Myoblast proliferation promoter, composition for promoting myoblast proliferation, muscle atrophy inhibitor, and composition for inhibiting muscle atrophy

Abstract

Provided are a myoblast proliferation promoter and a muscle atrophy inhibitor, each of which comprises a compound represented by structural formula (1).

Description

Technical Field

[0001] The present invention relates to a myoblast proliferation promoter and a composition for promoting myoblast proliferation, as well as a muscle atrophy inhibitor and a composition for inhibiting muscle atrophy. Background Art

[0002] The aging rate (the proportion of the population aged 65 and over) in Japan is increasing year by year. In an aging society, it is important to improve the quality of life and extend the healthy life expectancy.

[0003] Muscular atrophy refers to muscle atrophy. For example, it is known that disuse muscle atrophy may occur due to a sedentary lifestyle or bed rest, and age-related muscle atrophy may occur due to damage to the homeostasis of muscle physiological tissues due to aging.

[0004] Such reduced muscle function may sometimes lead to insufficient exercise or action or mental stress, which in turn may hinder other body functions, sometimes leading to a decrease in quality of life or a shortened healthy life span.

[0005] Therefore, it is very important to prevent muscle function from decreasing.

[0006] Hitherto, as a composition having an excellent effect of maintaining muscle fibers constituting muscles, a composition for maintaining muscle fibers containing theaflavins as an active ingredient has been proposed (for example, see Patent Document 1).

[0007] However, there is still a strong demand for raw materials that can prevent muscle function decline and are highly safe and can be widely used as ingredients for food and beverages, pharmaceuticals, research reagents, etc., and the current situation is that there is a demand for rapid development.

[0008] Prior art literature

[0009] Patent Literature

[0010] Patent Document 1: Japanese Patent Application Publication No. 2021-169430 Summary of the invention

[0011] Problems to be solved by the invention

[0012] The present invention aims to solve the above-mentioned various conventional problems and achieve the following objectives: That is, the present invention aims to provide a myoblast proliferation promoting agent and a myoblast proliferation promoting composition having excellent myoblast proliferation promoting effect and high safety.

[0013] Another object of the present invention is to provide a muscle atrophy inhibitor and a muscle atrophy inhibitor composition having an excellent muscle atrophy inhibitory effect and high safety.

[0014] Methods used to solve problems

[0015] The present inventors have conducted intensive studies to solve the above problems and have found that the compound represented by the following structural formula (1) has excellent myoblast proliferation promoting effect and muscle atrophy inhibiting effect and is highly safe and useful for myoblast proliferation promoting effect and muscle atrophy inhibiting effect.

[0016] [Chemistry 1]

[0017]

[0018] The present invention is based on the above-mentioned knowledge of the present inventors, and as a method for solving the above-mentioned problems, it is as follows. That is,

[0019] <1> A myoblast proliferation promoter, comprising a compound represented by the following structural formula (1).

[0020] [Chemistry 2]

[0021]

[0022] <2> A composition for promoting myoblast proliferation, comprising the myoblast proliferation promoting agent according to <1> above.

[0023] <3> A muscle atrophy inhibitor, comprising a compound represented by the following structural formula (1).

[0024] [Chemistry 3]

[0025]

[0026] <4> A composition for inhibiting muscle atrophy, comprising the muscle atrophy inhibitor according to <3> above.

[0027] Effects of the Invention

[0028] The myoblast proliferation promoter and myoblast proliferation promoting composition of the present invention can solve the above-mentioned various existing problems, and can provide a myoblast proliferation promoter and myoblast proliferation promoting composition that can achieve the above-mentioned objects, have excellent myoblast proliferation promoting effects, and are highly safe.

[0029] According to the muscle atrophy inhibitor and the muscle atrophy inhibitor composition of the present invention, the above-mentioned various existing problems can be solved, and a muscle atrophy inhibitor and a muscle atrophy inhibitor composition that can achieve the above-mentioned purpose, have excellent muscle atrophy inhibitory effect and are highly safe can be provided. DETAILED DESCRIPTION

[0030] (Myoblast proliferation promoter and muscle atrophy inhibitor)

[0031] The myoblast proliferation promoter of the present invention contains the compound represented by the following structural formula (1) as an active ingredient, and further contains other ingredients as necessary.

