WO2020027586A1 - Composition comprising gallic acid as active ingredient for accelerating muscle differentiation - Google Patents
Composition comprising gallic acid as active ingredient for accelerating muscle differentiation Download PDFInfo
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- WO2020027586A1 WO2020027586A1 PCT/KR2019/009565 KR2019009565W WO2020027586A1 WO 2020027586 A1 WO2020027586 A1 WO 2020027586A1 KR 2019009565 W KR2019009565 W KR 2019009565W WO 2020027586 A1 WO2020027586 A1 WO 2020027586A1
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- muscle
- gallic acid
- active ingredient
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- GGCZERPQGJTIQP-UHFFFAOYSA-N sodium;9,10-dioxoanthracene-2-sulfonic acid Chemical compound [Na+].C1=CC=C2C(=O)C3=CC(S(=O)(=O)O)=CC=C3C(=O)C2=C1 GGCZERPQGJTIQP-UHFFFAOYSA-N 0.000 description 1
- 229960002920 sorbitol Drugs 0.000 description 1
- 235000014347 soups Nutrition 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 150000005846 sugar alcohols Chemical class 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 235000012222 talc Nutrition 0.000 description 1
- 235000013616 tea Nutrition 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000008733 trauma Effects 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 208000016261 weight loss Diseases 0.000 description 1
- 230000004580 weight loss Effects 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
- 235000010447 xylitol Nutrition 0.000 description 1
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/192—Carboxylic acids, e.g. valproic acid having aromatic groups, e.g. sulindac, 2-aryl-propionic acids, ethacrynic acid
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P21/00—Drugs for disorders of the muscular or neuromuscular system
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/316—Foods, ingredients or supplements having a functional effect on health having an effect on regeneration or building of ligaments or muscles
Definitions
- the present invention relates to a composition for promoting muscle differentiation comprising gallic acid as an active ingredient.
- the muscles in our bodies attach to bones, which protect them and keep them in shape. Muscles also promote calcium influx, which in turn increases bone density. However, as the body ages, the redistribution of body fat and protein occurs as a result of changes in its composition.At 50 years of age, the synthesis rate of proteins in muscle cells is slower than the rate of decomposition, and muscles begin to rapidly degenerate. May be exposed.
- Muscle loss which is one of the muscle loss diseases, refers to a condition in which about 13 to 24% of the body's body mass is usually reduced. Easily injured and even serious injuries.
- One of the causes of muscular dystrophy can be attributed to the gradual reduction in the amount and quality of skeletal muscle as aging progresses, and the weight loss of fat and body fat due to inadequate dietary energy intake.
- Myoblasts formed through the division and determination of precursor cells differentiate into myocytes and fuse with adjacent cells to differentiate into myotubes. Differentiated root canal cells form muscle through the process of forming myofibril. Some of the myoblasts remain stationary as stem satellite cells, but when muscle tissue is exposed to diseases such as trauma or muscular dystrophy, muscle stem cells, known as satellite cells, become damaged muscles. Heals and regenerates. In diseases such as muscular dystrophy, these cells are particularly important for muscle stability and healing.
- Gallic acid is a 3,4,5-oxybenzoic acid and is also referred to as a molar asset.
- Colorless columnar or long acicular crystal odorless, tasting, slightly acidic. It has one molecule of crystallization water, and it depends on the heating rate at 100 ⁇ 120 °C, and it decomposes at the same time as melting and becomes carbon dioxide and pyrogallol.
- Converging but not tannins It is insoluble in cold water but soluble in hot water, alcohol and acetone, and produces dark blue precipitate by iron (III) salt.
- aqueous alkali solution shows strong reducing properties and becomes brown. It is used as a hemostatic agent in addition to the production of blue ink, as a raw material for the developer or dye in photographs. Since it will discolor upon contact with sunlight and air, it should be sealed in a brown bottle or can and stored away from sunlight.
- Such a gallic acid (gallic acid) related technology is disclosed in Korean Patent No. 1286153, an anticancer composition containing a gallic acid derivative as an active ingredient, and a novel gallic acid-conjugated linoleic acid fatty ester in Korean Patent No. 0665315, Although a preparation method thereof and a composition containing the same are disclosed, the composition for promoting muscle differentiation comprising the gallic acid of the present invention as an active ingredient is not disclosed.
