JP2024032369A - Composition for increasing muscle mass or muscle strength - Google Patents

Abstract

An object of the present invention is to provide a composition for increasing muscle mass or muscle strength. A composition for increasing muscle mass or muscle strength, which contains dihydroferulic acid and/or its metabolite as an active ingredient. [Selection diagram] None

Description

The present invention relates to a composition for increasing muscle mass or muscle strength.

In countries like Japan, where the proportion of elderly people in the population is high, extending healthy life expectancy and improving quality of daily life (QOL) are challenges. One of the factors contributing to the shortening of healthy life expectancy is a decrease in muscle mass and muscle strength associated with aging. Muscle mass also decreases due to lack of exercise, or when long periods of rest are required, such as after surgery or while recovering from an illness. Exercise (strength training, etc.) is an effective means of increasing muscle mass or strength, but it may be difficult to perform continuously due to time and physical reasons, or it may be physically taxing. .

In order to increase muscle mass or muscle strength, it is effective to promote protein synthesis. It has been reported that the mTOR (mechanistic target of rapamycin) pathway is important in protein synthesis. T (threonine) 389 residue of S6K (ribosomal protein S6 kinase), which is a downstream factor of mTOR signal, and 4E-BP1 (Eukaryotic translation initiation factor 4E-binding protein) T (threonine) 37/46 residues of ein 1) are phosphorylated protein synthesis is promoted.

β-hydroxy-β-methylbutyric acid (HMB), a metabolite of leucine, has been reported as a substance that promotes phosphorylation in the mTOR pathway (Non-Patent Document 1). It has also been reported that when resistance training is performed while ingesting HMB, muscle mass increases compared to when the training is performed and HMB is not ingested (Non-Patent Document 2).

Kimura et al. , NUTRITION RESEARCH 34 (2014) 368-374 Wilson et al. , European Journal of Applied Physiology (2014) 114:1217-1227

An object of the present invention is to provide a composition for increasing muscle mass or muscle strength.

In view of the above problems, the present inventors have repeatedly investigated components that activate the mTOR pathway, and have discovered dihydroferulic acid and its metabolites. Dihydroferulic acid and its metabolites have the effect of promoting phosphorylation of ribosomal protein S6 kinase (S6K), and are useful for increasing muscle mass.

That is, although not limited thereto, the present invention relates to the following compositions for increasing muscle mass or muscle strength.
[1] A composition for increasing muscle mass or muscle strength, containing dihydroferulic acid and/or its metabolite as an active ingredient.
[2] The composition according to [1] above, wherein the metabolite is a glucuronide conjugate of dihydroferulic acid.
[3] The composition according to [1] or [2] above, which is used to promote protein synthesis in muscle.
[4] The composition according to [3] above, which is used to promote protein synthesis by activating the mTOR pathway.
[5] The composition according to any one of [1] to [4] above, which is an oral composition.
[6] The composition according to any one of [1] to [5] above, which is a food or drink.
[7] 1 or 2 selected from the group consisting of "muscle increase", "muscle improvement", "muscle strength increase", "muscle strength improvement", "promotion of skeletal muscle hypertrophy", "supporting the ability to build muscle" The composition according to any one of [1] to [6] above, which has the above indications.
[8] The composition according to any one of [1] to [7] above, which is used in combination with exercise.

According to the present invention, a composition for increasing muscle mass or muscle strength is provided. By ingesting the composition of the present invention, protein synthesis can be promoted and the effect of increasing muscle mass or muscle strength can be obtained.

FIG. 1 is a graph showing the amount of phosphorylation of the T389 residue of S6K per protein concentration in cells to which dihydroferulic acid or a glucuronide conjugate of dihydroferulic acid was added. The vertical axis of the graph in FIG. 1 represents the relative value of the amount of phosphorylation when the amount of phosphorylation of the T389 residue of S6K in the negative control (C) is set as 100.

The composition for increasing muscle mass or muscle strength of the present invention contains dihydroferulic acid and/or its metabolite as an active ingredient.
The composition for increasing muscle mass or muscle strength of the present invention is hereinafter also referred to as the composition of the present invention. In the present invention, the muscle is preferably skeletal muscle.

