KR20170112808A - Composition for increasing muscle mass comprising dieckol and preperation method of the same - Google Patents

Abstract

The food composition for promoting muscle growth of the present invention contains diokor or a salt thereof to promote the differentiation and proliferation of myoblasts to grow into muscle cells, thereby promoting muscle growth. In addition, the pharmaceutical composition of the present invention can treat or prevent muscular dystrophy (Sacopenia) by including diecor or a pharmaceutically acceptable salt thereof as an active ingredient.

Description

TECHNICAL FIELD The present invention relates to a composition for accelerating muscle growth and a method for producing the same,

The present invention relates to a composition for promoting muscle growth and a method for producing the same, and more particularly, to a food or pharmaceutical composition containing Dieckol or a salt thereof and having a function of promoting muscle cell growth and a method for producing the composition.

Muscle is a very important body organ in energy metabolism and athletic performance. Athletes concentrate on improving body strength to increase performance.

In order to improve physical strength, supplementation with athletic performance is needed along with training and diet. In particular, nutritional energy supplements provide the necessary nutrients and energy quickly and efficiently, and are a positive factor in improving exercise performance by delaying energy accumulation and fatigue.

Doping is defined as athletes taking medications forbidden to increase performance or using prohibited methods.

In order to prevent doping, the World Anti-Doping Agenct (WADA) announces prohibited drugs and prohibition methods each year in September to create an international list of prohibited lists. WADA is not responsible for any activity that uses prohibited methods such as taking, inhaling, injecting, injecting, skin adhesion and blood products, blood transfusions, anthropogenic oxygen ingestion or forbidding, It is defined as an anti-doping rule violation even to try.

Anabolic steroids are a testosterone derivative of the male hormone, an anabolic effect that strengthens the muscle and an androgenic effect that acts as a male hormone. It is a synthetic steroid hormone.

Anabolic steroids help strengthen athletes' performance by strengthening muscles and bones through increased protein synthesis in the body. However, continuous administration of steroids has a slow and fatal side effect on health such as liver tumors, kidney diseases, endocrine disorders, hypertension, male mastication, depression, hair loss, and jaundice.

Korean Registered Patent No. 10-1091775 / December 12, 2011. Announcement Korean Registered Patent No. 10-1247239 / March 23, 2013. Announcement

It is an object of the present invention to provide a composition for promoting muscle growth. It has been confirmed that Dieckol stimulates the differentiation and growth of muscle cells by fractionating the extract of Ganoderma lucidum, and the composition for promoting muscle growth or muscle And to provide a pharmaceutical composition for treating or preventing Sacopenia. It is another object of the present invention to provide a method for extracting and fractionating diecor from menthol.

In order to achieve the above object, the food composition for promoting muscle growth according to an embodiment of the present invention contains Dieckol, a derivative thereof, or a salt thereof.

The diacol may be isolated from Ecklonia cava .

The diecol may be isolated from the ethyl acetate fraction of Ecklonia cava extract.

The food composition may be any one selected from the group consisting of a health functional food, a food additive and a beverage additive.

A pharmaceutical composition for the treatment or prevention of Sacopenia according to another embodiment of the present invention contains Dieckol, a derivative thereof or a pharmaceutically acceptable salt thereof.

The pharmaceutical composition may be one that activates the expression of p-Akt and p-FoxO, decreases the expression of Smad4, and activates MyoD expression.

The diacol may be isolated from Ecklonia cava .

The diecol may be isolated from the ethyl acetate fraction of Ecklonia cava extract.

According to another embodiment of the present invention, there is provided a method for producing a composition containing diacol , comprising the steps of: extracting Ecklonia cava with an alcohol-containing extraction solvent to obtain a perennial alcohol extract; A fractionating step of adding ethyl acetate to the chewy alcohol extract to obtain an ethyl acetate fraction; And separating the fraction containing the diacol component from the ethyl acetate fraction.

The composition containing the diacol may be a food composition for promoting muscle growth or a pharmaceutical composition for treating or preventing muscular dystrophy (Sacopenia).

