Composition for inhibiting muscle atrophy containing ergothioneine and composition for inhibiting Src tyrosine kinase

Abstract

The purpose of the present invention is to provide a composition for suppressing muscle atrophy. The present invention relates to a composition for suppressing muscle atrophy, characterized by containing ergothioneine or a salt thereof as an active ingredient.

Description

Technical Field

[0001] The present invention relates to a composition for suppressing muscle atrophy and a composition for inhibiting Src tyrosine kinase, etc., containing ergothioneine. Background Art

[0002] The global aging population is accelerating, and elderly individuals are facing a vicious cycle of muscle atrophy caused by prolonged bed rest and reduced physical activity, which in turn worsens with further physical activity. The only effective way to increase muscle mass is through continuous muscle training, but as metabolic function declines with aging, maintaining sufficient muscle mass through training alone is no longer feasible. Therefore, there is an urgent need for pharmaceuticals and dietary supplements that can prevent muscle atrophy.

[0003] Muscles maintain a balance between the synthesis and breakdown of muscle protein, each of which is related to different factors. Muscle protein synthesis requires raw materials such as amino acids and energy, necessitating a diet rich in muscle protein. However, the appetite of the elderly decreases compared to younger individuals. Therefore, inhibiting the breakdown of muscle protein is considered more important and effective in preventing muscle atrophy in the elderly.

[0004] The Src tyrosine kinase family is particularly important as intracellular signal transduction proteins associated with acute inflammatory responses. Its phosphorylation is one of the key factors determining the expression or production of inflammatory mediators. Src tyrosine kinase is expressed in muscle, and its inhibition can inhibit the breakdown of muscle cells.

[0005] Ergothioneine is an amino acid found in mushrooms and other organisms, and its L-form exists naturally. Patent Document 1 discloses a composition containing ergothioneine for promoting muscle differentiation. The examples describe that when C2C12 cells are cultured in a medium containing ergothioneine, differentiation from C2C12 cells into muscle cells is promoted, and the expression level of MyoD mRNA, which induces muscle differentiation, increases.

[0006] Patent Literature Patent Document 1: International Publication No. 2020 / 004014 Summary of the Invention

[0007] However, Patent Document 1 does not describe any effects on muscle atrophy or inhibition of Src tyrosine kinase.

[0008] The present invention aims to provide a composition for inhibiting muscle atrophy. Another object of the present invention is to provide a composition for inhibiting Src tyrosine kinase.

[0009] The present inventors conducted intensive studies to solve the above-mentioned problems and found that L-ergothioneine or a salt thereof has an effect of inhibiting muscle atrophy and can inhibit Src tyrosine kinase.

[0010] That is, although not limited to the following, the present invention relates to the following muscle atrophy suppressing composition and the like. [1] A composition for inhibiting muscle atrophy, characterized in that it contains ergothioneine or a salt thereof as an active ingredient. [2] The composition according to [1] above, wherein the muscle atrophy is caused by aging. [3] The composition according to [1] or [2] above, characterized in that it inhibits muscle atrophy via ATP-sensitive potassium channels. [4] A composition for inhibiting Src tyrosine kinase, characterized in that it contains ergothioneine or a salt thereof as an active ingredient. [5] The composition according to any one of [1] to [4] above, which is a composition for oral administration. [6] The composition according to any one of [1] to [5] above, which is a food or beverage. [7] The composition according to any one of [1] to [6] above, characterized in that it is marked with at least one function selected from the group consisting of "inhibiting muscle loss", "maintaining muscle", "improving muscle", "inhibiting muscle strength loss", "maintaining muscle strength", "improving muscle strength", "improving walking function", "maintaining walking function", "improving motor function", "maintaining motor function", "maintaining muscles that have aged due to aging", "maintaining muscle strength that has aged due to aging", and "maintaining physical strength". [8] A method for inhibiting muscle atrophy, characterized in that ergothioneine or a salt thereof is administered to a subject. [9] A method for inhibiting Src tyrosine kinase, characterized in that ergothioneine or a salt thereof is administered to a subject.

[10] An application characterized in that ergothioneine or a salt thereof is used to inhibit muscle atrophy.

[11] An application characterized in that ergothioneine or a salt thereof is used to inhibit Src tyrosine kinase.