In the present invention, the origin and production method of dihydroferulic acid (3-(4-hydroxy-3-methoxyphenyl)propionic acid, hereinafter also referred to as HMPA) are not particularly limited, and it may be a naturally derived product or a chemically synthesized product. , fermentation method, or enzyme method. Dihydroferulic acid is produced by a fermentation method from, for example, ferulic acid or a ferulic acid derivative such as ethyl ferulate, or a plant-derived raw material containing these (for example, a crushed product or extract of a plant containing ferulic acid). can do. In production by a fermentation method, ferulic acid, a ferulic acid derivative, or a plant-derived raw material containing these is fermented by a microorganism having phenolic acid reductase activity to convert ferulic acid to dihydroferulic acid. Dihydroferulic acid can be extracted or purified from the obtained fermented product. The method for extracting or purifying dihydroferulic acid from the fermented product is not particularly limited. A known method can be adopted. Examples of plants containing ferulic acid include coffee, rice, wheat, barley, rye, and corn. Examples of microorganisms having phenolic acid reductase include lactic acid bacteria such as Lactobacillus plantarum, Lactobacillus fermentum, and Lactobacillus gasseri. Dihydroferulic acid is commercially available, and commercially available products can also be used.

In the present invention, examples of metabolites of dihydroferulic acid include glucuronic acid conjugates and sulfuric acid conjugates of dihydroferulic acid. Among these, it is preferable to use a glucuronide conjugate of dihydroferulic acid because it provides a high effect.
Glucuronide conjugates of dihydroferulic acid are commercially available, and commercially available products can also be used.

In order to increase muscle mass or muscle strength, it is effective to promote protein synthesis.
Protein synthesis is controlled by the mTOR pathway, and protein synthesis is promoted by activating the mTOR pathway (eg, phosphorylating a signal factor of the mTOR pathway). For example, promoting phosphorylation of S6K, a downstream factor of mTOR, promotes protein synthesis. Therefore, promotion of S6K phosphorylation promotes protein synthesis in muscle, leading to an increase in muscle mass. By increasing muscle mass, the effect of increasing muscle strength can be obtained.

As shown in the Examples below, dihydroferulic acid and its metabolites promoted phosphorylation of the T389 residue of S6K in C2C12 cells, which are muscle cells. Therefore, dihydroferulic acid and its metabolites have the effect of activating the mTOR pathway and promote protein synthesis in muscle. Dihydroferulic acid and its metabolites have a muscle mass increasing effect or a muscle strength increasing effect and can be used as active ingredients for increasing muscle mass or muscle strength. Dihydroferulic acid and its metabolites can also be used to promote S6K phosphorylation and protein synthesis.

Since the composition of the present invention contains dihydroferulic acid and/or its metabolite, it can promote protein synthesis in muscle. The composition of the present invention can promote protein synthesis by activating the mTOR pathway, and can be used for such purposes. In one aspect, the compositions of the invention can be used to increase muscle mass or increase muscle strength by promoting protein synthesis in muscle.

The composition of the present invention exhibits an effect of increasing muscle mass or muscle strength. In addition, such effects can increase the effect of increasing muscle mass and muscle strength due to exercise, making it possible to prevent loss of muscle mass, prevention of muscle weakness, and the like. Therefore, the composition of the present invention can be used, for example, to prevent muscle mass loss, muscle weakness, and the like. In the present invention, increasing muscle mass is different from suppressing muscle atrophy (reducing or alleviating muscle mass loss due to muscle atrophy, etc.).
Muscle mass in animals such as humans can be measured, for example, by micro PET/CT (positron emission tomography/computed tomography, INVEON, Siemens, USA).

The composition of the present invention can be used to prevent or improve conditions or diseases that can be expected to be prevented or improved by increasing muscle mass or muscle strength. Such conditions or diseases include, for example, locomotive syndrome, sarcopenia, frailty, and cachexia (caused by debilitating diseases such as cancer or chronic diseases, resulting in severe atrophy of skeletal muscles and organs due to loss of appetite and impaired metabolic control mechanisms). (conditions indicating pathological conditions of functional dysfunction), etc. Frailty refers to physical frailty, including loss of muscle mass and/or muscle weakness. In one aspect, compositions of the invention may be used to prevent or ameliorate such conditions or diseases. In one embodiment, compositions of the invention may be used in subjects in need of prevention or amelioration of such conditions or diseases.
Prevention of a condition or disease as used herein refers to preventing the onset of the condition or disease, delaying the onset of the condition or disease, reducing the incidence of the condition or disease, reducing the risk of developing the condition or disease. It includes things such as alleviation. Ameliorating a condition or disease includes recovering a subject from the condition or disease, alleviating symptoms of the condition or disease, slowing or preventing progression of the condition or disease, and the like. Restoring includes at least partially restoring.

