KR101385191B1 - Use of Cichorium intybus extracts for preventing, treating improving muscular atrophy - Google Patents

Abstract

The present invention provides a use for the prevention, treatment or improvement of atrophy of chicory extract. The chicory extract according to the present invention shows excellent muscular atrophy inhibiting activity, and thus can be usefully used for the prevention, treatment or improvement of muscular atrophy.

Description

Use of Cichorium intybus extracts for preventing, treating improving muscular atrophy}

The present invention relates to a cichorium intybus ) The use of the extract for the prevention, treatment or amelioration of muscle damage.

Atrophy can be defined as the loss of muscle tissue that results from not using muscle or the disease of the muscle itself or damage to the nerves that control the muscle. In general, the most important cause of muscle atrophy is disuse atrophy caused by not using muscle. In other words, in the case of a socially declining activity, muscle tone itself decreases and gradually progresses to muscular atrophy. In addition, people living in places where there is no gravity will also have a drop in muscle strength due to a decrease in calcium and muscle strength.

In addition, muscle atrophy due to the disease of the muscle itself is myasthenia gravis, dystrophy: dystrophic dystrophy, myotonic dystrophy, duchenne type, Becker's type, belt type, facial scapula and inflammation of the muscle itself. Muscle atrophy due to damage to the nerves that dominate the muscles; spinal muscular amyotrophy: Berradnich-Hoffmann's type, Kugelberg-Belander's disease, amyotrophic lateral sclerosis (ALS) : Lou Gehrig's disease and spinobular muscular atrophy: Kennedy's disease.

The cause of muscular dystrophy has not yet been elucidated, and to date, the main causes of the development of muscular dystrophy have been suggested in terms of muscle cell damage caused by increased oxide quality, decreased muscle protein production-regeneration due to decreased stress protein expression, or activation of proteosomal ubiquitination. Accelerating myoprotein degradation, and the like.

Muscle atrophy is diagnosed through creatine kinase (CK), electromyography, muscle biopsy, molecular biological genetic testing, cytogenetic testing, etc. To date, the main treatment methods for muscle atrophy are physical therapy, Occupational therapy for the prevention of complications and malformations, dysfunction, combined with respiratory therapy and additional symptomatic and supportive therapy are the only responses.

1. Korea Registered Patent Publication No. 10-0899377 2. Korean Unexamined Patent Publication No. 10-2012-0047510

1. Yoo Shin-Soo and 9 others, The Korean Society of Medicinal Crop Science Conference 2011 Spring Conference, Page 286-287

To date, the treatment of muscle diseases is a supplementary therapy for symptoms, and it is difficult to suppress or cure the onset of muscle atrophy. Therefore, there is a need for the development of new materials that can treat muscular dystrophy. Accordingly, the present invention seeks to discover new materials for the prevention, treatment or improvement of muscular dystrophy and to provide new uses thereof.

In order to achieve the above object, the present inventors have conducted research to find a substance that inhibits muscle damage, necrosis and apoptosis from natural products. As a result, it was confirmed that the chicory extract can inhibit myocyte damage, necrosis and apoptosis by oxidative stress and consequently prevent, treat or improve muscle atrophy.

Accordingly, the present invention provides a pharmaceutical composition and a food composition for preventing, treating or improving muscle atrophy comprising chicory extract as an active ingredient.

Hereinafter, the configuration of the present invention will be described in detail.

Chicory, which is used as an active ingredient in the composition of the present invention, is a perennial herb belonging to the Asteraceae family, and is known to be effective in improving liver and restoring vision, and is known to be a beneficial bacterium among intestinal bacteria when chicory intake is increased. It is reported that the number of bacteria of Bifidobacteria and Lactobacillus increases significantly. It has also been reported that intake of chicory consumes almost no production of methane gas in the human intestine and doubles the production of organic acids such as lactic acid.

