JP2015193547A - Resveratrol heat treatment composition which has motion-like action - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a composition having motion-like action with safety and high practicality, a muscle-enhancing agent containing the composition, an HDL cholesterol increasing agent, and a feed for animals.SOLUTION: The invention provides a composition having motion-like action obtained by subjecting resveratrol to heat treatment under alkaline conditions, a muscle-enhancing agent containing the composition, an HDL cholesterol increasing agent and a feed for animals. The composition has the motion-like action, containing three kinds of resveratrol derivatives represented by the following formula.

Description

  The present invention relates to a composition having a motion-like action obtained by a completely new method of heat-treating resveratrol under alkaline conditions, a muscle strengthening agent containing the composition as an active ingredient, and an HDL cholesterol raising agent.

  Currently, highly functional ingredients are obtained using various methods such as fermentation and synthesis, and are used in a wide range of fields such as foods and pharmaceuticals. However, there are a limited number of things that are both useful and practical. Furthermore, in a composition containing a novel compound that has not yet been reported, it is more difficult to achieve both usefulness and practicality due to difficulty in production.

  The inventors of the present invention have found an epoch-making method for producing a composition containing a novel compound having high functionality by using a component derived from food in a simple and easy manner. (Patent Document 1). However, the application range is not wide, and it is assumed that there are many applications that have not been found.

  In the current social life, where excessive stress, food intake, and lack of exercise are prevalent, the metabolic syndrome caused by these has become a major social problem. Metabolic syndrome is a condition that has two or more of hyperglycemia, hypertension, and lipid abnormalities in addition to built-in fat obesity, increasing the risk of cardiovascular diseases such as cerebral infarction, arteriosclerosis, and myocardial infarction. State.

  Moderate exercise is most effective in preventing metabolic syndrome. However, in modern societies with limited time, it is often difficult to exercise regularly. In addition, continuous exercise often does not continue from the balance with daily life. In such a case, many simple exercise devices have been developed, but there are many problems in the effect and continuity as exercise.

Although the effectiveness of the exercise effect is recognized, in order to overcome the current situation that has a problem in the implementation of continuous exercise in real-life life, exercised without exercise by enhancing the exercise effect or ingesting Development of drugs that exert the effects obtained in some cases is underway. As such a drug, for example, catechin has been reported to improve basal metabolism and lipid metabolism (Patent Document 2). In this case, improvement of lipid metabolism and improvement of basal metabolism can be said to be exercise-like effects. However, its action is limited to lipids.
In addition, AICAR (5-aminoimidazole-4-carboxamide ribonucleotide), which is a compound, is said to exert a movement-like effect by activating AMPK (AMP-activated protein kinase). However, it cannot be used for healthy people because it is used only for pharmaceuticals. Treatment of metabolic syndrome and the like is important, but prevention becomes more important. Therefore, there is a demand for the development of a safe material that has a wider range of motion-like effects and uses.

  A physiological phenomenon that changes with exercise is suppression of myostatin secretion. Myostatin (GDF-8) is a muscle growth inhibitory factor and has been reported as a muscle development inhibitory hormone (Non-patent Document 1). It has been reported that the blood concentration of this hormone decreases with exercise (Non-patent Document 2). Therefore, suppressing myostatin secretion or inhibiting the action is useful for muscle strengthening and motor function improvement.

  In addition, an increase in HDL cholesterol, commonly called good cholesterol, is one of the physiological phenomena that changes with exercise. It is well known that HDL cholesterol has an action of extracting excess cholesterol from tissues. This action is said to be effective in preventing cardiovascular diseases such as arteriosclerosis. So far, the total cholesterol level has attracted attention as a risk factor for arteriosclerosis. Therefore, drugs that lower cholesterol levels, such as statin drugs, and foods for specified health use are sold, but in recent years, the difference between LDL cholesterol and HDL cholesterol (LH ratio) called bad cholesterol is high in the incidence of cardiovascular disease It is attracting attention as being correlated. It is also known that the value of HDL cholesterol decreases due to obesity and lack of exercise, and that the value is improved by exercise therapy for a certain period.

  In order to realize the effectiveness of these exercise-like actions, the present inventors have focused on resveratrol, which is famous as a functional ingredient that has both a certain effectiveness and high safety.