The compositions of the invention can be applied for either therapeutic (medical) or non-therapeutic (non-medical) uses. Non-therapeutic is a concept that does not include medical procedures, ie, surgery, treatment, or diagnosis of humans.
The composition of the present invention can be provided in the form of, for example, a food or drink, a drug, a quasi-drug, a feed, etc., but is not limited thereto. The composition of the present invention may itself be a food or drink, a drug, a quasi-drug, a feed, etc., or may be a preparation or material such as an additive used therein. The composition of the present invention can be provided in the form of a drug, for example, but is not limited to this form. The agent can be provided as a composition as it is or as a composition containing the agent. The composition of the present invention can also be referred to as an agent for increasing muscle mass or muscle strength.
In one embodiment, the composition of the invention is preferably an oral composition. According to the present invention, it is possible to provide an oral composition that has an excellent effect of increasing muscle mass or muscle strength. Oral compositions include foods and drinks, oral pharmaceuticals, and oral quasi-drugs, preferably food and drinks.

The composition of the present invention may contain any additives or components other than dihydroferulic acid and/or its metabolites, as long as they do not impair the effects of the present invention. These additives and components can be selected depending on the form of the composition, etc., and those that can generally be used in foods and drinks, pharmaceuticals, quasi-drugs, feeds, etc. can be used.

When the composition of the present invention is used as a food or drink, dihydroferulic acid and/or its metabolites are blended with ingredients that can be used in the food or drink (e.g., food materials, food additives used as necessary, etc.). It can be made into various food and drink products. Foods and drinks are not particularly limited, and include, for example, general foods and drinks, health foods, health drinks, foods with functional claims, foods for specified health uses, health supplements, foods and drinks for patients, and the like. The above health foods, foods with functional claims, foods for specified health uses, health supplements, etc. are, for example, fine granules, tablets, granules, powders, capsules, chewables, dry syrups, syrups, liquids, drinks, and liquids. It can be used in various formulations such as food.

When the composition of the present invention is used as a pharmaceutical or quasi-drug, dihydroferulic acid and/or its metabolites are mixed with pharmacologically acceptable excipients, etc., to produce pharmaceuticals in various dosage forms. be able to. From the viewpoint of obtaining the effects of the present invention more fully, the administration form of the drug or quasi-drug is preferably an oral administration form. The dosage form may be any dosage form suitable for administration. Dosage forms for oral administration include, for example, oral solid preparations such as tablets, coated tablets, fine granules, granules, powders, pills, capsules, dry syrups, and chewable preparations; oral liquid preparations, syrups, etc. liquid formulations for use in Dosage forms for parenteral administration include injections, drops, ointments, lotions, patches, suppositories, nasal preparations, and pulmonary preparations (inhalants). The drug may be a non-human veterinary drug.

When the composition of the present invention is used as feed, ingredients that can be used in feed can be blended with dihydroferulic acid and/or its metabolites. Examples of feed include livestock feed for cows, pigs, chickens, sheep, horses, etc.; small animal feed for rabbits, guinea pigs, rats, mice, etc.; pet food for dogs, cats, small birds, etc. .

When the composition of the present invention is used as food/drinks, pharmaceuticals, quasi-drugs, feeds, etc., the manufacturing method thereof is not particularly limited. can be manufactured.

