WO2015199516A1 - Composition for improving muscular function or enhancing exercise performance, containing kirenol or hui chum extract - Google Patents

Abstract

The present invention relates to a composition improving a muscular function or enhancing an exercise performance, the composition containing, as an active ingredient, kirenol, a Hui Chum (Siegesbeckia spp.) extract containing the same, or a fraction thereof. The kirenol, the Hui Chum (Siegesbeckia spp.) extract containing the same, or the fraction thereof, according to the present invention, increases the protein expression of p-mTOR, which is a main gene involved in the muscular function, thereby significantly enhancing the muscular function. The kirenol, the Hui Chum (Siegesbeckia spp.) extract containing the same, or the fraction thereof, according to the present invention, increases the protein expression of PGC-1α, which is a main gene involved in the exercise performance, thereby significantly enhancing the exercise performance. Furthermore, the present invention can be safely used without side effects since it is a natural material, and thus can be used as a medicinal product or food.

Description

Composition for improving muscle function or enhancing exercise performance, including chilenol or rare extract

The present invention relates to a composition for improving muscle function or enhancing exercise performance, containing a kyrenol, Sigesbeckia spp. Extract or a fraction thereof as an active ingredient.

Over the last 50 to 100 years, it has been known that a decrease in human physical activity is associated with an increased incidence of metabolic diseases such as type 2 diabetes, obesity, and cardiovascular disease. Lack of physical activity ranks fourth among the causes of death reported by the World Health Organization. Because of these phenomena, organizations such as the World Health Organization, the American Heart Association and the British Heart Foundation recommend a minimum of 30 minutes of aerobic exercise five days a week. Indeed, exercise reduces the incidence of diabetes, obesity, breast and colorectal cancer, and has good therapeutic effects on depression (Br. J. Pharmacol. 170: 1153-1166, 2013, Am. J. Cardiol. 110: 58B-68B, 2012).

One of the typical methods for improving energy performance by promoting energy consumption is to increase fatty acid oxidation by mitochondria to generate ATP energy. The number and ability of mitochondria to control this is regulated by a co-activator called peroxisome proliferator-activated receptor-gamma coactivator 1 alpha (PGC-1α), and the activity of PGC-1α is regulated by SIRT1 (sirtulin). (EMBO. J. 26: 1913-1923, 2007, Cell Metab. 1: 361-370, 2005).

Scarpulla and his co-workers identified a protein called nuclear respiratory factor (NRF) (J. Cell. Biochem. 97: 673-683, 2006). Proteins in the NRF family bind to the promoters of various genes of mitochondria in the nucleus to activate mitochondrial replication and transcription. It has been found that the production of mitochondria is induced by replication of mitochondria and activation of transcription. In addition, proteins in the NRF group have been shown to physically interact with PGC-1α coactivators to enhance expression (Cell 98: 115-24, 1999).

In addition, AMPK, p-AMPK, PPARδ ERRα and Tfam are known as factors related to the improvement of exercise performance.

Meanwhile, muscle atrophy is caused by a gradual decrease in muscle mass and refers to muscle weakness and degeneration (Cell 119: 90710, 2004). Muscular atrophy is promoted by inactivity, oxidative stress and chronic inflammation, weakening muscle function and motor capacity (Clin. Nutr. 26: 524-534, 2007). The most important factor in determining muscle function is muscle mass, which is maintained by the balance of protein synthesis and degradation. Muscular atrophy occurs when proteolysis occurs more than synthesis (Cell Biol. 37: 1985-1996, 2005).

Muscle size is controlled by intracellular signaling pathways that induce anabolism or catabolism in the muscle, leading to more signaling reactions that induce synthesis than muscle protein degradation. In this case, muscle protein synthesis is increased. An increase in muscle protein synthesis may be attributed to an increase in muscle size (hypertrophy) or muscle fiber number (hyperplasia) as muscle protein increases (The Korea Journal of Sports Science 30: 1551-1561, 2011).

Muscle hypertrophy inducers induce protein synthesis by phosphorylating downstream proteins from the stimulation of the phosphatidylinositol-3 kinase (PI3K) / Akt pathway in myocytes. The activity of mTOR (mammalian target of rapamycin) by PI3K / Akt signaling is recognized as a central growth signaling mechanism that integrates various growth signals in cells. Activation of mTOR contributes to muscle mass gain by inducing muscle protein synthesis by activating two downstream targets, 4E-BP1 (4E-binding protein) and p70S6K (phosphorylated 70-kDa ribosomal S6 kinase). Korea Journal of Sports Science 30: 1551-1561, 2011, J Biol Chem 278: 40717-40722, 2003).

Differentiation and muscle formation of muscle cells are regulated by various muscle regulatory factors (cell Mol Life Sci 70: 4117-4130, 2013). Among them, myoD initiates the expression of genes specific for muscle differentiation and induces mesenchymal stem cells to differentiate into myoblasts (myoblasts). Myogenin regulated by MyoD is the most important factor in the fusion of myoblasts and is involved in the formation of myotubes. Muscle fibers formed through this process are bundled to finally form muscles (Cell Mol Life Sci 70: 4117-4130, 2013; Sci Signal 6: re2, 2013).

Huicheom (aka huiryeom) is Shige's Becky Ashok plants (Siegesbeckia spp.) Of the Asteraceae in Shige's Becky Oh Mugla Brenna sense (Siegesbeckia glabrescens Mak.), Shigeru's Becky Oh Mfuwe sense (Siegesbeckia pubescens Mak.) Or Shige's Becky Ah The ground part of the Orientalis ( Siegesbeckia orientalis L.) is dried. Siegesvecchia glabreense, called Jindukchal, is an antibacterial agent (Int. J. Food Microbiol. 160: 260-266, 2013), anticancer (Oncol. Rep. 30: 221-226, 2013), antidiabetic (J. Enzyme). Inhib. Med. Chem. 21: 379-383, 2006), anti-inflammatory (Food Agric. Immunol. 22: 145-160, 2011) and the like have been reported. Siegesvecchia fuvesense, called hair decoction, has been shown to include anti-inflammatory and analgesic (Pak. J. Pharm. Sci. 21: 89-91, 2008), antioxidant and anti-obesity (Kor. J. Microbiol. Biotechnol. 41: 341-349 , 2013), wound healing (J. Ethnopharmacol. 134: 1033-1038, 2011), arthritis treatment (Phytomedicine 19: 882-889, 2012) and the like have been reported. Sigebecia orientalis, called Jeju Derivatives, is known as anti-cancer (Natural Product Radiance 6: 34-39, 2007), anti-inflammatory (Chem. Biodivers. 3: 754-761, 2006), and antioxidant (Korean J. Pharmacogn. 36: 150 -163, 2005) and the like have been reported.

Kirenol is a diterpenoid mainly found in rapeseed and has anti-inflammatory and analgesic effects (J. Ethnopharmacol. 137: 1089-1094, 2011), antibacterial effect (Pharmacogn. Mag. 8: 149-155 , 2012), arthritis therapeutic effects (Phytomedicine 19: 882-889, 2012), anti-obesity effects (BBRC 445, 433-438, 2014) and the like.

However, prior to the present invention, there is no known effect of improving muscle function or enhancing exercise performance of rare or kyrenol.

Thus the present inventors have superior oblique modulating activity or superior physical performance result of browsing the natural substances that can have an active safely applied, Kastrup play (kirenol) or huicheom (Hui Chum, Siegesbeckia spp.) Extracts or fractions thereof containing the same The present invention was completed by confirming that there are muscle function improving activity and exercise performance enhancing activity.

Accordingly, it is an object of the present invention to provide a composition for improving muscle function or enhancing exercise performance including a rare extract as an active ingredient.

Still another object of the present invention is to provide a composition for improving muscle function or enhancing exercise performance, comprising the compound of Formula 1 as an active ingredient:

[Formula 1]

As a means for solving the above problems, the present invention

It provides a pharmaceutical composition and food composition for improving muscle function or enhancing exercise performance, including a rare extract or a fraction thereof as an active ingredient.

As another means for solving the above problems, the present invention

It provides a pharmaceutical composition and food composition for improving muscle function or enhancing exercise performance, comprising the compound represented by the formula (1) as an active ingredient:

[Formula 1]

Kirenol according to the present invention, or a rare extract containing the same or fractions thereof increases the protein expression of p-mTOR, a major gene involved in improving muscle function, and has an excellent effect on increasing muscle mass. In addition, the kyrenol (kirenol), or a rare extract containing the same or a fraction thereof according to the present invention by increasing the protein expression of PGC-1α, a major gene involved in exercise performance, and excellently enhance the exercise performance It is effective.

In addition, since the present invention is a natural product can be used safely without side effects, it can be used as a medicine or food.

1 is a protein of p-mTOR according to the treatment of chirenol or Sigebexkia glabressenth ethanol extract, Sigebexkia fuvesense ethanol extract, Sigebexchia orientalis ethanol extract in L6 muscle cells The result of measuring the expression level is shown.

Figure 2 shows the results of measuring the protein expression level of p-mTOR according to the treatment of Sigebek Beckia orientalis ultra-high pressure extract containing the chilenol in L6 muscle cells.

Figure 3 shows the results of measuring the mRNA expression level of muscle differentiation regulating genes (myogenin and MyoD) according to the treatment of the Siegesvekchia orientalis ethanol extract containing chilenol in L6 muscle cells.

Figure 4 shows the results of measuring the mRNA expression level of muscle differentiation regulating genes (myogenin and MyoD) according to the chilenol treatment in L6 muscle cells.

FIG. 5 shows the major genes involved in protein catabolism (Atrogin-1 and MuRF1) in L6 muscle cells following the treatment of Shigeresvecchia orientalis ethanol extract containing chilenol. The result of measuring mRNA expression is shown.

Figure 6 shows the results of measuring the mRNA expression level of the major genes (Atrozine-1 and MuRF1) involved in the protein catabolism in muscle following the chilenol treatment in L6 muscle cells.

Figure 7 shows the results of measuring the level of muscle mass increase according to administration of Shigeresvecchia orientalis ethanol extract containing chilenol in a normal diet animal model.

Figure 8 shows the results of measuring the muscle volume increase level according to the administration of Sigestescchia orientalis ethanol extract containing chilenol in the normal diet animal model (left: PET / CT results, right: muscle volume measurement results).

Figure 9 shows the results of measuring the level of muscle mass increase according to administration of Shigeresvecchia orientalis ethanol extract containing chilenol in a high-fat diet animal model.

Figure 10 shows the results of measuring the muscle volume increase level according to the administration of Shigeresvecchia orientalis ethanol extract containing chilenol in a high-fat diet animal model (left: PET / CT results, right: muscle volume measurement results) .

FIG. 11 is a protein of PGC-1α according to the treatment of chirenol or Sigestesevikia glabressenth ethanol extract, Sigestesevikia fuvesens ethanol extract, Sigestesevichia orientalis ethanol extract in L6 muscle cells The result of measuring the expression level is shown.