  Resveratrol, which is famous for being contained in red wine, was reported in 1997 to have a high anticancer effect (Non-patent Document 3). This report attracts attention to resveratrol, and it is becoming clear that it is effective for many diseases (Non-patent Document 4). The phenomenon called “French paradox” in France, where the incidence of cardiovascular diseases is extremely low, is known to correlate with the intake of red wine, and is due to the antioxidant and anti-inflammatory effects of resveratrol. It is said that vascular protection is a factor.

  It has been reported in the prior art that said resveratrol also has a useful effect on motor function and muscle energy metabolism. For example, an endurance improver containing resveratrol as an active ingredient (Patent Document 3) and a composition that maintains or enhances mitochondrial biosynthesis containing resveratrol as an active ingredient (Patent Document 4) have been reported. In addition, resveratrol is reported to activate AMPK, which is an important factor for muscle tissue metabolism, in a human clinical study ingested 150 mg / day for 30 days (Non-patent Document 5), and is also an insulin resistance index. Improvements in HOMA-R have also been observed.

Japanese Patent Application No. 2013-267473 JP 2004-161616 A Japanese Patent No. 4739161 Special table 2012-524033 gazette

NATURE, 387, p83-90 (1997) Med. Sci. Sports Excrc. , 42 (11), p2023-2029 (2010) Science, 275 (10), p218-220 (1997). Drug Discovery, 5, p493-506 (2006) Cell, 134, p405-415 (2008)

  As a result of earnest examination of the effect of the composition containing the novel compound obtained by the technique invented by the present inventors, surprisingly, the composition obtained by heat-treating resveratrol under alkaline conditions has a myostatin secretion inhibitory action. The accompanying muscle strengthening action and the HDL cholesterol raising action were recognized, and it succeeded for the first time in discovering that it had a movement-like action, and came to complete this invention.

  Therefore, an object of the present invention is to provide a composition having a safe and highly practical exercise-like action, a muscle strengthening agent containing the composition, an HDL cholesterol raising agent, and an animal feed.

The gist of the present invention is as follows:
[1] A composition having a motion-like effect obtained by heat-treating resveratrol under alkaline conditions;
[2] Formula (1):

Formula (2):

And formula (3):

A composition having a motion-like action according to claim 1, comprising three types of resveratrol derivatives represented by:
[3] A muscle strengthening agent comprising the composition having the exercise-like action according to [1] or [2],
[4] The muscle enhancer according to [3], wherein the exercise-like action is a myostatin secretion inhibitory action,
[5] An HDL cholesterol raising agent comprising the composition having the exercise-like action according to [1] or [2],
[6] An animal feed comprising the composition having the exercise-like action according to [1] or [2].

The composition having a motion-like action of the present invention is useful as a composition having a novel motion-like action because it is highly practical and has a high practicality because of its simple manufacturing method.
Since the composition having a motion-like action of the present invention has a myostatin secretion inhibitory action, it is useful as an active ingredient of a muscle potentiator.
Moreover, since the composition which has the exercise | movement like effect | action of this invention has a HDL cholesterol raising effect, it is useful as a HDL cholesterol raising agent.
In addition, by combining the composition having the exercise-like action of the present invention with foods, pharmaceuticals, quasi-drugs and animal feeds, it has the functionality expected to increase muscle and raise HDL cholesterol in terms of exercise-like action. Foods, pharmaceuticals to treat muscle dystrophy and low HDL cholesterolemia, quasi-drugs for muscle enhancement and HDL cholesterol elevation, and animal feeds for growth promotion and meat quality improvement be able to.

FIG. 1 shows a chromatogram of the composition having the motion-like action of the present invention produced in Example 1. The numbers correspond to 1: UHA4002, 2: UHA4003, 3: UHA4022, respectively. FIG. 2 is a graph showing the muscle wet weight per body weight of the rat obtained in Example 2. As the muscle tissue, quadriceps, long extensor, anterior tibialis, soleus and plantar muscles were collected and weighed, and the wet muscle weight per body weight of each muscle tissue was calculated. In the figure, “control” indicates a high fat diet group, and “invention product” indicates a 1% invention product-containing high fat diet group. “*” Indicates a significant difference (p <0.05) from the high fat diet group in the t test. FIG. 3 is a graph showing the total muscle weight of the rat obtained in Example 2. The total muscle weight is the sum of the quadriceps, long extensor, anterior tibial, soleus and plantar muscle weights, and the total muscle weight per body weight was calculated from the total muscle weight. In the figure, “control” indicates a high fat diet group, and “invention product” indicates a 1% invention product-containing high fat diet group. “*” Indicates a significant difference (p <0.05) from the high fat diet group in the t test. FIG. 4 is a graph showing the blood myostatin concentration in rats obtained in Example 3. In the figure, “control” indicates the high fat diet group, and “0.1 to 1.0% invention product” indicates the 0.1%, 0.5%, and 1.0% invention product-containing high fat diet group, respectively. . “*” Indicates a significant difference (p <0.05) from the high fat diet group in the t test. FIG. 5 is a graph showing the blood cholesterol, LDL / VLDL cholesterol, and HDL cholesterol concentrations of the rat obtained in Example 4. In the figure, “control” indicates a high fat diet group, and “0.1 to 1.0% invention product” indicates a 0.1%, 0.5%, and 1.0% invention product-containing high fat diet group. “*” Indicates a significant difference (p <0.05) from the high fat diet group in the t test.

