KR20180000127A - Composition for improving metabolism containing extraction of Aconitum carmichaeli Debx - Google Patents

Abstract

The present invention relates to a composition for promoting body energy consumption or promoting muscle metabolism containing an Aconitum carmichaeli Debeaux extract as an active ingredient. The Aconitum carmichaeli Debeaux extract of the present invention has an effect of promoting PGC1- and the production of mitochondria, thereby promoting body energy consumption and muscle metabolism.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for stimulating body energy consumption or facilitating muscle metabolism containing an extract of Rich extract as an active ingredient.

The present invention relates to a composition for accelerating body energy consumption or muscle metabolism, which comprises a rich extract as an active ingredient.

Mitochondria play an important role in energy metabolism by activating glucose transport and fatty acid oxidation. Disparities in energy intake and consumption lead to mitochondrial disorders that contribute to the pathogenesis of obesity, insulin resistance, and aging-related diseases such as type 2 diabetes.

The skeletal muscle is an important tissue for mitochondrial dysfunction and insulin resistance. It is known that changes in mitochondrial function in skeletal muscles are associated with insulin resistance and second-line diabetes. In addition, insulin resistance is strongly associated with fat accumulation and damaged oxidative capacity of the skeletal muscle.

The peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1α) is a key component of mitochondrial function. PGC1α is a key regulator of mitochondrial production and is a co-activator of nuclear transcription factors such as nuclear respiratory factor-1 (NRF-1) and transcription factor A (TFAM), which are required for the expression of mitochondrial genes and replication of the mitochondrial genome. These mitochondria regulate the ATP energy and promote the energy consumption of the body. In this connection, mitochondria are abundant and the action of PGC1 α in muscle, a key organ of metabolic regulation of body energy, is particularly important.

In skeletal muscle, the metabolic sensor, AMP-activated protein kinase (AMPK), is known to directly affect the activity of PGC-1α through phosphorylation. The AMPK system is an important factor in the regulation of energy balance at all cellular and systemic levels and has received attention in the field of obesity, diabetes, and metabolic syndrome. In particular, activation of AMPK in skeletal muscle increases the expression of genes associated with these pathways, thereby increasing glucose uptake, fatty acid oxidation and mitochondrial biosynthesis.

Recently, the development of mitochondrial targeting drugs or nutrients such as metformin (AMPK agonist), thiazolidinedione (PPARγ agonist), resveratrol (SIRT1 activator) has been noted in the field of treatment of insulin resistance, obesity and type 2 diabetes. These drugs and natural nutrients have been shown to regulate mitochondrial neogenesis and reduce insulin resistance.

Meanwhile, Aconitum carmichaeli Debx) is a medicinal plant used in oriental medicine. Its pharmacological action is known to be central inhibition, sedation, analgesic, antipyretic, increase of coronary blood flow and inhibition of skin fungi.

The present inventors have studied PGC1-alpha (peroxisome proliferator-activated receptor gamma coactivator 1-alpha, peroxisome proliferator-activated receptor gamma coactivator 1-alpha) And promoting the body energy consumption and muscle metabolism through the action of promoting the production of mitochondria, the mechanism of which is confirmed and the present invention is completed.

Accordingly, an object of the present invention is to provide a composition for accelerating body energy consumption or muscle metabolism, which comprises a rich extract as an active ingredient.

Another object of the present invention is to provide a health food composition for accelerating body energy consumption or muscle metabolism, which comprises a rich extract as an active ingredient.

However, the technical problem to be solved by the present invention is not limited to the above-mentioned problems, and other matters not mentioned can be clearly understood by those skilled in the art from the following description.

In order to accomplish the object of the present invention as described above, the present invention provides a composition for accelerating body energy consumption or muscle metabolism, which comprises a rich extract as an active ingredient.

In one embodiment of the present invention, the rich extract is selected from water, C1 to C4 lower alcohols, n-hexane, ethyl acetate, acetone, butyl acetate, 1,3-butylene glycol, methylene chloride, And is extracted with at least one solvent selected from the group consisting of.