[0032] The muscle atrophy inhibitor of the present invention contains the compound represented by the following structural formula (1) as an active ingredient, and further contains other ingredients as necessary.

[0033] [Chemistry 4]

[0034]

[0035] In the present specification, the promotion of myoblast proliferation refers to promoting the proliferation of myoblasts. The degree of the proliferation is not particularly limited and can be appropriately selected according to the purpose.

[0036] The type of the myoblasts is not particularly limited and can be appropriately selected depending on the intended purpose. Examples thereof include skeletal muscle-derived myoblasts.

[0037] The source of the myoblasts is not particularly limited and can be appropriately selected depending on the intended purpose. Examples thereof include humans and animals other than humans (eg, mice, rats, hamsters, dogs, cats, cows, pigs, monkeys, etc.).

[0038] In this specification, the inhibition of muscle atrophy refers to the inhibition of muscle atrophy. Specifically, for example, it refers to the inhibition of the reduction of muscle mass, or the prevention of the inhibition of the increase of muscle mass. The degree of the inhibition of muscle atrophy is not particularly limited and can be appropriately selected according to the purpose.

[0039] The type of muscle atrophy is not particularly limited and can be appropriately selected depending on the intended purpose. Examples thereof include disuse muscle atrophy and aging muscle atrophy.

[0040] The compound represented by the structural formula (1) has excellent myoblast proliferation promoting action and muscle atrophy inhibiting action and is useful as a myoblast proliferation promoting agent or a muscle atrophy inhibiting agent, which has never been known before and is a new discovery by the present inventors.

[0041] <Compound represented by structural formula (1)>

[0042] The compound represented by the following structural formula (1) is named 3,4-dihydroxyhydrocinnamic acid (English name: 3,4-Dihydroxyhydrocinnamic acid).

[0043] [Chemistry 5]

[0044]

[0045] The compound represented by the above structural formula (1) is a known compound, and a commercially available product may be used, or a compound extracted from a plant or the like may be used.

[0046] The compound represented by the structural formula (1) has excellent myoblast proliferation promoting effect and muscle atrophy inhibiting effect, and can be used as an active ingredient of a myoblast proliferation promoter or a muscle atrophy inhibitor. The myoblast proliferation promoter and muscle atrophy inhibitor can be used in a wide range of applications such as pharmaceuticals, quasi-drugs, oral compositions, cosmetics, and research reagents.

[0047] The myoblast proliferation promoter and muscle atrophy inhibitor may be formed solely from the compound represented by the above structural formula (1), or the compound represented by the above structural formula (1) may be formulated.

[0048] The above-mentioned myoblast proliferation promoter and muscle atrophy inhibitor can use pharmaceutically acceptable carriers such as dextrin and cyclodextrin and other arbitrary auxiliary agents, and according to a conventional method, the formulation is powdered, granular, tableted, liquid and other arbitrary dosage forms. At this time, as auxiliary agents, for example, excipients, adhesives, disintegrants, lubricants, stabilizers, flavor-corrective agents, etc. can be used. The above-mentioned myoblast proliferation promoter and muscle atrophy inhibitor can be used in conjunction with other compositions (for example, myoblast proliferation promotion compositions described later, muscle atrophy inhibition compositions, etc.), or can also be used as tablets, capsules, etc.

[0049] The content of the compound represented by the above structural formula (1) in the formulated myoblast proliferation promoter and muscle atrophy inhibitor is not particularly limited and can be appropriately selected depending on the intended purpose.

[0050] Furthermore, the above-mentioned myoblast proliferation promoting agent may also contain other components having myoblast proliferation promoting effects as active ingredients together with the compound represented by the above-mentioned structural formula (1), if necessary.

[0051] Furthermore, the muscle atrophy inhibitor can also contain other components having muscle atrophy inhibitory effects as active ingredients together with the compound represented by the above structural formula (1), if necessary.

[0052] <Other ingredients>

[0053] As the above-mentioned other components, there are no particular restrictions as long as the effects of the present invention are not impaired, and they can be appropriately selected according to the utilization form of the above-mentioned myoblast proliferation promoter and muscle atrophy inhibitor, and for example, the components that can be used when the above-mentioned formulation is prepared can be mentioned. The above-mentioned other components can be used alone or in combination of two or more.

