Composition for Preventing or Treating Muscular disease containing Aralia Elata extract

Abstract

The present invention relates to a composition for the prevention or treatment of muscle diseases comprising an extract of Aralia elata. The Aralia elata extract of the present invention has the effect of enhancing grip strength, increasing muscle weight, and improving exercise performance.

Description

Technology Field

[0001] The present invention relates to a composition for the prevention or treatment of muscle diseases comprising Aralia elata extract. Background Technology

[0002] Muscles are broadly classified into skeletal muscle, cardiac muscle, and visceral muscle. Among these, skeletal muscle is the most abundant tissue in the human body, accounting for 40–45% of body weight. Skeletal muscle attaches to bones via tendons and plays a role in generating bone movement or force. A single muscle is composed of numerous muscle fibers, which in turn are made up of numerous myofibrils composed of actin and myosin. When actin and myosin overlap and move, the muscle shortens or lengthens, inducing overall muscle contraction and relaxation. An increase in the size of myofibrils implies an increase in the thickness of the muscle fibers, resulting in muscle growth.

[0003] Muscle diseases follow a course in which the weakening of skeletal muscles gradually leads to impaired walking and mobility functions, making activities of daily living (ADL) difficult and ultimately rendering independent living impossible. Furthermore, since they cause cardiopulmonary dysfunction and co-occur with other complications, it is important to accurately understand the characteristics of each muscle disease and approach them accordingly.

[0004] South Korea entered an aging society in 2000 when the elderly population accounted for 7.2% of the total population, and it is predicted to enter a super-aging society (over 20%) by 2050 (2013 Statistics on the Elderly, Statistics Korea). Human muscle mass decreases with age (by about 10–15% between the ages of 50 and 70, and by more than 30% between the ages of 70 and 80), leading to a weakening of muscle strength and function; this condition is known as senile sarcopenia. Senile sarcopenia causes mobility and gait disorders, serving as a major cause that limits the independent living of the elderly. Furthermore, sarcopenia lowers the anaplastic mortality rate, increasing insulin resistance and accelerating the development of type 2 diabetes, while increasing the risk of hypertension and cardiovascular diseases by 3 to 5 times. Currently, there are no approved drugs for the treatment of sarcopenia, and drug repositioning technology is under development to apply myostatin inhibitors or existing FDA-approved treatments for other diseases to sarcopenia.

[0005] The inventors of the present invention completed the present invention by experimentally confirming that Aralia elata extract is useful for improving muscle strength and exercise performance, as a result of efforts to develop a natural product with preventive or therapeutic effects for muscle diseases. Prior art literature

[0006] Republic of Korea Published Patent No. 10-2020-0104744 The problem to be solved

[0007] The objective of the present invention is to provide a pharmaceutical composition for the prevention or treatment of muscle diseases comprising Aralia elata extract as an active ingredient.

[0008] In addition, the objective of the present invention is to provide a food composition for preventing or improving muscle diseases comprising Aralia elata extract as an active ingredient.

[0009] The objective of the present invention is to provide a quasi-drug composition for preventing or improving muscle diseases, comprising Aralia elata extract as an active ingredient.

[0010] However, the technical problems that the present invention aims to solve are not limited to those mentioned above, and other unmentioned problems will be clearly understood by those skilled in the art from the description below. means of solving the problem

[0011] The present invention provides a food composition for preventing or improving muscle diseases, comprising Aralia elata extract as an active ingredient.

[0012] In addition, the present invention provides a quasi-drug composition for preventing or improving muscle diseases, comprising Aralia elata extract as an active ingredient.

[0013] In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of muscle diseases comprising Aralia elata extract as an active ingredient.

[0014] The above muscle disease may be one or more diseases selected from the group consisting of sarcopenia, muscular atrophy, myasthenia, muscular dystrophy, myotonia, hypotonia, muscular weakness, muscular dystrophy, atony, amyotrophic lateral sclerosis, and inflammatory myopathy.

[0015] The above composition can enhance grip strength.

[0016] The above composition can increase muscle weight.

[0017] The above muscles may include one or more selected from the group consisting of thigh muscles, calf muscles, shin muscles, and soleus muscles.

[0018] The above composition can be extracted with a solvent selected from the group consisting of water, organic solvents, and mixtures thereof.

[0019] The above organic solvent may be one or more solvents selected from the group consisting of lower alcohols having 1 to 6 carbon atoms, hexane, acetone, ethyl acetate, chloroform, and diethyl ether.

