KR102217264B1 - Composition for Preventing, Improving or Treating of muscular disease containing Codium SPP. algae extract - Google Patents

Abstract

The present invention relates to a composition for preventing, improving, or treating muscle diseases comprising an extract of seaweed in Cordium, and more particularly, a pharmaceutical composition for preventing or treating muscle diseases, comprising the extract of seaweed in Cordium as an active ingredient, It relates to a food composition for prevention or improvement. The composition containing the seaweed extract in Cordium of the present invention may be usefully used in preventing, improving, or treating muscle diseases, as well as as a food composition for improving exercise capacity or a composition for strengthening muscles.

Description

Composition for Preventing, Improving or Treating of muscular disease containing Codium SPP. algae extract}

The present invention relates to a composition for preventing, improving, or treating muscle diseases comprising an extract of seaweed in Cordium, and more particularly, a pharmaceutical composition for preventing or treating muscle diseases, comprising the extract of seaweed in Cordium as an active ingredient, It relates to a food composition for prevention or improvement.

Skeletal muscle is an organ that occupies the largest part of the human body and accounts for 40-50% of the total body weight, and plays an important role in various metabolic functions in the body, including energy homeostasis and heat generation. Muscle size is regulated by intracellular signaling pathways that induce anabolic or catabolism occurring within the muscle. When the signaling response that induces synthesis occurs more than the decomposition of muscle protein, muscle protein synthesis increases, resulting in hypertrophy or hyperplasia, which increases the size of the muscle (The Korea). Journal of Sports Science, 20(3): 1551-1561, 2011).

Human muscles decrease by more than 1% every year from the age of 40, and by the age of 80, the level of maximum muscle mass decreases by 50%, and the loss of muscles in old age is recognized as the most important factor in reducing overall physical function. As aging progresses, it is recognized that body shape changes such as muscle and fat content and skeletal distortion, and the prevalence of obesity due to muscle loss in old age is showing a continuous increasing trend at a level of over 30% worldwide. In the case of abnormal insulin secretion, it can cause muscle development disorders by not properly supplying energy to the cells, resulting in increased sarcopenia in diabetics than in the general population. In addition, muscle loss causes arthritis, back pain, and chronic pain to increase further, and can worsen urinary incontinence symptoms due to abdominal obesity, and injuries caused by fractures can increase depression in old age and lead to death. Sarcopenia is a major cause of poor quality of life associated with various diseases. It is known that sarcopenia is also closely related to senile chronic diseases such as osteoporosis, insulin resistance, and arthritis, and it is possible to suppress a decrease in physical activity due to aging through the prevention or improvement of sarcopenia.

The global advanced ataxia and muscle weakness treatment market reached about $14 billion in 2011, and after that, it is expected to grow at a compound annual growth rate of 9.4%, reaching about $23.5 billion in 2017. As treatments for muscle reduction, increased mitochondrial production, inhibition of muscle protein degradation, and anti-inflammatory drugs have been suggested, but there are no clear therapeutic drugs.

In addition, as a dietary method proposed to prevent sarcopenia, it was found that 25-30 g of high-quality protein should be consumed at each meal. This is because 4-5 eggs or about 120 g of chicken breast must be consumed at each meal, and it is difficult for ordinary people to practice daily life. Therefore, recently, many people are choosing protein supplements as an alternative, but protein supplements have a high possibility of side effects as they cause excessive protein intake. Moreover, if you have kidney disease, you cannot eat a high protein diet, and kidney function is also reduced with aging. Therefore, alternatives other than high protein intake are needed to prevent sarcopenia.

Accordingly, the present inventors conducted studies using various natural materials to develop a composition capable of strengthening muscles, and as a result, the auditory extract, a seaweed in Codium, showed the binding index of MHC and mRNA expression in myoblasts and myotubes. The present invention was completed by confirming that it has the effect of increasing and increasing the diameter of root canal cells.

Accordingly, it is an object of the present invention to provide a composition for preventing, improving or treating muscle diseases comprising an extract of seaweed in Codium as an active ingredient.

In addition, another object of the present invention is to provide a composition for strengthening muscles comprising a seaweed extract from Codium as an active ingredient.