The texture of the extraction step may be freeze-dried freeze-dried pulverized material.

The extraction solvent may be an aqueous alcohol solution, alcohol or alcohol containing 40 to 99% by volume of alcohol.

The cutaneous alcohol extract of the fractionation step may be a solution in which the pulverized cuttlefish alcohol extract is suspended in water.

The separation step may be performed using reverse phase high performance liquid chromatography reversed-phase HPLC.

Hereinafter, the present invention will be described in more detail.

The inventors of the present invention have continued to search for a fluorotannin-based active substance having an activity of inducing muscle growth, and found that Diokor, a kind of phlorotannin isolated from menthol extract, promotes muscle growth Specifically, C2C12 myocyte proliferation effect, myocyte differentiation effect, and creatine kinase activity effect, and completed the present invention.

A food composition according to one embodiment of the present invention contains Dieckol, a derivative thereof, or a salt thereof. The food composition may be a functional food composition, specifically, a functional food composition for promoting muscle growth, for enhancing exercise performance, for promoting muscle growth, or for promoting recovery of muscle fatigue.

Diekol, 4- [4- [6- (3,5-dihydroxyphenoxy) -4,7,9-trihydroxydibenzo-p-dioxin-2-yl] oxy-3,5-dihydroxyphenoxy] dibenzo-pdioxin- , 3,6,8-tetrol) is a kind of phlorotannin found in some algae.

In one embodiment of the present invention, the food composition for promoting muscle growth of the present invention can be used as a health functional food which can be ingested routinely for muscle augmentation, inhibition of muscle aging, or exercise function improvement.

In this specification, food refers to a natural product or a processed product containing one or more specific nutrients, and specifically refers to a state in which it can be directly eaten through a certain degree of processing. In general terms, , Functional foods, beverages, food additives, and beverage additives.

In this specification, food refers to a natural product or a processed product containing one or more nutrients. Specifically, it means that the product can be directly eaten through a certain degree of processing. In general terms, Functional foods, beverages, food additives, and beverage additives.

In the food composition, the amount of the diecor and the like may be 0.00001 wt% or more, specifically 0.1 wt% or more, 99 wt% or less, specifically 80 wt% or less, more specifically 60 wt% or less And when the food is a beverage, it may be contained in a proportion of not less than 0.001 g, specifically not less than 0.01 g, not more than 50 g, specifically not more than 10 g, more specifically not more than 2 g based on 100 ml of the whole volume of the food But is not limited thereto.

The food composition of the present invention may contain sweetening agents, flavors, physiologically active ingredients, minerals and the like in addition to the active ingredients thereof. Sweetening agents may be used in an amount that sweetens the food in a suitable manner, and may be natural or synthetic. Specifically, natural sweeteners are used. Examples of the natural sweeteners include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose and maltose. Flavors may be used to enhance taste or flavor, both natural and synthetic. Specifically, a natural one is used. When using natural ones, the purpose of nutritional fortification can be performed in addition to the flavor. Examples of natural flavoring agents include those obtained from apples, lemons, citrus fruits, grapes, strawberries, peaches, and the like, or those obtained from green tea leaves, Asiatica, Daegu, Cinnamon, Chrysanthemum leaves and Jasmine. Also, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, banks and the like can be used. The natural flavoring agent may be a liquid concentrate or a solid form of extract. Synthetic flavors may be used in some cases, and synthetic flavors may be esters, alcohols, aldehydes, terpenes, and the like. Examples of the physiologically active substance include catechins such as catechin, epicatechin, gallocatechin and epigallocatechin, and vitamins such as retinol, ascorbic acid, tocopherol, calciferol, thiamine and riboflavin. As the mineral, calcium, magnesium, chromium, cobalt, copper, fluoride, germanium, iodine, iron, lithium, magnesium, manganese, molybdenum, phosphorus, potassium, selenium, silicon, sodium, sulfur, vanadium and zinc can be used. Proteins that are applied to general protein supplements can also be included.