Hereinafter, the present invention will be described in detail.
The resveratrol heat treatment composition having a motion-like action of the present invention is obtained by heat-treating resveratrol under alkaline conditions.

In the present invention, the “composition having a movement-like action” means that it is administered to a human or non-human animal to promote the generation of a physiological phenomenon caused by exercise, or to perform the phenomenon without exercise. A composition that can be generated.
“Muscle enhancing agent” refers to a drug capable of increasing muscle mass in a human or non-human animal.
“Myostatin secretion inhibitory action” refers to an action of reducing blood myostatin concentration in a human or non-human animal.
The “HDL cholesterol raising agent” refers to a drug capable of raising the blood HDL cholesterol level in a human or non-human animal.
In the present invention, a composition having at least one of these actions is referred to as a “composition having a movement-like action”.
In addition, the movement-like action on a human or non-human animal can be specifically confirmed by measuring by a method described in Examples described later.

  As a result of intensive studies, the present inventors have found that heat-treating resveratrol can efficiently and safely produce a composition having a motion-like action without requiring harmful reagents and steps. I found it.

  In the present invention, resveratrol is used as a raw material.

  Resveratrol has structural isomers of trans form and cis form, but some conversion of trans form and cis form occurs by heating or ultraviolet rays. Therefore, the resveratrol used in the present invention may be a trans isomer, a cis isomer, or a mixture of a trans isomer and a cis isomer.

  Resveratrol also includes pharmaceutically acceptable salts. Examples of the pharmaceutically acceptable salt include alkali metal salts such as lithium salt, sodium salt and potassium salt; alkaline earth metal salts such as magnesium salt, calcium salt and barium salt; aluminum salt; aluminum hydroxide salt and the like Metal hydroxide salts of; amine salts such as alkylamine salts, dialkylamine salts, trialkylamine salts, alkylenediamine salts, cycloalkylamine salts, arylamine salts, aralkylamine salts, heterocyclic amine salts; α-amino acid salts And amino acid salts such as ω-amino acid salts; peptide salts or primary, secondary, tertiary or quaternary amine salts derived therefrom. These pharmaceutically acceptable salts can be used alone or in admixture of two or more.

The resveratrol may be a naturally-derived product extracted and purified from grape skin, or may be a chemically synthesized chemical product with high purity. When natural resveratrol is used, it does not need to be completely purified, and the desired production reaction proceeds as described later, and the composition having the motion-like action of the present invention is finally obtained. Therefore, a mixture containing components other than resveratrol can also be used.
However, since a certain amount of resveratrol is required to produce a composition that exhibits sufficient effectiveness, a mixture containing 1% by weight or more in terms of resveratrol is desirable as a raw material.
Examples of the mixture containing resveratrol include grape skins, wine, wine concentrated powder, meringo, lingonberry, peanuts, locust root or rhizome, extracts from raw materials such as passion fruit seeds, and freeze-drying of these extracts Products.

  In the present invention, resveratrol is dissolved or dispersed in a suitable solvent. In this case, the solvent is not particularly limited, and may be water, a mixed solution of water and an organic solvent, or an organic solvent. In the case of a mixed solution of water and an organic solvent, there are no particular restrictions on the blending ratio and the type of organic solvent. Among them, it is preferable from the viewpoint of safety and cost to use a solvent containing only water, methanol and ethanol, or a mixed solution of water and methanol, water and ethanol, or the like. When the composition obtained after the heating reaction is used in pharmaceuticals, quasi-drugs, animal feeds, etc. without fully applying final purification, safety and legal aspects Therefore, it is desirable to use water, ethanol or hydrous ethanol as a solvent.