- the present invention is derived from the above requirements, the present invention provides a composition for promoting muscle differentiation comprising gallic acid as an active ingredient, gallic acid as an active ingredient not only increases the survival rate of myoblasts, but also the source
- the present invention was completed by identifying differentiation of cells into muscle cells and changes in the expression level of muscle differentiation-related proteins.
- the present invention provides a differentiation promoter of myoblast (galolic acid) or a pharmaceutically acceptable salt thereof as an active ingredient.
- the present invention also provides a pharmaceutical composition for the prevention or treatment of muscle diseases containing the source cell differentiation promoter as an active ingredient.
- the present invention provides a health functional food composition for preventing or improving muscle growth or senile muscle loss, containing the source cell differentiation promoter as an active ingredient.
- the present invention relates to a composition for promoting muscle differentiation comprising gallic acid as an active ingredient, gallic acid as an active ingredient of the present invention not only enhances cell survival rate of myoblasts, but also induces and promotes differentiation into muscle cells. The effect is to regulate the expression level of muscle cell differentiation-related proteins.
- 1 is a result of the cell survival rate confirmed by treating the gallic acid according to the concentration by concentration.
- Figure 3 is the result of confirming the proliferation and differentiation state of myoblasts after treatment with the gallic acid according to the present invention to myoblasts 48 hours.
- the present invention relates to a differentiation promoter of myoblasts comprising gallic acid or a pharmaceutically acceptable salt thereof as an active ingredient.
- the gallic acid is a compound consisting of a structural formula represented by the following formula (1).
- the present invention also relates to a pharmaceutical composition for preventing or treating muscle diseases containing the myocyte differentiation promoter as an active ingredient.
- the muscle disease is any one selected from atony, muscular atrophy, muscular dystrophy, muscle degeneration, myopathy, muscular atrophy, ataxia, cachexia and sarcopenia Although it is preferable, it is not limited to this.
- Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are conventionally used in the preparation, and include saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, lactose, water Cross, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzo 8, talc, magnesium stearate and mineral oil, and the like.
- Suitable dosages of the pharmaceutical compositions of the present invention can be variously prescribed by such factors as the formulation method, mode of administration, age, weight, sex, morbidity, condition of food, time of administration, route of administration, rate of excretion and response to response of the patient. Can be.
- the route of administration of the pharmaceutical composition for the prophylaxis or treatment of the muscular diseases of the present invention may be administered via any route generally accepted as long as the target tissue can be reached.
- the pharmaceutical composition of the present invention is not particularly limited, but as desired, intramuscular administration, eye drop administration, intraperitoneal administration, intravenous administration, subcutaneous administration, intradermal administration, transdermal patch administration, oral administration, intranasal administration, pulmonary administration, It may be administered via a route such as rectal administration, and specifically may be administered through a route of intramuscular administration.
- the present invention also relates to a health functional food composition for preventing or improving muscle growth or senile muscle loss, containing the muscle cell differentiation promoter as an active ingredient.
- the health functional food composition is preferably made of any one selected from powder, granules, pills, tablets, capsules, candy, syrups and beverages, but is not limited thereto.
- the health functional food composition of the present invention may be prepared by adding gallic acid as it is or mixing with other food or food ingredients, and may be appropriately prepared according to a conventional method.
- Examples of foods to which the lenders extract may be added include dairy products including caramel, meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, various soups, beverages, tea , Drink, alcoholic beverages and vitamin complexes may be in any one form selected, and includes all of the health functional foods in the conventional sense. That is, there is no particular limitation on the kind of food.
- the health functional food composition includes various nutrients, vitamins, minerals (electrolytes), synthetic and natural flavors, colorants and enhancers (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners. , pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. It may also contain pulp for the production of natural fruit juices and vegetable drinks. The above components can be used independently or in combination.
- the health functional food composition of the present invention may contain a variety of flavors or natural carbohydrates, etc.
- the natural carbohydrates are monosaccharides such as glucose, fructose, maltose, sucrose, disaccharides, dextrin, cyclo Polysaccharides such as dextrin, sugar alcohols such as xylitol, sorbitol, and erythritol.
- the ratio of the said natural carbohydrate is not important, it is preferable that it is 0.01-0.04g with respect to 100g of the composition of this invention, More preferably, it contains 0.02-0.03g, but it is not limited to this.