In addition, another object of the present invention is to provide a food composition for enhancing exercise capacity, comprising an extract of seaweed in Codium as an active ingredient.

In order to achieve the above object, the present invention provides a composition for preventing, improving, or treating muscle diseases comprising a seaweed extract from Codium as an active ingredient.

In order to achieve another object of the present invention, the present invention provides a composition for strengthening muscles comprising a seaweed extract from Codium as an active ingredient.

In order to achieve another object of the present invention, the present invention provides a food composition for enhancing exercise capacity, comprising a seaweed extract in Codium as an active ingredient.

Hereinafter, the present invention will be described in detail.

The present invention provides a composition for preventing, improving, or treating muscle diseases, comprising an extract of seaweed in Codium as an active ingredient.

The composition according to the present invention may be a pharmaceutical composition or a food composition, and the pharmaceutical composition may be a composition comprising a seaweed extract from Cordium as an active ingredient, or a composition consisting of seaweed extract from Cordium as an active ingredient It may be, or it may be a composition consisting essentially of seaweed extract in Cordium as an active ingredient.

In the present specification, the term'comprising' is used in the same meaning as'including' or'characterized by', and in the composition or method according to the present invention, specifically Additional components or method steps, etc. not mentioned are not excluded. In addition, the term “consisting of” means excluding additional elements, steps, or ingredients that are not separately described. The term'essentially consisting of' means that, in the scope of the composition or method, it is possible to include substances or steps that do not substantially affect their basic properties, in addition to the substances or steps described. .

The'seaweed in a codium' of the present invention is widely distributed around the world as a green algae plant group, and is widely used for food in Korea, China, Japan, as well as the Philippines and Hawaii. It contains a lot of sugar, protein, and fiber, and contains a wider variety of minerals than land plants, so it is rich in calcium, potassium and iron. The height is 10~30cm, the thickness is 1.5~3mm, and the lower part is a bit thicker. Branches are split into deer antlers and grow straight, all reaching the same height to form a fan shape. The surface is smooth like a ridge, and when viewed under a microscope, a colorless, transparent thread-like tissue is irregularly entangled inside the branch. Young ones have hairs in the front, especially in the upper part, but fall off later, leaving traces of hair loss. The whole body is a structure in which several cell nuclei are contained in one cell, all connected without a membrane between cells. The pouch containing spores is club-shaped, and the length is 5-7 times the thickness, and the top is pointed. Since it has anthelmintic component, it was used as a roundworm medicine in the past, and water-soluble extracts are known to have a strong antibiotic action against bacteria.

In the present invention, the seaweed in the cordium is not limited thereto, but may be any one selected from the group consisting of a hearing (Codium fragile), a lumpy hearing (Codium contractum), a broad hearing (Codium latum), and a supine hearing (Codium coactum). .

In the present specification,'muscle' refers to tendons, muscles, and tendons generically, and'muscle function' refers to the ability to exert power by contraction of muscles, and the muscles can exert their contractile force as much as possible to overcome resistance. It includes muscle strength, which is the ability to be capable, muscle endurance, which is the ability to repeat contractions and relaxations for a given weight, for how long or many times, and quickness, which is the ability to exert strong power in a short time. These muscle functions are controlled by the liver and are proportional to muscle mass.

'Muscular disease' of the present invention means a disease caused by muscle wasting or degeneration. Examples of such diseases include sarcopenia, atony, muscular atrophy, muscular dystrophy, muscle degeneration, muscular stiffness, muscular dystrophy, amyotrophic axonal sclerosis, myasthenia and cachexia. Can be mentioned. Muscle wasting is characterized by gradual loss of muscle mass, weakness and degeneration of muscles, especially skeletal or voluntary and cardiac muscles. Muscle wasting and degeneration occurs due to genetic factors, acquired factors, and aging. The composition of the present invention has a muscle increasing effect, and the type of muscle is not limited. Muscle gain, among other body components, is to improve muscle performance, in particular, it can increase muscle mass through physical exercise and endurance improvement, and muscle mass can be increased by administering a substance having a muscle-increasing effect into the body.