  There is no restriction | limiting in particular about the density | concentration of the resveratrol in the solution (henceforth a resveratrol containing solution) containing the resveratrol obtained as mentioned above. Since the higher the resveratrol concentration, the smaller the amount of solvent used, the resveratrol does not need to be completely dissolved in the solution at a saturation concentration or higher.

  Next, the pH of the resveratrol-containing solution is adjusted to be alkaline. As an adjustment method, for example, after preparing a resveratrol-containing solution, an alkalizing agent may be added to adjust the pH, or the pH of the solvent may be adjusted in advance when preparing the solution. If the pH of the resveratrol-containing solution at the start of heating is 8.0 or more, it is preferable because the reaction for efficiently producing a plurality of useful resveratrol derivatives proceeds as described later. Simultaneously with the reaction, other reactions and decomposition of the target compound occur on the other hand, so that the final recovered amount of resveratrol derivative group tends to decrease. Therefore, the pH at the start of the reaction is desirably 8 to 13.0.

  The alkalinizing agent is not particularly limited, but sodium hydroxide, potassium hydroxide, sodium carbonate, sodium hydrogen carbonate and the like are desirable from the viewpoint of safety, efficiency and cost. When the case where the pH change during the reaction is suppressed as much as possible occurs, a buffer solution may be used, but this is not always necessary.

  Next, the resveratrol containing solution adjusted to alkalinity is heat-processed. By this heat treatment, a useful compound is produced by cyclizing the resveratrol. Such useful compounds include those represented by formula (1):

Formula (2):

Formula (3):

The resveratrol derivative of the kind shown by these is mentioned.

  In the resveratrol derivative, the carbon-carbon double bond may be a trans form or a cis form, and includes a mixture of a cis form and a trans form.

  The resveratrol derivative may be a pharmaceutically acceptable salt thereof.

  Examples of the pharmaceutically acceptable salt of the resveratrol derivative include alkali metal salts such as lithium salt, sodium salt and potassium salt; alkaline earth metal salts such as magnesium salt, calcium salt and barium salt; aluminum salt Metal hydroxide salts such as aluminum hydroxide salts; amines such as alkylamine salts, dialkylamine salts, trialkylamine salts, alkylenediamine salts, cycloalkylamine salts, arylamine salts, aralkylamine salts, heterocyclic amine salts Salts; amino acid salts such as α-amino acid salts and ω-amino acid salts; peptide salts or primary, secondary, tertiary, or quaternary amine salts derived therefrom. These pharmacologically acceptable salts can be used alone or in admixture of two or more.

In the present invention, the cyclization reaction for forming the resveratrol derivative refers to a reaction in which resveratrol forms a 6-membered ring by polymerization reaction.
In order to efficiently advance the cyclization reaction, the heating temperature of the resveratrol-containing solution adjusted to be alkaline is preferably adjusted to 110 ° C. or higher. Also, considering the boiling point of the solvent used, pressure heating is desirable. For example, the resveratrol-containing solution adjusted to alkalinity is placed in an open container, the container is heated at a high temperature exceeding the boiling point of the solvent, the resveratrol-containing solution adjusted to alkalinity is placed in a sealed container, and the container is The temperature of the resveratrol-containing solution adjusted to be at least partially alkaline, such as heating, pressure heating using a retort apparatus or autoclave, pressure heating using a supercritical apparatus, a pressure cooker or the like is 110. Heating is preferably performed so that the temperature reaches at least ° C. From the viewpoint of recovery efficiency of the resveratrol derivative and the like, it is more preferable that the temperature of the resveratrol-containing solution adjusted to be alkaline is 110 ° C. to 150 ° C. uniformly. The heating time is not limited as in the case of the heating temperature, and may be a time condition in which the target reaction efficiently proceeds. In particular, the heating time depends on the heating temperature, and it is desirable to set the heating time according to the heating temperature. For example, when heating near 130 ° C., a heating time of 30 to 120 minutes is desirable. Further, the heating may be performed once or may be repeated repeatedly in a plurality of times. In the case of heating in a plurality of times, it is preferable to add a new solvent to supplement the evaporated solvent.

The completion of the production reaction of the resveratrol derivative represented by the above formulas (1) to (3) by the heat treatment may be judged by confirming the production amount by component analysis by HPLC as described later, for example. . Specifically, it may be evaluated whether or not the generation is confirmed on the chromatogram.
The amount of the resveratrol derivative represented by the above formulas (1) to (3) is preferable because the amount of the resveratrol derivative represented by the formulas (1) to (3) increases in the composition having the exercise-like action of the present invention. Specifically, the total amount of each component may be 1% by weight or more in the composition having a motion-like action of the present invention.
Moreover, there is no limitation in particular about each content ratio of the resveratrol derivative represented by Formula (1), Formula (2), and Formula (3).