[0020] The above Aralia elata extract may include one or more compounds selected from the group consisting of compounds represented by the following chemical formulas 1 to 13. Effects of the invention

[0021] The Aralia elata extract of the present invention has the effect of enhancing grip strength, increasing muscle weight, and improving exercise performance. In addition, a composition containing the Aralia elata extract of the present invention as an active ingredient can be applied as a natural medicine to various products, such as pharmaceutical compositions, food compositions, and quasi-drug compositions, for the prevention, treatment, or improvement of muscle diseases. Brief explanation of the drawing

[0022] Figure 1 is a schematic diagram of an animal model design created to confirm the therapeutic or improving effects of Aralia elata extract on sarcopenia. Figure 2 shows the results confirming the effects of Aralia elata extract on body weight gain and grip strength improvement (Con: control group, Dex: Dexamethasone, S23: positive control group - anabolic drug 25 mg / kg, AE 100: Aralia elata extract 100 mg / kg, AE 50: Aralia elata extract 50 mg / kg, AE 10: Aralia elata extract 10 mg / kg; *, p <0.05; **, p <0.01; ***, p <0.001). Figure 3 shows the results confirming the effect of Aralia elata extract on improving exercise performance. a: Maximum running speed, b: Distance run until exhaustion, c: Time taken to exhaustion (Con: Control group, Dex: Dexamethasone, S23: Positive control group - anabolic drug 25 mg / kg, AE 100: Aralia elata extract 100 mg / kg, AE 50: Aralia elata extract 50 mg / kg, AE 10: Aralia elata extract 10 mg / kg; *, p <0.05). Figure 4 shows the results confirming the muscle weight increase effect of Aralia elata extract. a: Thigh muscle weight, b: Calf muscle weight, c: Leg muscle weight, d: Soleus muscle weight (Con: Control group, Dex: Dexamethasone, S23: Positive control-anabolic drug 25 mg / kg, AE 100: Aralia elata extract 100 mg / kg, AE 50: Aralia elata extract 50 mg / kg, AE 10: Aralia elata extract 10 mg / kg; *, p <0.05; ** p <0.01; ***, p <0.001). Figure 5 shows the results confirming the efficacy of 13 compounds isolated from Aralia elata extract in enhancing muscle cell viability reduced by dexamethasone (Con: control group, Dex: Dexamethasone, Catechol: catechol, 4-EC: 4-ethyl catechol, MD: methyl dihydroxycapate; *, p <0.05; ** p <0.01; ***, p <0.005). Figure 6 shows the results confirming the efficacy of three compounds isolated from Aralia elata extract on average muscle fiber enhancement in muscle cells (Con: control group, Dex: Dexamethasone, Catechol: catechol, 4-EC: 4-ethyl catechol, MD: methyl dihydroxycapate; *, p <0.05; ** p <0.01; ***, p <0.005). Figure 7 shows the results confirming the efficacy of three compounds isolated from Aralia elata extract in promoting muscle differentiation-related gene expression (Con: control group, Dex: Dexamethasone, Catechol: catechol, 4-EC: 4-ethyl catechol, MD: methyl dihydroxycapate; *, p <0.05; ** p <0.01; ***, p <0.005). Figure 8 shows the results confirming the efficacy of three compounds isolated from Aralia elata extract in improving oxidative stress and mitochondrial function (Con: control group, Dex: Dexamethasone, Catechol: catechol, 4-EC: 4-ethyl catechol, MD: methyl dihydroxycapate; *, p <0.05; ** p <0.01; ***, p <0.005). Figure 9 shows the results confirming the efficacy of three compounds isolated from Aralia elata extract in enhancing gene expression related to exercise performance (Con: control group, Dex: Dexamethasone, Catechol: catechol, 4-EC: 4-ethyl catechol, MD: methyl dihydroxycapate; *, p <0.05; ** p <0.01; ***, p <0.005). Specific details for implementing the invention

[0023] The inventors completed the present invention by experimentally confirming that an extract of Aralia elata, a natural product with few or no side effects, is effective in increasing muscle weight and improving grip strength and exercise performance.

[0024] The present invention will be described in detail below.

[0025] The present invention provides a composition for the prevention, improvement, or treatment of muscle diseases comprising Aralia elata extract as an active ingredient.

[0026] The above muscle disease may be one or more diseases selected from the group consisting of sarcopenia, muscular atrophy, myasthenia, muscular dystrophy, myotonia, hypotonia, muscular weakness, muscular dystrophy, atony, amyotrophic lateral sclerosis, and inflammatory myopathy.

[0027] The above composition can enhance grip strength.

[0028] The above composition can increase exercise performance.

[0029] The above composition can increase muscle weight.

[0030] The above muscles may include one or more selected from the group consisting of thigh muscles, calf muscles, shin muscles, and soleus muscles.

[0031] The above Aralia elata extract may include one or more compounds selected from the group consisting of compounds represented by the following chemical formulas 1 to 13.