In another embodiment of the present invention, the rich extract is characterized in that it is extracted from the root of the rich.

In another embodiment of the present invention, the composition is characterized by an increase in the expression of peroxisome proliferator-activated receptor gamma-assisting activator 1-alpha (PGC1-a).

In another embodiment of the present invention, the composition is characterized by promoting the production of mitochondria.

In addition, the present invention provides a health functional food composition for accelerating body energy consumption or muscle metabolism, which comprises a rich extract as an active ingredient.

The present invention also provides a method for stimulating body energy consumption or muscle metabolism, which comprises administering a rich extract to a subject.

In addition, the present invention provides the use of a rich extract for promoting body energy consumption or promoting muscle metabolism.

The rich extract according to the present invention has been shown to increase the expression of PGC1-alpha and promote the production of mitochondria, thus promoting body energy consumption and promoting muscle metabolism. Thus, it can have an effect of increasing body basic metabolism and strengthening muscular endurance. It can also be used for the prophylaxis and treatment of diseases that are closely related to decreased energy metabolism including obesity, diabetes, weakening of muscular function and muscle weakness.

FIG. 1 shows the results of confirming the expression level of PGC1-alpha in C2C12 cells treated with the extract of the present invention.
FIG. 2 shows the results of confirming the expression level of AMPK in C2C12 cells treated with the extract of the present invention.
FIG. 3 shows the result of confirming the glucose uptake regulating action in C2C12 cells treated with the extract of the present invention.
FIG. 4 shows the results of confirming the increase in production of ATP in C2C12 cells treated with the extract of the present invention.

The present invention provides a composition for accelerating body energy consumption or muscle metabolism, which comprises a rich extract as an active ingredient.

In addition, the present invention provides a health functional food composition for accelerating body energy consumption or muscle metabolism, which comprises a rich extract as an active ingredient.

The composition comprises a pharmaceutical composition and a food composition.

Hereinafter, the present invention will be described in detail.

The rich extract according to the present invention was extracted from the root of the bud, and the mouse muscle cells (C2C12) were cultured to induce differentiation into myotubes, and then treated with the supplement to obtain PGC1-alpha (peroxisome proliferator- -Activated receptor gamma-assisted activator 1-alpha, peroxisome proliferator-activated receptor gamma coactivator 1-alpha) and promotes the production of mitochondria, and thus the extract has an effect of promoting body energy consumption and muscle metabolism Respectively.

Therefore, the rich extract according to the present invention can be used as a medicament useful for a disease caused by mitochondrial dysfunction, that is, a disease caused by insufficient energy metabolism and muscle metabolism, and a health functional food for promoting body energy consumption or promoting muscle metabolism.

The extract of the present invention can be prepared using conventional solvents known in the art, that is, under the conditions of ordinary temperature and pressure, using conventional solvents. Examples of the extraction solvent that can be used for preparing the extract of the present invention include water, C1 to C4 lower alcohols, n-hexane, ethyl acetate, acetone, butyl acetate, 1,3-butylene glycol, methylene chloride, And a mixed solvent thereof may be used alone or in combination. The solvent is preferably water, but is not limited thereto. The suitable amount of the extraction solvent is about 1 to 10 times as much as the dry weight of the extract. Examples of the extraction method include a heat extraction method, a cold extraction method, a reflux cooling extraction method, and an ultrasonic extraction method. have. The extraction temperature is not particularly limited as long as the effective activity of the active ingredient is not removed, but is preferably extracted by immersion at room temperature.

The composition of the present invention may contain one or more known active ingredients having the effect of stimulating body energy consumption or promoting muscle metabolism together with the extract of the present invention.

The composition of the present invention may further comprise at least one pharmaceutically acceptable carrier in addition to the above-described effective ingredients for administration. The pharmaceutically acceptable carrier may be a mixture of saline, sterilized water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol and one or more of these components. If necessary, an antioxidant, , And other conventional additives such as a bacteriostatic agent may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Further, it can be suitably formulated according to each disease or ingredient according to a suitable method in the art.