Figure 12 shows the results of measuring the protein expression level of PGC-1α in accordance with the treatment of Sigebek Beckia orientalis ultra-high pressure extract containing chilenol in L6 muscle cells.

Figure 13 shows the results of measuring the change in exercise performance by administration of Shigeresvecchia orientalis ethanol extract containing chilenol in a normal diet animal model (a: exercise distance, b: exercise time).

Figure 14 shows the results of measuring the change in exercise performance by administration of Shigeresvecchia orientalis ethanol extract containing chilenol in a high fat diet-induced obese animal model (a: exercise distance, b: exercise time ).

FIG. 15 shows the results of measuring the expression levels of p-AMPK, SIRT1, PGC-1α, and PPARδ proteins, which are genes related to improvement of exercise ability, by administration of Shigeresvecchia orientalis ethanol extract containing chilenol in an animal model calf muscle. Shows.

FIG. 16 shows the results of measuring mRNA expression levels of PGC-1α, NRF-1, ERRα, and Tfam, genes related to mitochondrial biosynthesis by administration of Shigeresvecchia orientalis ethanol extract containing chilenol in an animal model calf muscle Shows.

EMBODIMENT OF THE INVENTION Hereinafter, the structure of this invention is demonstrated concretely.

The present invention is a rare extract or fractions thereof, or the use of the compounds represented by the formula (1) to improve muscle function or enhance the performance of exercise; Rare extract or fractions thereof, or a composition for improving muscle function or enhancing exercise performance, including a compound represented by the following formula (1); Or a rare extract or a fraction thereof, or a compound represented by the following Formula 1 is provided to a subject, thereby improving muscle function or enhancing exercise performance.

[Formula 1]

Herein from "huicheom" or "huiryeom 'is Shigeru's Becky subgenus plant of the Asteraceae (Siegesbeckia spp.) In Shigeru's Becky Oh Glidden breather sense (Siegesbeckia glabrescens Mak.), Shigeru's Becky Oh Mfuwe sense (Siegesbeckia pubescens Mak.) Or dry above ground of Siegesbeckia orientalis L. Shigebexkia Glabressen is also known as Jindeoksaeng, Sigebexkia Puvacese is known as Hairseam, and Siegevecchia Orientalis is called Jeju.

In the present specification, the `` extracted extract '' or the `` development extract '' is used interchangeably, and refers to an extract obtained by extracting the rare (excreted). The preparation method of the rare extract may be used without limitation conventional extraction methods known in the art, for example, water, organic solvents having 1 to 6 carbon atoms, and children from rare plants or parts of plants (leaves or roots) It can be obtained by extraction with one or more solvents selected from the group consisting of critical fluids and supercritical fluids.

As used herein, 'fraction' means a result obtained by the fractionation method of separating a specific component or a specific group from a mixture comprising various components. Methods for preparing fractions are well known in the art, and known methods can be used without limitation. For example, a specific fraction in which the active substance is concentrated can be prepared using a technique such as solvent fractionation, silica gel chromatography, and prep-HPLC.

In one embodiment, the fraction of the rare extract may be obtained by fractionating the rare extract with ethyl acetate, methanol or a mixed solvent thereof.

In the present specification, 'muscle' refers to tendons, muscles, and tendons, and 'muscle function' refers to the ability to exert strength by contraction of muscles, and muscles can exert maximum contraction force to overcome resistance. Muscle strength, which is the ability to be present, muscle endurance, which is how long or how many times a muscle can repeat contraction and relaxation, and quickness, which is the ability to exert a strong force in a short time. This muscle function is controlled by the liver and is proportional to muscle mass. The term 'improving muscle function' refers to improving muscle function better.

In the present specification, 'composition for improving muscle function' refers to a composition containing a substance having an effect on improving muscle function as an active ingredient, and includes a pharmaceutical composition or a food composition.

In the present specification, the 'exercise performance ability' is a fast, strong, accurate, long, skillfully when the physical movements seen in daily life or sports are divided into running, running, throwing, swimming, etc. The exercise performance is defined as factors such as muscle strength, agility and endurance. The term 'enhancing athletic performance' refers to improving or improving athletic performance.

In the present specification, the 'exercise performance enhancing composition' refers to a composition containing a substance having an athletic performance enhancing effect as an active ingredient, and includes a pharmaceutical composition or a food composition.

The composition for improving muscle function or enhancing exercise performance of the present invention may further contain at least one active ingredient exhibiting the same or similar function, in addition to the rare extract or fractions thereof. For example, it may include a known muscle function improving component or exercise performance enhancing component. Including the additional components will be able to further enhance the muscle function improvement effect or exercise performance enhancing effect of the composition of the present invention. When the ingredient is added, skin safety, ease of formulation, and stability of the active ingredients may be considered. In one embodiment of the present invention, the composition is a muscle function improving component or exercise performance enhancing component known in the art, the extract of Camperia Fabiflora extract, Piperretrofractum Vahl. Fruit, mycetin , cooker It may further comprise at least one component selected from the group consisting of bitine extract, panduratata extract, grape root extract, root root extract and red ginseng extract. Additional components may be included in at least 0.0001% and up to 10% by weight relative to the total composition weight. For example, at least 0.0001 wt% to at most 1 wt%, at least 0.0001 wt% to at most 0.1 wt%, at least 0.0001 wt% to 0.001 wt%, at least 0.001 wt% to 10 wt%, at least 0.001 wt% to 1 It may be up to 0.001% by weight or less, up to 0.1% by weight, 0.01% by weight to 10% by weight, 0.01% by weight to 1% by weight or less. The content range may be adjusted according to requirements such as skin safety, ease of formulating the compound of Formula 1, and the like.

The composition for improving muscle function or enhancing exercise performance including the rare extract or fractions thereof of the present invention as an active ingredient may be a pharmaceutical composition or a food composition.

The composition for improving muscle function or enhancing exercise performance, comprising the compound represented by Formula 1 of the present invention as an active ingredient, may be a pharmaceutical composition or a food composition:

[Formula 1]

The compound of Formula 1 may include all possible isomers, for example, may include a compound of Formula 2 below.

[Formula 2]

The compound of Formula 2 is referred to as kirenol.

The compound of Formula 1 or the compound of Formula 2 may be used by separating or synthesizing from a plant extract or using a commercially available compound.

In one embodiment, the compound represented by Formula 1 or the compound represented by Formula 2 may be isolated from the rare extract.

In one embodiment, the rare extract may be an extract of one or more plants selected from the group consisting of Siegesvecchia glabresense, Siegesvecchia fubesense, and Siegesvecchia orientalis. For example, ethanol extract, hot water extract, hexane extract, ethyl acetate extract, ultra high pressure extract using dried leaves and stems of Siegesvecchia glabresense, Sigebesvecchia fubense or Siegesvecchia orientalis have.

In one embodiment, the rare extract may be obtained by extracting the rare extract with one or more solvents selected from the group consisting of water, an organic solvent having 1 to 6 carbon atoms, a subcritical fluid and a supercritical fluid. For example, rare plants may be obtained by extraction under ultrahigh pressure conditions of 100 MPa or more. If necessary, it may be prepared by additionally including filtration and concentration steps according to methods known in the art.

In one embodiment, the organic solvent having 1 to 6 carbon atoms may be selected from alcohols having 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, It may be at least one selected from the group consisting of methylene chloride, hexane, cyclohexane, and petroleum ether.

In addition, the rare extract of the present invention can be obtained by extracting and purifying the dried rare extract using purified water, ethanol, subcritical water or supercritical carbon dioxide suitable for food processing, or extracted and purified using an ultra-high pressure extraction device Alternatively, the rare plant can be obtained by separation and purification from oil obtained by direct compression. For example, the extract can be obtained by extracting the rare oil under ultra-high pressure conditions of 100 MPa or more.

In one embodiment, the fraction of the rare extract may be obtained by fractionating the rare extract with ethyl acetate, methanol or a mixed solvent thereof.

When the composition of the present invention is a pharmaceutical composition, in order to improve muscle function, it may be used for the prevention or treatment of muscle diseases due to muscle wasting or degeneration. Muscle depletion and degeneration occur due to genetic factors, acquired factors, aging, etc., muscle depletion is characterized by a gradual loss of muscle mass, weakening and degeneration of muscles, especially skeletal or veterinary and cardiac muscles. Examples of related diseases include atony, muscular atrophy, muscular dystrophy, muscle degeneration, myasthenia, cachexia and sarcopenia. The composition of the present invention has an effect of increasing muscle mass, the type of muscle is not limited.

The pharmaceutical composition of the present invention may comprise a pharmaceutically acceptable salt of chilenol or a rare extract or fraction thereof containing it. As used herein, the term `` pharmaceutically acceptable '' refers to a physiologically acceptable and typically does not cause an allergic or similar reaction when administered to a human, wherein the salt is a pharmaceutically acceptable free acid. Acid addition salts formed by acid) are preferred.

Pharmaceutically acceptable salts of the chilenol or rare extracts or fractions thereof containing the same may be acid addition salts formed using organic or inorganic acids, for example, formic acid, acetic acid, propionic acid, lactic acid, butyric acid, Isobutyric acid, trifluoroacetic acid, malic acid, maleic acid, malonic acid, fumaric acid, succinic acid, succinic acid monoamide, glutamic acid, tartaric acid, oxalic acid, citric acid, glycolic acid, glucuronic acid, ascorbic acid, benzoic acid, phthalic acid, salicylic acid, anthranilic acid , Dichloroacetic acid, aminooxy acetic acid, benzenesulfonic acid, p-toluenesulfonic acid or methanesulfonic acid. Inorganic acids include, for example, hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid or boric acid. The acid addition salt may preferably be in the form of hydrochloride or acetate, more preferably in the form of hydrochloride.

The above-mentioned acid addition salts can be prepared by a) chirenol or its rare extracts or fractions containing them and acids, or b) by dissolving and mixing one of them in a solvent or a hydrous solvent, or c) Knoll or a rare extract containing the same or a fraction thereof is prepared by a general method for preparing a salt, which is placed in an acid in a solvent or an aqueous solvent and mixed with them.

In addition to the above, salts that can be additionally salted are gabar salt, gabapentin salt, pregabalin salt, nicotinate, adipate salt, hemimalonate, cysteine salt, acetylcysteine salt, methionine salt, arginine salt, lysine salt, ornithine salt or Aspartate.

In addition, the pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable carrier.

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

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

The pharmaceutical composition of the present invention may be formulated into a preparation for oral 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, extenders, binders, adjuvants (e.g. aluminum hydroxide) Side), suspending agents, thickening agents, wetting agents, disintegrants, surfactants, diluents or excipients.

Formulations for oral administration include tablets, pills, powders, granules, solutions, gels, syrups, slurries, suspensions or capsules and the like, and such solid preparations may be used in the pharmaceutical compositions of the present invention at least one excipient, for example , Starch (including corn starch, wheat starch, rice starch, potato starch, etc.), calcium carbonate, sucrose, lactose, dextrose, sorbitol, mannitol, xylitol, erythritol maltitol, cellulose , Methyl cellulose, sodium carboxymethyl cellulose and hydroxypropylmethyl-cellulose or gelatin can be prepared by mixing. For example, tablets or dragees can be obtained by combining the active ingredients with solid excipients and then grinding them and adding suitable auxiliaries and then processing them into granule mixtures.