[0032] More preferably, it may include one or more compounds selected from the group consisting of compounds represented by the following chemical formulas 3, 4, and 9.

[0033] [Chemical Formula 1]

[0034]

[0035] [Chemical Formula 2]

[0036]

[0037] [Chemical Formula 3]

[0038]

[0039] [Chemical Formula 4]

[0040]

[0041] [Chemical Formula 5]

[0042]

[0043] [Chemical Formula 6]

[0044]

[0045] [Chemical Formula 7]

[0046]

[0047] [Chemical Formula 8]

[0048]

[0049] [Chemical Formula 9]

[0050]

[0051] [Chemical Formula 10]

[0052]

[0053] [Chemical Formula 11]

[0054]

[0055] [Chemical Formula 12]

[0056]

[0057] [Chemical Formula 13]

[0058]

[0059] The above composition may be a food composition, a health functional food composition, a pharmaceutical composition, or a quasi-drug composition.

[0060] In this specification, "prevention" refers to any act of delaying the onset of muscle disease through the administration of a composition of the present invention, and "treatment" and "improvement" refer to any act of improving or beneficially altering the symptoms of muscle disease through the administration of a composition of the present invention.

[0061] The above Aralia elata extract can be extracted using a polar solvent or / and a non-polar solvent.

[0062] The above polar solvent may include one or more selected from the group consisting of (i) water, (ii) alcohol (preferably methanol, ethanol, propanol, butanol, normal-propanol, iso-propanol, normal-butanol, 1-pentanol, 2-butoxyethanol or ethylene glycol), (iii) acetic acid, (iv) DMFO (dimethyl-formamide), and (v) DMSO (dimethyl sulfoxide). The above non-polar solvent may include one or more selected from the group consisting of acetone, acetonitrile, ethyl acetate, methyl acetate, fluoroalkanes, pentane, hexane, 2,2,4-trimethylpentane, decane, cyclohexane, cyclopentane, diisobutylene, 1-pentene, 1-chlorobutane, 1-chloropentane, o-xylene, diisopropyl ether, 2-chloropropane, toluene, 1-chloropropane, chlorobenzene, benzene, diethyl ether, diethyl sulfide, chloroform, dichloromethane, 1,2-dichloroethane, aniline, diethylamine, ether, carbon tetrachloride, and THF.

[0063] More specifically, the extract may be extracted using water, an organic solvent, or a mixture thereof as a solvent, and according to a specific embodiment of the present invention, the extract of the present invention may be obtained by treating Aralia elata with water.

[0064] The above organic solvent may include one or more solvents selected from the group consisting of lower alcohols having 1 to 6 carbon atoms (methanol, ethanol, propanol, butanol, etc.), hexane, acetone, ethyl acetate, chloroform, and diethyl ether.

[0065] In addition, the above Aralia elata extract may be prepared as a fraction according to a method commonly used in the decomposition of the technology, and the solvent used to fractionate the Aralia elata extract may be one or more selected from the group consisting of water, C1 to C4 lower alcohols, n-hexane, ethyl acetate, acetone, acetonitrile, butyl acetate, 1,3-butylene glycol, methylene chloride, and mixed solvents thereof, but is not limited thereto. However, a butanol fraction may be more preferable.

[0066] As used in this specification, the term 'extract' has the meaning commonly understood in the art as a crude extract, as described above, but in a broader sense, it also includes fractions obtained by further fractionating the extract. That is, the Aralia elata extract includes not only that which is obtained using the extraction solvent described above, but also that which is obtained by additionally applying a purification process thereto. For example, fractions obtained through various additional purification methods, such as a fraction obtained by passing the extract through an ultrafiltration membrane having a certain molecular weight cut-off value, or separation by various chromatographs (designed for separation based on size, charge, hydrophobicity, or affinity), are also included in the natural product extract of the present invention.

[0067] The extract used in the present invention can be prepared in a powder state by additional processes such as vacuum distillation, freeze-drying, or spray-drying.

[0068] Specifically, the extract may be prepared by adding water, an organic solvent, or a mixture thereof to Aralia elata or Aralia elata leaves in an amount of 5 to 50 times the weight of the Aralia elata, and it is more preferable to add 10 to 30 times the amount, but is not limited thereto. The extraction temperature may be 10 to 150 ℃, and it is more preferable to be 15 to 120 ℃, but is not limited thereto. The extraction time is preferably 1 to 20 hours, and more preferably 2 to 8 hours, but is not limited thereto. The extraction method may be cold maceration, ultrasonic extraction, or reflux cooling extraction, but is not limited thereto. The number of extractions is preferably 1 to 5 times, and it is more preferable to repeat extraction 2 to 3 times, but is not limited thereto. Additionally, the extract may be diluted, concentrated, or purified and dried after dilution or concentration for use.

