JP2019026640A - Muscle activator - Google Patents

Abstract

To mainly provide a novel muscle activator.SOLUTION: The present invention relates to a muscle activator containing as active ingredients the extracts of Citrus bergamia, Eryngium foetidum, Artemisia morrisonensis, Pseudostellaria heterophylla, Annona muricata, Daemonorops, Persea americana, Carthamus tinctorius, Zea mays, Tetragonia tetragonioides, Fagopyrum esculentum, Aralia cordata, and Trichosanthes kirilowii. The muscle activator according to the present invention has myoblast activating action or p70S6K activating action, rpS6K activating action, and in particular, it is expected to be effective as a muscle enhancer, a muscle decay prophylactic agent and a skin ameliorating agent.SELECTED DRAWING: None

Description

  The present invention relates to one or more plant extracts selected from the group of plants consisting of bergamot, psyllium, sagebrush, scorpion, spruce, giraffe, crocodile, safflower, corn, tuna, buckwheat, udder, and scallop cucumber. It is related with the muscle activation agent containing a thing.

Muscles are composed of muscle fibers in which myoblasts are differentiated into myotube cells and myotube cells are fused. By activating these myoblasts and promoting cell proliferation, an effect can be expected in preventing muscle strengthening or muscle wasting. Furthermore, the same effect can be expected by activating (phosphorylating) p70S6K and rpS6, which are downstream factors of mTOR signal that promotes the synthesis of muscle protein. As described above, activation of myoblasts or activation of p70S6K and rpS6 is caused by muscular diseases such as myopathy, sarcopenia associated with aging, locomotive syndrome (motor organ syndrome), long-term cast fixation, It is considered useful for prevention and / or treatment of disuse atrophy due to lack of exercise, suppression of decrease in muscle protein synthesis during or after exercise. Furthermore, there are muscles under the skin that support the whole skin, but if the function of the muscles declines due to aging, etc., the ability to support the skin weakens, leading to reduced skin elasticity, sagging and wrinkle formation. Therefore, a skin improvement effect is also expected by activation of myoblasts or activation of p70S6K and rpS6.
Along with the prolongation of life expectancy and the declining birthrate and aging, it has excellent myoblast activation action, p70S6K activation action, rpS6 activation action and high safety function in order to live long while maintaining health Development of foodstuffs and cosmetic materials is eagerly desired.

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

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

  As a result of studying to solve the above problems, the present inventor has selected from a plant group consisting of bergamot, psyllium, sardine mugwort, waddows, spiders, giraffe, crocodile, safflower, corn, tuna, buckwheat, udo and kikarasuuri It is found that one or two or more kinds of plant extracts have an excellent myoblast activation action, and furthermore, a p70S6K activation action is found in bergamot, psyllium, niitaka mugwort, wadaso, kakirasuri, Completed the invention.

Examples of the present invention include the following.
(1) One or more plant extracts selected from the plant group consisting of bergamot, plantain, Japanese sagebrush, yellow-spotted swordfish, spruce, giraffe, crocodile, safflower, corn, tuna, buckwheat, udo A muscle activating agent containing as an active ingredient.
(2) The muscle activator according to (1), wherein the muscle activator is a muscle enhancer or a muscle attenuation preventive agent.
(3) The muscle activator according to (1), 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 enhancer, a muscle attenuation preventing agent, and a skin improving agent by promoting myoblast activation or muscle protein synthesis. The muscle activator according to the present invention is highly safe because it is derived from a component of an edible plant.

