KR20180000124A - Composition for improving metabolism containing extraction of Cinnamomum cassia Blume - Google Patents

Abstract

The present invention relates to a composition for promoting body energy consumption or promoting muscle metabolism containing a Cinnamomum cassia BLUME extract as an active ingredient. The Cinnamomum cassia BLUME 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 The present invention relates to a composition for accelerating body energy consumption or muscle metabolism containing an extract of broiler chickens as an active ingredient,

The present invention relates to a composition for promoting body energy consumption or muscle metabolism, which comprises a broiler chick 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.

Cinnamomum cassia Blume is a medicinal plant used in oriental medicine as diabetes and obesity treatment. Its pharmacological action is known to be central inhibition, sedation, pain, fever, increase of coronary blood flow and inhibition of skin fungus.

The present inventors investigated the metabolic activation effect of broiler chick extract and found that PGC1-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 promoting body energy consumption or muscle metabolism, which comprises a broiler chick extract as an active ingredient.

Another object of the present invention is to provide a health food composition containing a broiler chick extract as an active ingredient for promoting body energy consumption or promoting muscle metabolism.

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 achieve the above object, the present invention provides a composition for promoting body energy consumption or muscle metabolism, which comprises a broiler chick extract as an active ingredient.

In one embodiment of the present invention, the broiler chick extract is prepared by mixing 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 broiler chick extract is extracted from the stem shell of broiler chickens.

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 broiler chick extract as an active ingredient.

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

In addition, the present invention provides the use of broiler chick extract for promotion of body energy consumption or promotion of muscle metabolism.

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

The present invention provides a composition for accelerating body energy consumption or muscle metabolism comprising a broiler chick 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 broiler chick 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 broiler chick extract according to the present invention was extracted from the stem peel of broiler chickens. The mouse muscle cells (C2C12) were cultured to induce differentiation into myotubes, and broiler chickens were treated with PGC1-α (peroxisome proliferator- Gamma coactivator 1-alpha, peroxisome proliferator-activated receptor gamma coactivator 1-alpha) and promotes the production of mitochondria, thus confirming that the extract has an effect of promoting body energy consumption and muscle metabolism .

Accordingly, the broiler chick extract according to the present invention can be used as a medicament useful for diseases caused by mitochondrial dysfunction, that is, deficient in energy metabolism and muscle metabolism, and as a health functional food for promoting body energy consumption or promoting muscle metabolism.

The broiler chick 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 in the production of the broiler chick extract of the present invention include water, C1 to C4 lower alcohol, 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 the dry weight of the broiler. 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 broiler-derived useful component 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 promoting body energy consumption or promoting muscle metabolism in addition to broiler chick extract.

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 broiler chick extract is about 0.0001-500 mg / kg, preferably about 0.001-300 mg / kg, and is preferably administered once a day 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 broiler chick extract of the present invention is used as a food additive, the broiler chick extract can be used 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 broiler 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. 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 stimulating body energy consumption or muscle metabolism, which comprises administering a broiler 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. Broiler ( Cinnamomum cassia Blume) extract

Cinnamomum cassia Blume was purchased and used to produce Cinnamomum cassia Blume water extract (CCB) (Kwangmyungdang Medicinal Herbs, Ulsan, Korea). The extracts were filtered with a filter paper (Whatman NO. 1; Sigma-Aldrich, St. Louis, Mo., USA), and the extracts were filtered through a rotary vacuum evaporator , And lyophilized to give a water extract (yield: 6.7%).

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 broiler 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. Identification of expression of PGC1α by broiler

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

2.2. Identification of phosphorylation of AMPK by broiler

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

2.3. Identification of Glucose uptake control by broiler

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 broiler treatment group as compared with the control group.

2.4. Identification of changes in ATP amount by broiler

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 example, PGC1α, AMPK and glucose metabolism regulating effect of broiler chickens were confirmed, and the resultant increase in ATP production was analyzed. Broiler extracts showed a statistically significant increase in the total amount of 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

Broiler chick extract 200 mg

Lactose 100 mg

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

2. Preparation of tablets

Broiler chick 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

Broiler chick 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

Broiler chick 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 broiler chick extract of the present invention were prepared as follows.

1. Preparation of cooking seasoning

A broth for health promotion was prepared with 20-95% by weight of broiler chick extract.

2. Manufacture of tomato ketchup and sauce

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

3. Manufacture of flour food

A broth, a cake, a cookie, a cracker and a noodle were prepared by adding 0.5-5.0 wt% of broiler chick extract to wheat flour.

4. Manufacture of soups and gravies

0.1-5.0% by weight of broiler chick extract was added to soup and juice to prepare health promotion meat product, noodle soup and juice.

5. Manufacture of ground beef

10% by weight of broiler chick extract was added to ground beef to prepare ground beef for health promotion.

6. Manufacture of dairy products

5-10% by weight of broiler chick 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 broiler chick extract, 5-10% of sugar, 0.05-0.3% of citric acid, 0.005-0.02% of caramel and 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 0.5-0.82% carbon dioxide gas was injected to prepare a carbonated drink containing broiler chick extract.

2. Manufacture of health drinks

The extracts of broiler chickens (2.5%, 97.16%), jujube extract (65 brix, 2.67%), over-complex extracts (solid content 70%, 0.12%), vitamin C (0.02%), calcium pantothenate (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 broiler chick extract to 1,000 ml of tomato or carrot juice.

4. Manufacture of fruit juice

0.1 g of broiler chick extract was added to 1,000 ml of apple or grape juice to prepare health promotion fruit 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 accelerating body energy consumption or promoting muscle metabolism, comprising a broiler chick extract as an active ingredient.
The method according to claim 1,
Wherein the broiler chick extract comprises 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 broiler chick extract is extracted from the stem skin of broiler chickens.
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 accelerating body energy consumption or facilitating muscle metabolism, which comprises broiler chick extract as an active ingredient.
The method according to claim 6,
Wherein the broiler chick extract comprises 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 broiler chick extract is extracted from the stem skin of broiler chickens.

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