In addition, the food composition of the present invention may contain preservatives, emulsifiers, acidifiers, thickeners and the like as needed in addition to the sweeteners mentioned above. Such preservatives, emulsifiers and the like are preferably added in a very small amount as long as they can attain an application to which they are added. The term " trace amount " means, when expressed numerically, in the range of 0.0005% by weight to about 0.5% by weight based on the total weight of the food composition. Examples of the preservative which can be used include calcium sodium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate and EDTA (ethylenediaminetetraacetic acid). Examples of the emulsifier which can be used include acacia gum, carboxymethyl cellulose, xanthan gum, pectin and the like. Examples of the acidulant that can be used include acid, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, and phosphoric acid. Such an acidulant may be added so that the food composition has a proper acidity for the purpose of inhibiting the growth of microorganisms other than the purpose of enhancing the taste. Agents that may be used include suspending agents, sedimentation agents, gel formers, bulking agents and the like.

In particular, the food composition of the present invention can be manufactured in the form of a health functional food, a functional food (including a beverage), a food additive or a beverage additive.

In the present specification, the term "health functional food" refers to a food prepared by adding the diacol or the like to a food material such as a beverage, a tea, a spice, a gum or a confection, or an encapsulated, powdered or suspension liquid. However, unlike ordinary drugs, there is an advantage that there is almost no side effects that may occur when a drug is taken for a long time using a food as a raw material. For example, the food composition of the present invention, which is a health functional food, may be added in the form of a functional additive of a protein supplement.

The health functional food of the present invention is very useful because it can be ingested routinely. The health functional food may include a food-acceptable food-aid additive, and may further comprise suitable carriers, excipients and diluents conventionally used in the production of functional foods. The specific kind is the same as described above.

In addition, the food composition may have any one form selected from the group consisting of tablets, granules, powders, capsules, liquid solutions and rings, and the preparation method thereof may be applied to the manufacture of conventional food additives, health functional foods, and the like Can be applied.

A pharmaceutical composition according to another embodiment of the present invention contains Dieckol, a derivative thereof or a pharmaceutically acceptable salt thereof and has the use of Sacopenia treatment or prevention.

The hypopituitarism may be a proximal axis due to muscle atrophy due to loss of muscle tissue resulting from not using the muscle, muscle atrophy due to the muscle itself, or damage to the nerve that dominates the muscle.

The pharmaceutical composition may be one that activates the expression of p-Akt and p-FoxO, decreases the expression of Smad4, and activates MyoD expression.

The pharmaceutical composition according to the present invention may further comprise a carrier and a vehicle commonly used in the pharmaceutical field. Specifically, a buffering material (e.g., various phosphates, glycine, sorbic acid, potassium sorbate, a partial glyceride mixture of saturated vegetable fatty acids), water, salts or electrolytes (e.g., protamine sulfate, disodium hydrogen phosphate, But are not limited to, sodium chloride and zinc salts), colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose based substrates, polyethylene glycol, sodium carboxymethyl cellulose, polyarylate, wax or wool.

In addition, the pharmaceutical composition may be formulated in the form of granules, powders, coated tablets, tablets, capsules, suppositories, syrups, juices, suspensions, emulsions, drops, injections or sustained release formulations of active compounds. The pharmaceutical composition for the treatment or prevention of hypersensitivity can be administered orally or parenterally in various formulations. In the case of formulation, a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, . ≪ / RTI >

Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may be prepared by mixing the pharmaceutical composition of the present invention with at least one excipient such as starch, calcium carbonate, Sucrose, lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups and the like. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used. have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and the like may be used.

The dosage of the pharmaceutical composition according to the present invention may vary depending on factors such as the severity of the disease, the kind and amount of the active ingredient and other ingredients contained in the composition, the type of the formulation and the age, weight, general health status, sex and diet, The route and fraction of the composition, the duration of the treatment, the drug being co-administered, and the like. For example, in the case of an adult, the pharmaceutical composition of the present invention may be administered at a dose of 0.01 to 500 mg / kg once to several times a day. Preferably 0.1 to 200 mg / kg, more preferably 0.1 to 100 mg / kg, once or three times a day. In addition, the formulated unit dosage form can be administered several times at predetermined time intervals as needed.