Further, in the case where improvement in flavor and further enhancement of functionality are desired, the reactant is concentrated to increase the concentration of the resveratrol derivative represented by the formulas (1) to (3), or the reactant is Desalting and purification can be performed to increase the concentration of the resveratrol derivative represented by the formulas (1) to (3). The concentration and purification can be performed by a known method. For example, the resveratrol derivative represented by the formulas (1) to (3) can be concentrated by extraction using a solvent extraction method such as chloroform, ethyl acetate, ethanol, or methanol or a supercritical extraction method using carbon dioxide gas. It is also possible to perform concentration and purification using column chromatography. For example, it is possible to concentrate using a synthetic adsorbent as a carrier. For the concentration and purification, a membrane treatment method such as a recrystallization method or an ultrafiltration membrane can be applied.
Among them, by using a synthetic adsorbent, the resveratrol derivative represented by the above formulas (1) to (3) is adsorbed and then eluted and then can be easily concentrated and purified. Examples of the synthetic adsorbent include aromatic synthetic adsorbents such as Diaion (registered trademark) HP series and Sepabeads (registered trademark) SP series manufactured by Mitsubishi Chemical Corporation, and Amberlite (registered trademark) manufactured by Organo Corporation. ) Styrenic synthetic adsorbents such as XAD series.

  Moreover, a powdery solid can be obtained by removing the solvent from the concentrate or product by drying under reduced pressure or freeze drying as necessary.

  In addition, when the composition having the exercise-like action of the present invention is produced using only safe raw materials, it can be used as it is for foods, pharmaceuticals, quasi drugs or animal feeds. For example, when a naturally occurring resveratrol is dissolved in a water-containing ethanol solvent and an alkalizing agent such as sodium hydroxide, sodium carbonate, or sodium hydrogen carbonate is added and heat-treated, the resulting reaction solution can be used as a food or pharmaceutical product. It can be used as one of raw materials for quasi drugs or animal feeds.

  Moreover, you may improve the solubility of the obtained composition using a well-known method. For example, there is a method of adding cyclodextrin, but there is no particular limitation.

  The composition having the exercise-like action of the present invention containing the resveratrol derivative represented by the formulas (1) to (3) obtained as described above has an excellent exercise-like action. Therefore, the composition having the exercise-like action of the present invention is a composition having an excellent exercise-like action, and can be suitably used as a muscle enhancer, a myostatin secretion inhibitor, and an HDL cholesterol raising agent.

  The composition having a motion-like action of the present invention may comprise only the composition as an active ingredient, but it may be a solution obtained by dissolving the composition in a solvent such as ethanol or an ethanol-containing aqueous solution, or a known method Or an emulsion or suspension. The content of the composition having a motion-like action of the present invention in the solution, emulsion or suspension may be 0.001% by weight or more as the solid content.

  The composition having a motion-like action of the present invention may be formulated as a pharmaceutical product. There are no particular limitations on the form of the preparation, and examples include parenteral preparations such as injections, suppositories, eye drops, ointments, aerosols, tablets, coated tablets, powders, fine granules, granules, capsules, and liquids. , Pills, suspensions, emulsions, lozenges, chewable tablets, syrups and other oral preparations. For formulation, pharmaceutically acceptable carriers, excipients, lubricants, binders, disintegrants, diluents, stabilizers, tonicity agents, pH adjusters, buffers, etc. are used. It is done.

  Carriers and excipients include, for example, lactose, sucrose, sodium chloride, glucose, maltose, mannitol, erythritol, xylitol, maltitol, inositol, dextran, sorbitol, albumin, urea, starch, calcium carbonate, kaolin, crystalline cellulose , Silicic acid, methylcellulose, glycerin, sodium alginate, gum arabic and mixtures thereof.

Examples of the lubricant include purified talc, stearate, borax, polyethylene glycol, and a mixture thereof.
Examples of the binder include simple syrup, glucose solution, starch solution, gelatin solution, polyvinyl alcohol, polyvinyl ether, polyvinyl pyrrolidone, carboxymethyl cellulose, shellac, methyl cellulose, ethyl cellulose, water, ethanol, potassium phosphate, and a mixture thereof. Can be mentioned.