In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid formulations may include various excipients, such as wetting agents, sweeteners, fragrances or preservatives, in addition to the commonly used simple diluents, water or liquid paraffin.

In addition, in some cases, crosslinked polyvinylpyrrolidone, agar, alginic acid or sodium alginate may be added as a disintegrant, and may further include an anticoagulant, a lubricant, a humectant, a perfume, an emulsifier, a preservative, and the like. .

When administered parenterally, the pharmaceutical compositions of the present invention may be formulated according to methods known in the art in the form of injections, transdermal and nasal inhalants with suitable parenteral carriers. Such injections must be sterile and protected from contamination of microorganisms such as bacteria and fungi. Examples of suitable carriers for injections include, but are not limited to, solvents or dispersion media comprising water, ethanol, polyols (e.g., glycerol, propylene glycol and liquid polyethylene glycols, etc.), mixtures thereof and / or vegetable oils Can be. More preferably, suitable carriers include Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) containing triethanol amine or sterile water for injection, 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 parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like may be further included. In addition, the injection may in most cases further comprise an isotonic agent such as sugar or sodium chloride.

In the case of transdermal administrations, ointments, creams, lotions, gels, external preparations, pasta preparations, linen preparations, air rolls and the like are included. As used herein, 'transdermal administration' means that the pharmaceutical composition is locally administered to the skin so that an effective amount of the active ingredient contained in the pharmaceutical composition is delivered into the skin.

In the case of inhaled dosages, the compounds used according to the invention may be pressurized packs or by means of suitable propellants, for example dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. It can be delivered conveniently from the 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 to deliver a metered amount. For example, gelatin capsules and cartridges for use in inhalers or blowers can be formulated to contain a 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 prescription generally known in all pharmaceutical chemistries.

The pharmaceutical composition of the present invention may provide a desirable muscle function improving effect or exercise performance enhancing effect when it contains an effective amount of chilenol or a rare extract or a fraction thereof. In the present specification, the 'effective amount' refers to an amount that exhibits a higher response than the negative control, and preferably refers to an amount sufficient to improve muscle function or improve exercise performance. The pharmaceutical composition of the present invention may contain 0.01 to 99.99% of chilenol or a rare extract containing the same, and the remaining amount may be occupied by a pharmaceutically acceptable carrier. The effective amount of the chilenol or rare extract or fractions thereof included in the pharmaceutical composition of the present invention will vary depending on the form in which the composition is commercialized and the like.

The total effective amount of the pharmaceutical composition of the present invention may be administered to a patient in a single dose, or may be administered by a fractionated treatment protocol that is administered in multiple doses for long periods of time. The pharmaceutical composition of the present invention may vary the content of the active ingredient depending on the extent of the disease. When administered parenterally, the amount is preferably administered in an amount of 0.01 to 50 mg, more preferably 0.1 to 30 mg per kg of body weight per day based on the kyrenol or the rare extract containing the same, and on oral administration It may be administered in portions of 1 to several times so as to be administered in an amount of preferably 0.01 to 100 mg, more preferably 0.01 to 10 mg per kg of body weight per day, based on the knol or the rare extract containing the same. However, since the dose of the kyrenol or the rare extract containing the same is determined in consideration of various factors such as the age, weight, health condition, sex, severity of the disease, diet and excretion rate, as well as the route of administration and the number of treatments, In view of this, one of ordinary skill in the art will be able to determine the appropriate effective dosage of the kyrenol or rare extract containing the same according to the use for improving muscle function or enhancing exercise performance. The pharmaceutical composition according to the present invention is not particularly limited to its formulation, route of administration and method of administration as long as the effect of the present invention is shown.

The pharmaceutical composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy or biological response modifiers.

The pharmaceutical composition of the present invention may also be provided in the formulation of a topical preparation containing chilenol or a rare extract or a fraction thereof as an active ingredient.

When the pharmaceutical composition of the present invention is used as an external preparation for skin, it is additionally used for fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants. Skin external preparations such as water, ionic emulsifiers, nonionic emulsifiers, fillers, metal ion sequestrants, chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic active agents, lipophilic active agents or lipid vesicles It may contain adjuvants commonly used in the field of dermatology, such as any other ingredients commonly used in the art. The ingredients may also be introduced in amounts generally used in the field of dermatology.

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

The composition of the present invention may also be a food composition. When the composition for improving muscle function or enhancing exercise performance of the present invention is a food composition, it may be used for preventing or improving muscle diseases due to muscle wasting or degeneration. Muscle depletion and degeneration occur due to genetic factors, acquired factors, aging, etc., muscle depletion is characterized by a gradual loss of muscle mass, weakening and degeneration of muscles, especially skeletal or veterinary and cardiac muscles. Examples of related diseases include atony, muscular atrophy, muscular dystrophy, muscle degeneration, myasthenia, cachexia and sarcopenia. The composition of the present invention has an effect of increasing muscle mass, the type of muscle is not limited.

The food composition of the present invention includes all forms such as functional food, nutritional supplement, health food, food additives and feed, and includes animals including humans or livestock. It is for eating.

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 (e.g. canned fruit, canned foods, jams, marmalade, etc.), fish, meat and processed foods (e.g. hams, sausages) Cornbread, etc.), breads and noodles (e.g. udon, soba noodles, ramen, spagate, macaroni, etc.), fruit juices, various drinks, cookies, malts, dairy products (e.g. butter, cheese), edible vegetable oils, margarine In addition, the vegetable protein, retort food, frozen food, various seasonings (eg, miso, soy sauce, sauce, etc.) may be prepared by adding the above-mentioned chilenol or a rare extract containing the same. In addition, the nutritional supplements are not limited thereto, and may be prepared by adding the chilenol or a rare extract containing the same to capsules, tablets, pills, and the like. In addition, as a health functional food, but is not limited to, for example, the liquefied, granulated, encapsulated so that the chilenol or a rare extract containing the same itself is produced in the form of tea, juice and drinks for drinking (health drink) And it can be prepared by powdering. In addition, the chilenol or a rare extract containing the same may be prepared and used in the form of a powder or a concentrate for use in the form of a food additive. In addition, it can be prepared in the form of a composition by mixing with the chienol or a rare extract containing the same and known active ingredients known to have an effect of improving muscle function or enhancing exercise performance.

When the composition for improving muscle function or enhancing exercise performance of the present invention is used as a health beverage composition, the health beverage composition may contain various flavors or natural carbohydrates, etc. as additional ingredients, as in general beverages. The above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; Polysaccharides such as dextrin, cyclodextrin; Sugar alcohols such as xylitol, sorbitol, and erythritol. Sweeteners include natural sweeteners such as taumartin, stevia extract; Synthetic sweeteners such as saccharin and aspartame; The proportion of said 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.

Kirenol or a rare extract containing the same may be contained as an active ingredient of a food composition for improving muscle function or enhancing exercise performance, the amount of which is particularly limited to an amount effective to achieve the effect of improving muscle function or enhancing exercise performance. It is preferably, but it is preferably from 0.01 to 100% by weight relative to the total weight of the composition.

In addition to the above, the health food of the present invention is various nutrients, vitamins, electrolytes, flavors, coloring agents, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, Glycerin, alcohol or carbonation agent, and the like. In addition, the health food of the present invention may contain a flesh for preparing natural fruit juice, fruit juice beverage, or vegetable beverage. These components can be used independently or in combination. The proportion of such additives is not critical but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

The present invention also provides a method for improving muscle function or improving exercise performance, comprising administering to a subject a composition comprising a rare extract or a fraction thereof as an active ingredient.

In one embodiment, the subject refers to a subject in need of improving muscle function or enhancing exercise performance, but may be, but is not limited to, a mammal including a human.

In one embodiment, the rare extract may be one or more selected from the group consisting of Siegesvecchia glabresense, Siegesvecchia fubesense, and Siegesvecchia orientalis.

In one embodiment, the rare extract may be obtained by extracting the rare extract with one or more solvents selected from the group consisting of water, an organic solvent having 1 to 6 carbon atoms, a subcritical fluid and a supercritical fluid.

In one embodiment, the organic solvent having 1 to 6 carbon atoms may be selected from alcohols having 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, It may be at least one selected from the group consisting of methylene chloride, hexane, cyclohexane, and petroleum ether.

In one embodiment, the rare extract may be obtained by extracting the rare earth under ultra-high pressure conditions of 100 MPa or more.

In one embodiment, the fraction of the rare extract may be obtained by fractionating the rare extract with ethyl acetate, methanol or a mixed solvent thereof.

In one embodiment, the method of improving muscle function or enhancing motor performance consists of atony, muscular atrophy, muscular dystrophy, muscle degeneration, work sickness, cachexia and sarcopenia. By treating, preventing or ameliorating one or more diseases selected from the group.

The present invention also provides a method for improving muscle function or improving exercise performance, comprising administering to a subject a composition comprising a compound represented by Formula 1 as an active ingredient:

[Formula 1]

In one embodiment, the compound represented by Formula 1 may be a chilenol represented by Formula 2:

[Formula 2]

In one embodiment, the subject refers to a subject in need of improving muscle function or enhancing exercise performance, but may be, but is not limited to, a mammal including a human.

In one embodiment, the compound represented by Formula 1 may be isolated from one or more extracts or fractions thereof selected from the group consisting of Siegesvecchia glabresense, Siegesvecchia fubesense, and Siegesvecchia orientalis. have.

In one embodiment, the method of improving muscle function or enhancing motor performance consists of atony, muscular atrophy, muscular dystrophy, muscle degeneration, work sickness, cachexia and sarcopenia. By treating, preventing or ameliorating one or more diseases selected from the group.

All other matters described above regarding the composition for improving muscle function or improving exercise performance may be applied or mutatis mutandis to a method for improving muscle function or a method for enhancing exercise performance.

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are provided only for the purpose of illustration in order to help the understanding of the present invention, but the scope of the present invention is not limited thereto. The experimental results of the following examples were expressed as the mean ± standard deviation, statistical analysis was evaluated to be significant if the p value is less than 0.05 or 0.01 using the t test.

Reference Example 1: Kirenol Information

Designation: Kirenol; Kirel; (1R, 3S, 4aS, 4bS, 7S, 10aS) -1,2,3,4,4a, 4b, 5,6,7,9,10,10a-Dodecahydro-3-hydroxy-7-[(R) -1,2-dihydroxyethyl] -1,4a, 7-trimethylphenanthrene-1-methanol

CAS No .: 52659-56-0

Example 1 Preparation of Sigesvecchia Glabresense Extract

Example 1-1 Preparation of Siegesvecchia glabresense Ethanol Extract

The dried leaves and stems of Siegesvecchia glabresense were pulverized with a mixer, and then 100 g of the ground Siegesvecchia glabresense sample was placed in 1 L of ethanol and extracted with stirring at 50 ° C. for 60 minutes. The extracted sample was filtered using Whatman No. 2 filter paper, and the filtered extract was concentrated using a vacuum rotary concentrator to remove the solvent component, thereby obtaining an extract of Sigebexkia glabressen ethanol.

