used as it is, or if necessary, it may be subjected to further purification treatment within a range that does not affect its activity. Such purification treatment may be performed by a conventional method, for example, by filtering the extract by a conventional method. The obtained filtrate can also be subjected to concentration under reduced pressure, freeze drying or spray drying.

Muscle Activator The muscle activator according to the present invention (hereinafter referred to as the "muscle activator of the present invention") has myoblast activating action, p70S6K activating action, or rpS6 activating action, and is used to increase muscle mass. It is useful for strengthening and suppressing muscle atrophy, promoting recovery from exercise-induced muscle damage, and preventing and treating sarcopenia and locomotive syndrome associated with lack of exercise and aging. In addition, the muscle activator of the present invention activates myoblasts existing in the vicinity of the skin area and further promotes the synthesis of muscle proteins, thereby improving skin elasticity, sagging, wrinkles, etc. of the surrounding skin. It is also expected to have an improvement effect, an effect of alleviating fatigue, and an effect of alleviating stiff shoulders and eyestrain by alleviating muscle fatigue. The muscle activator according to the present invention is particularly expected to be effective as a muscle strengthening agent, a muscle wasting preventive agent, and a skin improving agent.
The muscle activator of the present invention can be obtained by mixing the extract of the present invention obtained as described above as it is or with a material that can be used as a carrier, and then using a conventional method in various forms such as powder, lumps, and liquid. It can be manufactured by processing to

The content of the extract of the present invention in the muscle activator of the present invention is not particularly limited as long as the myoblast activating action, p70S6K activating action, or rpS6 activating action can be confirmed, but for example, 0.01% by weight or more. , preferably 0.05% by weight or more and 20% by weight or less, particularly preferably 0.1% by weight or more and 5% by weight or less.

The muscle activator of the present invention may optionally contain pharmaceutically or food-acceptable additives. Examples of such additives include binders, disintegrants, lubricants, dispersants, suspending agents, emulsifiers, buffers, antioxidants, excipients, surfactants, UV inhibitors, and sequestering agents. , thickeners, preservatives, antibacterial agents, humectants, and pigments, and one or more of these can be used. The muscle activator of the present invention may further contain a component known to have a muscle-enhancing action or skin-improving action. Examples of muscle-enhancing components include creatine, amino acids such as BCAA and leucine, and HMB (3-hydroxyisovaleric acid), and skin-improving components include collagen, hyaluronic acid, elastin, proteoglycan, ceramide, and vitamin C. be done.
The muscle activator of the present invention can be appropriately prepared into dosage forms such as tablets, powders, granules, capsules, liquids, syrups, drop tablets, tonics, lotions, ointments, creams and the like by conventional methods.
The intake amount of the muscle activator of the present invention varies depending on the symptoms, dosage form, age, body weight, etc. of the subject of administration, but is usually in the range of 0.01 to 100 g of the extract of the present invention per day for adults. , preferably in the range of 0.03 g to 60 g, more preferably in the range of 0.1 g to 10 g, but not necessarily limited to this range. Such a daily intake may be taken in one time, or may be taken in 2 to 4 divided doses.

The present invention will be described in further detail below with reference to examples and test examples. However, it goes without saying that the present invention is not limited to the following examples.

Production Example 1 Production of Methanol Extract of Each Plant 15 g of the dried product of each plant listed in Table 1 was pulverized, immersed in 75 g of methanol for 7 days, filtered, and then concentrated and dried using a rotary evaporator. Then, it was adjusted to a final concentration of 50 mg/mL with dimethyl sulfoxide (DMSO) to obtain a methanol extract of each plant.
Production Example 2 Production of Ethanol Extract of Each Plant 15 g of the dried product of each plant listed in Table 1 was pulverized, immersed in 75 g of ethanol for 7 days, filtered, and then concentrated and dried using a rotary evaporator. Then, it was adjusted to a final concentration of 50 mg/mL with dimethyl sulfoxide (DMSO) to obtain an ethanol extract of each plant.
Production Example 3 Production of hot water extract of each plant After grinding 10 g of the dried product of each plant listed in Table 1, add it to 150 mL of hot water at 100 ° C., heat for 30 minutes, filter and concentrate with a rotary evaporator. Drying was performed. Then, it was adjusted to a final concentration of 50 mg/mL with dimethyl sulfoxide (DMSO) to obtain a hot water extract of each plant.