[0069] In this specification, the term "containing as an active ingredient" means containing an amount sufficient to achieve the efficacy or activity of Aralia elata extract. The present invention is a composition extracted from Aralia elata, a natural plant material, and the upper limit of the quantity of Aralia elata extract included in the composition of the present invention may be selected by a person skilled in the art within an appropriate range.

[0070] Pharmaceutical composition for the prevention or treatment of muscle diseases containing Aralia elata extract as an active ingredient

[0071] The composition of the present invention can be prepared as a pharmaceutical composition.

[0072] When the composition of the present invention is prepared as a pharmaceutical composition, the pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier.

[0073] According to a preferred embodiment of the present invention, the composition of the present invention may be a pharmaceutical composition comprising (a) a pharmaceutically effective amount of the Aralia elata extract of the present invention as described above; and (b) a pharmaceutically acceptable carrier. In this specification, the term “pharmaceutically effective amount” means an amount sufficient to achieve the efficacy or activity of the Aralia elata extract described above.

[0074] Pharmaceutically acceptable carriers are those commonly used in formulations and include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. In addition to the above components, the pharmaceutical composition of the present invention may further include lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, etc. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

[0075] The pharmaceutical composition of the present invention can be administered orally or parenterally.

[0076] Suitable dosages of the pharmaceutical composition of the present invention may be prescribed in various ways depending on factors such as the formulation method, mode of administration, patient's age, body weight, sex, pathological condition, food, time of administration, route of administration, excretion rate, and response sensitivity. The general dosage of the active ingredient included in the pharmaceutical composition of the present invention is within the range of 0.001-100 mg / kg / day, preferably 0.01-35 mg / kg / day, based on adults. Administration may be performed once a day or divided into several doses. However, the scope of the present invention is not limited by the above dosages.

[0077] The pharmaceutical composition of the present invention may be prepared in a unit volume form or contained in a multi-dose container by formulation using a pharmaceutically acceptable carrier and / or excipient, according to a method that can be easily carried out by a person skilled in the art to which the invention belongs. In this case, the formulation may be in the form of a solution, suspension, syrup, or emulsion in an oil or aqueous medium, or may be in the form of an extract, powder, powder, granule, tablet, or capsule, and may additionally include a dispersant or a stabilizer.

[0078] A quasi-drug composition for the prevention or improvement of muscle diseases containing Aralia elata extract as an active ingredient

[0079] The composition of the present invention may be provided as a quasi-drug composition.

[0080] The above Aralia elata extract may be added as is, used in combination with other ingredients of quasi-drugs, etc., and may be used appropriately according to conventional methods. The amount of the active ingredient can be appropriately determined according to the purpose of use (prevention, health, or therapeutic treatment). The above quasi-drug composition may be used in the manufacture of external preparations, patches, ointments, etc., but is not limited thereto.

[0081] Food composition for preventing or improving muscle diseases containing Aralia elata extract as an active ingredient

[0082] The composition of the present invention may be provided as a food composition or a health functional food composition. When a composition for the prevention, improvement, or treatment of muscle diseases containing the Aralia elata extract of the present invention as an active ingredient is prepared as a food composition, it includes not only the Aralia elata extract as an active ingredient but also ingredients that are typically added during food preparation, such as, for example, proteins, carbohydrates, fats, nutrients, seasonings, and flavorings. Examples of the carbohydrates described above include monosaccharides, e.g., glucose, fructose, etc.; disaccharides, e.g., maltose, sucrose, oligosaccharides, etc.; and polysaccharides, such as dextrin, cyclodextrin, etc., which are conventional sugars, and sugar alcohols such as xylitol, sorbitol, erythritol, etc. As flavorings, natural flavorings [taumatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.)] and synthetic flavorings (saccharin, aspartame, etc.) may be used. For example, when the food composition of the present invention is manufactured as a drink, in addition to the natural product extract of the present invention, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, Eucommia ulmoides extract, jujube extract, licorice extract, etc. may be additionally included.

[0083] The formulation of the above food composition or health functional food composition can be in the form of powder, granule, pill, tablet, or capsule, as well as any form of general food or beverage.

[0084] There are no specific restrictions on the types of food mentioned above, and examples of food to which the substance may be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, chewing gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and may include all food in the conventional sense.

[0085] Generally, when manufacturing food or beverages, the above Aralia elata extract may be added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, per 100 parts by weight of raw materials. However, in the case of long-term consumption for the purpose of health and hygiene or health control, the above amount may be less than the above range, and furthermore, since the present invention uses natural substances, there is no problem in terms of safety, so it may be used in an amount greater than the above range.