- natural sweeteners such as tautin, stevia extract, synthetic sweeteners such as saccharin, aspartame, and the like can be used.
- the cell line used in the examples of the present invention was obtained by separating muscle tissue from the hind limbs of mice 3 days old as mouse primary myoblasts.
- the cell line was added with 10% cosmic calf serum, 10ng / ml basic fibroblast growth factor, and 1% antibiotic in Ham's F-10 nutrient mixture (F-10) medium.
- the cells were cultured in a culture dish coated with poly-L-ornithin and collagen at 37 ° C., 5% CO 2 .
- MTS (3- (4,5-dimethylthiazol-2-yl) -5- (3-carboxymethoxyphenyl) -2- (4-sulfophenyl) -2H-tetrazolium, inner salt) assay was performed.
- Cytotoxicity (%) [(absorbance of the sample addition group-absorbance of the sample itself) / absorbance of the control group] ⁇ 100
- gallic acid increased the cell activity of myoblasts, and especially gallic acid showed the greatest cell activity of 178.32% at 40 ⁇ g / ml.
Abstract
The present invention relates to a composition for accelerating muscle differentiation, comprising gallic acid as an active ingredient, wherein the active ingredient of the present invention, gallic acid, has the effects of increasing cell viability of myogenic cells, inducing and accelerating myogenic cells to muscle cells, and regulating the expression of proteins involved in muscle cell differentiation. Accordingly, the gallic acid of the present invention can be beneficially used in a myogenic differentiation accelerator, a medication for muscle diseases, or a functional health food for promoting increase in muscle or preventing or alleviating age-induced sarcopenia.
Description
본 발명은 갈산을 유효성분으로 포함하는 근 분화 촉진용 조성물에 관한 것이다.The present invention relates to a composition for promoting muscle differentiation comprising gallic acid as an active ingredient.
우리 몸의 근육은 뼈에 붙어 뼈를 보호하고 체형을 바르게 유지시켜 주는 등의 여러 가지 기능을 한다. 또한, 근육은 칼슘 유입을 촉진시켜 골 밀도를 높여 주기도 한다. 그러나 신체는 노화하면서 구성성분의 변화로써 체지방과 체단백질의 재분포가 일어나며, 약 50세가 되면 근 세포 내 단백질의 합성 속도가 분해속도보다 느려져 근육이 급격하게 퇴화를 시작하게 되며, 근육 감소 질환에 노출될 수 있다.The muscles in our bodies attach to bones, which protect them and keep them in shape. Muscles also promote calcium influx, which in turn increases bone density. However, as the body ages, the redistribution of body fat and protein occurs as a result of changes in its composition.At 50 years of age, the synthesis rate of proteins in muscle cells is slower than the rate of decomposition, and muscles begin to rapidly degenerate. May be exposed.
근육 감소 질환의 하나인 근육 감소증은 평소 자기 체질량의 약 13~24%가 감소한 상태를 말하는 것으로, 근육 감소증이 있으면 행동량이 현격하게 줄어 정신건강을 해칠 뿐만 아니라 생활의 만족도도 떨어지며, 용이한 일상 생활에서도 쉽게 부상을 입고 심각한 중상에 이르기도 한다. 근육 감소증의 원인 중 하나는 노화가 진행됨에 따라 일어나는 골격근의 양과 질의 점진적 감소 및 부적절한 식이 에너지 섭취에 따른 지방과 체지방성분을 포함하는 체중감소 등을 원인으로 꼽을 수 있다.Muscle loss, which is one of the muscle loss diseases, refers to a condition in which about 13 to 24% of the body's body mass is usually reduced. Easily injured and even serious injuries. One of the causes of muscular dystrophy can be attributed to the gradual reduction in the amount and quality of skeletal muscle as aging progresses, and the weight loss of fat and body fat due to inadequate dietary energy intake.