The composition of the present invention may be administered orally or parenterally (for example, intravenously, subcutaneously, intraperitoneally or topically) depending on the intended method, and the dose may be appropriately determined depending on the patient's weight, age, , Diet, administration time, method of administration, excretion rate, and severity of the disease. The daily dose of the above extract is about 0.0001-500 mg / kg, preferably about 0.001-300 mg / kg, and it is preferable to administer the extract once or several times a day.

The composition of the present invention can be used alone or in combination with methods using surgery, hormone therapy, drug therapy and biological response modifiers to promote body energy consumption or muscle metabolism.

The composition of the present invention may be added to a health functional food for the purpose of stimulating body energy consumption or promoting muscle metabolism. When the extract of the present invention is used as a food additive, the extract of the present invention can be added as it is or can be used together with other food or food ingredients, and can be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment). Generally, the rich extract of the present invention is added in an amount of not more than 15% by weight, preferably not more than 10% by weight based on the raw material, in the production of food or beverage. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of health control, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

There is no particular limitation on the kind of the food. Examples of the food to which the above substances can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include health functional foods in a conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. The above-mentioned natural carbohydrates are monosaccharides such as glucose and fructose, polysaccharides such as disaccharides such as maltose and sucrose, dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01-0.20 g, preferably about 0.04-0.10 g, per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, A carbonating agent used in a carbonated beverage, and the like. In addition, the composition of the present invention may contain flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not critical, but is generally selected in the range of 0.01-0.20 parts by weight per 100 parts by weight of the composition of the present invention.

In addition, the present invention provides a method for promoting body energy consumption or muscle metabolism, which comprises administering a rich extract to a subject.

The subject is a mammal, including a human or non-human, and non-human mammals include, but are not limited to, mice, rats, dogs, cats, horses, cows, sheep, goats, pigs, rabbits and the like.

Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the examples.

Example  1: Preparation for experiment

1.1. Wealthy( Aconitum carmichael Debx ) Preparation of extract

Aconitum carmichael In order to produce water extracts (ACD), toxicants were purchased and used (Kwangmyungdang Medicinal Herbs, Ulsan, Korea). The resulting extract was filtered with a filter paper (Whatman No. 1; Sigma-Aldrich, St. Louis, Mo., USA), and the filtrate was concentrated under reduced pressure To obtain a water extract (yield: 22.5%).

1.2. Cell culture

The mouse-derived C2C12 cell line was used from the American Type Culture Collection (ATCC) (CRL-1772; Manassas, VA, USA). C2C12 muscle cells were seeded at 1 × 10 ⁶ cells per 60 mm petridish and then 100% confluence in high glucose Dulbecco's modified eagle's medium (DMEM) containing fetal bovine serum (10%) and penicillin streptomycin (1% . Then, horseserum (2%) induced differentiation of muscle cells. After differentiating for 4 days while exchanging the medium every 24 hours, the supplement was treated at a concentration of 0.5 mg / ml for 24 hours.

1.3. Western blot

Western blot. Cells were collected, centrifuged at 5,000 rpm for 5 minutes, washed twice with phosphate buffered saline, and lysed in 50 mM HEPES, pH 7.4, 150 mM NaCl, 1% deoxycholate, 1 mM ethylenediaminetetraacetic acid, 1 mM phenylmethanesulfonylfluoride, 1 μg / ml aprotinin) and allowed to react on ice for 30 minutes. 20 μg of protein was mixed with 2 × sample buffer (100 mM Tris-HCl, pH 6.8, 200 mM dithithreitol, 4% sodium dodecyl sulfate [SDS], 0.2% bromophenol blue and 20% glycerol) SDS-polyacrylamide gel electrophoresis. The separated gel proteins were electrophoretically transferred to a nitrocellulose membrane at 4 ° C and 30 V for 16 hours. Membrane was blocked with 10% skim milk at room temperature for 1 hour, and reacted with antibodies of each protein for 2 hours at room temperature. Anti-pAMPK, AMPK (cell signaling) and anti-β actin (Sigma-Aldrich, St. Louis, MO) were washed three times with Tris-buffered saline and Tween 20. Louis, MO, USA), reacted at room temperature for 1 hour, and subjected to substrate reaction with enhanced chemiluminoesence solution to sensitize them to X-ray film. The bands on the X-ray film were expressed as the ratio of expression of PGC1α to β actin and the expression ratio of pAMPK to AMPK using an image (Image-J) program.