According to another embodiment of the present invention, there is provided a method for producing a composition containing diecor , comprising the steps of: extracting Ecklonia cava with an alcohol-containing extraction solvent to obtain a perch alcohol extract; A fractionating step of adding ethyl acetate to the chewy alcohol extract to obtain an ethyl acetate fraction; And separating the fraction containing the diacol component from the ethyl acetate fraction.

The texture of the above-mentioned extraction step may be freeze-dried freeze-dried matter, and the freeze-dried freeze-dried matter may be a milled powder prepared in the form of powder. In this way, when the seaweeds are collected, they are washed and then dried and stored in a freeze-drying method, the seaweeds can be stored for a relatively long period without deterioration of the seaweed, which is a seaweed, while minimizing the loss of various useful ingredients including organic matters. The efficiency of extraction can be improved.

The extraction solvent in the extraction step is preferably water, alcohol or a mixed solvent thereof. Examples of the alcohol include methanol, ethanol, propanol, isopropanol, (butanol), and combinations thereof, and the alcohol may be applied.

Preferably, the extraction solvent may be an alcohol aqueous solution containing alcohol in an amount of 40 to 99% by volume, alcohol or alcohol, and most preferably an aqueous solution of ethanol in an amount of 70 to 90% by volume. When such an extraction solvent is used, the extraction of the fluorotentin component such as diacol, which is an effective component, can be efficiently performed.

The extraction may be carried out by applying an extracting solvent having a volume of 1 to 7 times the concentration to the sensation, followed by cooling at room temperature, cold-cooling, or heat extraction, but the present invention is not limited thereto.

The sensory alcohol extract of the extracting step may be one obtained by removing the solid matter from the extract obtained in the extraction step and concentrating under reduced pressure or by concentration under reduced pressure.

The cutaneous alcohol extract of the fractionation step may be applied in the form of a solution in which the pulverized cuttleberry alcohol extract is suspended in water.

It is preferable to apply ethyl acetate as a fraction solvent to the above fraction. When ethyl acetate is applied in various organic solvents used for the fractionation, it is advantageous to efficiently perform deoxidization as an active ingredient.

The separation step may be performed using reversed-phase high performance liquid chromatography reversed-phase HPLC, which separates the active ingredient diacol from the ethyl acetate fraction, and specifically, the celite fraction, the sephadex fraction and the HPLC And separating and purifying the diacol compound of the present invention through a separation process.

The diecor is safe in the body isolated from the gentian extract and promotes muscle growth, and can be used as a food or pharmaceutical composition for a new use of a diacol compound as a composition for muscle growth.

INDUSTRIAL APPLICABILITY The food composition for promoting muscle growth of the present invention promotes the growth of muscle cells to differentiate and proliferate into muscle cells to promote muscle growth and has a safe and excellent muscle growth effect on the human body, Those who need to increase muscle strength and exercise capacity can get excellent effects on ingestion. In addition, the pharmaceutical composition of the present invention can treat or prevent muscular dystrophy (Sacopenia) by including diocor as an active ingredient. When the muscles are lost due to various causes or the muscle growth is required, the composition of the present invention may be applied to promote excellent muscle growth.

FIG. 1 is a graph showing the results of measurement of creatine kinase activity of Diekol in Example 1 of the present invention. FIG.
FIG. 2 is a graph showing the results of experiments in which the effect of diercourse on C2C12 myocyte proliferation and hyperplasia in 1) of Example 2 of the present invention.
FIG. 3 is a graph showing the results of measurement of the cell proliferation promoting effect of Diokor in Example 2-2) of the present invention. FIG.
FIG. 4 shows Western blotting results obtained by analyzing the effect of diacol on expression of a muscle growth regulator in Example 3 of the present invention.
FIG. 5 is a graph showing quantitative results of p-Akt and p-FoxO among the results of FIG.
FIG. 6 is a graph showing the results of p-Smad2 / 3 and Smad4 quantified from the results of FIG.
FIG. 7 is a graph showing the result of quantifying MyoD among the results of FIG. 4 above. FIG.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings so that those skilled in the art can easily carry out the present invention. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

Preparation Example: Isolation of Dieckol (DK) from the Fentanyl Extract

Ecklonia cava , a brown algae, was collected from March to June 2013 on the coast of Jeju Island and was washed 3 times to remove foreign matter on the surface and stored at -20 ℃.