[0011] According to the present invention, there is provided a composition for inhibiting muscle atrophy. Also according to the present invention, there is provided a composition for inhibiting Src tyrosine kinase. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 It is a graph showing the change in the lean body mass ratio (Δ lean body mass ratio (%)) in the test food group and the control food group. Figure 2 This is a graph showing the evaluation of the inhibitory activity of Src tyrosine kinase by L-ergothioneine. Figure 3 This is a graph showing the evaluation of the substrate binding inhibitory activity of L-ergothioneine-based ATP-sensitive potassium channels. DETAILED DESCRIPTION

[0013] The muscle atrophy-inhibiting composition of the present invention contains ergothioneine or a salt thereof as an active ingredient. Ergothioneine is a type of sulfur-containing amino acid. In the present invention, ergothioneine is preferably L-ergothioneine. As the salt of ergothioneine, as long as it is a salt allowed in pharmacology or a salt allowed in a diet, it is not particularly limited and can be any of an acidic salt and a basic salt. As an acidic salt, for example, inorganic acid salts such as hydrochloride, sulfate, nitrate, phosphate can be listed; organic acid salts such as acetate, citrate, maleate, malate, oxalate, lactate, succinate, fumarate, propionate, etc. can be listed. As a basic salt, for example, alkali metal salts such as sodium salt, potassium salt can be listed; alkaline earth metal salts such as calcium salt, magnesium salt, etc.

[0014] Ergothioneine or its salt is not limited by its form or production method. Ergothioneine or its salt can be chemically synthesized or extracted and purified from natural sources. L-ergothioneine is abundant in the golden top mushroom (Golden / Yellow Oyster mushroom) (scientific name: Pleurotus cornucopiae var. citrinopileatus), which belongs to the genus Pleurotus of the family Pleurotaceae. L-ergothioneine is also found in mushrooms such as Agaricus bisporus (scientific name: Agaricus bisporus), white button mushrooms, portabella mushrooms, and gray oyster mushrooms (scientific name: Pleurotus ostreatus); shiitake mushrooms (scientific name: Lentinula edodes); maitake mushrooms (scientific name: Grifola Frondosa); Ganoderma lucidum (scientific name: Ganoderma lucidum); Hericium erinaceus (scientific name: Hericium erinaceus); Agrocybe aegerita (scientific name: Agrocybe aegerita); Chanterelles (scientific name: Cantharellus cibarius); Boletus edulis (scientific name: Boletus edulis); and Morchella esculenta (scientific name: Morchella esculenta). When obtaining L-ergothioneine from natural sources, extraction from Pleurotus eryngii is preferred. Ergothioneine or its salts can also be produced by microbial fermentation. Ergothioneine or its salts can also be isolated.

[0015] L-ergothioneine or its salt is contained in natural products or food and drink, is the compound with edible experience.Therefore, from the viewpoint of safety, think that ergothioneine or its salt even if for example long-term intake also has few problems.According to the present invention, can provide a kind of safe high muscle atrophy inhibition composition.

[0016] So-called muscle atrophy is the reduction of muscle volume. Muscle atrophy is divided into the muscle atrophy (myogenic muscle atrophy) caused by the disease of muscle itself and the muscle atrophy (neurogenic muscle atrophy) caused by the obstacle of the motor nerve that directly conveys movement instruction to muscle. Even if muscle or nerve do not have such disease, muscle will also atrophy if not used, and this state is called disuse muscle atrophy. In addition, the muscle mass of whole body also can reduce (muscle atrophy caused by age increase) with age increase. In one embodiment, the composition of the present invention is preferably used for suppressing the muscle atrophy caused by age increase, and is more preferably used for preventing the muscle atrophy caused by age increase.

[0017] As shown in the Examples described below, when L-ergothioneine is taken, a decrease in the lean body mass ratio is suppressed compared to when no L-ergothioneine is taken. Lean body mass (LBM) refers to the weight obtained by subtracting the weight of fat from the body weight, and is an indicator for clarifying the proportion of muscle. By investigating the increase or decrease in lean body mass, the increase or decrease in muscle weight can be clarified. The so-called lean body mass ratio is the ratio of lean body mass to total body weight. A decrease in the lean body mass ratio indicates a decrease in muscle mass. Therefore, suppressing the decrease in the lean body mass ratio indicates suppressing the decrease in muscle mass. Therefore, ergothioneine can be used as an active ingredient for suppressing muscle atrophy.