1.4. Glucose uptake assay

To the differentiated skeletal muscle cells, the medium was added to glucose-free DMEM containing 2- [N- (7-160 nitrobenz-2-oxa-1,3-diazol-4-yl) amino] -2-deoxy- 150 μg / ml, and treated with each drug. After incubation for 4 hours, the medium was removed and 200 μl of cell-based assay buffer was added. The amount of 2-NBDG uptake was measured by fluorescence microscopy (Leica Biosystems, Wetzlar, Germany).

1.5. ATP measurement

Skeletal muscle cells were differentiated according to the cell culture method described above, treated with each drug extract, and cultured for 24 hours. Absorbance was measured at 570 nm after the experiment using the adenosine triphosphate (ATP) calorimetric assay kit (BioVision Inc., Milpitas, Calif., USA).

1.6. Statistical analysis

StatPad Prism (GraphPad Software Inc., San Diego, Calif., USA). The results of each of the three replicates were expressed as mean ± standard errors of mean and analyzed using analysis of variance (Tukey's test). Statistical significance was accepted when the P-value was less than 0.05.

Example  2: Results

2.1. Rich Of PGC1a  Identification of expression changes

PGC1α is an important factor in mitochondrial neogenesis and plays an important role in fatty acid oxidation and heat generation in skeletal muscle. In order to confirm the expression of PGC1? By the rich in C2C12 cells, the protein level of PGC1? Was measured by Western blotting. When 0.5 mg / mL of rich was treated, the level of PGC1? Protein was increased (FIG. 1).

2.2. Rich AMPK's  Identification of phosphorylation change

Phosphorylation of AMPK increases mitochondrial neogenesis, glucose transport, and fatty acid oxidation in skeletal muscle. In particular, AMPK directly phosphorylates PGC1a. Thus, in the present invention, the amount of protein expression of AMPK and pAMPK was measured after treatment with the extract. As a result, as shown in Fig. 2, treatment of the rich in C2C12 myotubes was shown to increase phosphorylation of AMPK.

2.3. Confirmation of Glucose uptake control by rich

The glucose uptake capacity of the drug was analyzed by measuring the expression level of 2-NBDG by fluorescence microscopy. As a result, as shown in FIG. 3, glucose uptake was increased in the rich treatment group compared to the control group.

2.4. Confirmation of change of ATP amount by rich

Increases in enzymes involved in regulation of body energy, such as PGC1α and AMPK, lead to an increase in sugar and fatty acid metabolism and ultimately an increase in ATP production. In the previous examples, PGC1α, AMPK and glucose metabolism controlling activity of the rich were confirmed, and the resulting increase in ATP production was analyzed. Rich extract showed a statistically significant increase in total ATP compared to the control (Fig. 4).

Examples of formulations for the composition of the present invention are illustrated below.

Formulation example  1: Preparation of pharmaceutical preparations

1. Manufacturing of powder

Rich extract 200 mg

Lactose 100 mg

The above components were mixed and packed in airtight bags to prepare powders.

2. Preparation of tablets

Rich extract 200 mg

100 mg of corn starch

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

3. Preparation of capsules

Rich extract 200 mg

100 mg of corn starch

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, the capsules were filled in gelatin capsules according to the conventional preparation method of capsules.

4. Preparation of injections

Rich extract 200 mg

100 mg mannitol

Na ₂ HPO _{4 12} H ₂ O 2 mg

Sterile sterilized water for injection

Injection was prepared by mixing the above components per ampoule (2 mL) according to the usual injection preparation method.