The extract was filtered by adding 80% aqueous ethanol solution to the dried gum powder, and the solid content was removed. The dried gum powder was vacuum-dried to obtain powdery The ethanol extract of Ganoderma lucidum was obtained. Ethanol extract was suspended in water, and ethyl acetate was added to obtain an ethylacetate fraction (EtOAc, 46.27 g). This ethyl acetate fraction was separated by reverse-phase HPLC using a Shephadex LH-20 column, and a diacol compound (DK) was prepared and applied to the following experiments.

Preparation for experiment: C2C12 Myoblast  culture

C2C12 myofibroblast was used from ATCC. Dulbecco's Modified Eagle Medium (DMEM; Gibco, CA, USA) medium was used as a growth medium. Penicillin (100 U / ml) and streptomycin Respectively. During the experiment, the cell lines were cultured in a 37 ° C environment containing 5% CO ₂ .

Example 1: Measurement of creatine kinase activity of diecor

In muscle growth, creatine is converted to phosphocreatine (PCr) and adenosine triphosphate (ATP) is converted to adenosine diphosphate (ADP). Creatine kinase (CK) Is activated. Whether Diocor regulates the activity of creatine kinase was confirmed by the following method.

C2C12, which was normally grown, was added to a 6-well plate in an amount of 5 × 10 ⁴ / mL and adhered for 48 hours in a growth medium containing 10% FBS at 37 ° C and 5% CO ₂ . Two days later, the cells were washed twice with Dulbecco's Phosphate-Buffered Saline (DPBS) and differentiation was induced by maintaining 5% CO ₂ at 37 ° C in a medium containing 2% horse serum (HS). Two days later, the diacol compound was added in concentration by re-exchanging with a medium containing 2% HS. After 48 hours of treatment with the diacol compound, the cells were recovered and the activity of creatine kinase enzyme was measured. The results are shown in Fig.

Referring to the results of FIG. 1, it was confirmed that the creatine kinase activity was significantly higher than that of the negative control group but lower than that of the positive control group DHT (10 nM).

Example 2 Measurement of the Effect of Diekol on Muscle Cell Proliferation

1) Cell proliferation effect of diecor

C2C12, which was growing normally, was added to a 12-well plate in an amount of 5 × 10 ⁴ / mL and adhered for 48 hours in a growth medium containing 10% FBS at 37 ° C. and 5% CO ₂ . Two days later, the cells were washed twice with DPBS (A, day 0), and the diacol compound was added at different concentrations while changing to a medium containing 2% horse serum (HS).

After 48 hours (B, day 2) of treatment with the diacol compound, the cells were washed twice with DPBS. The degree of cell proliferation was confirmed through an Olympus microscope equipped with a CoolSNAP-Pro color digital camera in A and B, and the results are shown in Fig.

Cell proliferation and hyperplasia should occur in order to differentiate into C2C12 myoblasts. 2, cell proliferation and hyperplasia were observed when differentiation of C2C12 myofibroblasts into myocytes was carried out while treating the dioecol compound together.

2) Measurement of cell proliferation effect of diecor

To confirm the cell proliferation effect, C2C12, which was normally grown, was added to a 12-well plate at an amount of 5 × 10 ⁴ / mL. The cells were maintained in a growth medium containing 10% FBS at 37 ° C. and 5% CO ₂ for 48 hours Respectively. Two days later, the cells were washed twice with DPBS, and then the diacol compound was added in concentration by changing to a medium containing 2% horse serum (HS). After 48 hours of treatment with the diacol compound, MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) was added and incubated for 4 hours. Dimethylsulfoxide (DMSO) was added to dissolve the insoluble formazan crystals. The absorbance at 540 nm was measured to confirm the degree of cell proliferation. The results are shown in FIG.