[0018] Furthermore, as shown in the Examples described below, L-ergothioneine inhibits Src tyrosine kinase. Furthermore, L-ergothioneine inhibits the binding of substrates to ATP-sensitive potassium (KATP) channels. Src tyrosine kinase is known to be expressed in muscle, and inhibition of this kinase can suppress the breakdown of muscle cells. Therefore, ergothioneine inhibits the breakdown of muscle cells by inhibiting Src tyrosine kinase, thereby exhibiting a muscle atrophy inhibitory effect. KATP channels are known to be expressed in muscle and are involved in regulating muscle atrophy. Therefore, it is believed that ergothioneine has a muscle atrophy inhibitory effect through ATP-sensitive potassium channels.

[0019] The composition of the present invention is used to inhibit muscle atrophy. Inhibiting muscle atrophy includes preventing muscle atrophy, delaying its progression, and preventing the progression of muscle atrophy. Preventing muscle atrophy includes preventing symptoms, delaying their onset, reducing their incidence, and alleviating their risk. Inhibiting muscle atrophy can achieve effects such as inhibiting muscle mass loss and maintaining muscle mass.

[0020] The composition of the present invention may be either an oral composition or a parenteral composition, but is preferably an oral composition. As oral composition, specifically can enumerate food and drink, oral pharmaceutical product, quasi-medicine, feed etc., be preferably food and drink or oral pharmaceutical product, more preferably food and drink.Composition of the present invention also can be for being deployed in the material or preparation etc. used in food and drink, pharmaceutical product, quasi-medicine, feed etc.

[0021] The composition of the present invention can be used for either therapeutic (medical) or non-therapeutic (non-medical) applications. The term "non-therapeutic" refers to a concept that does not include medical procedures, i.e., human surgery, treatment, or diagnosis.

[0022] The composition of the present invention, as long as not to damage the effect of the present invention, except thioneine or its salt, can contain any additive, any composition.These additives and composition can be selected according to the form of composition etc. When the composition of the present invention is made into oral composition, the additive etc. that can be generally used for oral composition such as food and beverage, pharmaceuticals, quasi-drugs, feed can be used.

[0023] When the composition of the present invention is prepared as a food or beverage, a pharmaceutical product, a quasi-drug, a feed, or the like, the production method is not particularly limited and the composition can be produced by a general method. The form of the oral composition of the present invention is not particularly limited, and may be solid (powder, granule, tablet, etc.), liquid, paste, or the like.

[0024] When such as compositions of the present invention are made into food and drink, various food and drink can be made in thioneine or its salt, allocating spendable composition (such as, food material, the food additive used as required etc.) in food and drink.Food and drink are not particularly limited, and such as general food and drink, health food, health supplement food, health drink, functional label food, specific health food, patient food and drink etc. can be enumerated.Above-mentioned health food, health supplement food, functional label food, specific health food etc. such as can be used in the various preparation forms such as fine granule, tablet, granule, powder, capsule, chewable, dry syrup, syrup, liquid, beverage, oral liquid, liquid food.

[0025] When pharmaceuticals or quasi-medicines are made into compositions of the present invention, for example, the carrier allowed in pharmacology can be allotted in thioneine or its salt, the additive etc. added as needed, make pharmaceuticals or quasi-medicines of various dosage forms.Such carrier, additive etc., for the material allowed in the pharmacology that can be used for pharmaceuticals or quasi-medicines, for example, one or more than two kinds of excipients, binding agents, disintegrants, lubricants, antioxidants, coloring agents etc. can be enumerated. As the mode of administration (intake) of pharmaceuticals or quasi-medicines, the mode of administration orally or parenterally (percutaneous, through mucosa, enteral, injection etc.) can be enumerated. When pharmaceuticals or quasi-medicines are made into compositions of the present invention, it is preferably made into oral pharmaceuticals or oral quasi-medicines. As the dosage form for oral administration of pharmaceuticals or quasi-medicines, liquid, tablet, powder, fine granules, granules, sugar-coated tablets, capsules, suspensions, emulsions, chewing agents etc. can be enumerated. Examples of dosage forms for parenteral administration include injections, drops, ointments, lotions, patches, suppositories, nasal preparations, pulmonary preparations (inhalants), etc. The pharmaceutical product may be a pharmaceutical for non-human animals.