[0086] The present invention will be explained in more detail below through examples. The purpose, features, and advantages of the present invention will be easily understood through the following examples. The present invention is not limited to the examples described herein and may be embodied in other forms. The examples introduced herein are provided to ensure that the concept of the present invention is sufficiently conveyed to those skilled in the art to which the present invention pertains. Therefore, the present invention should not be limited by the following examples.

[0087] [Preparation Example 1] Preparation of Aralia elata extract

[0088] Aralia elata Leaves were purchased from Herb Village Co., Ltd. and chopped into appropriate sizes. 2 kg of Aralia elata leaves and 10 L of water were added to an extraction container and cold-infused for 3 days. The precipitate was removed using a centrifuge, and the impurities were filtered to obtain the filtrate. Subsequently, the solvent was concentrated under reduced pressure and dried to obtain 310 g of extract.

[0089] [Preparation Example 2] Isolation of compounds derived from Aralia elata extract

[0090] The extract obtained according to Preparation Example 1 was separated into compounds after solvent fractionation. Specifically, distilled water (10 L x 3 times) was added to the extract of Preparation Example 1 to suspend it, and then an equal amount of butanol was added to separate it into a butanol layer and a water layer. The mixture was then filtered and concentrated under reduced pressure to obtain a butanol fraction (40 g) and a water fraction (260 g). Subsequently, the butanol fraction was separated using a silica gel open column under the conditions MC:MeOH = 15:1-0:15, separated by molecular weight using a Sephadex open column under 50% methanol conditions, and 13 compounds were obtained using prep-MPLC.

[0091] Compounds derived from Aralia elata extract No. compound 1 (Chemical Formula 1) 1-ethyl-3,4,6-trihydroxy-5H-benzocyclohepten-5-one 2 (Chemical Formula 2) 3,4,6-trihydroxy-5H-benzocyclohepten-5-one 3 (Chemical Formula 3) Catechol 4 (Chemical Formula 4) 4-ethyl catechol 5 (Chemical Formula 5) Protocatechuic acid 6 (Chemical Formula 6) Caffeic acid 7 (Chemical Formula 7) Dihydroxycaffeic acid 8 (Chemical Formula 8) Phloretic acid 9 (Chemical Formula 9) Methyl dihydroxycaffeate 10(Chemical Formula 10) Junipediol A 11(Chemical Formula 11) Chlorogenic acid 12(Chemical Formula 12) Neochlorogenic acid 13(Chemical Formula 13) Quercetin 7-O-neohesperidoside

[0092] [Chemical Formula 1]

[0093]

[0094] [Chemical Formula 2]

[0095]

[0096] [Chemical Formula 3]

[0097]

[0098] [Chemical Formula 4]

[0099]

[0100] [Chemical Formula 5]

[0101]

[0102] [Chemical Formula 6]

[0103]

[0104] [Chemical Formula 7]

[0105]

[0106] [Chemical Formula 8]

[0107]

[0108] [Chemical Formula 9]

[0109]

[0110] [Chemical Formula 10]

[0111]

[0112] [Chemical Formula 11]

[0113]

[0114] [Chemical Formula 12]

[0115]

[0116] [Chemical Formula 13]

[0117]

[0118] [Example 1] Confirmation of changes in grip strength and body weight in mice with sarcopenia induced by administration of Dexamethasone with Aralia elata extract

[0119] The inventors constructed a sarcopenia mouse model to confirm the therapeutic or improving effect of the Aralia elata extract prepared according to the aforementioned manufacturing example, and measured grip strength using a control mouse and a sarcopenia mouse model.

[0120] Mice used to verify the efficacy of Aralia elata extract were prepared through the following process. As shown in Figure 1, C57BL / 6J mice (male, 12 weeks old) were obtained from Dooyul Biotech Co., Ltd. and, after a 2-week acclimatization period, were divided into 6 test groups (n=10 per group) according to body weight. The diet, dosage, and administration method for the experimental groups are shown in Table 1 below. The test substance was prepared in the form of a liquid suspension using 0.5% CMC and administered for 4 weeks. To compensate for the placebo effect of 0.5% CMC and the weight loss effect caused by administration stress, 0.5% CMC was orally administered daily to the vehicle control group. Two weeks after the administration of the test substance, sarcopenia was induced in the experimental group by intraperitoneal injection of Dexamethasone for 2 weeks, while the normal control group was administered saline solution by intraperitoneal injection. All mice were fed the same AIN76 diet (Research Diets), and the test substance was administered for an additional 2 weeks, for a total of 4 weeks. The dark:light cycle was maintained at 12-hour:12-hour intervals, and water was available for free intake. As shown in Figure 2, the experimental results confirmed that grip strength and body weight decreased over time in the Dexamethasone (Dex) group, while grip strength and body weight increased in the positive control group of mice fed an anabolic drug and Aralia elata extract compared to the group treated with Dexamethasone alone (Dex: Dexamethasone sarcopenia-inducing agent (25 mg / kg), AE: Aralia elata extract (10, 50, or 100 mg / kg)).