Ingestion of chicory results in a 50% or more reduction in cholesterol absorption in the intestine, which acts as a factor in lowering the total cholesterol content in the liver or blood by preventing chicory extracts from re-absorbing cholesterol and excreting it into feces. . In addition, as a result of examining the effect of chicory extract on the intake of glucose, the total glycated hemoglobin content used as an indicator of the average blood glucose level was significantly lower in the chicory intake. In other words, ingesting chicory can reduce the absorption of glucose can be confirmed to be effective in the prevention or treatment of diabetes.

Chicory has been used as a herb for a long time, and has been reported to greatly affect liver and digestive function recovery. Chicory has been used as an herbal remedy for clearing blood, relieving emotional excitement, and for many diseases. It is effective for loss of appetite, jaundice and cholelithiasis in the body, and leaves are used for inflammation and swelling of the skin in vitro, and it is also used for the treatment of gout and rheumatism by promoting the discharge of unnecessary moisture in the body as a diuretic. The roots and leaves of chicory are currently used as an appetite stimulant, bile stimulant for the secretion of bile, purifying agent to remove toxins and wastes in the body, digestive agent, hypoglycaemia and laxative to help defecation. In addition, chicory has been reported to be effective in improving blood circulation by facilitating the exchange of blood flow and fluid through veins and arteries. The roots of chicory are either used directly or dried, and are widely used medically. Root extracts have been tested experimentally for heart disease and abnormalities by making them have a weak pulse rate (heart rate).

Chicory is also used as a kind of Baja flower therapy to treat diseases with flowers or plants in response to various emotions and heart conditions such as strong possessiveness, self-pity and narcissism. Chicory stem extract is used for skin diseases such as warts, which also heal or eliminate it. Treatment guides for German herbs report that chicory is effective in improving anorexia and indigestion.

In addition, chicory has potential as a biomass resource that could be useful for the industry. Chicory contains many inulins, which can be easily converted to alcohol. In addition, the leaves are extracted with dyes, and the flowers are dried, powdered, mixed with other herbs, and added to fertilizers, which can be used to make compost in a short time.

However, no examples of using chicory for use in the prevention, treatment or amelioration of muscular dystrophy have been reported. Accordingly, the present invention is characterized by providing novel uses for the prevention, treatment or amelioration of chicory atrophy.

Chicory extract used as an active ingredient of the pharmaceutical composition or the food composition of the present invention may be prepared according to a conventional method for producing a plant extract.

In one embodiment, the chicory extract may be extracted from the flowers, leaves, branches, fruits, roots, stems, seeds or mixtures of chicory with a conventional solvent.

In one embodiment of the invention, the chicory extract may be a root extract of chicory.

The extraction solvent used for the preparation of the chicory extract may be, but is not limited to, a solvent selected from water, an organic solvent or a mixture thereof. The organic solvent may be a polar solvent such as alcohol having 1 to 5 carbon atoms, ethyl acetate, acetone, a nonpolar solvent such as ether, chloroform, benzene, hexane, dichloromethane, or a mixed solvent thereof. For example, the alcohol having 1 to 5 carbon atoms may be selected from methanol, ethanol, propanol, butanol, isopropanol and the like.

In the present invention, the chicory extract includes a fraction obtained by re-fractionation of the primary extract extracted using the extraction solvent using an extraction solvent having a different polarity. For example, the chicory extract may be a fraction obtained by extracting chicory with an alcohol having 1 to 5 carbon atoms and then re-dividing the chicory with a solvent having a different polarity such as ether, benzene, and hexane.

Two or more solvents may be used in the fractionation, and each solvent extract may be prepared using sequentially or mixed according to the polarity of the solvent, but is not limited thereto.

The thus-prepared extract or the fraction obtained by performing the fractionation process may be filtered, concentrated or dried to remove the solvent, and may be subjected to both filtration, concentration and drying. Specifically, the filtration can be performed using a filter paper or a vacuum filter, and the concentration can be reduced by using a vacuum concentrator, for example, a rotary evaporator. The drying can be performed by, for example, freeze drying.