  Examples of the disintegrant include dry starch, sodium alginate, agar powder, laminaran powder, sodium hydrogen carbonate, calcium carbonate, polyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate, stearic acid monoglyceride, starch, lactose and mixtures thereof. Is mentioned.

Examples of the diluent include water, ethyl alcohol, macrogol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, polyoxyethylene sorbitan fatty acid esters, and mixtures thereof.
Examples of the stabilizer include sodium pyrosulfite, ethylenediaminetetraacetic acid, thioglycolic acid, thiolactic acid, and a mixture thereof.

  Examples of the isotonic agent include sodium chloride, boric acid, glucose, glycerin and a mixture thereof.

  Examples of the pH adjuster and buffer include sodium citrate, citric acid, sodium acetate, sodium phosphate, and mixtures thereof.

  Furthermore, the composition having a motion-like action of the present invention contains a bulking agent, a solubilizer, a dispersant, a suspending agent, an emulsifier, an antioxidant, a bacteria inhibitor, a coloring agent, a corrigent, a corrigent and the like. Also good.

  In addition, tablets, pills, granules, and capsules containing granules may be sugar-coated with sugars such as sucrose and sugar alcohols such as maltitol, gelatin, hydroxypropylcellulose, hydroxypropylmethylcellulose as necessary. It can also be applied with a coating. Moreover, you may coat | cover with the film of a gastric or enteric substance. Moreover, in order to improve the solubility of a formulation, a well-known solubilization process can also be performed.

  When using the composition having the exercise-like action of the present invention, for example, the amount of intake of the composition having the exercise-like action of the present invention is not particularly limited as long as the desired effect can be obtained. Although it is appropriately selected according to the form, age, sex, constitution and other conditions of the patient, the type of disease and its degree, etc., for example, about 0.1 mg to 1,000 mg / kg as a solid content per adult day It can be taken and divided into 1 to 4 times a day.

  Further, the composition having a motion-like action of the present invention may be formulated into a quasi-drug form. Although it does not specifically limit as a quasi-drug, For example, nutritional supplement quasi-drugs, such as a drink, are preferable. In this case, the content of the extract as an active ingredient in the quasi drug may be about 0.001 to 30% by weight as the solid content.

  In addition, since the composition having the exercise-like action of the present invention is excellent in safety, not only for humans, for example, non-human animals such as rats, mice, guinea pigs, rabbits, sheep, You may mix | blend with mammals, such as a pig, a cow, a horse, a cat, a dog, a monkey, a chimpanzee, birds, amphibians, a reptile, etc., or feed. As feed, for example, livestock feed used for sheep, pigs, cattle, horses, chickens, etc., feed for small animals used for rabbits, rats, mice, etc., feed for seafood used for eel, Thailand, yellowtail, shrimp, etc., dogs, Pet foods used for cats, small birds, squirrels, etc. are listed.

  EXAMPLES Next, although this invention is demonstrated in detail based on an Example, this invention is not limited only to this Example.

(Example 1: Preparation of resveratrol alkaline heat treatment composition)
The resveratrol alkaline heat treatment composition was performed according to the following procedure. To 40 g of trans-resveratrol (manufactured by TECNO SCIENCE), 400 mL of 10% aqueous sodium carbonate (manufactured by Wako Pure Chemical Industries, Ltd.) is added to obtain a resveratrol-containing aqueous solution (pH 9.9). It was. This resveratrol-containing solution was heated at 128 ° C. for 120 minutes in a pressure cooker (zero vitality pan, manufactured by Asahi Light Metal Industry Co., Ltd.) to prepare a resveratrol derivative-containing solution.

  Next, the resveratrol derivative-containing solution was supplied to 4.0 kg of synthetic adsorbent Diaion (registered trademark) HP-20 (manufactured by Mitsubishi Chemical Corporation). The synthetic adsorbent was washed with 24 L of distilled water and eluted with 16 L of 100% ethanol, and the solvent was removed from the resulting eluate by drying under reduced pressure to obtain 22 g of a solid resveratrol alkaline heat treatment composition. .

  The obtained resveratrol alkaline heat treatment composition was dissolved in methanol at a concentration of 2 mg / mL, 10 μL of which was analyzed by HPLC.