The composition for preventing, improving, or treating muscle diseases of the present invention may contain one or more active ingredients having a function similar to that of the seaweed extract in Cordium or containing the seaweed extract in Cordium. If additional components are included, the effect of improving muscle disease of the composition of the present invention may be further enhanced. When the above ingredients are added, skin safety, ease of formulation, and stability of active ingredients can be considered in combination with use.

The seaweed extract in Cordium of the present invention may include a pretreatment process such as a drying process for the raw material in order to increase the extraction efficiency when the extract is prepared.

The seaweed extract in Cordium of the present invention can be extracted by a known natural product extraction method. Preferably, it may be extracted with one or more solvents selected from the group consisting of water, an organic solvent having 1 to 6 carbon atoms, and a subcritical or supercritical fluid. The organic solvent having 1 to 6 carbon atoms is alcohol, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, and 1 to 6 carbon atoms. chloride), hexane, cyclohexane, and petroleum ether, but is not limited thereto.

In addition, the seaweed extract from Cordium of the present invention can be obtained by extracting and purifying the seaweed from Cordium using purified water suitable for food processing, ethanol and subcritical water, or supercritical carbon dioxide, or by directly compressing the seaweed from Cordium. It can be obtained by separation and purification from the obtained oil. For example, it is possible to obtain an extract by extracting seaweed in Cordium under ultra-high pressure conditions of 100 Mpa or more. Preferably, it may be an ultra-high pressure condition of 100 MPa to 1000 MPa, but is not limited thereto.

The extract of the present invention may be filtered and/or concentrated to be used as a liquid, and may be solidified through a conventional drying process such as spray drying or freeze drying. In the drying process, dextrin or the like may be mixed and dried before spray drying or freeze drying.

Since the seaweed extract in Cordium of the present invention has excellent effect of increasing the diameter of muscle cells, it can be used as an effective ingredient in a pharmaceutical composition.

In addition, the seaweed extract in Cordium of the present invention has an excellent effect of increasing the diameter of muscle cells, and thus can be used as an effective ingredient in food compositions.

The pharmaceutical composition for preventing and treating muscle diseases of the present invention may further include a pharmaceutically acceptable carrier.

The pharmaceutically acceptable carrier may further include, for example, a carrier for oral administration or a carrier for parenteral administration. Carriers for oral administration may include lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like. In addition, the carrier for parenteral administration may include water, suitable oil, saline, aqueous glucose and glycol. In addition, it may further comprise a stabilizer and a preservative. Suitable stabilizers include antioxidants such as sodium hydrogen sulfite, sodium sulfite or ascorbic acid. Suitable preservatives are benzalkonium chloride, methyl- or propyl-paraben and chlorobutanol. Other pharmaceutically acceptable carriers may be referred to as those described in the following literature (Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, PA, 1995).

The pharmaceutical composition of the present invention can be administered to mammals including humans by any method. For example, it can be administered orally or parenterally, and parenteral administration methods are not limited thereto, but intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal , Intranasal, intestinal, topical, sublingual or rectal administration.

The pharmaceutical composition of the present invention can be formulated into a formulation for oral administration or parenteral administration according to the route of administration as described above. When formulated, one or more buffers (e.g., saline or PBS), antioxidants, bacteriostatic agents, chelating agents (e.g., EDTA or glutathione), fillers, bulking agents, binders, adjuvants (e.g., aluminum hydroxide). Side), suspending agents, thickening agents, wetting agents, disintegrants or surfactants, diluents or excipients.

Solid preparations for oral administration include tablets, pills, powders, granules, liquids, gels, syrups, slurries, suspensions or capsules, and the like, and such solid preparations include at least one excipient in the pharmaceutical composition of the present invention. For example, starch (including corn starch, wheat starch, rice starch, potato starch, etc.), calcium carbonate, sucrose, lactose, dextrose, sorbitol, mannitol, xylitol, erythritol maltitol, It may be prepared by mixing cellulose, methyl cellulose, sodium carboxymethylcellulose and hydroxypropylmethyl-cellulose or gelatin. For example, tablets or dragees can be obtained by blending the active ingredient with a solid excipient, pulverizing it, adding a suitable auxiliary, and processing into a granule mixture.