[0054] The content of the other components in the myoblast proliferation promoter and muscle atrophy inhibitor is not particularly limited and can be appropriately selected depending on the intended purpose.

[0055] The administration method of the myoblast proliferation promoter and muscle atrophy inhibitor is not particularly limited and can be appropriately selected depending on the purpose. For example, oral administration and the like are mentioned.

[0056] The dosage, administration period, and administration interval of the myoblast proliferation promoter and muscle atrophy inhibitor are not particularly limited and can be appropriately selected depending on the intended purpose.

[0057] <Application>

[0058] The use of the myoblast proliferation promoting agent is not particularly limited and can be appropriately selected depending on the purpose. For example, it can be suitably used as an active ingredient of a composition for promoting myoblast proliferation described below.

[0059] The use of the muscle atrophy inhibitor is not particularly limited and can be appropriately selected depending on the purpose. For example, it can be suitably used as an active ingredient of the muscle atrophy inhibitory composition described later.

[0060] The myoblast proliferation promoter can promote myoblast proliferation by the action of the compound represented by the structural formula (1) as an active ingredient. Therefore, the myoblast proliferation promoter of the present invention can be used for all significant uses that exert a myoblast proliferation promoting action.

[0061] The muscle atrophy inhibitor can inhibit muscle atrophy by the action of the compound represented by the structural formula (1) as an active ingredient. Therefore, the muscle atrophy inhibitor of the present invention can be used for all purposes that have a significant effect of inhibiting muscle atrophy.

[0062] The myoblast proliferation promoter and muscle atrophy inhibitor have excellent myoblast proliferation promoting effect or muscle atrophy inhibiting effect, and can be formulated in oral compositions such as food and beverages, for example. In this case, the compound represented by the above structural formula (1) can be formulated directly, or the myoblast proliferation promoter or muscle atrophy inhibitor formulated from the compound represented by the above structural formula (1) can be formulated.

[0063] The myoblast proliferation promoter and muscle atrophy inhibitor of the present invention are suitable for use in humans, and can be applied to animals other than humans (eg, mice, rats, hamsters, dogs, cats, cows, pigs, monkeys, etc.) as long as their effects are exerted.

[0064] The usage of the myoblast proliferation promoter and muscle atrophy inhibitor is not particularly limited and can be appropriately selected depending on the purpose. Examples of the usage include oral, parenteral, and external administration.

[0065] The application site, dosage, and dosage form of the myoblast proliferation promoter and muscle atrophy inhibitor can be appropriately selected according to the intended use.

[0066] There is no particular limitation on the usage method such as the amount, period, and interval of use of the myoblast proliferation promoter and muscle atrophy inhibitor, and they can be appropriately selected depending on the intended purpose.

[0067] Furthermore, the myoblast proliferation promoter of the present invention can be used as a myoblast proliferation promoter, and the muscle atrophy inhibitor of the present invention can also be used as a reagent for studying the mechanism of action of the muscle atrophy inhibitory effect.

[0068] (Composition for promoting myoblast proliferation and composition for inhibiting muscle atrophy)

[0069] The composition for promoting myoblast proliferation of the present invention contains the myoblast proliferation promoter of the present invention and further contains other components as necessary.

[0070] The muscle atrophy inhibitory composition of the present invention contains the muscle atrophy inhibitor of the present invention and further contains other components as necessary.

[0071] <Myoblast proliferation promoter and muscle atrophy inhibitor>

[0072] The above-mentioned myoblast proliferation promoter is the above-mentioned myoblast proliferation promoter of the present invention.

[0073] The above-mentioned muscle atrophy inhibitor is the muscle atrophy inhibitor of the above-mentioned present invention.

[0074] The content of the myoblast proliferation promoter in the myoblast proliferation promoting composition is not particularly limited and can be appropriately adjusted according to the form of the myoblast proliferation promoting composition, etc., and is preferably 0.0001% by mass to 20% by mass, more preferably 0.0001% by mass to 10% by mass, calculated as the amount of the compound represented by the structural formula (1). The myoblast proliferation promoting composition may be formed solely from the myoblast proliferation promoter.