[0121] Test group feedstuff sarcopenia-inducing substances Oral substances Dosage and Method of Administration Administration period 1(Con) AIN76 diet Saline Vehicle - 2 weeks of oral administration of the test substance followed by 2 weeks of administration of the test substance and inducer (total 4 weeks) 2(Dex) Dexamethasone (25 mg / kg, once daily, for 2 weeks) Vehicle - 3(Dex+S23) anabolic drug(S-23) 25 mg / kg, once daily, 4(Dex+AE 100) Aralia elata extract 100 mg / kg, once daily 5(Dex+AE 50) 50 mg / kg, once daily 6(Dex+AE 10) 10 mg / kg, once daily

[0122] [Example 2] Confirmation of the efficacy of Aralia elata extract in improving exercise performance in mice with sarcopenia induced by Dexamethasone administration

[0123] After the administration of Aralia elata extract for a total of 4 weeks in [Example 1] above, exercise performance was evaluated for all administration groups. Specifically, maximum running speed, distance run until exhaustion, and time taken to exhaustion were evaluated, and the results are shown in Figure 3. As shown in Figure 3, it was confirmed that exercise performance significantly increased in the group administered with Aralia elata extract.

[0124] [Example 3] Confirmation of weight increase by muscle type in mice with sarcopenia induced by Dexamethasone administration using Aralia elata extract

[0125] After the administration of Aralia elata extract for a total of 4 weeks in [Example 1] above, the weight of muscle tissue by type in all administration groups was checked.

[0126] As a result, as shown in Figure 4, compared to the control group administered saline solution, it was confirmed that the weight of the thigh muscles, calf muscles, shin muscles, and soleus muscles decreased in the group administered Dexamethasone, but it was confirmed that the weight of these muscles significantly increased when Aralia elata extract was administered.

[0127] [Example 4] Confirmation of the efficacy of a compound derived from Aralia elata extract in enhancing myotube cell viability

[0128] Cell culture and differentiation of myoblasts: Myoblasts derived from C2C12 mice (CRL1772; American type cell collection, USA) were cultured in DMEM (Dulbecco's Modified Eagle's Medium) supplemented with 10(v / v)% FBS, and after reaching 90% confluence, were transferred to DMEM medium containing 2(v / v)% horse serum and cultured for 7 days to differentiate into myotubes.

[0129] After differentiating C2C12 myoblasts into myotubes, myoatrophy was induced by treating with 100 μM of Dexamethasone for 24 hours, and the myoatrophy alleviation effect was confirmed by treating with 20 μM of 13 compounds isolated from Aralia elata extract (see Table 1). As a result, as shown in Figure 5, it was confirmed that the viability of myotubes reduced by Dexamethasone treatment was restored in the groups treated with compounds isolated from Aralia elata extract. In particular, it was confirmed that the groups treated with Compound 3 (catechol), Compound 4 (4-ethyl catechol), and Compound 9 (methyl dihydroxycapate) showed significantly superior effects.

[0130] [Example 5] Confirmation of the efficacy of a compound derived from Aralia elata extract in enhancing myotube thickness

[0131] In the same manner as in Example 4, C2C12 myoblasts were differentiated into myotubes, and myoatrophy was induced by treating with 100 μM of Dexamethasone for 24 hours. Then, changes in myotube thickness were confirmed by treating with 20 μM of three compounds (catechol, 4-ethyl catechol, methyl dihydroxycapate) isolated from Aralia elata extract.

[0132] Changes in myotube thickness were confirmed by the following method. Cells were washed twice with phosphate buffered saline (PBS) and fixed with 4% formaldehyde for 15 minutes. Subsequently, the cells were permeated with 0.4% Triton-X100 in DPBS for 10 minutes, blocked for 1 hour with 10% donkey serum prepared with 0.1% Triton-X100 and 1% BSA / DPBS, and then stained with 1:100 total MHC (MF20, Developmental Research Hybrid Species Bank) overnight at 4°C. After washing three times with 0.1% Triton-X100 / DPBS, the cells were treated with FITC fluorophore secondary antibody (1:100) prepared with 0.1% Triton-X100 and 1% BSA / DPBS Hoechst 1 uM and incubated at room temperature for 1 hour. After washing three times with 0.1% Triton-X100 / DPBS, C2C12 myotube immunofluorescence was observed using an Operetta Machine 20 at 10x magnification and muscle fibers were quantified (3 repetitions / Group).