Example 1-2 Preparation of Siegesvecchia Glabresense Hot Water Extract

The dried leaves and stems of Siegesvecchia glabresense were ground with a mixer, and then 100 g of the ground Siegesvecchia glabresense sample was put in 1 L of water and extracted with stirring at 100 ° C. for 4 hours. The extracted sample was filtered through Whatman No. 2 filter paper, and the filtrate was concentrated with a vacuum rotary concentrator to remove the solvent component, thereby obtaining Sigebexkia glabresense hydrothermal extract.

Example 1-3 Preparation of Siegesvecchia Glabresense Hexane Extract

The dried leaves and stems of Siegesvecchia glabresense were pulverized with a mixer, and then 100 g of the ground Siegesvecchia glabresense sample was placed in 1 L of hexane and extracted with stirring at 50 ° C. for 60 minutes. The extracted sample was filtered through Whatman No. 2 filter paper, and the filtered extract was concentrated with a vacuum rotary concentrator to remove the solvent component, thereby obtaining Sigebexkia glabresense hexane extract.

Example 1-4 Preparation of Siegesvecchia Glabresense Ethyl Acetate Extract

The dried leaves and stems of Siegesvecchia glabresense were pulverized with a mixer, and 100 g of the ground Siegesvecchia glabresense sample was placed in 1 L of ethyl acetate and extracted with stirring at 50 ° C. for 60 minutes. The extracted sample was filtered using Whatman No. 2 filter paper, and the filtered extract was concentrated with a vacuum rotary concentrator to remove the solvent component, thereby obtaining Sigebexkia glabressen ethyl acetate extract.

Example 1-5 Preparation of Sigesvecchia Glabresense Ultra High Pressure Extract

The dried leaves and stems of Siegesvecchia glabresense were pulverized with a mixer, and then 1 g of the ground Siegesvecchia glabresense sample and 76 mL of 18% ethanol were put in a polyethylene pack and sealed, followed by ultra high pressure Extraction was performed using an extractor (Frescal MFP-7000; Mitsubishi Heavy Industries, Tokyo, Japan). The ultrahigh pressure extraction conditions were 320 MPa extraction time and 5 min extraction time. The extracted sample was filtered using Whatman No. 2 filter paper, and the filtered extract was concentrated with a vacuum rotary concentrator to remove the solvent component, thereby obtaining Sigebexkia glabresense ultrahigh pressure extract.

Example 1-6 Preparation of Sigesvecchia Glabresense Supercritical Fluid Extract

The dried leaves and stems of Siegesvecchia glabresense were ground with a mixer, and then 1 g of the ground Siegesvecchia glabresense sample was charged into a sample cartridge and a supercritical fluid extraction device (SFX 3560, Isco Inc. , Lincoln, NE, USA). Supercritical fluid extraction conditions were 20 MPa extraction pressure, 60 ° C extraction temperature, 60 mL / min flow rate of supercritical carbon dioxide, 60 min extraction time. When the supercritical fluid extraction was completed, the pressure of the extraction device was lowered to release the supercritical fluid state to obtain a Siegesvecchia glabresense supercritical fluid extract.

Example 1-7 Preparation of Sigesvecchia glabresense Subcritical Fluid Extract

The dried leaves and stems of Siegesvecchia glabresense were ground with a mixer, and then 1 g of the ground Siegesvecchia glabresense sample was placed in 10 mL of distilled water, and a subcritical fluid extraction device (DIONEX Accelerated Solvent Extractor 100, DIONEX co., USA). Subcritical fluid extraction conditions were performed under the conditions of extraction pressure 0.5 MPa, extraction temperature 240 ℃, extraction time 20 minutes. The extracted sample was filtered through Whatman No. 2 filter paper, and the filtered extract was freeze-dried at -40 ° C to obtain a Siegesvecchia glabresense subcritical fluid extract.

Example 2 Preparation of Siegesvekchia fuvesense Extract

Example 2-1 Preparation of Sigestesvecchia fuvesense Ethanol Extract

The dried leaves and stems of Siegesvecchia glabresense were pulverized with a mixer, and then 100 g of the ground Siegesvecchia glabresense sample was placed in 1 L of ethanol and extracted with stirring at 50 ° C. for 60 minutes. The extracted sample was filtered using Whatman No. 2 filter paper, and the filtered extract was concentrated with a vacuum rotary concentrator to remove the solvent component, thereby obtaining an extract of Sievesbeckia fuvesense ethanol.

Example 2-2 Preparation of Siegesvekchia fuvesense Hot Water Extract

The leaves and stems of dried Siegesvecchia fuveycens were ground with a mixer, and then 100 g of the ground Siegesvecchia fuveysense sample was placed in 1 L of water and extracted with stirring at 100 ° C for 4 hours. The extracted sample was filtered through Whatman No. 2 filter paper, and the filtered extract was concentrated with a vacuum rotary concentrator to remove the solvent component, thereby obtaining Sigebexkia fuvacense hot water extract.

Example 2-3 Preparation of Sigesvecchia fuvesense Hexane Extract

The leaves and stems of dried Siegesvecchia fuveycens were pulverized with a mixer, and then 100 g of the ground Siegesvecchia fuveysense sample was placed in 1 L of hexane and extracted with stirring at 50 ° C. for 60 minutes. The extracted sample was filtered through Whatman No. 2 filter paper, and the filtered extract was concentrated with a vacuum rotary concentrator to remove the solvent component, thereby obtaining Sigebexkia fuvacense hexane extract.

Example 2-4 Preparation of Siegesvecchia fuvesense Ethyl Acetate Extract

The leaves and stems of dried Siegesvecchia fuveycens were pulverized with a mixer, and 100 g of the ground Siegesvecchia fuveysense sample was put in 1 L of ethyl acetate and extracted with stirring at 50 ° C. for 60 minutes. The extracted sample was filtered through Whatman No. 2 filter paper, and the filtered extract was concentrated using a vacuum rotary concentrator to remove the solvent component, thereby obtaining Sigebexkia fuvacense ethyl acetate extract.

Example 2-5 Preparation of Sigesvecchia fuveycens Ultra High Pressure Extract

The dried leaves and stems of Siegesvecchia fuveysense were pulverized with a mixer, and then 1 g of ground Siegesvecchia fuescens samples and 76 mL of 18% ethanol were put in a polyethylene pack and sealed, and then the ultra-high pressure extractor Extracted using (Frescal MFP-7000; Mitsubishi Heavy Industries, Tokyo, Japan). The ultrahigh pressure extraction conditions were 320 MPa extraction time and 5 min extraction time. The extracted sample was filtered through Whatman No. 2 filter paper, and the extract was concentrated with a vacuum rotary concentrator to remove the solvent component, thereby obtaining a Siegesvecchia fuvacense ultrahigh pressure extract.

Example 2-6 Preparation of Sigesvecchia fuveycens Supercritical Fluid Extract

The dried leaves and stems of Siegesvecchia fuveysense are ground with a mixer, and then 1 g of the ground Siegesvecchia fuveysense sample is charged into a sample cartridge and a supercritical fluid extraction device (SFX 3560, Isco Inc., Lincoln , NE, USA). Supercritical fluid extraction conditions were 50 MPa extraction pressure, 40 ℃ extraction temperature, 60 mL / min flow rate of supercritical carbon dioxide, 60 min extraction time. When the supercritical fluid extraction was completed, the pressure of the extraction device was lowered to release the supercritical fluid state to obtain a Siegesvecchia fuvesense supercritical fluid extract.

Example 2-7 Preparation of Sigesvecchia fuveycens Subcritical Fluid Extract

After grinding the dried leaves and stems of Siegesvecchia fuveysense with a mixer, 1 g of the ground Siegesvecchia fuveysense sample was added to 10 mL of distilled water, and a subcritical fluid extraction device (DIONEX Accelerated Solvent Extractor 100, DIONEX co , USA). Subcritical fluid extraction conditions were carried out under the conditions of extraction pressure 5 MPa, extraction temperature 90 ℃, extraction time 15 minutes. The extracted sample was filtered through Whatman No. 2 filter paper, and the filtered extract was lyophilized at -40 ° C to obtain a Siegesbeckia fuvacense subcritical fluid extract.

Example 3 Preparation of Sigesvecchia Orientalis Extract

Example 3-1 Preparation of Siegesvecchia Orientalis Ethanol Extract

The dried leaves and stems of Siegesvecchia orientalis were ground with a mixer, and then 100 g of the Siegesvecchia orientalis sample was put in 1 L of ethanol and extracted with stirring at 50 ° C. for 60 minutes. The extracted sample was filtered using Whatman No. 2 filter paper, and the filtered extract was concentrated with a vacuum rotary concentrator to remove the solvent component, thereby obtaining an extract of Siegesvecchia orientalis ethanol.

Example 3-2 Preparation of Siegesvecchia Orientalis Hot Water Extract

The dried leaves and stems of Siegesvecchia orientalis were ground with a mixer, and then 100 g of the ground Siegesvecchia orientalis sample was added to 1 L of water and extracted with stirring at 100 ° C for 4 hours. The extracted sample was filtered through Whatman No. 2 filter paper, and the filtrate was concentrated with a vacuum rotary concentrator to remove the solvent component, thereby obtaining Sigebexkia orientalis hydrothermal extract.

Example 3-3 Preparation of Sigesvecchia Orientalis Hexane Extract

The leaves and stems of dried Siegesvecchia orientalis were ground with a mixer, and then 100 g of the Siegesvecchia orientalis sample was added to 1 L of hexane and extracted with stirring at 50 ° C. for 60 minutes. The extracted sample was filtered through Whatman No. 2 filter paper, and the filtered extract was concentrated with a vacuum rotary concentrator to remove the solvent component, thereby obtaining an extract of Sigebecia orientalis hexane.

Example 3-4 Preparation of Sigesvecchia Orientalis Ethyl Acetate Extract

The dried leaves and stems of Siegesvecchia orientalis were ground with a mixer, and then 100 g of the Siegesvecchia orientalis sample was added to 1 L of ethyl acetate and extracted with stirring at 50 ° C. for 60 minutes. The extracted sample was filtered through Whatman No. 2 filter paper, and the filtrate was concentrated with a vacuum rotary concentrator to remove the solvent component, thereby obtaining an extract of Sigestescchia orientalis ethyl acetate.

Example 3-5 Preparation of Sigesvecchia Orientalis Ultra High Pressure Extract

The dried leaves and stems of Siegesvecchia orientalis were ground with a mixer, and then 1 g of the ground Siegesvecchia orientalis sample and 76 mL of 18% ethanol were put in a polyethylene pack and sealed, and then the ultra-high pressure extractor Extracted using (Frescal MFP-7000; Mitsubishi Heavy Industries, Tokyo, Japan). The ultrahigh pressure extraction conditions were 320 MPa extraction time and 5 min extraction time. The extracted sample was filtered through Whatman No. 2 filter paper, and the filtered extract was concentrated with a vacuum rotary concentrator to remove the solvent component, thereby obtaining an extract of Sigesvecchia orientalis ultrahigh pressure.