[0075] The content of the muscle atrophy inhibitor in the muscle atrophy inhibitor composition is not particularly limited and can be appropriately adjusted according to the form of the muscle atrophy inhibitor composition, and is preferably 0.0001% by mass to 20% by mass, more preferably 0.0001% by mass to 10% by mass, calculated as the amount of the compound represented by the structural formula (1). The muscle atrophy inhibitor composition may be composed solely of the muscle atrophy inhibitor.

[0076] <Other ingredients>

[0077] There are no particular restrictions on other components in the composition for promoting myoblast proliferation and the composition for inhibiting muscle atrophy, and they can be appropriately selected according to the use form of the composition for promoting myoblast proliferation and the composition for inhibiting muscle atrophy, and examples thereof include the same components as those described in the items of the myoblast proliferation promoter and the muscle atrophy inhibitor, etc. These can be used alone or in combination of two or more.

[0078] The content of the other components in the composition for promoting myoblast proliferation and the composition for inhibiting muscle atrophy is not particularly limited and can be appropriately selected depending on the intended purpose.

[0079] <Form>

[0080] The form of the composition for promoting myoblast proliferation and the composition for inhibiting muscle atrophy is not particularly limited and can be appropriately selected depending on the purpose. Examples thereof include pharmaceuticals, quasi-drugs, and foods and beverages.

[0081] The composition for promoting myoblast proliferation and the composition for inhibiting muscle atrophy of the present invention can be used daily and can extremely effectively exert various physiological activities represented by myoblast proliferation promoting action and muscle atrophy inhibiting action due to the action of the compound represented by the above structural formula (1) as an active ingredient.

[0082] The composition for promoting myoblast proliferation and the composition for inhibiting muscle atrophy of the present invention are suitable for use in humans, but can be applied to animals other than humans (eg, mice, rats, hamsters, dogs, cats, cows, pigs, monkeys, etc.) as long as their respective effects are exerted.

[0083] The usage of the composition for promoting myoblast proliferation and the composition for inhibiting muscle atrophy of the present invention is not particularly limited and can be appropriately selected depending on the intended purpose. Examples thereof include oral, parenteral, and external administration, and oral administration is preferred.

[0084] As the composition used for above-mentioned oral use, for example, oral administration agent, diet can be enumerated.Here, so-called diet refers to that the possibility of causing harm to people's health is small, and in common social life, by oral or digestive tract casting and the food taken in, is not limited to the distinction of food, pharmaceuticals, quasi-drugs etc. on administrative division.Therefore, above-mentioned diet refers to the diet of the diet of the general food, health food (functional diet), health function food (specific health food, nutrition function food, functional labeling food), quasi-drugs, pharmaceuticals etc. that widely comprise oral intake.

[0085] There is no particular limitation on the type of the composition for oral administration, and it can be appropriately selected according to the purpose. For example, beverages such as tea beverages, refreshing drinks, carbonated drinks, nutritional drinks, fruit drinks, lactic acid drinks, alcoholic drinks, coffee drinks, and refreshing drinks containing coffee (including concentrated original solutions of these beverages and powders for adjustment); ice cream, smoothies, shaved ice and other ice creams; noodles such as soba noodles, udon noodles, vermicelli noodles, dumpling wrappers, shumai wrappers, Chinese noodles, and instant noodles; desserts such as sugar, candy, chewing gum, chocolate, hard candy, snacks, biscuits, jelly, jam, cream, baked snacks, and bread; marine products such as crabs, salmon, clams, tuna, sardines, shrimps, bonito, Spanish mackerel, whales, oysters, saury, squid, ark shells, scallops, abalone, sea urchins, fish roe, and shellfish; fish cakes, ham, sausages, etc. Processed foods from aquatic and livestock products; dairy products such as processed milk and fermented milk; oils and fats such as salad oil, tempura oil, margarine, mayonnaise, puff pastry, fresh cream, and sauces, and foods processed with oils and fats; seasonings such as sauces and dressings; steamed and cooked foods such as curry, stew, chicken and egg rice, porridge, porridge with stew, Chinese rice, fried pork cutlet rice, tempura rice, eel rice, beef rice, oden, mapo tofu, beef rice, meat sauce, egg soup, omelette rice, dumplings, steamed buns, hamburgers, and meatballs; home-cooked dishes such as salads and pickles; various forms of health, beauty, and nutritional supplements; pharmaceuticals and quasi-pharmaceuticals such as tablets, powders, capsules, granules, extracts, syrups, oral liquids, lozenges, and gargles; oral fresheners and toothpastes used in the mouth such as mouth fresheners and bad breath preventers.