In one embodiment of the present invention, the pharmaceutical composition for preventing and treating muscle atrophy according to the present invention dominates muscle atrophy due to the loss of muscle tissue caused by not using the muscle, muscle atrophy due to the disease of the muscle itself, muscle It can be used for the prevention and treatment of muscular atrophy caused by nerve damage.

As can be seen in the following examples, the chicory extract can suppress myocyte damage, necrosis and apoptosis, and because it is a natural material has no side effects and very high safety, it can safely and effectively prevent and treat muscle atrophy.

The pharmaceutical composition according to the present invention may further include a carrier and a vehicle commonly used in the pharmaceutical field. Particularly glyceride mixtures of ion exchange resins, alumina, aluminum stearate, lecithin, serum proteins (e.g. human serum albumin), buffer substances (e.g. various phosphates, glycine, sorbic acid, potassium sorbate, saturated vegetable fatty acids ), Water, salts or electrolytes (e.g., protamine sulfate, disodium hydrogen phosphate, hydrogen carbonate, sodium chloride and zinc salts), colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose based substrates, polyethylene glycols, sodium carboxy Methylcellulose, polyarylate, wax or wool, and the like, but are not limited thereto.

The pharmaceutical compositions may also be in the form of granules, powders, coated tablets, tablets, capsules, suppositories, syrups, juices, suspensions, emulsions, drops, injectables or sustained release formulations of the active compounds. Pharmaceutical compositions for the prevention and treatment of muscular atrophy can be administered in a variety of oral or parenteral formulations, when formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, etc. commonly used in the pharmaceutical field It can be prepared using.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient in the pharmaceutical composition of the present invention, for example, starch, calcium carbonate, It may be prepared by mixing sucrose or lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Examples of the liquid preparation for oral administration include suspensions, solutions, emulsions, and syrups. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used .

Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the suppository base, witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like can be used.

On the other hand, the dosage of the pharmaceutical composition according to the present invention, the type of disease, the severity of the disease, the type and amount of the active ingredient and other ingredients contained in the composition, the type of formulation and the age, weight, general state of health, sex and It can be adjusted according to various factors including the diet, the time of administration, the route of administration and the rate of secretion of the composition, the duration of treatment, and the drugs used concurrently. For example, in adults, the chicory extract of the present invention may be administered at a dose of 0.01 to 500 mg / kg, once or several times daily. Specifically, 0.1 to 200 mg / kg, more specifically 0.1 to 100 mg / kg may be administered 1 to 3 times daily. The unit dosage form thus formulated may be administered several times at regular time intervals as needed.

The present invention also relates to a food composition for muscular atrophy comprising chicory extract as an active ingredient.

In one embodiment of the present invention, the food composition for muscle atrophy including chicory extract can be used as a health functional food that can be ingested daily to inhibit muscle aging or improve motor function.

In the present specification, 'health functional food' is a food prepared by adding chicory extract to food materials such as beverages, teas, spices, gums, confectionery, encapsulated, powdered, suspensions, etc. Means to bring the effect, unlike the general medicine has the advantage that there are no side effects that can occur when taking a long-term use of the drug as a raw material. The health functional food of the present invention thus obtained is very useful because it can be ingested routinely.

In addition, in one embodiment of the present invention, the food composition for muscle atrophy including chicory extract can be used as a space food prepared by adding to various foods for inhibiting muscle aging due to space life or improving exercise function. In the present specification, 'space food' is made by adding chicory extract to various foods as a food prepared to supplement the problems caused by space life. The chicory extract may be added as it is or used in combination with other food or food ingredients, and the mixed amount of the chicory extract may be appropriately determined according to conventional methods depending on the purpose of use thereof.