It is a figure which shows the myoblast activation effect | action of a plantain extract and a Bergamot extract. The vertical axis represents the cell activation activity (%) when the control group is 100. The horizontal axis represents the treatment concentration (ppm) of psyllium extract and bergamot extract. It is a figure which shows the myoblast activation effect | action of Togebanreishi extract and giraffe butt extract. The vertical axis represents the cell activation activity (%) when the control group is 100. The abscissa represents the treatment concentration (ppm) of Togebanreishi extract and Giraffe extract. It is a figure which shows the myoblast activation effect | action of a crocodile extract. The vertical axis represents the cell activation activity (%) when the control group is 100. The horizontal axis represents the treatment concentration (ppm) of the crocodile extract. It is a figure which shows the myoblast activation effect | action of a safflower extract. The vertical axis represents the cell activation activity (%) when the control group is 100. The horizontal axis represents the treatment concentration (ppm) of safflower extract. It is a figure which shows the myoblast activation effect | action of a Niitakamugi extract. The vertical axis represents the cell activation activity (%) when the control group is 100. The abscissa represents the treatment concentration (ppm) of Niitakamugi extract. It is a figure which shows the myoblast activation effect | action of a corn extract. The vertical axis represents the cell activation activity (%) when the control group is 100. The horizontal axis represents the treatment concentration (ppm) of corn extract. It is a figure which shows the myoblast activation effect | action of a kilya cucumber extract, a tsuruna extract, a waddo extract, and a buckwheat extract. The vertical axis represents the cell activation activity (%) when the control group is 100. The abscissa represents the treatment concentration (ppm) of the red pepper extract, the tsuruna extract, the wad extract, and the buckwheat extract. It is a figure which shows the myoblast activation effect | action of a wood extract. The vertical axis represents the cell activation activity (%) when the control group is 100. The horizontal axis represents the treatment concentration (ppm) of the Udo extract. It is a figure which shows the p70S6K activation effect | action of a bergamot extract. The vertical axis represents the relative value of the phosphorylation amount of p70S6K when the control group is 1. The horizontal axis represents the treatment concentration (ppm) of bergamot extract. It is a figure which shows the p70S6K activation effect | action of the plantain extract. The vertical axis represents the relative value of the phosphorylation amount of p70S6K when the control group is 1. The horizontal axis represents the treatment concentration (ppm) of psyllium extract. It is a figure which shows the p70S6K activation effect | action of a Niitakamugi extract. The vertical axis represents the relative value of the phosphorylation amount of p70S6K when the control group is 1. The abscissa represents the treatment concentration (ppm) of Niitakamugi extract. It is a figure which shows the p70S6K activation effect | action of a Wadder extract. The vertical axis represents the relative value of the phosphorylation amount of p70S6K when the control group is 1. The abscissa represents the treatment concentration (ppm) of the Wadder extract. It is a figure which shows the p70S6K activation effect | action of a Kurikarauri extract. The vertical axis represents the relative value of the phosphorylation amount of p70S6K when the control group is 1. The horizontal axis represents the treatment concentration (ppm) of the red pepper extract.

Plant extract bergamot is a plant belonging to the genus Citrus mandarin. As the bergamot extract according to the present invention, extracts of various parts of bergamot, for example, leaves, stems, roots, fruits, fruit skins, seeds, seed coats, above-ground parts, underground parts, or whole grasses can be used. Preferably, it is a fruit skin extract.
Psyllium (Eryngium foetidum) is a plant belonging to the genus Syphaceae. For the psyllium extract according to the present invention, extracts of psyllium, such as leaves, stems, roots, fruits, pericarps, seeds, seed coats, above-ground parts, underground parts, or whole plants can be used. Preferably, it is a whole plant extract.
Nitta hawk mugwort (Artemisia campestris) is a plant belonging to the genus Artemisia mugwort. For the Niitake sagebrush extract according to the present invention, it is possible to use each part of Niitake sagebrush, for example, an extract of leaves, stems, branches, roots, fruits, peels, seeds, seed coats, aerial parts, underground parts or whole grasses. it can. Preferably, it is a twig and leaf extract.
Wadder (Pseudostellaria heterophylla) is a plant belonging to the genus Lacidae. As the Wadder extract according to the present invention, an extract of each part of Wadder, for example, leaf, stem, root, fruit, fruit skin, seed, seed coat, above-ground part, underground part or whole plant can be used. Preferably, it is a root extract.
Togebanreishi (Annona muricata) is a plant belonging to the genus Vanleci. For the spider banci extract according to the present invention, extracts of the spider basilis, for example, leaves, stems, roots, fruits, pericarps, seeds, seed coats, above-ground parts, underground parts, or whole grasses can be used. Preferably, it is a fruit extract.
The giraffe (Daemonorops draco) is a plant belonging to the genus Hymenoptera. For the giraffe extract according to the present invention, it is possible to use extracts of various parts of the giraffe, for example, leaves, stems, roots, fruits, pericarps, seeds, seed coats, above-ground parts, underground parts or whole grasses. Preferably, it is a fruit extract.
Crocodile (Persea americana) is an extract of a plant belonging to the genus Allenaceae. For the crocodile extract according to the present invention, extracts of each part of the crocodile, for example, leaves, stems, roots, fruits, pericarps, seeds, seed coats, above-ground parts, underground parts or whole grasses can be used. Preferably, it is a fruit extract.
Safflower (Carthamus tinctorius) is an extract of a plant belonging to the genus Asteraceae. For the safflower extract according to the present invention, extracts of safflower parts, such as flowers, leaves, stems, roots, fruits, pericarps, seeds, seed coats, above-ground parts, underground parts, or whole grasses, can be used. Preferably, it is a flower extract.
Maize (Zea mays) is a plant belonging to the genus Cornaceae. For the corn extract according to the present invention, extracts of various parts of corn, for example, pistil (beard), stem, root, fruit, peel, seed, seed coat, above-ground part, underground part or whole plant can be used. . Preferably, it is a pistil extract.
The tuna (Tetragonia tetragonioides) is a plant belonging to the genus Tsuruna. For the tsuruna extract according to the present invention, extracts of tsuruna parts such as leaves, stems, roots, fruits, fruit skins, seeds, seed coats, above-ground parts, underground parts or whole plants can be used. Preferably, it is a whole plant extract.
Buckwheat (Fagopyrum esculentum) is a plant belonging to the genus Buckwheat. For the buckwheat extract according to the present invention, the extract of each part of buckwheat, for example, leaf, stem, root, fruit, fruit skin, seed, seed coat, above-ground part, underground part or whole plant can be used. Preferably, it is a seed extract.
Udo (Aralia cordata) is a plant belonging to the genus Arachnaceae. As the udo extract according to the present invention, extracts of various parts of udo, for example, leaves, flowers, stems, roots, fruits, fruit skins, seeds, seed coats, above-ground parts, underground parts, or whole grasses can be used. Preferably, it is a stem and leaf extract.
Kikarasuri (Trichosanthes kirilowii var. Japonica) is a plant that belongs to the genus Lilyaceae. As the extract of citrus cucumber according to the present invention, an extract of each part of citrus cucumber, for example, leaf, stem, root, fruit, pericarp, seed, seed coat, above-ground part, underground part or whole plant can be used. Preferably, it is a seed extract.