In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include suspensions, liquid solutions, emulsions, or syrups, but may include various excipients, such as wetting agents, sweetening agents, fragrances, or preservatives, in addition to water or liquid paraffin, which are simple diluents commonly used. .

In addition, in some cases, cross-linked polyvinylpyrrolidone, agar, alginic acid, or sodium alginate may be added as a disintegrant, and an anti-coagulant, a lubricant, a wetting agent, a fragrance, an emulsifier and a preservative may be additionally included. .

When administered parenterally, the pharmaceutical composition of the present invention may be formulated according to a method known in the art in the form of an injection, a transdermal administration, and a nasal inhalation agent together with a suitable parenteral carrier. In the case of such injections, they must be sterilized and protected from contamination by microorganisms such as bacteria and fungi. Examples of suitable carriers for injections include, but are not limited to, water, ethanol, polyol (eg, glycerol, propylene glycol and liquid polyethylene glycol, etc.), a mixture thereof and/or a solvent or dispersion medium containing vegetable oil. I can. More preferably, suitable carriers include Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) containing triethanolamine or sterile water for injection, isotonic solutions such as 10% ethanol, 40% propylene glycol and 5% dextrose. Etc. can be used. In order to protect the injection from microbial contamination, various antibacterial and antifungal agents such as paraben, chlorobutanol, phenol, sorbic acid, thimerosal, and the like may be additionally included. In addition, the injection may further include an isotonic agent such as sugar or sodium chloride in most cases.

In the case of transdermal administration, ointments, creams, lotions, gels, external solutions, pasta, liniment, and air rolls are included. In the above, "transdermal administration" means that an effective amount of the active ingredient contained in the pharmaceutical composition is delivered into the skin by topically applying the pharmaceutical composition to the skin.

In the case of an inhaled dosage form, the seaweed extract of cordium of the present invention is used in a pressurized pack using a suitable propellant, for example dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. Alternatively, it can be conveniently delivered from a nebulizer in the form of an aerosol spray. In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve that delivers a metered amount. For example, gelatin capsules and cartridges for use in an inhaler or insufflator can be formulated to contain a powder mixture of the compound and a suitable powder base such as lactose or starch. Formulations for parenteral administration are described in Remington's Pharmaceutical Science, 15th Edition, 1975. Mack Publishing Company, Easton, Pennsylvania 18042, Chapter 87: Blaug, Seymour, a formula generally known for all pharmaceutical chemistry.

The pharmaceutical composition for preventing and treating muscle diseases of the present invention can provide a desirable muscle disease prevention and treatment effect when it contains an effective amount of a seaweed extract from Cordium. In the present specification, the term "effective amount" refers to an amount that exhibits a higher response compared to the negative control group, and preferably refers to an amount sufficient to increase muscle mass or improve muscle function. The pharmaceutical composition of the present invention may contain 0.01 to 99.99% of the seaweed extract in Cordium, and the balance may be occupied by a pharmaceutically acceptable carrier. The effective amount of the seaweed extract in Cordium contained in the pharmaceutical composition of the present invention will vary depending on the form in which the composition is commercialized.

In the present specification, "treatment" refers to a clinical procedure to change the natural process of an individual or cell to be treated, and may be performed to prevent clinical pathology. Desirable effects of treatment include suppression of the occurrence or recurrence of the disease, alleviation of symptoms, reduction of any direct or indirect pathological consequences of the disease, reduction of the rate of disease progression, improvement of the disease state, improvement, alleviation or improved prognosis, etc. Include. Also, the term'prevention' refers to any action that suppresses the onset or delays the progression of a disease.

The total effective amount of the pharmaceutical composition of the present invention may be administered to a patient in a single dose, and may be administered by a fractionated treatment protocol that is administered for a long time in multiple doses. . The pharmaceutical composition of the present invention may vary the content of the active ingredient according to the severity of the disease. When parenterally administered, preferably 0.01 to 50 mg, more preferably 0.1 to 30 mg per 1 kg of body weight per day based on the seaweed extract in Cordium, and when administered orally, seaweed extract in Cordium Based on 1 kg of body weight per day, preferably 0.01 to 100 mg, more preferably 0.01 to 10 mg per day, so that it may be administered in divided doses 1 to several times. However, the dose of the seaweed extract in the cordium is effective for the patient in consideration of various factors such as the age, weight, health condition, sex, disease severity, diet and excretion rate of the patient, as well as the administration route and number of treatments of the pharmaceutical composition. Since the dosage is determined, considering these points, those of ordinary skill in the art will be able to determine an appropriate effective dosage for the specific use of the seaweed extract in Cordium for the prevention and treatment of muscle diseases. The pharmaceutical composition according to the present invention is not particularly limited in its formulation, route of administration, and method of administration as long as it exhibits the effects of the present invention.