HPLC analysis was performed under the following conditions.
Column: CAPCELL PAK UG80 column (4.6 mm ID × 250 mm, manufactured by Shiseido Co., Ltd.)
Mobile phase A: 0.1% trifluoroacetic acid (TFA, manufactured by Wako Pure Chemical Industries, Ltd.) / H ₂ O
Mobile phase B: 0.1% TFA / acetonitrile (manufactured by Wako Pure Chemical Industries, Ltd.)
Gradient (volume%): 100% A / 0% B to 0% A / 100% B for 33 minutes, 0% A / 100% B for 7 minutes (all straight lines)

  The obtained chromatogram is shown in FIG. In the resveratrol alkaline heat treatment composition, a plurality of peaks different from the resveratrol peak (elution time of 15 minutes) could be detected.

  Next, among the obtained reactants, the compounds contained in the peaks shown in (1), (2) and (3) of FIG. 1 were isolated by preparative HPLC and dried by a conventional method. ), The compound contained in the peak shown in (2) was a light brown powdery substance, and the compound contained in the peak shown in formula (3) was a brown powdery substance.

  Next, the molecular weights of the three kinds of compounds were measured by high resolution negative-FAB-MS (fast atom bombardment mass spectrometry) according to the method described in Example 2 of JP2011-251914A, and nuclear magnetic resonance was performed. As a result of (NMR) measurement, the compound contained in the peak of (1) is a compound represented by the above formula (1) (hereinafter referred to as UHA4002), and the compound contained in the peak of (2) is the above formula. It was found that the compound contained in the peak of the compound represented by (2) (hereinafter referred to as UHA4003) (3) was the compound represented by formula (3) (hereinafter referred to as UHA4022).

  Further, UHA4002, UHA4003, and UHA4022 are contained in the same manner as described above except that a resveratrol-containing material (trade name “Vineatrol 20M” manufactured by Sunbright Co., Ltd.) is used instead of resveratrol. A resveratrol alkaline heat treatment composition could be prepared.

  In the following Examples, it investigated using the resveratrol alkali heat processing composition manufactured from the resveratrol.

(Example 2: Examination of muscle strengthening action)
The muscle strengthening action was examined as the exercise-like action of the resveratrol alkaline heat treatment composition (hereinafter also referred to as the present invention) obtained in Example 1.
For verification of the muscle strengthening action of the product of the present invention, a 9-week-old SD rat male (manufactured by CLEA Japan, Inc.) was used. Specifically, 8 weeks old SD rats were preliminarily raised (feed: normal diet) for 7 days, and then each group was divided into 2 groups (high fat diet) so that the average body weight of each group was almost equal. Group, high fat diet group containing 1% of the product of the present invention), and each was allowed to freely ingest the test feed for 7 days.
After the test, the animals were fasted for 16 hours and then necropsied, and the quadriceps, long extensor, anterior tibial, soleus and plantar muscles were collected. Each muscle weight was measured, and the muscle weight per body weight (g%) was calculated. Statistical analysis was performed using Microsoft's Excel t-test, with a significance level of less than 5%.

  The results are shown in FIGS. As shown in FIG. 2, a significant increase in muscle was observed with the product of the present invention, and a significant increase in total muscle weight was also observed as shown in FIG. Therefore, it can be seen that the product of the present invention is useful as a novel muscle enhancer.

(Example 3: Examination of myostatin secretion inhibitory effect)
The myostatin secretion inhibitory action was examined as the exercise-like action of the product of the present invention. For verification of the myostatin secretion inhibitory action of the product of the present invention, 9-week-old SD rat male (manufactured by CLEA Japan, Inc.) was used. Specifically, 8 weeks old SD rats were preliminarily raised (feed: normal diet) for 7 days, and then 4 groups (high fat diet) with 5 animals in each group so that the average body weight of each group was almost equal. Group, 0.1% high fat diet containing the product of the present invention, 0.5% high fat diet containing the product of the present invention, and 1% high fat diet containing the product of the present invention) I let you.
After the test, the blood was collected after fasting for 16 hours, and serum and plasma were prepared. The myostatin concentration in plasma obtained using a myostatin ELISA kit (manufactured by USCN) was quantified. The method was performed according to the protocol attached to the kit. Statistical analysis of the results obtained was performed using Microsoft's Excel t-test, with a significance level of less than 5%.

  The results are shown in FIG. As shown in FIG. 4, a significant decrease in myostatin concentration was observed with the product of the present invention. Therefore, it can be seen that the product of the present invention is useful as a novel muscle enhancer accompanied by suppression of myostatin secretion.