[0026] When the composition of the present invention is prepared into feed, ergothioneine or its salt is mixed into the feed. Feed also includes feed additives. Examples of feed include feed for livestock such as cattle, pigs, chickens, sheep, and horses; feed for small animals such as rabbits, rats, and mice; and pet food for dogs, cats, and birds.

[0027] The subject to which the composition of the present invention is ingested or administered (also referred to as the subject of administration) is not particularly limited, but is preferably a human or a non-human mammal, and more preferably a human. In one embodiment, subjects who need or want to suppress muscle atrophy can be listed as subjects. Examples of such subjects include middle-aged and elderly people. In one embodiment, the composition of the present invention is preferably used as a composition for suppressing muscle atrophy in middle-aged and elderly people. Middle-aged and elderly people include elderly people. Middle-aged and elderly people may be, for example, humans over 40 years old. Elderly people may be, for example, humans over 60 years old or over 65 years old. The subject of administration may be a healthy person. The composition of the present invention can be used on healthy people for the purpose of suppressing muscle atrophy, for example. In the present invention, particularly preferred subjects are healthy humans aged 50 to 65 years old, and the most preferred subjects are healthy humans aged 50 to 55 years old.

[0028] The content of ergothioneine or a salt thereof contained in the composition of the present invention is not particularly limited and can be set according to its form and the like. The content of ergothioneine or a salt thereof contained in the composition of the present invention is preferably 0.0001% by weight or more, more preferably 0.001% by weight or more, and is preferably 90% by weight or less, more preferably 50% by weight or less, for example, calculated as ergothioneine. In one embodiment, the content of thioneine or its salt contained in the composition of the present invention is preferably 0.0001 to 90% by weight, more preferably 0.001 to 50% by weight, calculated as thioneine. The content of thioneine or its salt can be measured by high performance liquid chromatography (HPLC).

[0029] In this specification, the amount converted to ergothioneine or an expression similar thereto refers to the amount of ergothioneine, and refers to the value obtained by multiplying the number of moles of the salt by the molecular weight of ergothioneine in the case of a salt of ergothioneine.

[0030] The composition of the present invention can be taken or administered in a suitable manner adapted to its form. In one embodiment, the composition of the present invention is preferably taken orally (orally administered). The dosage (also referred to as intake) of the composition of the present invention is not particularly limited, and may be appropriately set according to the dosage, administration method, and weight of the subject as long as the dosage is sufficient to achieve an inhibitory effect on muscle atrophy.

[0031] In one way, when the mankind (adult) is taken orally or the composition of the present invention is administered, its dosage is in the dosage (ergothioneine conversion) of thioneine or its salt, and is preferably more than 2mg per 1 day, more preferably more than 5mg, more preferably more than 7mg, in addition, is preferably less than 50mg, more preferably less than 25mg, more preferably less than 20mg. In one way, when the dosage of the composition of the present invention is in the dosage (ergothioneine conversion) of thioneine or its salt, for the mankind (adult), is preferably 2~50mg per 1 day, more preferably 5~25mg, more preferably 5~20mg, more preferably 7~20mg. Preferably, the above amount is divided into more than 1 time per day, for example, 1 time per day or multiple (for example 2~3 times) are taken or administered. In one way, preferably, the mankind is taken orally or the thioneine or its salt of the above amount is administered thereto. The above-mentioned dosage can be the dosage of every body weight 60kg. In one embodiment, the composition of the present invention can be used to allow humans, for example, to ingest or administer the above-mentioned amount of ergothioneine or a salt thereof per 60 kg body weight per day.

[0032] The content of ergothioneine or a salt thereof in the composition of the present invention is preferably in an adult's daily intake, calculated as ergothioneine, of 2 to 50 mg, more preferably 5 to 25 mg, further preferably 5 to 20 mg, and particularly preferably 7 to 20 mg.