The pharmaceutical composition for preventing and treating muscle diseases of the present invention may be used alone or in combination with surgery, radiation therapy, hormone therapy, chemotherapy, or methods using a biological response modifier.

The pharmaceutical composition for preventing and treating muscle diseases of the present invention may also be provided in the form of an external preparation comprising an extract of seaweed in Cordium as an active ingredient.

When the pharmaceutical composition for preventing and treating muscle diseases of the present invention is used as an external preparation for skin, additionally fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents agent), fragrance, surfactant, water, ionic emulsifier, nonionic emulsifier, filler, sequestering agent, chelating agent, preservative, vitamin, blocker, wetting agent, essential oil, dye, pigment, hydrophilic activator, lipophilic It may contain an adjuvant commonly used in the field of dermatology such as an active agent or any other component commonly used in skin external preparations such as lipid vesicles. In addition, the above ingredients may be introduced in amounts generally used in the field of dermatology.

When the pharmaceutical composition for preventing or treating muscle diseases of the present invention is provided as an external preparation for skin, it may be a formulation such as an ointment, patch, gel, cream, or spray, but is not limited thereto.

The composition for preventing or improving muscle disease of the present invention may also be a food composition. When the composition for preventing or improving muscle disease of the present invention is a food composition, it may be used to prevent or improve muscle disease due to muscle wasting or degeneration. Muscle wasting and degeneration occurs due to genetic factors, acquired factors, aging, etc., and muscle wasting is characterized by a gradual loss of muscle mass, weakness and degeneration of muscles, especially skeletal or voluntary muscles and heart muscles. Examples of related diseases include sarcopenia, atony, muscle atrophy, muscle dystrophy, muscle degeneration, muscle stiffness, muscular dystrophy, amyotrophic axonal sclerosis, myasthenia and cachexia. ), etc. These muscles do not limit their type.

The food composition of the present invention includes all forms such as functional food, nutritional supplement, health food, food additives and feed, and contains humans or animals including livestock. It is targeted for eating. Food compositions of this type can be prepared in various forms according to conventional methods known in the art.

Food compositions of this type can be prepared in various forms according to conventional methods known in the art. General foods include, but are not limited to, beverages (including alcoholic beverages), fruits and processed foods thereof (e.g., canned fruit, canned food, jam, marmalade, etc.), fish, meat and processed foods thereof (e.g. ham, sausage) Corn beef), bread and noodles (e.g. udon, buckwheat noodles, ramen, spagate, macaroni, etc.), fruit juice, various drinks, cookies, sweets, dairy products (e.g. butter, cheese, etc.), edible vegetable oil, margarine , Vegetable protein, retort food, frozen food, various seasonings (eg, soybean paste, soy sauce, sauce, etc.), etc., can be prepared by adding the seaweed extract of the genus Cordium. In addition, as a nutritional supplement, it is not limited thereto, but may be prepared by adding seaweed extract from Cordium to capsules, tablets, pills, and the like. In addition, as a health functional food, but not limited to this, for example, the seaweed extract in the cordium itself is prepared in the form of tea, juice, and drink, and liquefied, granulated, encapsulated and powdered so that it can be consumed (healthy beverage). It can be consumed. In addition, in order to use the seaweed extract in the cordium in the form of a food additive, it may be prepared and used in the form of a powder or a concentrate. In addition, it can be prepared in the form of a composition by mixing the seaweed extract from Cordium with known active ingredients known to have the effect of preventing muscle disease and improving muscle function.