The manufacturing method of the plant extract concerning this invention is not specifically limited, It can manufacture by the method used normally. For example, it can be produced by cutting each part of a plant as it is or by cutting it into an appropriate size, drying it if necessary, and extracting it with juice or a solvent.
Although it does not specifically limit as an extraction solvent, For example, lower alcohols, such as water, methanol, ethanol, propanol, isopropanol, n-butanol, polyhydric alcohols, such as 1, 3- butylene glycol, propylene glycol, and glycerol, ethyl ether, Mention may be made of ethers such as propyl ether, esters such as ethyl acetate, and ketones such as acetone and ethyl methyl ketone. These solvents can be used alone or in combination. Among the extraction solvents, water, methanol, ethanol, and water-containing ethanol are particularly preferable.

The amount of the solvent used for the extraction varies depending on the extraction conditions and the like, but the weight ratio is suitably in the range of 1: 2 to 1:30 (plant: solvent), and in the range of 1: 3 to 1:20. The range of 1: 5 to 1:10 is more preferable. The extraction time is suitably in the range of 1 hour to 15 days. The extraction temperature is suitably in the range of 5 to 100 ° C, and preferably in the range of 60 to 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, further purification treatment may be added within a range that does not affect its activity. Such purification treatment may be performed by a usual method, for example, by filtering the extract by a conventional method. Moreover, the obtained filtrate can also be used for vacuum concentration, freeze-drying, or a spray dryer.

Muscle Activator The muscle activator according to the present invention (hereinafter referred to as “the muscle activator of the present invention”) has a myoblast activation action, a p70S6K activation action, and an rpS6 activation action, and increases muscle mass. It is useful for enhancing and preventing muscle atrophy, promoting recovery from muscle damage caused by exercise, preventing and treating sarcopenia and locomotive syndrome associated with lack of exercise and aging. Further, the muscle activating agent of the present invention activates myoblasts existing in the vicinity of the skin region, and further promotes the synthesis of muscle protein, thereby improving the skin elasticity, sagging and wrinkles around the skin. It is also expected to have an improving effect, a fatigue-reducing action, a stiff shoulder by reducing muscle fatigue, and an eye strain-reducing action. The muscle activator according to the present invention is expected to be particularly effective as a muscle strengthening agent, a muscle attenuation preventing agent, and a skin improving agent.
The muscle activating agent of the present invention is prepared by mixing the extract of the present invention obtained as described above with a raw material that can be used as it is or as a carrier, and then in various forms such as powder, lump, and liquid by conventional methods. It can manufacture by processing.

  The content of the extract of the present invention in the muscle activating agent of the present invention is not particularly limited as long as the myoblast activation action, the p70S6K activation action, and the rpS6 activation action can be confirmed. For example, 0.01% by weight or more Preferably, they are 0.05 weight% or more and 20 weight% or less, Most preferably, they are 0.1 weight% or more and 5 weight% or less.