[0033] The composition of the present invention may be provided with, for example, an indication of its function of inhibiting muscle atrophy or an indication of its function exerted by inhibiting muscle atrophy. The composition of the present invention may also be accompanied by an indication of at least one function selected from, for example, "inhibiting muscle loss", "maintaining muscle", "improving muscle", "inhibiting muscle strength decline", "maintaining muscle strength", "improving muscle strength", "improving walking function", "maintaining walking function", "improving motor function", "maintaining motor function", "maintaining muscles that age", "maintaining muscle strength that age" and "maintaining physical strength". In one embodiment, the composition of the present invention is preferably a food or beverage with the above-mentioned label. In addition, the above-mentioned label may be a label indicating the purpose of obtaining the above-mentioned function. The above-mentioned label may be attached to the composition itself or to the container or packaging of the composition.

[0034] As mentioned above, ergothioneine or its salt has Src tyrosine kinase inhibitory effect. Ergothioneine or its salt can be used as the active ingredient for inhibiting Src tyrosine kinase. The composition of the present invention can be used as a Src tyrosine kinase inhibitory composition. The Src tyrosine kinase inhibitory composition containing ergothioneine or its salt as an active ingredient is also one of the present invention. The active ingredient, content and dosage of the active ingredient, subject of administration, and other aspects of the composition for inhibiting Src tyrosine kinase are the same as those of the composition for inhibiting muscle atrophy.

[0035] As shown above, thioneine or its salt hinders the combination of substrate and KATP channel.Therefore, thioneine or its salt can be used as the active ingredient for hindering substrate from being combined with KATP channel and use.Containing thioneine or its salt as active ingredient for the substrate combination for KATP channel hindering composition is also one of the present invention.

[0036] The present invention also includes the following method. A method for inhibiting muscle atrophy, characterized in that thioneine or a salt thereof is administered to a subject. A method for inhibiting Src tyrosine kinase, characterized in that ergothioneine or a salt thereof is administered to a subject. The above methods may be therapeutic methods or non-therapeutic methods.

[0037] The present invention also includes the following applications. An application is characterized in that ergothioneine or a salt thereof is used to inhibit muscle atrophy. An application is characterized in that ergothioneine or a salt thereof is used to inhibit Src tyrosine kinase. The above applications may be therapeutic applications or non-therapeutic applications. By administering ergothioneine or a salt thereof to a subject, muscle atrophy can be suppressed.

[0038] In the above-mentioned method and application, the preferred mode of thioneine or its salt is identical with the above-mentioned composition of the present invention. As thioneine or its salt, 1 kind of thioneine or its salt can be used, and 2 or more kinds can also be used. In the above-mentioned method and application, preferably more than 1 time a day, for example, 1 time a day~multiple times (for example 2~3 times) are administered to the object (making it take in). In the above-mentioned method and application, preferably oral administration (take in) thioneine or its salt. The above-mentioned application is preferably the application in human or non-human mammals, more preferably the mankind.

[0039] In said method and application, use the thioneine or its salt that can obtain the amount (also can be called effective dose) of desired effect.The preferred dosage of thioneine or its salt or the object of administration etc. are identical with above-mentioned composition of the present invention.Thioneine or its salt can directly be administered, and also can be administered as the composition containing it.For example, above-mentioned composition of the present invention can also be used.

[0040] The present invention also includes use of ergothioneine or a salt thereof for producing a composition for inhibiting muscle atrophy. The present invention also includes use of ergothioneine or a salt thereof for producing a composition for inhibiting Src tyrosine kinase. The present invention also provides ergothioneine or a salt thereof for inhibiting muscle atrophy.

[0041] In this specification, numerical ranges expressed with lower and upper limits, i.e., "lower limit to upper limit," include both lower and upper limits. For example, a range expressed with "1 to 2" means from 1 to 2, inclusive. In this specification, upper and lower limits may be used as ranges based on any combination. Example

[0042] Hereinafter, the present invention will be described in further detail with reference to examples, but the scope of the present invention is not limited thereto.

[0043] <Example 1> (Evaluation of muscle atrophy in humans) To evaluate the effect of a supplement formulated with ergothioneine on human lean body mass rate, a placebo-controlled, randomized, double-blind, parallel intergroup comparative trial was conducted in which healthy men and women aged 50 to 55 years old (12 in the tested food group and 11 in the control food group, a total of 23) were selected to take a capsule formulated with 8 mg of ergothioneine (tested food) or a capsule without ergothioneine (control food) once a day for 12 weeks.