Referring to the results shown in FIG. 3, when the diacol compound was treated with C2C12 myofibroblast differentiation, it was confirmed that C2C12 cells proliferated in a concentration-dependent manner except for the concentration of 40 nM there was.

Example 3: Expression analysis of muscle growth regulator of diacol compound

Myostatin (MSTN), a cell growth differentiation factor, binds to the activin type II receptor and Smad receives signal transduction. The intracellular Smad2 / 3 signaling is phosphorylated and p-Smad2 / 3 (phosphorylation-Smad2 / 3) binds to Smad4 to inhibit MyoD expression.

Insulin-like growth factor 1 (IGF-1), an insulin-like growth factor, inhibits signal transduction of muscle growth-negative regulators through phosphorylation of the phosphoatidylonositol 3-kinase (PI3K) / Akt pathway as a positive muscle growth regulator, . In addition, IGF-1 binds to the IGF-1 receptor to transmit signals. IGF-1, which binds to the receptor, induces overexpression of Akt and FoxO via PI3K pathway and increases muscle mass.

It is known that diecor compounds regulate the expression of p-Smad2 / 3 and Smad4, which are negative regulators of muscle growth in C2C12 cells, and the expression of p-Akt and p-FoxO, which are positive regulators of muscle growth in C2C12 cells Were confirmed by the following method.

C2C12, which was normally grown, was added to a 6-well plate at an amount of 5 × 10 ⁴ cells / mL and adhered for 48 hours in a growth medium containing 10% FBS at 37 ° C and 5% CO ₂ . Two days later, the cells were washed twice with Dulbecco's Phosphate-Buffered Saline (DPBS) and differentiation was induced by maintaining 5% CO ₂ at 37 ° C in a medium containing 2% horse serum (HS). Two days later, the diacol compound was added in concentration by re-exchanging with a medium containing 2% HS. Forty-eight hours after treatment with the diacrole compound, the cells were recovered and the expression of p-Smad2 / 3 and Smad4, the muscle growth negative regulators, was confirmed by western blotting analysis.

Specifically, the recovered cells were resuspended in lysis buffer [50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 1% Triton X-100, 0.1% SDS and 1 mM EDTA] ) Was centrifuged at 14,000 x g for 20 minutes at 4 째 C, and the protein content of the supernatant was measured using a BCA ^占 protein analysis kit and confirmed to contain 30 ^占 퐂 of protein. The supernatant was transferred to a membrane (nitrocellulose membrane, Bio-Rad, Hercules, Calif.) Using gel electrophoresis (10% sodium dodecyl sulfate-polyacrylamide gel) and blocked with 5% skim milk and 0.2% Tween 20 in TBS buffer . Membranes were incubated with primary antibodies (rabbit polyclonal anti-rabbit p-Smad2 / 3, p-Akt, p-FoxO1, MyoD Ab, 1: 500, Santa Cruz Biotechnology, (1: 500, Santa Cruz Biotechnology, Inc.) overnight at 4 ° C, washed with TTBS, and incubated with secondary antibody (goat anti-rabbit or anti-mouse IgG HRP conjugated secondary antibody, 1: 3,000 dilution) After the reaction, the results were observed using ECL Western blotting detection kit and FusionCapt Advance FX7 program (Vilber Lourmat, Australia). The results are shown in FIGS. 4 to 7, respectively.

Referring to the results of FIGS. 4 and 6, it was confirmed that the diacrole compound decreased the expression of p-Smad2 / 3 and decreased the expression of Smad4 significantly. In addition, referring to the results of FIGS. 4 and 5, it was confirmed that the diacol compound exhibited significant activation of p-Akt and p-FoxO expression.

In addition, since the expression of the protein of the muscle growth target gene MyoD is activated through the inhibition of the muscle growth negative regulator and the activity of the positive regulator, the effect of Diokor on the protein expression of the muscle growth target gene MyoD was also analyzed. Referring to the results of FIG. 4 and FIG. 7, it was confirmed that the diacol compound exhibited significant activation of MyoD expression.