[0133] As shown in Figure 6, it was confirmed that the thickness of myotubes reduced by Dexamethasone treatment increased in the group treated with compounds isolated from Aralia elata extract.

[0134] [Example 6] Confirmation of the efficacy of a compound derived from Aralia elata extract in enhancing gene expression related to muscle cell differentiation

[0135] C2C12 mouse-derived myotubes treated with 10 μM dexamethasone were washed twice with PBS, and total RNA was extracted according to the user-recommended protocol of the RNA extraction kit (GeneAll RNA isolation kit, Seoul, South Korea). cDNA was synthesized from the extracted RNA, and real-time quantitative polymerase chain reaction (RT-qPCR) was performed. PCR analysis was conducted according to the user-recommended cycling conditions of the analyzer (Applied Biosystems 7900 HT thermal cycler, Applied Biosystems). To confirm the expression levels of each mRNA, beta-actin was used as an endogenous control, and the fold change values ​​relative to the control group were calculated and compared to the relative expression levels. The primer sequences are as follows:

[0136] gene Types of primers order Sequence number Mstn Forward direction TGG CTC CTA CTG GAC CTC TC Sequence No. 1 reverse direction AAG ATG CAG CAG TCA CTC CC Sequence No. 2 Myd1 Forward direction CAT AGA CTT GAC AGG CCC CG Sequence No. 3 reverse direction CGG GTC CAG GTC CTC AAA AA Sequence No. 4 Myog Forward direction AGC TAT CCG GTT CCA AAG CC Sequence No. 5 reverse direction GCA CAG GAG ACC TTG GTC AG Sequence number 6 Myf5 Forward direction TCC AGG TAT TCC CAC CTG CT Sequence No. 7 reverse direction TCA GCT TTG TGT GCT CCG AA Sequence No. 8

[0137] As a result of the examination, it was confirmed that the expression of the Mstn gene, a muscle growth inhibitory marker increased by Dexamethasone treatment, was reduced by treatment with compounds derived from Aralia elata extract (catechol, 4-ethyl catechol, methyl dihydroxycapate). In addition, it was confirmed that the expression of Myd1, Myog, and Myf5 genes, muscle cell differentiation markers reduced by Dexamethasone treatment, was increased by treatment with compounds derived from Aralia elata extract (catechol, 4-ethyl catechol, methyl dihydroxycapate) (Fig. 7).

[0138] [Example 7] Confirmation of the efficacy of compounds derived from Aralia elata extract in improving oxidative stress and mitochondrial function

[0139] In the same manner as in Example 4, C2C12 myoblasts were differentiated into myotubes, and oxidative stress or mitochondrial dysfunction was induced by treating with 100 μM of Dexamethasone for 24 hours. Then, the efficacy of improving oxidative stress and mitochondrial dysfunction was confirmed by treating with 20 μM of three compounds (catechol, 4-ethyl catechol, and methyl dihydroxycapate) isolated from Aralia elata extract. As a result, it was confirmed that the content of glutathione (GSH), which has antioxidant activity, decreased due to Dexamethasone treatment, but the glutathione (GSH) content increased again upon treatment with compounds derived from Aralia elata extract. In addition, it was confirmed that the mitochondrial dysfunction induced by Dexamethasone treatment was improved by treatment with compounds derived from Aralia elata extract (Fig. 8).

[0140] [Example 8] Confirmation of the efficacy of a compound derived from Aralia elata extract in enhancing gene expression related to exercise performance

[0141] C2C12 mouse-derived myotubes treated with 10 μM dexamethasone were washed twice with PBS, and total RNA was extracted according to the user-recommended protocol of the RNA extraction kit (GeneAll RNA isolation kit, Seoul, South Korea). cDNA was synthesized from the extracted RNA, and real-time quantitative polymerase chain reaction (RT-qPCR) was performed. PCR analysis was conducted according to the user-recommended cycling conditions of the analyzer (Applied Biosystems 7900 HT thermal cycler, Applied Biosystems). To confirm the expression levels of each mRNA, beta-actin was used as an endogenous control, and the fold change values ​​relative to the control group were calculated and compared to the relative expression levels. The primer sequences are as follows:

[0142] gene Types of primers order Sequence number Ppargc1a Forward direction GTT GCC TGC ATG AGT GTG TG Sequence number 9 reverse direction CAC ATG TCC CAA GCC ATC CA Sequence number 10 Ucp3 Forward direction GTT TTG CGG ACC TCC TCA CT Sequence number 11 reverse direction CTC TGT GCG CAC CAT AGT CA Sequence No. 12 Tomm20 Forward direction TGT GCG GTG TGT TGT CTG TT Sequence No. 13 reverse direction TAA GTG CCC AGA GCA CAG GA Sequence No. 14 Tfam Forward direction GGG AAT GTG GAG CGT GCT AA Sequence number 15 reverse direction TGA TAG ACG AGG GGA TGC GA Sequence number 16

[0143] Confirmation results showed that the exercise performance marker reduced by Dexamethasone treatment Ppargc1a , UCp3 , Tomm20 and Tfam It was confirmed that gene expression was increased by treatment with compounds derived from Aralia elata extract (catechol, 4-ethyl catechol, methyl dihydroxycapate) (Fig. 9).

Claims

Claim 1 A food composition for the prevention or treatment of muscle diseases comprising, as an active ingredient, one or more compounds selected from the group consisting of a compound represented by Chemical Formula 3 (catechol), a compound represented by Chemical Formula 4 (4-ethyl catechol), and a compound represented by Chemical Formula 9 (methyl dihydroxycapate), wherein the muscle disease is one or more diseases selected from the group consisting of sarcopenia, muscular atrophy, muscular dystrophy, hypotonia, muscular weakness, muscular dystrophy, and amyotrophic lateral sclerosis. [Chemical Formula 3] [Chemical Formula 4] [Chemical Formula 9] Claim 2 A food composition according to claim 1, wherein the compound regulates the expression of one or more muscle differentiation-related genes selected from the group consisting of Mstn, Myd1, Myog, and Myf5. Claim 3 A food composition according to claim 1, characterized in that the compound has an effect of improving mitochondrial function through the enhancement of glutathione (GSH). Claim 4 A food composition according to claim 1, characterized in that the compound is derived from Aralia elata extract. Claim 5 A quasi-drug composition for preventing or improving muscle diseases, comprising as an active ingredient one or more compounds selected from the group consisting of a compound represented by Chemical Formula 3 (catechol), a compound represented by Chemical Formula 4 (4-ethyl catechol), and a compound represented by Chemical Formula 9 (methyl dihydroxycapate), wherein the muscle disease is one or more diseases selected from the group consisting of sarcopenia, muscular atrophy, muscular dystrophy, hypotonia, muscular weakness, muscular dystrophy, and amyotrophic lateral sclerosis. [Chemical Formula 3] [Chemical Formula 4] [Chemical Formula 9] Claim 6 A pharmaceutical composition for the prevention or treatment of muscle diseases comprising, as an active ingredient, one or more compounds selected from the group consisting of a compound represented by Chemical Formula 3 (catechol), a compound represented by Chemical Formula 4 (4-ethyl catechol), and a compound represented by Chemical Formula 9 (methyl dihydroxycapate), wherein the muscle disease is one or more diseases selected from the group consisting of sarcopenia, muscular atrophy, muscular dystrophy, hypotonia, muscular weakness, muscular dystrophy, and amyotrophic lateral sclerosis. [Chemical Formula 3] [Chemical Formula 4] [Chemical Formula 9] Claim 7 A food composition for preventing or improving muscle diseases comprising a butanol fraction of Aralia elata leaves as an active ingredient, wherein the muscle disease is one or more diseases selected from the group consisting of sarcopenia, muscular atrophy, muscular dystrophy, hypotonia, muscular weakness, muscular dystrophy, and amyotrophic lateral sclerosis. Claim 8 A food composition according to claim 7, characterized in that the butanol fraction of Aralia elata leaves comprises one or more selected from the group consisting of compounds represented by the following chemical formulas 1 to 13. [Chemical Formula 1] [Chemical Formula 2] [Chemical Formula 3] [Chemical Formula 4] [Chemical Formula 5] [Chemical Formula 6] [Chemical Formula 7] [Chemical Formula 8] [Chemical Formula 9] [Chemical Formula 10] [Chemical Formula 11] [Chemical Formula 12] [Chemistry Formula 13] Claim 9 A quasi-drug composition for preventing or improving muscle diseases comprising a butanol fraction of Aralia elata leaves as an active ingredient, wherein the muscle disease is one or more diseases selected from the group consisting of sarcopenia, muscular atrophy, muscular dystrophy, hypotonia, muscular weakness, muscular dystrophy, and amyotrophic lateral sclerosis. Claim 10 A pharmaceutical composition for the prevention or treatment of muscle diseases comprising a butanol fraction of Aralia elata leaves as an active ingredient, wherein the muscle disease is one or more diseases selected from the group consisting of sarcopenia, muscular atrophy, muscular dystrophy, hypotonia, muscular weakness, muscular dystrophy, and amyotrophic lateral sclerosis.

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