[0044] Lean body mass percentage was assessed using a DC-430A split-type body composition analyzer (manufactured by TBLITA Co., Ltd.). Before the start of the experiment (before ingestion of the test or control food), the subjects' lean body mass percentage was assessed using the body composition analyzer. Furthermore, after 12 weeks of ingestion of the test or control food, the subjects' lean body mass percentage was measured again (12-week assessment).

[0045] (Food for evaluation) Two types of food were used for evaluation, and they were indistinguishable in appearance, flavor, etc. Food tested: Capsules containing the substance tested (8 mg of L-ergothioneine) Control food: capsules that do not contain the substance being tested The raw materials of each evaluation food included dextrin, calcium stearate, and hydroxypropyl methylcellulose in addition to the test substance. A control food was made using the same raw materials as the test food, except that the test substance (L-ergothioneine) was not added.

[0046] The average lean body mass percentage (%) of the subjects in the test food group was used as the lean body mass percentage of the test food group. The average lean body mass percentage (%) of the subjects in the control food group was used as the lean body mass percentage of the control food group. Furthermore, for each group, the change in lean body mass percentage (Δ lean body mass percentage (%)) was calculated by subtracting the lean body mass percentage at the pre-test from the lean body mass percentage at the 12th week test. The change in lean body mass percentage (Δ lean body mass percentage (%)) at the pre-test was set to zero.

[0047] Figure 1 It is a graph showing the change in fat-free body mass ratio (Δ fat-free body mass ratio (%)) in the tested food group and the control food group. As a result, compared to the control food group, the reduction in fat-free body mass ratio was suppressed in the tested food group. It is clear from this result that L-ergothioneine inhibits muscle atrophy. Figure 1 In the table, black squares (■) represent the tested food group, and white squares (□) represent the control food group. "0W" on the horizontal axis represents the time of inspection before the start of the experiment, and "12W" represents the time of inspection at the 12th week.

[0048] <Example 2> (Evaluation of Src tyrosine kinase inhibitory activity) To evaluate the molecular target of L-ergothioneine in maintaining lean mass, an in vitro assay targeting Src tyrosine kinase was performed. The in vitro assay was commissioned by Eurofins Panlabs and performed as follows.

[0049] Human transgenic Src tyrosine kinase expressed in insect cells was used. 0.023 μg / mL of Src tyrosine kinase was dissolved in HEPES buffer (pH 7.4) and pre-incubated with L-ergothioneine (L-ergothioneine concentrations: 10 μM, 1000 μM) at 37°C for 15 minutes. 0.2 mg / mL Ploy (Glu:Tyr), 10 μM ATP, and 0.25 μM Ci[γ 32 The reaction was started by adding 3% H₃PO₄ after incubation for 30 minutes. The amount of [₃₂P]Ploy(Glu:Tyr) formed in the reaction solution was used as an indicator to evaluate the inhibitory activity of L-ergothioneine. The less amount of [₃₂P]Ploy(Glu:Tyr) formed, the more the Src tyrosine kinase activity was inhibited.

[0050] Figure 2 This is a graph showing the evaluation of the Src tyrosine kinase inhibitory activity by L-ergothioneine. The concentration (μM) on the horizontal axis represents the concentration of L-ergothioneine. The ratio (%) of the inhibitory activity of Src tyrosine kinase was determined by the following steps. 32 The amount of P]Ploy (Glu:Tyr) was taken as 100% of Src tyrosine kinase activity and the amount of [ 32 The relative activity (%) of Src tyrosine kinase relative to the control was determined by measuring the amount (%) of P]Ploy(Glu:Tyr). The inhibitory activity (%) of Src tyrosine kinase was determined by subtracting the relative activity (%) of Src tyrosine kinase when L-ergothioneine was added from the Src tyrosine kinase activity (100%) of the control. L-ergothioneine shows hindering activity for Src tyrosine kinase.Src tyrosine kinase is expressed in muscle, and it is known that can suppress the decomposition of muscle cell by hindering its activity.Therefore, it is inferred that L-ergothioneine suppresses the decomposition of muscle cell by hindering Src tyrosine kinase, shows above-mentioned muscle atrophy inhibitory effect.