Example  4: Diekol  Receptor binding assay of compounds in silico  molecular docking analysis)

1) muscle growth negative regulator receptor binding assay

The Smad signaling pathway, the muscle growth regulator, is signaled by the MSTN binding to the activin type II receptor. Inhibitors of MSTN signaling include follistatin, follistatin-like 3 protein, and growth and differentiation factor-associated serum protein-1 (GASP-1). These factors inhibit signal transduction by preventing active MSTN from binding to receptors .

The molecular docking simulation confirmed whether the diacol compound had a similar action to the inhibitory factors, and confirmed the binding ability of MSTN to the four active sites (active sites 1 to 4) of MSTN, and the results are shown in Table 1 below .

Ligand
Ligand Site Location
Site position Bonding energy
Binding energy
(kcal / mol) CDOCK action energy
CDOCK interaction energy (kcal / mol) DK Active site 1 -146.06 -50.77 Active site 2 -118.99 -40.01 Active site 3 -126.49 -47.71 Active site 4 -108.64 -38.27 DHT Active site 1 -50 -24.07 Active site 2 -48.16 -18.28 Active site 3 -51.51 -39 Active site 4 -40.08 -18.76

The results of Table 1 indicate that the diacrole compound binds to all four active stances of MSTN, particularly active site 1 (binding energy, -146.06 kcal / mol, CDOCK interaction energy, -50.77 kcal / mol) The in silico results strongly confirmed binding to site 3 (binding energy, -126.49 kcal / mol; CDOCK interaction energy, -47.71 kcal / mol).

2) muscle growth regulatory factor receptor binding assay

IGF-1 binds to the IGF-1 receptor, and Akt and FoxO, which regulate muscle growth, are signaled. The binding activity of IGF-1 receptor to the IGF-1 receptor was confirmed by a molecular docking simulation to confirm whether the diacrole compound had a similar action to IGF-1. The results are shown in Table 2 below.

Ligand
Ligand Bonding energy
Binding energy
(kcal / mol) CDOCK action energy
CDOCK interaction energy (kcal / mol) DK -118.71 -50.66 DHT -60.25 -20.33

Referring to the results in Table 2, the call diethoxy compound is IGF-1 receptor and has bonded to the active site (Binding energy, -118.71 kcal / mol; CDOCK interaction energy, -50.66 kcal / mol) in silico to The results were confirmed.

<Statistics Processing>

All experiments were performed independently 3 times (n = 3) and analyzed using the variance (ANOVA) test (using SPSS 12.0 statistical software) (one-way analyis). Significant differences were assessed using the Duncan and Tukey test, and P <0.05 or P <0.01 was considered statistically significant.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, Of the right.

Claims (6)

A food composition for promoting muscle growth, comprising Dieckol, a derivative thereof, or a salt thereof.
The method according to claim 1,
Wherein the food composition is any one selected from the group consisting of a health functional food, a food additive, and a beverage additive.
A pharmaceutical composition for the treatment or prophylaxis of diabetes mellitus (Sacopenia), which contains Dieckol, a derivative thereof or a pharmaceutically acceptable salt thereof.
The method of claim 3,
Wherein said pharmaceutical composition activates the expression of p-Akt and p-FoxO, reduces the expression of Smad4, and activates MyoD expression.
An extracting step of obtaining a perennial alcohol extract by adding an extractive solvent containing alcohol to Ecklonia cava ;
A fractionating step of adding ethyl acetate to the chewy alcohol extract to obtain an ethyl acetate fraction; And
And separating the fractions containing the diacol component from the ethyl acetate fraction to produce a composition having muscle growth promoting activity.
6. The method of claim 5,
Wherein the extract is a freeze-dried pulverized product and the extraction solvent is an alcohol aqueous solution, alcohol or alcohol containing 40 to 99% by volume of alcohol,
The cutaneous alcohol extract of the fractionation step is a solution in which the pulverized cuttlefish alcohol extract is suspended in water,
Wherein said separating step is carried out using reversed-phase high performance liquid chromatography reversed-phase HPLC.

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