The muscular activator of the present invention can optionally contain a pharmaceutically or food acceptable additive. 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, moisturizers, and pigments, and one or more of these can be used. The muscle activator of the present invention may further contain components known to have muscle strengthening action or skin improvement action. Examples include muscle strengthening components such as creatine, amino acids such as BCAA and leucine, HMB (3-hydroxyisovaleric acid), and skin improving effects such as collagen, hyaluronic acid, elastin, proteoglycan, ceramide, and vitamin C. It is done.
The muscle activator of the present invention can be suitably prepared into dosage forms such as tablets, powders, granules, capsules, solutions, 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 of the administration subject, body weight, etc., but usually within the range of 0.01 to 100 g as the weight of the extract of the present invention per adult day. It is suitable to be within the range of 0.03 g to 60 g, and preferably within the range of 0.1 g to 10 g, but is not necessarily limited to this range. Such daily intake may be taken once, or may be taken divided into 2 to 4 times.

  Hereinafter, the present invention will be described in more detail 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 shown in Table 1 was pulverized and then immersed in 75 g of methanol for 7 days, and after filtration, concentrated and dried by a rotary evaporator. Thereafter, the final concentration was adjusted to 50 mg / mL with dimethylsulfoxide (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 shown in Table 1 was pulverized and then immersed in 75 g of ethanol for 7 days, and after filtration, concentrated and dried by a rotary evaporator. Thereafter, the final concentration was adjusted to 50 mg / mL with dimethylsulfoxide (DMSO) to obtain an ethanol extract of each plant.
Production Example 3 Production of hot water extract of each plant 10 g of the dried product of each plant listed in Table 1 was pulverized, then added to 150 mL of hot water at 100 ° C., heated for 30 minutes, filtered and concentrated on a rotary evaporator. Drying was performed. Then, it adjusted to final concentration 50mg / mL with dimethylsulfoxide (DMSO), and the hot water extract of each plant was obtained.

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

Test Example 2 Measurement of phosphorylation of p70S6K in the plant extract according to the present invention The mouse myoblast cell line (C2C12 cell, manufactured by DS Pharma Biomedical Co., Ltd.) was used. C2C12 cells are dispersed in DMEM (Dulbecco's Modified Eagle Medium, Gibco) containing 10% FBS (Fetal Bovine Serum, Gibco) (hereinafter referred to as “10% FBS / DMEM medium”). Cell culture flasks (bottom area: 25 cm ² , manufactured by Nippon Biosciences Co., Ltd.) were seeded with 5.0 × 10 ⁴ cells, and CO ₂ incubator (manufactured by Sanyo Biomedica Co., Ltd .; CO ₂ concentration 5%, humidity 90%) And cultured at 37 ° C. for 3 days. It was confirmed by a microscope that it was 80% confluent and used for the experiment. C2C12 cells that became 80% confluent were detached from the flask using Trypsin-EDTA (manufactured by Gibco) and collected. These cells were suspended in 10% FBS / DMEM medium and seeded at 12 wellplates at 1.7 × 10 ⁵ cells / well. After 24 hours from seeding, 1 mL of 2% HS (Horse Serum, manufactured by Gibco) was substituted to differentiate into myotube cells. The cells were replaced with 2% HS once every two days from the start of differentiation, and myotubes on the third day of differentiation were used in the experiment. After replacing myotube cells on the third day of differentiation with serum-free medium, No. 9 described in Table 1 prepared in Production Example 1 was prepared as a test substance 9 hours later. The sample treatment was performed by replacing each of the methanol extracts 1 to 4 and 13 with 1 mL of serum-free medium containing 1, 10, and 50 ppm. After 2 hours from the start of treatment, the cells were washed with PBS (Phosphate buffered saline), and then the cells were collected with 20 μL of RIPA buffer (Fuji Film Wako Pure Chemical Industries, Ltd.). Using the obtained protein solution, it was added to SDS-PAGE sample buffer, subjected to reduction treatment at 100 ° C. for 3 minutes, 50 μg of protein was separated by 10% acrylamide gel in the subsequent Western blot, and then p70S6K phosphorous. Quantification of oxidation was performed. Anti-p70S6K and Anti-phospho-p70S6K (manufactured by Cell Signaling Technology) were used as antibodies. The results are shown in FIGS.
As is clear from FIGS. 9 to 13, the bergamot extract, psyllium endosperm extract, Niitake sagebrush extract, Wadder extract and red pepper extract according to the present invention promoted phosphorylation of p70S6K.

Test Example 3 rpS6 Phosphorylation Measurement of 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. Thus, phosphorylation of rpS6K can be quantified.

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

Claims (3)

One or more plant extracts selected from the group consisting of bergamot, psyllium, Japanese mugwort, sagebrush, spinach, giraffe, crocodile, safflower, corn, tsuruna, buckwheat, udo, and red-crowned cucumber as an active ingredient As a muscle activator. The muscle activator according to claim 1, wherein the muscle activator is a muscle enhancer or a muscle attenuation preventive agent. The muscle activator according to claim 1, wherein the muscle activator is a skin improving agent.

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