[0051] <Example 3> (Evaluation of substrate binding inhibition activity of ATP-sensitive potassium channels) To evaluate the molecular targets of L-ergothioneine in maintaining fat-free mass, an in vitro assay targeting ATP-sensitive potassium channels was conducted. The in vitro assay was commissioned by Eurofins Panlabs and performed as follows.

[0052] 400 μg of rat cerebral cortical membrane fraction (containing KATP channels, manufactured by Eurofins Cerep) was dissolved in 50 mM Tris-HCl buffer (pH 7.4) and 0.1 nM [ 3 [3H]Glyburide and L-ergothioneine (L-ergothioneine concentration: 100 μM or 1000 μM) were incubated together at 22°C for 60 minutes. Glyburide is a KATP channel inhibitor. Nonspecific binding was determined in the presence of 1 μM gliburide (without L-ergothioneine). When gliburide was present in excess, [3H]gliburide could not bind to the KATP channel, and the radioactivity detected at this time was nonspecific binding. The membrane fraction was washed and filtered through a filter membrane, and the substrate binding inhibition activity of L-ergothioneine was evaluated using radioactivity as an indicator. [ 3 The more [H] glibenclamide is present, the stronger the radioactivity of the membrane fraction.

[0053] Figure 3 A graph for evaluating the substrate binding inhibition activity of the ATP-sensitive potassium channel based on L-ergothioneine. The concentration (μM) on the horizontal axis is the concentration of L-ergothioneine. The substrate binding inhibition activity (%) of the KATP channel is obtained according to the following steps. The radioactivity of the membrane fraction in the reaction solution (control) that does not contain L-ergothioneine is set to 100%. Based on the radioactivity of the membrane fraction in the reaction solution with L-ergothioneine, the relative activity (%) relative to the control is obtained when L-ergothioneine (100 μM or 1000 μM) is added. When L-ergothioneine hinders the binding of glibenclamide to the KATP channel, the radioactivity of the membrane fraction decreases. Therefore, by deducting the relative activity (%) when L-ergothioneine is added from the radioactivity (100%) of the membrane fraction of the control, the substrate binding inhibition activity (%) for the KATP channel is obtained.

[0054] L-ergothioneine shows the combination of glibenclamide as substrate to ATP-sensitive potassium channel and hinders activity. Known ATP-sensitive potassium channel is also expressed in muscle and is relevant to the regulation of muscle atrophy process. Therefore, it is speculated that L-ergothioneine shows above-mentioned muscle atrophy inhibitory effect by ATP-sensitive potassium channel.

Claims

1. A composition for inhibiting muscle atrophy, characterized in that Contains ergothioneine or its salt as an active ingredient.

2. The composition according to claim 1, characterized in that Muscle atrophy is the loss of muscle mass due to aging.

3. The composition according to claim 1 or 2, characterized in that Inhibits muscle atrophy through ATP-sensitive potassium channels.

4. A composition for inhibiting Src tyrosine kinase, characterized in that: Contains ergothioneine or its salt as an active ingredient.

5. The composition according to claim 1 or 4, characterized in that It is a composition for oral use.

6. The composition according to claim 5, characterized in that For food and drink.

7. The composition according to claim 1 or 4, characterized in that The product is labeled with at least one function selected from the group consisting of "suppressing muscle loss", "maintaining muscle", "improving muscle", "suppressing muscle strength loss", "maintaining muscle strength", "improving muscle strength", "improving walking function", "maintaining walking function", "improving motor function", "maintaining motor function", "maintaining muscles that deteriorate with age", "maintaining muscle strength that deteriorates with age", and "maintaining physical strength".

8. A method for inhibiting muscle atrophy, characterized in that: Ergothioneine or a salt thereof is administered to the subject.

9. A method for inhibiting Src tyrosine kinase, characterized in that: Ergothioneine or a salt thereof is administered to the subject.

10. An application, characterized in that: Ergothioneine or its salt is used for inhibiting muscle atrophy.

11. An application, characterized in that: Ergothioneine or its salt is used to inhibit Src tyrosine kinase.

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