When the composition for preventing or improving muscle diseases of the present invention is used as a health drink composition, the health drink composition may contain various flavoring agents or natural carbohydrates as an additional ingredient, such as a conventional beverage. The natural carbohydrates described above include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; Polysaccharides such as dextrin and cyclodextrin; It may be a sugar alcohol such as xylitol, sorbitol, and erythritol. Sweeteners include natural sweeteners such as taumatin and stevia extract; Synthetic sweeteners such as saccharin and aspartame can be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 mL of the composition of the present invention.

When the seaweed extract from Cordium is contained as an active ingredient in a food composition for preventing or improving muscle disease, the amount is not particularly limited to an amount effective to achieve the action for preventing or improving muscle disease, but the total weight of the composition. It is preferably 0.01 to 100% by weight based on. The food composition of the present invention may be prepared by mixing with the seaweed extract from Cordium with other active ingredients known to be effective in preventing muscle disease and improving muscle function.

In addition to the above, the health food of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, It may contain glycerin, alcohol or a carbonating agent. In addition, the health food of the present invention may contain flesh for the manufacture of natural fruit juice, fruit juice beverage or vegetable beverage. These ingredients may be used independently or in combination. The ratio of these additives is not very important, but it is generally selected from 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

In addition, the present invention provides a composition for strengthening muscles comprising the seaweed extract from Codium as an active ingredient.

In the present specification, the term "muscle strengthening" means that muscle mass is increased or muscle loss is suppressed.

Since the seaweed extract in Cordium of the present invention has excellent effect of increasing the binding of MHC (Myosin heavy chain), it can be used as an effective ingredient of a pharmaceutical composition.

In addition, the seaweed extract in Cordium of the present invention has excellent effect of increasing the binding of MHC (Myosin heavy chain), and thus can be used as an effective ingredient of a food composition.

The preparation method of the seaweed extract in Cordium is as described above, and the carrier that can be included in the pharmaceutical composition containing the seaweed extract in Cordium as an active ingredient, formulation of the pharmaceutical composition, administration route and dosage, etc. The administration method and the like are as described above.

The pharmaceutical composition according to the present invention may be a composition comprising a seaweed extract from Cordium as an active ingredient, or may be a composition consisting of seaweed extract from Cordium as an active ingredient, or as an active ingredient. It may be a composition consisting essentially of this.

The seaweed extract in Cordium for preparing the food composition is as described above. In addition, the examples and contents of the food composition are as described above.

In addition, the present invention provides a food composition for improving exercise capacity, comprising the seaweed extract in Codium as an active ingredient.

In the present specification, the term'exercise ability enhancement' refers to an effect of improving muscle function such as muscle strength and muscle endurance, and preferably may be an increase in muscle strength or endurance, and degenerative diseases, mitochondrial disorders, hypotension, and spontaneity It may be to prevent or improve one or more symptoms selected from the group consisting of hypothyroidism, lethargy, muscle discard and depression.

The seaweed extract in Cordium for preparing the food composition is as described above. In addition, the examples and contents of the food composition are as described above.

Therefore, the present invention confirmed the effect of the seaweed extract in Cordium on muscle cells, and the seaweed extract in Cordium increased the binding index and mRNA expression of MHC in myoblasts and myotubes, and the effect of increasing the diameter of myotubes. As shown, it can be usefully used as a composition for preventing, improving or treating muscle diseases, a food composition for improving exercise capacity, and a composition for strengthening muscles.

1 is a graph showing the fusion index and mRNA expression of the myosin heavy chain (MHC) according to the concentration when the myoblasts are treated with an auditory extract.
2 is a result of confirming changes in cell diameter and MHC (myosin heavy chain) mRNA expression according to concentration when the auditory extract was treated after differentiating myoblasts into myotubes and inducing atrophy.
3 is a result of confirming the diameter of the cells after treatment of the auditory extract on the root canal cells induced muscle atrophy.
4 is a result of confirming the change in MHC mRNA expression after treatment of the auditory extract on the root canal cells induced muscle atrophy.
5 is a graph showing the binding index of MHC when treated with extracts of auditory, monguri auditory, broad auditory, and nueun auditory gak in the process of differentiating myoblasts into root canal cells.

Hereinafter, the present invention will be described in detail.