(Example 4: Examination of HDL cholesterol elevation action)
As an exercise-like action of the product of the present invention, the HDL cholesterol raising action was examined. Examination of the HDL cholesterol raising action of the product of the present invention was performed using the rat serum obtained in Example 3. HDL cholesterol and LDL / VLDL cholesterol were measured using HDL and LDL / VLDL quantification kit (manufactured by Biovision). The method was performed according to the protocol attached to the kit. Statistical analysis of the results obtained was performed using Microsoft's Excel t-test, with a significance level of less than 5%.

  The results are shown in FIG. As shown in FIG. 5, a significant increase in HDL cholesterol was observed with the product of the present invention. Therefore, it can be seen that the product of the present invention is useful as a novel HDL cholesterol raising agent.

  Note that the present inventors have described the resveratrol derivatives represented by the above formulas (1) to (3) as “a white adipocyte brown-like cell differentiation promoter” (Japanese Patent Application No. 2013-157659) and “slow muscle formation”. Although "accelerator" (Japanese Patent Application No. 2013-157660) has been reported, it has not been reported that the resveratrol derivative is related to the motility-like action such as muscle strengthening action, myostatin secretion inhibiting action, and HDL cholesterol raising action. Therefore, exercise-like effects such as muscle strengthening action and HDL cholesterol raising action are new uses found for the first time in the present invention.

(Example 5: Food containing the product of the present invention)
1 g of the product of the present invention obtained in the same manner as in Example 1 was dissolved in 100 mL of ethanol in advance, and 500 g of sugar and 400 g of starch syrup were mixed and dissolved therein, and 100 g of fresh cream, 20 g of butter, 70 g of condensed milk, and 1.0 g of emulsifier were added. After mixing, the pressure was reduced by −550 mmHg in a vacuum kettle and concentrated under a condition of 115 ° C. to obtain a milk hard candy having a moisture value of 3.0% by weight. This milk hard candy is not only easy to eat as a confectionery, but also can be used as a functional food that is expected to increase muscle and raise HDL cholesterol.

(Example 6: Drug containing the product of the present invention)
1 g of the product of the present invention obtained in the same manner as in Example 1 was dissolved in ethanol, and the resulting solution was adsorbed on microcrystalline cellulose and dried under reduced pressure. 10 parts by weight of the product of the present invention adsorbed on the microcrystalline cellulose, 23 parts by weight of corn starch, 12 parts by weight of lactose, 8 parts by weight of carboxymethyl cellulose, 32 parts by weight of microcrystalline cellulose, 4 parts by weight of polyvinylpyrrolidone, 3 parts by weight of magnesium stearate Then, 8 parts by weight of talc was mixed and tableted to obtain tableted tablets containing the product of the present invention. This tablet can also be used as a therapeutic agent for muscular dystrophy with weakened muscles and low HDL cholesterolemia.

(Example 7: Quasi-drug containing the product of the present invention)
10 g of an ethanol solution obtained by dissolving 1.2 g of the product of the present invention obtained in the same manner as in Example 1 in ethanol, 20 g of taurine, 0.12 g of vitamin B1 nitrate, 0.6 g of sodium benzoate, 4 g of citric acid, 60 g of sugar and 10 g of polyvinylpyrrolidone were dissolved in purified water, and the total amount was made up to 1000 mL. The pH was adjusted to 3.2 using dilute hydrochloric acid. 50 ml of 1000 ml of the obtained solution was filled in a glass bottle and sterilized at 80 ° C. for 30 minutes to obtain a drink as a quasi drug. This drink can also be used as a quasi-drug for the purpose of strengthening muscles and raising HDL cholesterol.

(Example 8: Animal feed containing the product of the present invention)
50 g of the product of the present invention obtained in the same manner as in Example 1, powdered soybean meal, fish mince, rice bran, wheat and corn are mixed, and soybean oil, vitamins and minerals are added to this, and then molded. 1 kg of feed was obtained. The obtained animal feed can also be used as an animal feed for the purpose of promoting growth and improving meat quality.

Claims (6)

  A composition having a motion-like action, which is obtained by heat-treating resveratrol under alkaline conditions. Formula (1):

Formula (2):

And formula (3):

The composition which has the exercise-like effect | action of Claim 1 containing three types of resveratrol derivatives shown by these.   A muscle strengthening agent comprising the composition having exercise-like action according to claim 1 or 2.   The muscle strengthening agent according to claim 3, wherein the exercise-like action is a myostatin secretion inhibitory action.   An HDL cholesterol-elevating agent comprising the composition having exercise-like action according to claim 1 or 2.   An animal feed comprising the composition having exercise-like action according to claim 1 or 2.

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