The content of dihydroferulic acid and its metabolites in the composition of the present invention can be appropriately set depending on the form of the composition. For example, the content of dihydroferulic acid and/or its metabolites may be 0.0025 to 50% by weight in the composition. In one embodiment, when the composition of the present invention is used as an oral composition such as a food or drink, a drug, a quasi-drug, etc., the content of dihydroferulic acid and/or its metabolite is 0. The content may be 0.025% by weight or more, and may be 50% by weight or less, preferably 10% by weight or less, and more preferably 1% by weight or less. In one embodiment, the content of dihydroferulic acid and/or its metabolites in the composition of the present invention is preferably 0.0025 to 10% by weight, more preferably 0.0025 to 1% by weight. The content of dihydroferulic acid and/or its metabolites can be measured according to a known method, for example, an HPLC method or the like can be used.

The composition of the present invention can be ingested or administered in an appropriate manner depending on its form. The composition of the invention is preferably administered orally orally.
The intake amount (also referred to as dosage) of the composition of the present invention is not particularly limited, and may be any amount (effective amount) that can obtain the effect of increasing muscle mass or muscle strength, and the dosage form and administration method It may be set as appropriate depending on the situation. For example, when administering or ingesting dihydroferulic acid and/or its metabolites to humans (adults), the intake amount of dihydroferulic acid and/or its metabolites is preferably 1 mg or more in terms of dihydroferulic acid per 60 kg of body weight per day. More preferably 7.5 mg or more, even more preferably 10 mg or more, even more preferably 20 mg or more, particularly preferably 22.5 mg or more, and preferably 1000 mg or less, more preferably 375 mg or less, even more preferably 200 mg or less, and 80 mg or less is even more preferred, and 75 mg or less is particularly preferred. In one embodiment, when administered orally to a human (adult) subject, the intake amount of dihydroferulic acid and/or its metabolite is 1 to 1000 mg per 60 kg of body weight per day in terms of dihydroferulic acid. is preferred, 7.5 to 375 mg is more preferred, 10 to 200 mg is even more preferred, 20 to 80 mg is even more preferred, and 22.5 to 75 mg is particularly preferred. It is preferable to orally administer or ingest the above amount, for example, once a day or in 2 to 3 divided doses. When ingesting the composition of the present invention for the purpose of increasing muscle mass or muscle strength in humans (adults), the intake amount of dihydroferulic acid and/or its metabolites should be within the above range. Preferably, the composition of the present invention is orally ingested or administered to a subject.

In one embodiment, the composition of the present invention contains dihydroferulic acid and/or its metabolite in an amount that achieves the desired effect of the present invention, taking into account its dosage form, administration method, etc. It is preferable to contain. For example, in one embodiment, when the composition of the present invention is an oral composition such as a food or drink or an oral pharmaceutical, dihydroferulic acid is added to the intake amount per 60 kg body weight per adult per day of the composition. and/or its metabolite content is preferably 1 to 1000 mg, more preferably 7.5 to 375 mg, even more preferably 10 to 200 mg, even more preferably 20 to 80 mg, and even more preferably 22.5 to 1000 mg in terms of dihydroferulic acid. 75 mg is particularly preferred.

Continuous ingestion (administration) of dihydroferulic acid and/or its metabolites is expected to enhance the effect of increasing muscle mass or muscle strength. In a preferred embodiment, the compositions of the invention are to be taken continuously. In one embodiment of the present invention, the composition of the present invention is preferably continuously ingested for 4 weeks or more, more preferably 8 weeks or more, and still more preferably 12 weeks or more.

The subject to whom the composition of the present invention is administered or ingested (hereinafter also simply referred to as the subject) is preferably a mammal (human or non-human mammal), and more preferably a human. Furthermore, the subject to be administered in the present invention is preferably a subject who requires or desires an increase in muscle mass or muscle strength. For example, suitable subjects include subjects who wish to prevent or improve the above-mentioned condition or disease in which muscle mass has decreased or muscle strength has decreased. In one embodiment, the subject to whom the composition of the present invention is administered is preferably a middle-aged or elderly person, and more preferably an elderly person. In the present invention, a middle-aged person may be, for example, a person over 40 years old. An elderly person may be, for example, a person over 60 years old or over 65 years old. In one embodiment, the composition of the present invention can be used on a healthy subject for the purpose of preventing or ameliorating a condition or disease that can be expected to be prevented or ameliorated by increasing muscle mass or muscle strength. In one embodiment, the composition of the present invention can be used on subjects who exercise on a daily basis for the purpose of enhancing the effects of exercise. In one embodiment, the composition of the invention is preferably used in conjunction with exercise. For example, compositions of the invention can be administered to a subject before, during, or after exercise. By ingesting the composition of the present invention in combination with exercise, a greater effect of increasing muscle mass or muscle strength can be obtained.