The food composition according to the present invention includes all foods such as beverages, meat, chocolate, foods, confectionary, pizza, ramen, other noodles, gums, ice creams, alcoholic beverages, vitamin complexes and the like. Although it cannot be prescribed | regulated uniformly according to the kind of food, what is necessary is just to add in the range which does not impair the original taste of food, and it is the range of 0.01-50 weight% normally, specifically 0.1-20 weight% with respect to the target food. In the case of food in the form of granules, tablets or capsules, it is usually added in the range of 0.1 to 100% by weight, specifically 5 to 100% by weight.

Chicory extract can be usefully used as an active ingredient for preventing, treating or improving muscle atrophy by inhibiting myocyte damage, necrosis and apoptosis.

1 is a graph showing the effect of chicory extract (root extract and stem extract) in the concentration-dependent prevention and recovery of muscle cells necrosis in muscle cells induced muscle cell necrosis with H ₂ O ₂ .
Figure 2 is a graph showing the cytotoxicity when cultured in muscle cells after treatment with chicory extract (root extract and stem extract) 5, 10, 25 and 50 μg / ml.
Figure 3 is a graph showing the effect of inhibiting apoptosis in muscle cells after 25 and 50 μg / ml of chicory root extract.

BRIEF DESCRIPTION OF THE DRAWINGS The present invention will become more apparent by describing in detail exemplary embodiments thereof with reference to the attached drawings, in which: FIG. The present invention may, however, be embodied in many different forms and should not be construed as being limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art. Is provided to fully convey the scope of the invention to those skilled in the art, and is defined only by the scope of the claims of the invention.

[Example]

Example 1: Preparation of Chicory Extract

Chikery was cultivated by the Rural Devel- opment Administration (RDA) Good Agricultural Practice (GAP) and was harvested in Chungcheongbuk-do Voice (GPS: E 128 ° 62'N 36 ° 56 ') in 2009. Extraction of the sample was performed as follows. 50 g of chicory root sample and 50 g of chicory stem sample were added to 150 ml of 99% ethanol, respectively, and reacted for 48 hours at 150 rpm. The resulting mixture was concentrated under reduced pressure and lyophilized powder was used as a sample of this experiment.

Experimental Example Confirmation of Biological Activity

Experimental Example 1: Improvement of survival rate through antioxidant in muscle cell culture system

(1) Cell culture

Normal muscle cells (C ₂ C ₁₂ ) were cultured in DMEM medium containing 10% fetal bovine serum (FBS). In a 75 cm 2 plastic flask (Falcon Co., England), normal skeletal muscle cells were loaded with 10% FBS, 7.5% NaHCO ₃ 150 μg / ml, 58.4 μg / ml glutamine and 4.4 μl / antibiotic / antimycotics. The cells were incubated at 37 ° C. and 5% CO ₂ in DMEM medium containing ml. Cell lines were maintained by secondary culture once every 2-3 days.

(2) cell quantification

Remove the medium solution from the grown 75 cm 2 plastic flask, wash with CMF-PBS (calcium magnesium free-phosphate buffered saline, pH 7.2), remove the cells from the bottom of the flask by treatment with 0.25% trypsin / EDTA. It was neutralized with cell culture and centrifuged (1200 rpm, 5 min). Culture medium was added to the pellets of the remaining cells, and then aspirated repeatedly with a sterile pipette to make a single cell suspension, and measured using a hemocytometer on an optical microscope.

(3) Effect of chicory extract on H ₂ O ₂ induced muscle cell necrosis

In order to observe the chicory extract concentrations effect of (root extract and stem extract) for the cell death induced by H ₂ O _2, placed such that the first muscle cells 5 × 10 ⁴ cells / well in 96 well plates CO ₂ incubator Incubated at. Then, add chicory extract at concentrations of 0, 5, 10, 25 and 50 μg / ml, respectively, and adjust the total volume to 100 μl with 10% FBS-DMEM medium to obtain 37 ° C. and 5% CO ₂ . Incubated for 24 hours at. H ₂ O ₂ was added to the culture solution at a concentration of 1 mM so that the total volume was 200 μl, which was then incubated for 60 minutes. After incubation, 10 μl of EzCytox kit was added, and after 1 hour, the absorbance was measured to determine the effect.