Example 3-6 Preparation of Sigesvecchia Orientalis Supercritical Fluid Extract

The dried leaves and stems of Siegesvek orientalis are pulverized with a mixer, and then 1 g of the ground Siegesvecchia orientalis sample is filled into a sample cartridge and a supercritical fluid extraction device (SFX 3560, Isco Inc., Lincoln) , NE, USA). Supercritical fluid extraction conditions were 40 MPa extraction pressure, 50 ℃ extraction temperature, 60 mL / min flow rate of supercritical carbon dioxide, 60 min extraction time. When the supercritical fluid extraction was completed, the pressure of the extraction device was lowered to release the supercritical fluid state to obtain a Siegesvecchia orientalis supercritical fluid extract.

 Example 3-7 Preparation of Sigesvecchia Orientalis Subcritical Fluid Extract

The leaves and stems of the dried Sigestescchia orientalis were ground using a mixer, and then 1 g of the ground Sigesvecchia orientalis sample was added to 10 mL of distilled water, and a subcritical fluid extraction unit (DIONEX Accelerated Solvent Extractor 100, DIONEX co , USA). Subcritical fluid extraction conditions were carried out under the conditions of 2.5 MPa, extraction temperature 150 ℃, extraction time 15 minutes. The extracted sample was filtered through Whatman No. 2 filter paper, and the filtered extract was lyophilized at -40 ° C to obtain a Siegexvek orientalis subcritical fluid extract.

Example 4 Isolation and Structure Determination of Kirenol

Example 4-1 Separation of Kirenol

The concentrated Sigestescchia orientalis ethanol extract obtained in Example 3-1 was loaded on a column filled with silica gel, and ethyl acetate and methanol were mixed at a ratio of 10: 0.5 (v / v) using a solvent system. Aliquoted. The fractions were divided into a total of seven fractions according to the aliquots and concentrated to dry each fraction. Fraction 6 of 7 fractions (fraction 6) were aliquoted into 10% ethylacetate of developing solvent using RP-18 reverse phase column chromatography (Lichroprep RP-18 25-40um, Merck & Co., Whitehouse Station, NJ, USA). . Each fraction was concentrated to dryness by dividing into a total of two fractions according to the aliquot sequence. Fraction 2 of the two fractions (fraction 6-2) was concentrated to dryness and again fractionated with 20% ethyl acetate in developing solvent using RP-18 reverse phase column chromatography. Each fraction was concentrated to dryness by dividing into a total of three fractions according to the preparative sequence. Finally fraction 2 of the three fractions (fraction 6-2-2) was concentrated to dryness to separate a single pure active substance.

Example 4-2 Structure Determination of Kirenol

In order to determine the structure of the single active material separated in Example 4-1, ¹ H-NMR spectrum and ¹³ C-NMR spectrum were measured at 500 MHz and 125 MHz (solvent: MeOH), respectively. The ¹ H ¹ H COSY spectrum and ¹ H ¹³ C HSQC spectra in order to obtain the ¹ H-NMR measurement of correlation between the spectrum and the ¹³ C-NMR correlation of the ¹ H ¹ H on the basis of the result of the spectrum related to the ¹ H ¹³ C relationships Each carbon signal was distinguished by the wavelength emitted through the carbon resonance, and the result was measured.

In addition, FAB-MS was measured for the mass spectrometry of the isolated single material. This compound had a molecular weight of 338.48 as [M] was observed at m / z 338.48 in FAB-MS, and has the molecular formula C ₂₀ H ₃₄ O ₄ .

The results of the above ¹ H-NMR, ¹³ C-NMR, and FAB-MS and previously published research reports (Wang JP et al., Pharmacogn. Mag., 8: 149-155, 2012) were identified and compared. As a result, the isolated single substance was identified as a kirenol compound represented by the following Chemical Formula 2.

[Formula 2]

Example 5 Protein Expression Increase Effect of P-mTOR, a Major Gene Involved in Muscle Function

[Example 5-1] Effects of improving the muscle function of chirenol or Sigestesevikia glabressenth ethanol extract, Siegesbeckia fuvecens ethanol extract, Sieves Beckia orientalis ethanol extract

Muscle cells, L6 myoblast (American Type Culture Collection, Manassas, VA, USA), were treated with 10% fetal bovine serum (FBS; Hyclone, Logan, UT, USA) and 100 U / mL penicillin, 100 μg / mL streptomycin. Cultured in Dulbecco's modified Eagle's media (DMEM; Hyclone) containing (Gibco, Grand Island, NY, USA). Sigebexkia glabracens ethanol extract prepared in Example 1-1, Sigebexkia fuvesense ethanol extract prepared in Example 2-1, Sigebexki prepared in Example 3-1 on L6 cells Orientalis ethanol extract was treated to 10 ppm concentration. In addition, L6 cells were treated with the chilenol prepared in Example 4 at a concentration of 10 μM. At this time, the group treated with 0.01% DMSO instead of the sample was used as a control. After 24 hours, L6 cells were lysed with RIPA (ELPIS-Biotech, Daejeon, Korea) buffer solution containing a proteinase inhibitor cocktail (Sigma-Aldrich, St. Louis, MO, USA). After boiling the sample for 5 minutes, the same amount of protein (20 μg) was separated by electrophoresis with 10% SDS-PAGE. After electrophoresis, the separated proteins were transferred to nitrocellulose membrane and Western blot was performed. After reacting with the primary antibody, it was washed three times for 10 minutes using Tris-buffered saline containing 0.1% Tween 20 (TBST). At this time, the type and dilution ratio of the primary antibody used in the present invention is 1: 1000. In the secondary antibody reaction, a secondary antibody (anti-rabbit horseradish) was added to the membrane subjected to the primary antibody reaction and reacted at room temperature for 2 hours. At this time, the dilution ratio of the secondary antibody was 1: 5000. The protein band was developed using ECL western blotting detection reagents (Amersham, Tokyo, Japan) to confirm the protein expression of the major gene p-mTOR involved in muscle function, and α-tubulin. It is shown that the protein loading is constant.

As a result, as shown in Fig. 1, chilenol or Sigebec kia glabressenth ethanol extract, Sigebec kia fuvecens ethanol extract, Sige beckia orientalis ethanol extract treatment according to the major involved in improving muscle function Protein expression level of the gene p-mTOR was found to increase.

These results indicate that the chilenol of the present invention, the rare extract containing the same or fractions thereof have the activity of increasing muscle mass through the increase of the protein expression level of p-mTOR.

[Example 5-2] Muscle Function Improvement Effect of Sigesvecchia Orientalis Ultra High Pressure Extract

Sigebexchia orientalis ultra-high pressure extract prepared in Example 3-5 was treated to muscle cells in the same manner as in Example 5-1 at a concentration of 10, 20, 40 ppm.

As a result, as shown in FIG. 2, it was found that the protein expression level of the main gene p-mTOR involved in muscle function improvement was increased by treatment with Sigebekicia orientalis ultra high pressure extract containing chilenol.

These results indicate that the chilenol of the present invention, the rare extract containing the same or fractions thereof have the activity of increasing muscle mass through the increase of the protein expression level of p-mTOR.

[Example 5-3] Muscle Function Improvement Effect of Rare Ethyl Acetate Extract

Siege Beckia glavesense ethyl acetate extract prepared in Example 1-4, Siege Beckia fuvesense ethyl acetate extract prepared in Example 2-4, Siegece prepared in Example 3-4 Beckia orientalis ethyl acetate extract was treated to muscle cells in the same manner as in Example 5-1 at a concentration of 10 ppm.

The p-mTOR protein band was developed using ECL western blotting detection reagents (Amersham, Tokyo, Japan) and the protein band developed using G; BOX EF imaging system (Syngene, Cambridge, UK). The density was measured. In this case, the density of the control protein band was 100%, and the density of the relative protein band of the experimental group treated with the sample was expressed as a percentage (%).

The results are shown in Table 1 below.

As shown in Table 1, the Siege Beckia glabreense ethyl acetate extract, the Siege Beckia fuvesense ethyl acetate extract, and the Siege Beckia orientalis ethyl acetate extract are proteins of the main gene p-mTOR involved in muscle function improvement. It was found to increase the amount of expression.

These results indicate that the chilenol of the present invention, the rare extract containing the same or fractions thereof have the activity of increasing muscle mass through the increase of the protein expression level of p-mTOR.

[Example 5-4] Muscle Function Improvement Effect of Rare Supercritical and Subcritical Extracts

Sigebexkia glabresense supercritical and subcritical extracts prepared in Examples 1-6 and 1-7, Sigebexkia fuvesense prepared in Examples 2-6 and 2-7 Supercritical and subcritical extracts, the Siegesvecchia orientalis supercritical and subcritical extracts prepared in Examples 3-6 and 3-7 at 20 ppm concentration in the same manner as in Example 5-1 The cells were treated. Next, the density of the control protein band was 100% in the same manner as in Example 5-3, and the density of the relative protein band of the experimental group treated with the sample was expressed as a percentage (%).

The results are shown in Table 2 below.

As shown in Table 2, the Siege Beckia glabreense, Siege Beckia fuubesense, Siege Beckia orientalis supercritical and subcritical extracts express the protein expression level of the main gene p-mTOR involved in muscle function improvement. It was found to increase.

These results indicate that the chilenol of the present invention, the rare extract containing the same or fractions thereof have the activity of increasing muscle mass through the increase of the protein expression level of p-mTOR.

[Example 5-5] Effect of Muscle Fraction of Fractions of Sigeresvecchia Orientalis Ethanol Extract

The fraction 6 of the 10: 0.5 (v / v) mixed solvent of ethyl acetate and methanol of the Siegesvecchia orientalis ethanol extract prepared in Example 4-1 was the same as Example 5-1 at 20 ppm. Muscle cells were treated by the method. Next, the density of the control protein band was 100% in the same manner as in Example 5-3, and the density of the relative protein band of the experimental group treated with the sample was expressed as a percentage (%).

As a result, it was found that the relative density of the control group of the extract of Siegesvecchia orientalis ethanol extract was 126%, increasing the protein expression level of the main gene p-mTOR involved in muscle function improvement.

These results indicate that the chilenol of the present invention, the rare extract containing the same or fractions thereof have the activity of increasing muscle mass through the increase of the protein expression level of p-mTOR.