The composition of the present invention may be labeled with one or more of the intended use, type of active ingredient, the above-mentioned effects, method of use (e.g., method of ingestion, method of administration), etc. on the package, container, or instruction manual. . The composition of the present invention may be labeled to the effect that it has an effect of increasing muscle mass or muscle strength, or an effect based on these effects. The composition of the present invention may include, for example, ``increase muscle'', ``improve muscle'', ``increase muscle strength'', ``improve muscle strength'', ``promote skeletal muscle hypertrophy'', ``support the ability to build muscle'', etc. Alternatively, two or more indications may be attached.
In one aspect of the present invention, the composition of the present invention is preferably a food or drink labeled with the above label. Moreover, the above-mentioned display may be a display to the effect that it is used to obtain the above-mentioned function. The label may be attached to the composition itself or to the container or packaging of the composition.

As mentioned above, dihydroferulic acid and its metabolites have an effect of increasing muscle mass or muscle strength, so they can be used in a method for increasing muscle mass or strength, which involves administering dihydroferulic acid and/or its metabolites. Can be done. A method for increasing muscle mass or muscle strength, which involves administering dihydroferulic acid and/or its metabolite, is also encompassed by the present invention.
The above methods may be therapeutic or non-therapeutic.
Dihydroferulic acid and its metabolites can also be used to increase muscle mass or strength, and may be used therapeutically or non-therapeutically.

By administering dihydroferulic acid and/or its metabolites to a subject, the mTOR pathway can be activated to promote protein synthesis in muscle. In one embodiment, dihydroferulic acid and/or its metabolites can be used to promote protein synthesis in muscle to increase muscle mass or muscle strength. In one embodiment, dihydroferulic acid and/or its metabolites can be used to prevent or ameliorate conditions or diseases such as locomotive syndrome, sarcopenia, frailty, cachexia, etc. by increasing muscle mass or strength.

In the above methods and uses, dihydroferulic acid and/or its metabolites are administered (ingested) to the subject at least once a day, for example, once to several times a day (for example, 2 to 3 times). is preferred. In the above methods and uses, it is preferred that dihydroferulic acid and/or its metabolites be orally administered (ingested). The above uses are preferably in humans or non-human mammals, more preferably in humans.

In the above methods and uses, dihydroferulic acid and/or its metabolites may be used in an amount (effective amount) that provides the desired effect. The preferred dosage, administration target, etc. of dihydroferulic acid and/or its metabolite are the same as those for the composition of the present invention described above. Dihydroferulic acid and/or its metabolites may be administered as such, or as a composition containing dihydroferulic acid and/or its metabolites. For example, the compositions of the invention described above can be administered.

In one aspect, the present invention also encompasses the use of dihydroferulic acid and/or its metabolites to produce a composition for increasing muscle mass or strength. The composition for increasing muscle mass or muscle strength and its preferred embodiments are the same as the composition of the present invention and its preferred embodiments described above. The present invention also includes dihydroferulic acid and/or its metabolites for use in increasing muscle mass or strength. The present invention also includes dihydroferulic acid and/or its metabolites for use in the prevention or amelioration of locomotive syndrome, sarcopenia, frailty or cachexia.

EXAMPLES Hereinafter, the present invention will be explained in more detail with reference to Examples, but the scope of the present invention is not limited thereby.

<Example 1>
A mouse myoblast cell line (C2C12 cells, manufactured by ATCC) was used. C2C12 cells were seeded on a 96-well plate at 0.2×10 ⁴ cells/well, and supplemented with 10% FBS (fetal bovine serum, manufactured by Sigma-Aldrich) and 1% antibiotic (Nacalai Tesque Co., Ltd.). The cells were cultured at 37° C. for 3 days in high glucose normal DMEM (manufactured by Sigma-Aldrich) containing a fungal agent mixed solution). After confirming that the C2C12 cells were confluent, subsequent experiments were conducted.