However, the following examples are only illustrative of the present invention, and the contents of the present invention are not limited to the following examples.

<Example 1>

Effect of Auditory Extract on Differentiation of Muscle Cells

Codium fragile extract was purchased from the Korea Plant Extract Bank and used as a sample, and each sample is an extract extracted with 95% ethanol or water.

After treatment of the auditory extract, the binding index (fusion index, number of nuclei in MHC/total number of nuclei) of the myosin heavy chain (MHC) involved in the process of muscle contraction or relaxation by decomposing ATP in myosin in cells was measured.

MHC binding index was measured as follows. C2C12 myoblasts were treated with 1 or 2 μg/ml of auditory extract, cultured for 2 days, and then fixed with 4% formaldehyde. After permeabilization with 0.1% triton X-100 PBS, blocking with 5% bovine serum albumin for 1 hour, overnight at 4°C with total MHC antibody, 2 A secondary antibody (Alexa Fluor 488- secondary antibody, Cell Signaling Biotechnology) was attached at room temperature for 2 hours. After DAPI staining, an image was obtained under a microscope (Olympux IX71). The fusion index was calculated by dividing the number of nuclei in the MHC by the total number of nuclei.

As a result, as shown in Fig. 1, when the auditory extract was treated compared to the control without the auditory extract, the binding index of MHC was found to be high, and the binding index was increased according to the concentration of the extract. .

<Example 2>

Effect of Auditory Extract on MHC Expression in Muscle Cells

After culturing C2C12 muscle cells, the auditory extract was treated with 1, 5, and 10 μg/mL and incubated for 24 hours. After washing the cells with PBS, cells were obtained, followed by RNA extraction according to the manufacturer's instructions. Total RNA was extracted using a kit (NucleoSpin RNA plus, MN). Then, after reverse transcription of 1 μg of RNA using 1 mM dNTP and oligo dT (0.5 μg/μl) to synthesize cDNA, 1 unit of Tag DNA polymerase, 0.2 mM dNTP, X10 reaction buffer, and 100 pmol of primer were mixed to obtain the total volume. Was made to be 25 μl. PCR was performed with the mixed samples to measure the mRNA expression level of MHC (myosin heavy chain).

As a result, as shown in FIG. 2, when the auditory extract was treated compared to the control without the auditory extract, the expression level of MHC mRNA was increased, and the expression level of the mRNA increased according to the concentration of the extract. Appeared.

Primer sequence Base sequence Sequence number MHC2a Sense AAG CGA AGA GTA AGG CTG TC One Anti-sense GTG ATT GCT TGC AAA GGA AC 2 18s RNA Sense GTA ACC CGT TGA ACC CCA TT 3 Anti-sense CCA TCC AAT CGG TAG TAG CG 4

<Example 3>

Effect of Auditory Extract on Root Canal Cells

After the muscle cells were differentiated into root canal cells, the auditory extract was treated thereto, and changes in cell size and MHC mRNA expression were confirmed. To this end, first, C2C12 myoblasts were differentiated with DMEM containing 2% horse serum for 4 days, and then TNFα 25 ng/ml was treated for 24 hours to induce senescent myoatrophy, and at the same time, the auditory extract was 1.0, 5.0. And treatment with a concentration of 10.0 μg / ml confirmed the effect of relieving muscle atrophy.

In more detail, it was carried out as follows. Myoblasts derived from C2C12 mice (CRL1772; American type cell collection, USA) were cultured in DMEM (Dulbecco's Modified Eagle's Medium) medium supplemented with 10(v/v)% FBS, and 95% in 6-well plates. After reaching the growth level (confluence), it was transferred to a DMEM medium containing 2(v/v)% horse serum, and cultured for 4 days to differentiate into myotubes. Thereafter, 25 ng/ml of TNFα was treated for 24 hours to induce myoatrophy, and at the same time, the auditory extract was treated at 1.0, 5.0 and 10.0 μg/ml concentrations, and then fixed with 4% formaldehyde. After permeabilization with 0.1% triton X-100 PBS, blocking with 5% bovine serum albumin for 1 hour, overnight at 4°C with total MHC antibody, 2 A secondary antibody (Alexa Fluor 488- secondary antibody, Cell Signaling Biotechnology) was attached at room temperature for 2 hours. Thereafter, DAPI staining was performed and an image was acquired under a microscope (Olympux IX71), and the diameter of the myotube was calculated using iSolution software.