Example 6 Muscle Differentiation, Increased Assimilation, and Reduced Catabolism

[Example 6-1] Effect of Muscle Differentiation and Increased Assimilation

Muscle cells, L6 myoblasts (American Type Culture Collection, Manassas, VA, USA) were cultured in DMEM (10% FBS, 100 U / mL penicillin, 100 g / mL streptomycin). When the cell density reached about 80-85%, the cell medium was replaced with DMEM growth medium containing 2% FBS, followed by differentiation for 6 days, and then experimented. After treatment with TNF-α for 24 hours in order to induce muscle reduction in differentiated L6 cells, Siegesvecchia orientalis ethanol extract (10, 40 ppm) prepared in Example 3-1 and Example 4 The prepared chilenol (10, 40 μM) was treated. RT-PCR was performed to determine the mRNA expression levels of myogenin and myoD, which are muscle differentiation regulators. Total RNA was harvested and reverse-transcribed using TRIzol reagent (Invitrogen, Carlsbad, Calif., USA) from differentiated cells, followed by RT-PCR analysis. The RNA was first reverse transcribed with reverse transcriptase for cDNA synthesis. RT-PCR was performed with the following specific primers, indicating that mRNA loading was constant with β-actin.

Myogenin:

SEQ ID NO: 5'-TGGGCTGCCACAAGCCAGAC-3 '(Forward primer)

SEQ ID NO: 5'-CAGCCCAGCCACTGGCATCA-3 '(Reverse primer)

MyoD:

SEQ ID NO: 5'-GGATGGTGCCCCTGGGTCCT-3 '(Forward primer)

SEQ ID NO: 5'-TGGCCTTCGCTGTGAGTCGC-3 '(Reverse primer)

β-actin:

SEQ ID NO: 5'-AGCCATGTACGTAGCCATCC-3 '(Forward primer)

SEQ ID NO: 5'-CTCTCAGCTGTGGTGGTGAA-3 '(Reverse primer)

As a result, it can be seen that as shown in Figures 3 and 4 Siegesvekchia orientalis ethanol extract or Kirenol increases the mRNA expression levels of myogenin and myoD which are muscle differentiation regulating factors.

These results indicate that the chilenol of the present invention, the rare extract containing the same or fractions thereof increase muscle differentiation and assimilation, thereby contributing to muscle formation.

Example 6-2 Muscle Catabolism Reducing Effect

Total RNA was harvested and reverse transcribed from cells differentiated in the same manner as in Example 3-1, followed by RT-PCR. At this time, the Siegesvecchia orientalis ethanol extract (10, 40 ppm) prepared in Example 3-1 and the chilenol (10, 40 μM) prepared in Example 4 were treated. RT-PCR was performed with the following specific primers.

Art rosin-1:

SEQ ID NO: 5'-CCCTGAGTGGCATCGCCCAA-3 '(Forward primer)

SEQ ID NO: 5'-AGGTCCCGCCCATCGCTCA-3 '(Reverse primer)

MuRF1:

SEQ ID NO: 5'-TCTACTCGGCCACAGGCGCT-3 '(Forward primer)

SEQ ID NO: 10'-CTTGACAGCTCCCGCCGCAA-3 '(Reverse primer)

As a result, mRNA expression levels of atherosine-1 and MuRF1, which are major ligase involved in catabolism of muscle protein of Siegesvecchia orientalis ethanol extract or Kirenol, as shown in FIGS. 5 and 6 It can be seen that the decrease.

This result means that the kyrenol of the present invention, a rare extract containing the same or a fraction thereof inhibits the muscle catabolism involved in muscle loss, thereby contributing to the maintenance of muscle mass.

Example 7 Confirmation of Muscle Mass Increase Effect in Normal Dietary Animal Model

Example 7-1 Experimental Animal Diet

Four-week-old C57BL / 6 male mice were acclimated for one week and the diet provided a normal diet (Research Diets D12450B, 10% kcal from fat, New Brunswick, NJ, USA). Five dogs each were divided into three groups and used for the experiment. In the experimental group, the Siege Beckia orientalis ethanol extract prepared in Example 3-1 was suspended in 0.25% carboxymethyl cellulose, and the Siege Beckia orientalis ethanol extract 250 mg / day / kg Once a day for 6 weeks orally was administered orally at a constant time, the control group was orally administered only the same amount of 0.25% carboxymethyl cellulose. The diet and the weight of the subjects were measured every week.

Example 7-2 Effect of Muscle Mass Increase

Both calf muscles were extracted from the normal diet mice of Example 7-1 and weighed by microbalance (Mettler PE160, USA). The test result was verified by the t-test between the normal diet group and the Siegesbeckia orientalis ethanol extract administration group, and showed a statistically significant difference (** p <0.01).

As a result, as shown in Figure 7, compared to the control group compared to the control group, the weight of the muscles increased by 23% in the group administered with the Siegesvecchia orientalis ethanol extract.

These results indicate that the chilenol of the present invention, the rare extract containing the same or a fraction thereof have an effect on increasing muscle mass.

Example 7-3 Muscle Volume Increase Effect

Final muscle volume was measured using micro PET / CT (positron emission tomography / computed tomography, INVEON, Siemens, USA) from the normal diet of Example 7-1.

As a result, as shown in Figure 8, compared to the control group compared to the control group was administered 11.4% of the volume of muscle increased in the group administered with ethanol extract Siegesvecchia orientalis.

These results indicate that the chilenol of the present invention, the rare extract containing the same, or a fraction thereof has an effect on increasing muscle mass by increasing muscle volume.

Example 7-4 Effect of Muscle Mass Increase on Rare Hot Water Extract

Siege Beckia glabracens hydrothermal extract prepared in Example 1-2 in the same manner as in Example 7-1 and Example 7-2, Siege Beckia fuvacense prepared in Example 2-2 The muscle mass weights of the hydrothermal extracts and the Siegesvecchia orientalis hydrothermal extracts prepared in Example 3-2 were measured.

The results are shown in Table 3 below.

As shown in Table 3, compared to the control group, it was found that the muscle mass of the Siege Beckia glabracens hydrothermal extract, Siege Beckia fuubessen hydrothermal extract, and Siege Beckia orientalis hydrothermal extract were significantly increased. .

These results indicate that the chilenol of the present invention, the rare extract containing the same or a fraction thereof have an effect on increasing muscle mass.

Example 8 Confirmation of the Muscle Mass Increase Effect in the High Fat Diet Animal Model

Example 8-1 Experimental Animal Diet

Four-week-old C57BL / 6 male mice were acclimated for 1 week and fed with a high-fat diet (Research Diets D12492, 60% kcal from fat) for 6 weeks to induce obesity. In the same manner as in Example 7-1 was administered orally to the obese animal model.

Example 8-2 Effect of Muscle Mass Increase

Both calf muscles were extracted from the high fat diet mouse of Example 8-1, and the weight was measured in the same manner as in Example 7-2. The experimental results were tested by the t-test of the high-fat diet group and the high-fat diet Siegesvecchia orientalis ethanol extract group, and the significance was verified (** p <0.01).

As a result, as shown in Figure 9, compared to the control group compared to the control group, the weight of the muscles increased by 23% in the group administered with the Siegesvek orientalis ethanol extract.

This means that the chilenol of the present invention, the rare extract containing the same or a fraction thereof have an effect on increasing muscle mass.

Example 8-3 Muscle Volume Increase Effect

The high-fat diet of Example 8-1 was used to measure muscle volume in the same manner as in Example 7-3.

As a result, as shown in Figure 10, compared to the control group compared to the control group, the volume of muscle increased 6% in the group administered with ethanol extract Siegesvecchia orientalis.

These results indicate that the chilenol of the present invention, the rare extract containing the same, or a fraction thereof has an effect on increasing muscle mass by increasing muscle volume.

Example 9 Protein Expression Increase Effect of PGC-1α, a Major Gene Involved in Exercise Performance

Example 9-1 Effect of Enhancing Exercise Performance of Cyrenol or Sigestesckkia Glabressenth Ethanol Extract, Sievesbeckia Fuvesense Ethanol Extract, Siesbekschia Orientalis Ethanol Extract

Muscle cells, L6 myoblast (American Type Culture Collection, Manassas, VA, USA), were treated with 10% fetal bovine serum (FBS; Hyclone, Logan, UT, USA) and 100 U / mL penicillin, 100 μg / mL streptomycin. Cultured in Dulbecco® modified Eagle® media (DMEM; Hyclone) containing (Gibco, Grand Island, NY, USA). Sigebexkia glabracens ethanol extract prepared in Example 1-1, Sigebexkia fuvesense ethanol extract prepared in Example 2-1, Sigebexki prepared in Example 3-1 on L6 cells Orientalis ethanol extract was treated to 10 ppm concentration. In addition, L6 cells were treated with the chilenol prepared in Example 4 at a concentration of 10 μM. At this time, the group treated with 0.01% DMSO instead of the sample was used as a control. After 24 hours, L6 cells were lysed with RIPA (ELPIS-Biotech, Daejeon, Korea) buffer solution containing a proteinase inhibitor cocktail (Sigma-Aldrich, St. Louis, MO, USA). After boiling the sample for 5 minutes, the same amount of protein (20 μg) was separated by electrophoresis with 10% SDS-PAGE. After electrophoresis, the separated proteins were transferred to nitrocellulose membrane and Western blot was performed. After reacting with the primary antibody, it was washed three times for 10 minutes using Tris-buffered saline containing 0.1% Tween 20 (TBST). At this time, the type and dilution ratio of the primary antibody used in the present invention is 1: 1000. In the secondary antibody reaction, a secondary antibody (anti-rabbit horseradish) was added to the membrane subjected to the primary antibody reaction and reacted at room temperature for 2 hours. At this time, the dilution ratio of the secondary antibody was 1: 5000. Protein bands were developed using ECL western blotting detection reagents (Amersham, Tokyo, Japan) to confirm the protein expression of the major gene PGC-1α involved in motor performance, and α-tubulin (α- tubulin) showed a constant protein loading.

As a result, as shown in FIG. 11, chirenol or Sigebec kia glabressen ethanol extract, Sige beckia fuvesense ethanol extract, and Sige beckia orientalis ethanol extract were involved in exercise performance. Protein expression of the major gene PGC-1α was found to increase.

These results indicate that the chilenol of the present invention, the rare extract containing the same or fractions thereof have the activity of enhancing exercise performance through the increase in the amount of PGC-1α protein expression.

Example 9-2 Exercise Performance Enhancement Effect of Sigesvecchia Orientalis Ultra High Pressure Extract

Sigebexkia orientalis ultra-high pressure extract prepared in Example 3-5 was treated to muscle cells in the same manner as in Example 9-1 at a concentration of 10, 20, 40 ppm.

As a result, as shown in FIG. 12, it was found that the expression level of the protein of PGC-1α, which is a major gene involved in exercise performance, was increased according to the treatment of Siegveskchia orientalis ultra high pressure extract containing chilenol.

These results indicate that the chilenol of the present invention, the rare extract containing the same or fractions thereof have the activity of enhancing exercise performance through the increase in the amount of PGC-1α protein expression.

Example 9-3 Exercise Performance Enhancement Effect of Rare Ethyl Acetate Extract

Siege Beckia glavesense ethyl acetate extract prepared in Example 1-4, Siege Beckia fuvesense ethyl acetate extract prepared in Example 2-4, Siegece prepared in Example 3-4 Beckia orientalis ethyl acetate extract was treated to muscle cells in the same manner as in Example 5-1 at a concentration of 10 ppm.