The above FBS-containing DMEM medium was replaced with high glucose normal DMEM (manufactured by SIGMA-Aldrich) (hereinafter referred to as FBS-free medium) containing no FBS and 1% antibiotic, and cultured for an additional 20 hours. .

The FBS-free medium was replaced with FBS supplemented with dihydroferulic acid (manufactured by Tokyo Kasei Kogyo Co., Ltd.), a glucuronide conjugate of dihydroferulic acid (manufactured by Toronto Research Chemicals), or IGF-1 (Recombinant Analog human, manufactured by Cell Sciences). The medium was replaced with non-supplemented medium. The final concentration of dihydroferulic acid was 30 μM, 100 μM, 300 μM, or 1000 μM. The final concentration of the glucuronide conjugate of dihydroferulic acid was 300 μM or 1000 μM. IGF-1 was used as a positive control, and the final concentration of IGF-1 in the medium was 100 ng/mL. In addition, dimethyl sulfoxide (DMSO) was added to the FBS-free medium at a final concentration of 0.1% (v/v) in addition to the medium supplemented with dihydroferulic acid, the glucuronide conjugate of dihydroferulic acid, or IGF-1. A medium (control medium) was prepared, and the medium without FBS was replaced with the control medium (negative control).

FBS-free medium supplemented with dihydroferulic acid, FBS-free medium supplemented with dihydroferulic acid glucuronide conjugate, FBS-free medium supplemented with IGF-1 (positive control), or FBS-free medium supplemented with DMSO After culturing C2C12 cells in (negative control) for 1 hour, cell lysis buffer (manufactured by Cell Signaling Technology) was added and the cells were collected. Phenylmethylsulfonyl fluoride (PMSF) was added to the cell lysis buffer at a final concentration of 1 mM.

Using ELISA Kits (manufactured by Cell Signaling Technology), the amount of phosphorylation of T (Thr) 389 residue of S6K (ribosomal protein S6 kinase) was measured by ELISA method according to the attached protocol. The amount of phosphorylated S6K (T389) per concentration was calculated. Significance test was performed by Dunnett's test (significance level: 0.01, 0.05 or 0.1, relative to negative control).

The results are shown in Figure 1. FIG. 1 is a graph showing the amount of phosphorylation of the T389 residue of S6K per protein concentration in cells to which dihydroferulic acid or a glucuronide conjugate of dihydroferulic acid was added. The vertical axis of the graph in FIG. 1 represents the relative value of the amount of phosphorylation when the above amount of phosphorylation in the negative control (C) is taken as 100.
In FIG. 1, "C" is the negative control, "IGF" is the positive control IGF-1, "HMPA" is dihydroferulic acid, "HMPA-GlcA" is the glucuronide conjugate of dihydroferulic acid, show. The results in FIG. 1 are all mean values ± standard error of n=4 (*: p<0.05, **: p<0.01, relative to the negative control). 30, 100, 300, and 1000 in HMPA indicate that the dihydroferulic acid concentration in the medium is 30 μM, 100 μM, 300 μM, and 1000 μM, respectively. 300 and 1000 in HMPA-GlcA indicate that the concentrations of the glucuronide conjugate of dihydroferulic acid in the medium are 300 μM and 1000 μM, respectively.

As is clear from FIG. 1, dihydroferulic acid and its metabolites promoted S6K phosphorylation. These results revealed that dihydroferulic acid and its metabolites have the effect of promoting protein synthesis.

Claims (8)

A composition for increasing muscle mass or muscle strength, comprising dihydroferulic acid and/or its metabolite as an active ingredient. The composition according to claim 1, wherein the metabolite is a glucuronidate of dihydroferulic acid. The composition according to claim 1 or 2, which is used to promote protein synthesis in muscle. 4. A composition according to claim 3, which is used to promote protein synthesis by activating the mTOR pathway. The composition according to claim 1 or 2, which is an oral composition. The composition according to claim 1 or 2, which is a food or drink. One or more indications selected from the group consisting of "muscle increase", "muscle improvement", "muscle strength increase", "muscle strength improvement", "promotion of skeletal muscle hypertrophy", and "supporting the ability to build muscle" The composition according to claim 1 or 2. 3. A composition according to claim 1 or 2 for use in combination with exercise.

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