[0086] The method for producing the composition for promoting myoblast proliferation and the composition for inhibiting muscle atrophy is not particularly limited and can be appropriately selected according to the form of use of the composition for promoting myoblast proliferation and the composition for inhibiting muscle atrophy.

[0087] The amount, period of use, interval of use, etc. of the composition for promoting myoblast proliferation and the composition for inhibiting muscle atrophy are not particularly limited and can be appropriately selected depending on the intended purpose.

[0088] As described above, the myoblast proliferation promoting agent and the myoblast proliferation promoting composition of the present invention have an excellent myoblast proliferation promoting effect.

[0089] Therefore, the present invention also relates to a method for promoting myoblast proliferation, characterized in that at least one selected from the group consisting of the above-mentioned myoblast proliferation promoting agent and myoblast proliferation promoting composition is administered to an individual.

[0090] As described above, the muscle atrophy inhibitor and the muscle atrophy inhibitory composition of the present invention have excellent muscle atrophy inhibitory effects.

[0091] Therefore, the present invention also relates to a method for inhibiting muscle atrophy, characterized in that at least one selected from the group consisting of the above-mentioned muscle atrophy inhibitor and the composition for inhibiting muscle atrophy is administered to an individual.

[0092] Example

[0093] Hereinafter, test examples and formulation examples of the present invention will be described, but the present invention is not limited to these test examples and formulation examples.

[0094] (Test Example 1: Myoblast Proliferation Promotion Effect Test)

[0095] The compound represented by the above structural formula (1) (manufactured by Sigma-Aldrich) was tested for its myoblast proliferation promoting effect by the following test method.

[0096] <Test method>

[0097] Mouse skeletal muscle-derived myoblasts (C2C12) were cultured in Dulbecco's modified Eagle's medium (DMEM) containing 10% FBS and then collected by trypsin treatment. The collected cells were plated at 2.0×10 4 After diluting with DMEM containing 5% FBS at a concentration of cells / mL, 100 μL was inoculated into each well of a 96-well plate and cultured for 6 hours. After the culture was completed, 100 μL of the test sample dissolved in DMEM containing 5% FBS (final concentration is shown in Table 1 below) was added to each well and cultured for 2 days. In addition, even when the test sample was not added, the culture was carried out in the same manner except that the test sample was not added.

[0098] The myoblast proliferation promoting effect was determined using the MTT assay. After the above culture was completed, the culture medium was removed and 100 μL of MTT dissolved in PBS buffer (PBS(-)) at a final concentration of 0.4 mg / mL was added to each well. After 2 hours of culture, the blue formazan generated in the cells was extracted with 100 μL of 2-propanol. After extraction, the absorbance at a wavelength of 570 nm was measured. At the same time, the absorbance at a wavelength of 650 nm was measured as turbidity, and the difference between the two was used as the amount of blue formazan generated.

[0099] From the obtained results, the myoblast proliferation promotion rate (%) was calculated by the following formula. The results are shown in Table 1.

[0100] Myoblast proliferation promotion rate (%) = A / B × 100

[0101] A to B in the above formula are respectively represented as follows.

[0102] A: Amount of blue formazan produced in cells to which the test sample was added

[0103] B: Amount of blue formazan produced in cells without the addition of the test sample

[0104] [Table 1]

[0105]

[0106] From the results in Table 1, it was confirmed that the compound represented by the above structural formula (1) exhibited an excellent myoblast proliferation promoting effect.

[0107] (Test Example 2: Muscle atrophy inhibitory effect test)

[0108] The compound represented by the above structural formula (1) (manufactured by Cayman Chemical) was orally administered to a rat disuse muscle atrophy model for 14 days, and whether muscle atrophy was suppressed was tested by the following test method.