The PGC-1α protein band was developed using ECL western blotting detection reagents (Amersham, Tokyo, Japan), and the protein band developed using G; BOX EF imaging system (Syngene, Cambridge, UK). The density was measured. In this case, the density of the control protein band was 100%, and the density of the relative protein band of the experimental group treated with the sample was expressed as a percentage (%).

The results are shown in Table 4 below.

As shown in Table 4, the Siege Beckia glabreense ethyl acetate extract, the Siege Beckia fuvesense ethyl acetate extract, and the Siege Beckia orientalis ethyl acetate extract are involved in the exercise performance of PGC-1α. It was found to increase the amount of protein expression.

These results indicate that the chilenol of the present invention, the rare extract containing the same or fractions thereof have the activity of enhancing exercise performance through the increase in the amount of PGC-1α protein expression.

Example 9-4 Exercise Performance Enhancement Effects of Rare Supercritical and Subcritical Extracts

Sigebexkia glabresense supercritical and subcritical extracts prepared in Examples 1-6 and 1-7, Sigebexkia fuvesense prepared in Examples 2-6 and 2-7 Supercritical and subcritical extracts, the Siegesvecchia orientalis supercritical and subcritical extracts prepared in Examples 3-6 and 3-7 at 20 ppm concentration in the same manner as in Example 5-1 The cells were treated. Next, the density of the control protein band was 100% in the same manner as in Example 9-3, and the density of the relative protein band of the experimental group treated with the sample was expressed as a percentage (%).

The results are shown in Table 5 below.

As shown in Table 5, the Shigebexkia glabresense, Sigebexkia fuebense, Sigebexkia orientalis supercritical and subcritical extracts are expressed in the amount of protein expression of the main gene PGC-1α involved in exercise performance It can be seen that increases.

These results indicate that the chilenol of the present invention, the rare extract containing the same or fractions thereof have the activity of enhancing exercise performance through the increase in the amount of PGC-1α protein expression.

[Example 9-5] The Exercise Performance Enhancement Effect of the Fractions of Siegesbeckia Orientalis Ethanol Extract

The fraction 6 of the 10: 0.5 (v / v) mixed solvent of ethyl acetate and methanol of the Siegesvecchia orientalis ethanol extract prepared in Example 4-1 was the same as Example 5-1 at 20 ppm. Muscle cells were treated by the method. Next, the density of the control protein band was 100% in the same manner as in Example 5-3, and the density of the relative protein band of the experimental group treated with the sample was expressed as a percentage (%).

As a result, the relative density of the control group of the extract of Siegesvek orientalis ethanol extract was 168%. It was found to increase.

These results indicate that this result, the chilenol of the present invention, a rare extract containing the same or a fraction thereof has the activity to enhance the exercise performance through the increase in the amount of PGC-1α protein expression.

Example 10 Checking the Effect of Improving Exercise Performance in the Normal Dietary Animal Model

Example 10-1 Experimental Animal Diet

Four-week-old C57BL / 6 male mice were acclimated for one week and the diet provided a normal diet (Research Diets D12450B, 10% kcal from fat, New Brunswick, NJ, USA). Five dogs each were divided into three groups and used for the experiment. In the experimental group, the chirenol prepared in Example 4 and the Siegesvecchia orientalis ethanol extract prepared in Example 3-1 were suspended in 0.25% carboxymethylylcellulose, respectively, and the Siegesvecchia orientalis ethanol. The extract was administered orally once a day for 6 weeks at a concentration of 250 mg / day / kg body weight, and the control group was orally administered the same amount of 0.25% carboxymethylcellulose. The diet and the weight of the subjects were measured every week.

[Example 10-2] Exercise Performance Improvement Effect

After administering the sample for 6 weeks, the exercise performance was evaluated using a treadmill. The performance evaluation was performed 20 minutes at a speed of 25 cm / sec at a slope of 5 °, 20 minutes at a speed of 30 cm / sec, 20 minutes at a speed of 33 cm / sec, 20 minutes at a speed of 36 cm / sec, 36 cm / sec. The maximum exercise performance of the test animals was measured by running 15 minutes at a slope of 10 °, 15 minutes at a speed of 38 cm / sec, and 41 cm / sec. The point of determination of the maximum exercise performance was defined as the time when the experimental animals could not catch up with the treadmill speed for more than 10 seconds after the start of exercise, or the cumulative number of electric shocks exceeded 100 times in 5 minutes. Maximum exercise performance was measured by running distance and running time of the experimental animals. T-test was performed between the control group and the Sigebaccus orientalis ethanol extract-administered group to verify the significance of the experimental results. There was a statistically significant difference (** p <0.01).

As a result, as shown in Figure 13, compared to the control group was administered exercise exercise ability was increased in the group administered with Shigeresvecchia orientalis ethanol extract (a: exercise distance, b: exercise time ).

These results indicate that the chilenol of the present invention, the rare extract containing the same or fractions thereof have the effect of effectively enhancing the exercise performance.

[Example 10-3] Effect of Exercise Performance on Rare Hot Water Extract

Sigebexkia glabresense hydrothermal extract prepared in Example 1-2 in the same manner as in Example 10-1 and Example 10-2, Sigebexkia fuvacense prepared in Example 2-2 The exercise time for the hydrothermal extract, Siegesvecchia orientalis hydrothermal extract prepared in Example 3-2 was measured.

The results are shown in Table 6 below.

As a result, as shown in Table 6, the exercise time was significantly increased by Siege Beckia glabracens hydrothermal extract, Siege Beckia fuubessen hydrothermal extract, and Siege Beckia orientalis hydrothermal extract, respectively. It was found (p <0.01).

These results indicate that the chilenol of the present invention, the rare extract containing the same or fractions thereof have the effect of effectively enhancing the exercise performance.

Example 11 Confirmation of the Improvement of Exercise Performance in the Obese Animal Model Induced by High Fat Diet

Example 11-1 Experimental Animal Diet

Four-week-old C57BL / 6 male mice were acclimated for 1 week and fed with a high-fat diet (Research Diets D12492, 60% kcal from fat) for 6 weeks to induce obesity. Divided into and administered orally to the obese animal model in the same manner as in Example 10-1.

Example 11-2 Improvement of Exercise Performance Ability

The exercise performance was evaluated in the same manner as in Example 10-2 using the obese animal model induced by the high fat diet. The experimental results were tested by the t-test of the high-fat diet control group and the high-fat diet Siegesvecchia orientalis ethanol extract group, and the significance was verified (** p <0.01).

As a result, as shown in Figure 14, the high-fat diet compared to the control group was confirmed that the exercise performance increased in the group administered the Sigebes Beckia orientalis ethanol extract (a: exercise distance, b: exercise time).

These results indicate that the chilenol of the present invention, the rare extract containing the same or fractions thereof have the effect of effectively enhancing the exercise performance in the obesity model.

Example 12 Protein Expression Enhancement Effects of Genes Related to Motor Performance Enhancement in Muscle Tissues of Animal Models

Calf muscles were extracted from the normal diet mouse of Example 10-1 and the high-fat diet mouse of Example 11-1, and then, p-AMPK, SIRT1, PGC-1α, Protein expression of PPARδ was confirmed, indicating that protein loading was constant with α-tubulin.

As a result, as shown in Fig. 15, Sigesvecchia orientalis extract increased the protein expression levels of p-AMPK, SIRT1, PGC-1α, and PPARδ, which are genes related to the improvement of exercise ability in both the normal diet model and the high-fat diet model. Sikkim was known.

These results indicate that the chilenol of the present invention, the rare extract containing the same or fractions thereof have the effect of effectively increasing the exercise performance by increasing the protein expression of genes related to the improvement of the exercise ability in the animal model.

Example 13 Gene Expression Enhancement Effects Related to Mitochondrial Biosynthesis in Muscle Tissues of Animal Models

Calf muscles were extracted from the normal diet mice of Example 10-1 and the high-fat diet mice of Example 11-1, and then mRNA of PGC-1α, NRF-1, ERRα, and Tfam through RT-PCR. Expression was confirmed. Total RNA was harvested and reverse transcribed from calf muscle using TRIzol reagent (Invitrogen, Carlsbad, CA, USA), followed by RT-PCR analysis. The RNA was first reverse transcribed with reverse transcriptase for cDNA synthesis. RT-PCR was performed with the following specific primers:

PGC-1α:

SEQ ID NO: 5'-ATGTGTCGCCTTCTTGCTCT-3 '(Forward primer)

SEQ ID NO: 12'-ATCTACTGCCTGGGGACCTT-3 '(Reverse primer)

ERRα:

SEQ ID NO: 13: 5'-GCTGGTGGTTGAACCTGAGA-3 '(Forward primer)

SEQ ID NO: 14 5'-GGGCTAACACCCTATGCCAG-3 '(Reverse primer)

NRF-1:

SEQ ID NO: 15'-TGGACCCAAGCATTACGGAC-3 '(Forward primer)

SEQ ID NO: 5'-GGTCATTTCACCGCCCTGTA-3 '(Reverse primer)

Tfam:

SEQ ID NO: 17'5-TCGCCTGTCAGCCTTATCTG-3 '(Forward primer)

SEQ ID NO: 18'5-CCGGGCTTCCTTCTCTAAC-3 '(Reverse primer)

β-actin:

SEQ ID NO: 19: 5'-TAACCAACTGGGACGATATG-3 '(Forward primer)

SEQ ID NO: 20'-ATACAGGGACAGCACAGCCT-3 '(Reverse primer)

As a result, as shown in Figure 16, Sigebecia orientalis extract increased the mRNA expression levels of mitochondrial biosynthetic genes PGC-1α, NRF-1, ERRα, Tfam in the normal diet and high-fat diet model Could.

These results indicate that the chilenol of the present invention, the rare extract containing the same or fractions thereof increase mitochondrial biosynthesis, thereby effectively enhancing exercise performance.

Hereinafter, the preparation examples of the foods and medicines for improving muscle function or enhancing exercise performance, containing the chilenol or a rare extract containing the same according to the present invention as an active ingredient, but the present invention is not intended to be limited thereto. To explain. Food and pharmaceutical composition for improving muscle function or enhancing exercise performance of Preparation Examples 1 to 2 according to the composition components and the composition ratios as described below having a keenol or a rare extract containing the same excellent muscle function improving effect or exercise performance enhancing effect Was prepared according to conventional methods.