As can be seen in Figure 1, as a result of measuring the muscle cell survival rate of the chicory extract for muscle cell necrosis induced by 1 mM H ₂ O ₂ , the concentration of the chicory root extract is 50 from 0 μg / ml As the concentration increased to μg / ml, it was confirmed that cell viability was recovered in a concentration-dependent manner. Accordingly, it can be seen that the effect of preventing muscle cell necrosis is increased depending on the concentration of chicory extract.

 (4) cytotoxicity measurement

Toxicity of chicory extract was measured using EzCytox kit. That is, normal muscle cells (C ₂ C ₁₂ ) were dispensed to 96 × plate 5 × 10 ⁴ cells / well. After incubation in an incubator at 37 ° C., 5% CO ₂ , chicory extracts (root extract and stem extract) of various concentrations of 0, 5, 10, 25, and 50 μg / ml were added and incubated for 48 hours. The cytotoxicity of chicory extract was determined using EzCytox kit on normal muscle cells cultured for 48 hours.

The results of measuring cytotoxicity in cultured normal muscle cells to which chicory extract was added are shown in FIG. 2. When chicory extract was not added, the survival rate was 100%, and in the addition group to which chicory extract was added, the survival rate was similar to that of the non-addition group. Therefore, it can be seen that the chicory extract used in this experiment does not show toxicity to cultured muscle cells.

(5) Effect of chicory extract on myocyte apoptosis by H ₂ O ₂

In order to observe the effect of chicory extract on apoptosis according to the concentration of H ₂ O ₂ , firstly put the muscle cells in the cover glass to 5 × 10 ⁴ cells / well and incubated in a CO ₂ incubator, and then the concentration of chicory extract 25 , 50 μg / ml each was added, followed by incubation for 24 hours at 37 ° C. and 5% CO ₂ in 10% FBS-DMEM medium. H ₂ O ₂ was added to the culture solution at a concentration of 1 mM, followed by another 60 minutes of incubation, and then the effect was determined after DAPI staining.

As shown in FIG. 3, apoptosis of normal muscle cells according to H ₂ O ₂ was measured, and it was found that H ₂ O ₂ induced apoptosis of muscle cells. As a result of measuring the ability of chicory extract to prevent muscle cell apoptosis, it can be seen that the chicory extract added group prevents apoptosis.

Claims (9)

Pharmaceutical composition for preventing and treating muscle atrophy comprising chicory extract as an active ingredient.
The method according to claim 1,
Chicory extract is a pharmaceutical composition for preventing and treating muscular atrophy which is an extract of chicory flowers, leaves, branches, berries, roots, stems, seeds or mixtures thereof.
The method according to claim 1,
Chicory extract is a pharmaceutical composition for preventing and treating muscular atrophy that is extracted with a solvent selected from water, organic solvents or mixtures thereof.
The method according to claim 1,
Muscular atrophy is a pharmaceutical composition for preventing and treating muscular atrophy, which is muscle atrophy due to loss of muscle tissue caused by not using muscle, muscle atrophy due to a disease of the muscle itself, or muscle atrophy due to nerves that dominate the muscle.
Food composition for muscle atrophy comprising chicory extract as an active ingredient.
6. The method of claim 5,
Chicory extract is a food composition for improving muscle atrophy which is an extract of chicory flowers, leaves, branches, berries, roots, stems, seeds or mixtures thereof.
6. The method of claim 5,
Chicory extract is a food composition for improving muscle atrophy which is extracted with a solvent selected from water, organic solvents or mixtures thereof.
6. The method of claim 5,
Muscle atrophy is a muscle composition caused by the loss of muscle tissue caused by not using the muscle, muscle atrophy due to the disease of the muscle itself or muscle atrophy due to damage to the nerves that control the muscle.
6. The method of claim 5,
Food composition is a food composition for muscle atrophy improvement that is contained in space foods for inhibiting muscle aging or motor function due to space life.

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