As a result, as shown in FIG. 3, when the TNF-α was treated, the diameter of the cells decreased. When treated with the auditory extract, it was found that the diameter of the cells increased.

In addition, as a result of confirming the change in the expression of MHC mRNA in the same manner as in Example 2, as shown in FIG. 4, it was found that the expression level of MHC mRNA was increased when the auditory extract was treated compared to the case where TNF-α was treated. It was found that the expression level of mRNA increased according to the concentration of the extract.

<Example 4>

Effect of Algae Extract from Cordium on Root Canal Cells

In addition to hearing, the effects of the extracts of Codium contractum, Codium latum, and Codium coactum, seaweeds in the same codium, on the binding index of MHC in root canal cells were confirmed. Each extract was purchased from Korea Plant Extract Bank and used as a sample, and each sample was an extract extracted with 95% ethanol or water.

The binding index was measured in the same manner as in Example 1 using muscle cells treated for 48 hours by adding each extract to DMEM containing 2% horse serum at a concentration of 10 μg/mL.

As a result, as shown in FIG. 5, it was confirmed that the binding index of MHC was increased when the auditory, munguri auditory, broad auditory auditory extract, and supine auditory extract were treated compared to the untreated control group.

As described above, since the seaweed extract in Cordium shows the effect of increasing the binding index and mRNA expression of MHC in myoblasts and myotubes, and increasing the diameter of myotube cells, the composition comprising the seaweed extract in Cordium of the present invention is In addition to being useful for preventing, improving, or treating muscle diseases, it can also be usefully used as a food composition for improving exercise capacity or a composition for strengthening muscles.

<110> KOREA FOOD RESEARCH INSTITUTE <120> Composition for Preventing, Improving or Treating of muscular disease containing Codium SPP. algae extract <130> NP17-0117 <160> 4 <170> KoPatentIn 3.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> MHC2a sense primer <400> 1 aagcgaagag taaggctgtc 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> MHC2a anti-sense primer <400> 2 gtgattgctt gcaaaggaac 20 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> 18sRNA sense primer <400> 3 gtaacccgtt gaaccccatt 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> 18sRNA anti-sense primer <400> 4 ccatccaatc ggtagtagcg 20

Claims (10)

A composition for preventing, improving, or treating muscle diseases, comprising a seaweed extract in Codium as an active ingredient.
The composition of claim 1, wherein the extract is an extract obtained by at least one solvent selected from the group consisting of water, an organic solvent having 1 to 6 carbon atoms, a subcritical fluid, a supercritical fluid, and a mixture thereof.
The method of claim 1, wherein the muscle disease is sarcopenia, atony, muscular atrophy, muscular dystrophy, muscle degeneration, muscular stiffness, muscular dystrophy, amyotrophic axonal sclerosis, myasthenia gravis. And cachexia (cachexia) composition, characterized in that at least one selected from the group consisting of.
A composition for strengthening muscles comprising the seaweed extract from Codium as an active ingredient.
The composition of claim 4, wherein the muscle strengthening means increasing muscle mass or inhibiting muscle loss.
The composition according to any one of claims 1 to 5, wherein the composition is a pharmaceutical composition or a food composition.
The method of claim 1 or 4, wherein the seaweed in the Codium is selected from the group consisting of Codium fragile, Codium contractum, Codium latum and Codium coactum. Composition, characterized in that any one.
Exercise ability that contains the extract of seaweed from Codium, which is one selected from the group consisting of Codium Fragile, Codium contractum, Codium latum, and Coactum, as an active ingredient Food composition for promotion.
The food composition according to claim 8, wherein the improvement in exercise capacity means that muscle strength or muscular endurance is increased.
The food according to claim 8, wherein the improvement of exercise capacity is to prevent or improve one or more symptoms selected from the group consisting of degenerative diseases, mitochondrial abnormal diseases, hypotension, hypotonia, lethargy, muscle abolition and depression. Composition.

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