Production Example 1 Preparation of Food

Production Example 1-1 Preparation of Health Food

Examples 1 to 3 of the rare extract or Kirenol 1000 mg, Vitamin A Acetate 70 ug, Vitamin E 1.0 mg, Vitamin B1 0.13 mg, Vitamin B2 0.15 mg, Vitamin B6 0.5 mg, Vitamin B12 0.2 ug, Vitamin C 10 mg Biotin 10 ug, nicotinic acid amide 1.7 mg, folic acid 50 ug, calcium pantothenate 0.5 mg, ferrous sulfate 1.75 mg, zinc oxide 0.82 mg, magnesium carbonate 25.3 mg, potassium monophosphate 15 mg, dicalcium phosphate 55 mg, 90 mg of potassium citrate, 100 mg of calcium carbonate, and 24.8 mg of magnesium chloride may be mixed, and the compounding ratio may be arbitrarily modified. The above ingredients are mixed according to a conventional health food manufacturing method, and then granulated. It can be prepared and used in the manufacture of health food compositions according to conventional methods.

Preparation Example 1-2 Preparation of Health Beverage

Example 1 to 3 of the rare extract or Kirenol 1000 mg, citric acid 1000 mg, oligosaccharide 100 g, plum concentrate 2 g, taurine 1 g of purified water was added to the above ingredients according to the general 900 ml conventional health beverage production method After mixing and heating at 85 for about 1 hour after stirring, the resulting solution is filtered, obtained in a sterilized 2 L container, sealed and sterilized, and then refrigerated and then used for preparing a healthy beverage composition.

Preparation Example 1-3 Chewing Gum

Chewing gum was prepared in a conventional manner by combining 20% by weight of gum base, 76.9% by weight of sugar, 1% by weight of perfume, and 2% by weight of water, and 0.1% by weight of the rare extract or chilenol of Examples 1 to 3.

[Production Example 1-4] Candy

Candy was prepared in a conventional manner by combining 60% by weight of sugar, 39.8% by weight of starch syrup, and 0.1% by weight of fragrance, and 0.1% by weight of the rare extract or chilenol of Examples 1 to 3.

Preparation Example 1-5 Biscuits

Force 25.59% by weight, 22.22% by weight of gravity, 4.80% by weight, white salt 0.73% by weight, 0.78% by weight, palm shortening 11.78% by weight, ammonium 1.54% by weight, 0.17% by weight sodium bisulfite 0.16% by weight , 1.45 wt% rice flour, 0.0001 wt% vitamin B, 0.04 wt% milk, 20.6998 wt% water, 1.16 wt% whole milk powder, 0.29 wt% substitute milk powder, 0.03 wt% calcium monophosphate, 0.29 wt% spray salt Biscuits were prepared in a conventional manner by combining 7.27 wt% of oil and the rare extract or chilenol wt% of Examples 1 to 3.

Preparation Example 2-Medicines

Production Example 2-1 Powder

Powders were prepared by mixing the rare extracts of Examples 1 to 3 or 50 mg of chilenol and 2 g of crystalline cellulose and then filling them in airtight cloths according to a conventional powder preparation method.

Production Example 2-2 Tablet

The tablets were prepared by mixing the rare extracts of Examples 1 to 3 or 50 mg of chilenol, 400 mg of crystalline cellulose, and 5 mg of magnesium stearate, followed by compression according to a conventional tablet preparation method.

Preparation Example 2-3 Capsule

To prepare a capsule by mixing the rare extract of Example 1 to 3 or 30 mg of wheyol, 100 mg of whey protein, 400 mg of crystalline cellulose, 6 mg of magnesium stearate and filling it into gelatin capsules according to a conventional capsule preparation method It was.

Production Example 2-4 Injection

The active ingredient was dissolved in distilled water for injection according to a conventional injection method, and the pH was adjusted to about 7.5. Then, 100 mg of chilenol, distilled water for injection, and a pH adjusting agent of Example 3-5 were mixed to prepare an ampoule of 2 ml. Injections were made by filling and sterilizing.

Claims (33)

1. A pharmaceutical composition for improving muscle function or enhancing athletic performance, comprising a rare extract or a fraction thereof as an active ingredient.
2. The pharmaceutical composition for improving muscle function or enhancing exercise performance according to claim 1, wherein the rare extract is one or more extracts selected from the group consisting of Siege Beckia glabreense, Siege Beckia fuvesense, and Siege Beckia orientalis. Composition.
3. The method of claim 1, wherein the rare extract is obtained by extracting rare pigment with at least one solvent selected from the group consisting of water, an organic solvent having 1 to 6 carbon atoms, a subcritical fluid, and a supercritical fluid. Pharmaceutical composition for enhancing ability.
4. The organic solvent of claim 3, wherein the organic solvent having 1 to 6 carbon atoms is alcohol, acetone, ether, ether, benzene, chloroform, ethyl acetate. , Methylene chloride (hexane), hexane (hexane), cyclohexane (cyclohexane) and petroleum ether (petroleum ether) is one or more selected from the group consisting of a pharmaceutical composition for improving muscle function or enhancing performance.
5. The pharmaceutical composition of claim 1, wherein the rare extract is obtained by extracting rare extract under ultra-high pressure conditions of 100 MPa or more.
6. The method of claim 1, wherein the fraction of the rare extract is obtained by fractionation of the rare extract with ethyl acetate, methanol or a mixed solvent thereof for improving muscle function or enhancing exercise performance.
7. The method of claim 1, wherein the disease comprises one or more diseases selected from the group consisting of atony, muscular atrophy, muscular dystrophy, muscle degeneration, myasthenia, cachexia, and sarcopenia. A pharmaceutical composition for improving muscle function or enhancing exercise performance that is to be treated or prevented.
8. A food composition for improving muscle function or enhancing athletic performance, comprising a rare extract or a fraction thereof as an active ingredient.
9. The food for improving muscle function or improving exercise performance according to claim 8, wherein the rare extract is one or more extracts selected from the group consisting of Siege Beckia glabreense, Siege Beckia fuvesense and Siege Beckia orientalis. Composition.
10. The method of claim 8, wherein the rare extract is obtained by extracting the rare earth with at least one solvent selected from the group consisting of water, an organic solvent having 1 to 6 carbon atoms, a subcritical fluid and a supercritical fluid. Food composition for enhancing ability.
11. The organic solvent of claim 10, wherein the organic solvent having 1 to 6 carbon atoms is alcohol, acetone, ether, ether, benzene, chloroform, ethyl acetate. , Methylene chloride, hexane (hexane), cyclohexane (cyclohexane) and petroleum ether (petroleum ether) is at least one selected from the group consisting of food composition for improving muscle function or enhancing performance.
12. The food composition for improving muscle function or enhancing exercise performance of claim 8, wherein the rare extract is obtained by extracting rare extract under ultra-high pressure conditions of 100 MPa or more.
13. The method of claim 8, wherein the fraction of the rare extract is obtained by fractionation of the rare extract with ethyl acetate, methanol or a mixed solvent thereof, the composition for improving muscle function or enhancing exercise performance.
14. The method of claim 8, wherein the disease comprises one or more diseases selected from the group consisting of atony, muscular atrophy, muscular dystrophy, muscle degeneration, myasthenia, cachexia and sarcopenia. Food composition for muscle function improvement or exercise performance enhancement that is to prevent or improve.
15. A pharmaceutical composition for improving muscle function or enhancing exercise performance, comprising the compound represented by Formula 1 as an active ingredient:

    [Formula 1]

    
16. The pharmaceutical composition of claim 15, wherein the compound represented by Chemical Formula 1 is a chilenol represented by Chemical Formula 2 below.

    [Formula 2]

    
17. 16. The compound of claim 15, wherein the compound represented by Formula 1 is isolated from one or more extracts or fractions thereof selected from the group consisting of Siegesvecchia glabresense, Siegesvecchia fuvesense and Sigebesvecchia orientalis. Pharmaceutical composition for improving muscle function or enhancing exercise performance.
18. 16. The method of claim 15, wherein the at least one disease is selected from the group consisting of atony, muscular atrophy, muscular dystrophy, muscle degeneration, myasthenia, cachexia and sarcopenia. A pharmaceutical composition for improving muscle function or enhancing exercise performance that is to be treated or prevented.
19. Food composition for improving muscle function or enhancing exercise performance comprising the compound represented by the formula (1) as an active ingredient:

    [Formula 1]

    
20. The food composition of claim 19, wherein the compound represented by Chemical Formula 1 is a chilenol represented by Chemical Formula 2 below.

    [Formula 2]

    
21. 20. The compound of claim 19, wherein the compound represented by Formula 1 is isolated from one or more extracts or fractions thereof selected from the group consisting of Siegesvecchia glabresense, Siegesvecchia fubesense, and Siegesvecchia orientalis Food composition for improving muscle function or enhancing exercise performance.
22. 20. The method of claim 19, wherein the at least one disease is selected from the group consisting of atony, muscular atrophy, muscular dystrophy, muscle degeneration, myasthenia, cachexia and sarcopenia. Food composition for muscle function improvement or exercise performance enhancement that is to prevent or improve.
23. Muscle function improvement or exercise performance enhancement method comprising administering to the subject a composition comprising a rare extract or fractions thereof as an active ingredient.
24. 24. The method of claim 23, wherein the rare extract is one or more extracts selected from the group consisting of Siegesvecchia glabresense, Siegesvecchia fubesense, and Siegesvecchia orientalis.
25. 24. The muscle function improving or athletic performance according to claim 23, wherein the rare extract is obtained by extracting rare extract with at least one solvent selected from the group consisting of water, an organic solvent having 1 to 6 carbon atoms, a subcritical fluid and a supercritical fluid. Augmentation method.
26. The organic solvent of claim 25, wherein the organic solvent having 1 to 6 carbon atoms is alcohol, acetone, ether, ether, benzene, chloroform, ethyl acetate. , Methylene chloride (hexane), hexane (hexane), cyclohexane (cyclohexane) and petroleum ether (petroleum ether) at least one selected from the group consisting of muscle function improvement or exercise performance enhancement method.
27. 24. The method of claim 23, wherein the rare extract is obtained by extracting rare extract under ultra-high pressure conditions of 100 MPa or more.
28. 24. The method of claim 23, wherein the fraction of the rare extract is obtained by fractionating the rare extract with ethyl acetate, methanol or a mixed solvent thereof.
29. The method of claim 23, wherein the disease is one or more diseases selected from the group consisting of atony, muscular atrophy, muscular dystrophy, muscle degeneration, myasthenia, cachexia, and sarcopenia. Method of improving muscle function or exercise performance that is made by treatment, prevention or improvement.
30. Method for improving muscle function or improving exercise performance, comprising administering to a subject a composition comprising a compound represented by Formula 1 as an active ingredient:

    [Formula 1]

    
31. 31. The method of claim 30, wherein the compound represented by Formula 1 is a chilenol represented by Formula 2 below.

    [Formula 2]

    
32. 31. The compound of claim 30, wherein the compound represented by Formula 1 is isolated from one or more extracts or fractions thereof selected from the group consisting of Siegesvecchia glabresense, Siegesvecchia fubesense, and Siegesvecchia orientalis. How to improve muscle function or increase exercise performance.
33. 31. The method of claim 30, wherein the at least one disease is selected from the group consisting of atony, muscular atrophy, muscular dystrophy, muscle degeneration, myasthenia, cachexia, and sarcopenia. Method of improving muscle function or exercise performance that is made by treatment, prevention or improvement.

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