Formulation example  2: Manufacturing of food

Foods containing the rich extract of the present invention were prepared as follows.

1. Preparation of cooking seasoning

A cooked sauce for health promotion was prepared from 20 to 95% by weight of a rich extract.

2. Manufacture of tomato ketchup and sauce

0.2-1.0% by weight of the rich extract was added to tomato ketchup or sauce to make healthy tomato ketchup or sauce.

3. Manufacture of flour food

0.5-5.0 wt.% Of the extract was added to wheat flour, and the mixture was used to prepare bread, cake, cookies, crackers and noodles to produce health promotion foods.

4. Manufacture of soups and gravies

0.1-5.0 wt.% Of the rich extract was added to the soup and juice to prepare a health improvement meat product, noodle soup and juice.

5. Manufacture of ground beef

A ground beef for health promotion was prepared by adding 10% by weight of the rich extract to ground beef.

6. Manufacture of dairy products

5-10% by weight of the rich extract was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

Formulation example  3: Manufacturing of beverages

1. Manufacture of carbonated beverages

Syrup was prepared by mixing 10-15% of rich extract, 5-10% of sugar, 0.05-0.3% of citric acid, 0.005-0.02% of caramel, 0.1-1% of vitamin C and 75-80% of purified water . The syrup was sterilized at 85-98 ° C for 20-180 seconds, mixed with cooling water at a ratio of 1: 4, and then 0.5-0.82% carbon dioxide gas was injected to prepare a carbonated drink containing the rich extract.

2. Manufacture of health drinks

(50%), extract of overdose (70%, 0.12%), vitamin C (0.02%), calcium pantothenate (0.02%), licorice extract (Solid content: 65%, 0.01%) were uniformly blended, instant sterilized, and packaged in small containers such as glass bottles and plastic bottles to produce health drinks.

3. Manufacture of vegetable juice

Healthy vegetable juice was prepared by adding 0.5 g of rich extract to 1,000 ml of tomato or carrot juice.

4. Manufacture of fruit juice

Healthy fruit juice was prepared by adding 0.1 g of the rich extract to 1,000 ml of apple or grape juice.

It will be understood by those skilled in the art that the foregoing description of the present invention is for illustrative purposes only and that those of ordinary skill in the art can readily understand that various changes and modifications may be made without departing from the spirit or essential characteristics of the present invention. will be. It is therefore to be understood that the above-described embodiments are illustrative in all aspects and not restrictive.

Claims (8)

A composition for stimulating body energy consumption or promoting muscle metabolism, comprising a rich extract as an active ingredient.
The method according to claim 1,
Wherein the rich extract is at least one selected from the group consisting of water, C1 to C4 lower alcohols, n-hexane, ethyl acetate, acetone, butyl acetate, 1,3-butylene glycol, methylene chloride, A composition for stimulating body energy consumption or promoting muscle metabolism, which is extracted with a solvent.
The method according to claim 1,
Wherein the rich extract is extracted from the root of the rich. ≪ RTI ID = 0.0 > 20. < / RTI >
The method according to claim 1,
Wherein the composition increases the expression of peroxisome proliferator-activated receptor gamma-assisting activator 1-alpha (PGC1-alpha).
The method according to claim 1,
Wherein said composition promotes the production of mitochondria.
A health-functional food composition for stimulating body energy consumption or promoting muscle metabolism, comprising a rich extract as an active ingredient.
The method according to claim 6,
Wherein the rich extract is at least one selected from the group consisting of water, C1 to C4 lower alcohols, n-hexane, ethyl acetate, acetone, butyl acetate, 1,3-butylene glycol, methylene chloride, Wherein the composition is extracted with a solvent, wherein the health functional food composition is used for stimulating body energy consumption or promoting muscle metabolism.
The method according to claim 6,
Wherein the rich extract is extracted from the root of the rich. 20. A health functional food composition for promoting body energy consumption or muscle metabolism.

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