Test Example 1 Measurement of myoblast activating activity of plant extract according to the present invention A mouse myoblast cell line (C2C12 cells, manufactured by DS Pharma Biomedical Co., Ltd.) was used as cells. C2C12 cells are dispersed in DMEM (Dulbecco's Modified Eagle Medium, manufactured by Gibco) containing 10% FBS (Fetal Bovine Serum, manufactured by Gibco) (hereinafter referred to as "10% FBS/DMEM medium"), 5.0 × 10 ⁴ cells were seeded in an adherent cell culture flask (bottom area: 25 cm ² , manufactured by Nippon Bioscience) and placed in a CO ₂ incubator (manufactured by Sanyo Electric Biomedica; CO ₂ concentration 5%, humidity 90%). ) at 37° C. for 3 days. It was confirmed with a microscope that it became 80% confluent and used for experiments. 80% confluent C2C12 cells were detached from the flask using Trypsin-EDTA (manufactured by Gibco) and collected. The cells were suspended in a 10% FBS/DMEM medium and seeded in 100 μL portions on a 96-well plate (manufactured by Corning) at a cell density of 3.0×10 ⁴ cells/mL. After culturing for 24 hours at 37° C. and 5% CO ₂ conditions, the medium was removed with an aspirator and replaced with 200 μL of sample-containing medium prepared with 2% FBS/DMEM medium. The samples were No. 1 listed in Table 1 prepared in Production Example 1. Each methanol extract from 1 to 13 was adjusted with DMSO at 1000-fold concentration of the final concentration, then diluted 1000-fold with 2% FBS/DMEM medium, sample concentrations of 0.5, 1, 5, 10, 20 ppm media were prepared and used. In addition, DMSO was used as a control, and IGF-1 (Insulin-like Growth Factor-1, manufactured by Life Technologies), which is known to have a myoblast activation effect, was adjusted to 0.05 ppm as a positive control by the same sample preparation method. and used for the experiment. 48 hours after addition of the sample, the medium was replaced with 100 μL of serum-free DMEM medium, and 10 μL of WST-1 reagent (manufactured by Roche Diagnostics) was added thereto. After culturing for 30 minutes, the cell activating activity was determined by measuring absorbance at 450 nm with a microplate reader (MTP-300, manufactured by CORONA ELECTRIC). The results are shown in Figures 1-8.
As is clear from FIGS. 1 to 8, an increase in cell activating activity was observed when each of the plant extracts was added, compared to the case where the cell activating activity of the control group to which no drug was added was set at 100%.

Test Example 2 Measurement of Phosphorylation of p70S6K in Plant Extract According to the Present Invention A mouse myoblast cell line (C2C12 cells, manufactured by DS Pharma Biomedical) was used. C2C12 cells were dispersed in DMEM (Dulbecco's Modified Eagle Medium, manufactured by Gibco) containing 10% FBS (Fetal Bovine Serum, manufactured by Gibco) (hereinafter referred to as "10% FBS/DMEM medium") and adhered. 5.0 × 10 ⁴ cells were seeded in a culture flask for lineage cells (bottom area: 25 cm ² , manufactured by Nippon Bioscience) and placed in a CO ₂ incubator (manufactured by Sanyo Electric Biomedica; CO ₂ concentration 5%, humidity 90%). was cultured at 37°C for 3 days. It was confirmed with a microscope that it became 80% confluent and used for experiments. 80% confluent C2C12 cells were detached from the flask using Trypsin-EDTA (manufactured by Gibco) and collected. These cells were suspended in 10% FBS/DMEM medium and seeded in 12 wellplates at 1.7×10 ⁵ cells/well. 24 hours after seeding, the cells were replaced with 1 mL of 2% HS (Horse Serum, manufactured by Gibco) and differentiated into myotube cells. Myotube cells on the 3rd day of differentiation were used in the experiment by substituting with 2% HS once every 2 days from the start of differentiation. Myotube cells on the third day of differentiation were replaced with a serum-free medium, and after 9 hours, No. 1 shown in Table 1 prepared in Production Example 1 was used as a test substance. Samples were treated by substituting 1 mL of serum-free medium containing 1, 10 and 50 ppm of methanol extracts 1 to 4 and 13, respectively. Two hours after the start of the treatment, the cells were washed with PBS (Phosphatebuffered saline) and collected with 20 μL of RIPA buffer (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.). The resulting protein solution was added to an SDS-PAGE sample buffer, subjected to reduction treatment at 100° C. for 3 minutes, and subjected to Western blotting. Quantification of oxidation was performed. The antibodies used were Anti-p70S6K and Anti-phospho-p70S6K (manufactured by Cell Signaling Technology). The results are shown in Figures 9-13.
As is clear from FIGS. 9 to 13, the bergamot extract, Psyllium coriander extract, wormwood extract, crocodile extract, and gourd extract according to the present invention promoted the phosphorylation of p70S6K.

Test Example 3 Measurement of Phosphorylation of rpS6 in Plant Extract According to the Present Invention The same method as in Test Example 2 was used, except that Anti-rpS6K and Anti-phospho-rpS6K (manufactured by Cell Signaling Technology) were used as antibodies. can be used to quantify phosphorylation of rpS6K.

Since the muscle activator according to the present invention has an excellent muscle activating effect, for example, sarcopenia associated with lack of exercise or aging, locomotive syndrome, disuse atrophy due to various environments (casting, etc.), or locomotive It can be used for prevention and improvement of syndrome, enhancement of muscle building by exercise, reduction of skin elasticity, and suppression of wrinkles and sagging.

Claims (6)

A muscle activator containing a seed extract of gourd as an active ingredient. 2. The muscle activator according to claim 1, wherein the muscle activator is a muscle enhancer or a muscle wasting preventive agent. 3. The muscle activator according to claim 1 or 2, wherein the muscle activator is a skin improving agent or fatigue reducing agent. The muscle activator according to any one of claims 1 to 3 , wherein the extraction solvent for the plant extract is one or more selected from the group consisting of water, methanol, ethanol and hydrous ethanol. . A muscle activator according to any one of claims 1 to 4 , wherein the weight ratio of said extraction solvent to said dry weight of said plant is from 1:2 to 1:30. The muscle activator according to any one of claims 1 to 5 , which is a food.

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