[0109] <Test method>

[0110] Male Wistar (Crlj: WI) rats aged 7 weeks were purchased from Charles River, Japan, and housed in an environment with a temperature of 23±3°C, a humidity of 55±15%, and a light-dark cycle of 12 hours each. The rats were acclimated for 1 week before use and were allowed to freely consume solid feed (CRF-1, manufactured by Oriental Yeast Industry Co., Ltd.) and tap water.

[0111] After acclimation for one week, Wistar rats were randomly divided into groups of 8 rats each. According to the Morey-Holton ER method, the rats were kept for 14 days with their tails clamped and their hind limbs off the floor. During the feeding period, the rats were allowed to freely take water and food by moving around the cage with their forelimbs.

[0112] The experimental groups were (1) normal group (no hind limb suspension), (2) control group (hind limb suspension + 0 mg / kg of the compound represented by structural formula (1)), and (3) compound group represented by structural formula (1) (hind limb suspension + 125 mg / kg of the compound represented by structural formula (1)).

[0113] The compound represented by the structural formula (1) was dissolved in water and orally administered daily for 14 days. The control group was administered with water.

[0114] At the end of the experiment, the rats were bled to death under isoflurane anesthesia, and the soleus muscle was removed to measure the muscle weight. The results are shown in Table 2. In Table 2, the soleus muscle weight shows the average value.

[0115] [Table 2]

[0116]

[0117] As shown in Table 2, the compound group represented by structural formula (1) showed an increase in soleus muscle weight compared with the control group. This showed that the compound represented by structural formula (1) inhibited muscle atrophy in the rat disuse muscle atrophy model.

[0118] (Matching Example 1)

[0119] Tablets having the following composition were produced by a conventional method.

[0120] 5.0 mg of the compound represented by the above structural formula (1)

[0121] ·Dolomite 83.4mg

[0122] (Contains Calcium 20%, Magnesium 10%)

[0123] Casein phosphono peptide 16.7 mg

[0124] Vitamin C 33.4mg

[0125] Maltitol 136.8mg

[0126] ·Collagen 12.7mg

[0127] Sucrose fatty acid esters 12.0 mg

[0128] (Matching Example 2)

[0129] An oral liquid preparation having the following composition was produced by a conventional method.

[0130] <Composition in 1 ampoule (1 bottle 100mL)>

[0131] ·0.3% by mass of the compound represented by the above structural formula (1)

[0132] ·Sorbitol 12.0 mass%

[0133] ·Sodium benzoate 0.1 mass%

[0134] · Fragrance 1.0 mass%

[0135] Calcium sulfate 0.5 mass%

[0136] · Refined water residue (combination example 3)

[0137] By a conventional method, a coffee beverage having the following composition was produced.

[0138] ·0.1% by mass of the compound represented by the above structural formula (1)

[0139] Coffee extract 40% by mass

[0140] (L(lightness)=20, Brix=3)

[0141] ·Maltitol 2% by mass

[0142] ·Spices to taste

[0143] · The remaining water (Formulation Example 4)

[0144] A capsule having the following composition was produced by a conventional method. In addition, a No. 1 hard gelatin capsule was used as the capsule.

[0145] <Composition of 1 capsule (200 mg per capsule)>

[0146] ·30.0 mg of the compound represented by the above structural formula (1)

[0147] Corn starch 70.0 mg

[0148] Lactose 80.0 mg

[0149] Calcium lactate 10.0mg

[0150] ·Hydroxypropylcellulose (HPC-L) 10.0mg

[0151] This application claims the priority based on Japanese Patent Application No. 2022-095282 filed on June 13, 2022, and all contents of Japanese Patent Application No. 2022-095282 are hereby incorporated by reference into this application.

Claims

1. A myoblast proliferation promoter, characterized in that Contains a compound represented by the following structural formula (1), [Chemistry 1] 2. A composition for promoting myoblast proliferation, characterized in that: The invention comprises the myoblast proliferation promoter according to claim 1.

3. A muscle atrophy inhibitor, characterized in that: Contains a compound represented by the following structural formula (1), [Chemistry 2] 4. A composition for inhibiting muscle atrophy, characterized in that: Contains the muscle atrophy inhibitor according to claim 3.

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