전구세포(precursor cell)의 분열과 결정 과정을 통해 형성된 근원세포(myoblast)는 근세포(myocyte)로 분화하고, 인접한 세포들과 융합하여 근관세포(myotube)로 분화한다. 분화된 근관세포는 myofibril(근원섬유)를 형성하는 과정을 통해 근육이 형성된다. 근원세포의 일부는 줄기 위성세포(satellite cell)로서 정지 상태로 있다가, 근육조직이 외상 또는 근육위축병(muscular dystrophy)과 같은 질병에 노출되면, 위성세포로 알려진 근육 줄기세포들은 손상된 부위의 근육을 치유하고 재생시킨다. 근육위축병과 같은 질환에 있어서, 이들 세포들은 근육의 안정성과 치유에 있어 특히 중요하다.Myoblasts formed through the division and determination of precursor cells differentiate into myocytes and fuse with adjacent cells to differentiate into myotubes. Differentiated root canal cells form muscle through the process of forming myofibril. Some of the myoblasts remain stationary as stem satellite cells, but when muscle tissue is exposed to diseases such as trauma or muscular dystrophy, muscle stem cells, known as satellite cells, become damaged muscles. Heals and regenerates. In diseases such as muscular dystrophy, these cells are particularly important for muscle stability and healing.
한편, 갈산은 3,4,5－옥시벤존산으로, 몰식자산이라고도 한다. 무색의 주상 또는 긴 침상 결정으로, 냄새는 없고 맛이 떫으며, 약간 산성을 띤다. 1분자의 결정수를 가지며, 100~120℃로 가열속도에 따라 다르며, 융해와 동시에 분해하여 이산화탄소 및 피로갈롤이 된다. 금염 ·은염 ·펠링용액을 환원시킨다. 수렴성이 있으나, 타닌산만은 못하다. 찬물에는 잘 녹지 않으나, 뜨거운 물, 알코올, 아세톤에는 녹으며, 철(Ⅲ)염에 의해서 암청색 침전을 생성한다. 식물의 잎, 줄기, 뿌리 및 몰식자, 오배자(五倍子) 등에 유리상태로 존재하고, 또는 타닌의 구성분자로 분포한다. 알칼리 수용액은 강력한 환원성을 보이며, 갈색이 된다. 청색 잉크의 제조를 비롯하여 사진의 현상액 또는 염료의 원료로 사용하는 외에 지혈제로 사용된다. 햇빛 및 공기와 접촉하면 변색하므로, 갈색병 또는 깡통에 밀봉하여 햇빛을 받지 않도록 저장해야 한다.Gallic acid, on the other hand, is a 3,4,5-oxybenzoic acid and is also referred to as a molar asset. Colorless columnar or long acicular crystal, odorless, tasting, slightly acidic. It has one molecule of crystallization water, and it depends on the heating rate at 100 ~ 120 ℃, and it decomposes at the same time as melting and becomes carbon dioxide and pyrogallol. Reduce the gold salt, silver salt, and pelleting solution. Converging but not tannins. It is insoluble in cold water but soluble in hot water, alcohol and acetone, and produces dark blue precipitate by iron (III) salt. It exists in a free state in the leaves, stems, roots and gluttony of the plant, gallastia (五倍子), or is distributed as a constituent molecule of tannin. The aqueous alkali solution shows strong reducing properties and becomes brown. It is used as a hemostatic agent in addition to the production of blue ink, as a raw material for the developer or dye in photographs. Since it will discolor upon contact with sunlight and air, it should be sealed in a brown bottle or can and stored away from sunlight.
이와 같은 갈산(gallic acid) 관련 기술로는 한국등록특허 제1286153에 갈산 유도체를 유효성분으로 함유하는 항암 조성물이 개시되어 있고, 한국등록특허 제0665315호에 신규한 갈산-콘쥬게이티드 리놀레산 지방 에스테르, 이의 제조방법 및 이를 함유하는 조성물이 개시되어 있으나, 본 발명의 갈산을 유효성분으로 포함하는 근 분화 촉진용 조성물은 개시된 바 없다.Such a gallic acid (gallic acid) related technology is disclosed in Korean Patent No. 1286153, an anticancer composition containing a gallic acid derivative as an active ingredient, and a novel gallic acid-conjugated linoleic acid fatty ester in Korean Patent No. 0665315, Although a preparation method thereof and a composition containing the same are disclosed, the composition for promoting muscle differentiation comprising the gallic acid of the present invention as an active ingredient is not disclosed.
본 발명은 상기와 같은 요구에 의해 도출된 것으로서, 본 발명은 갈산을 유효성분으로 포함하는 근 분화 촉진용 조성물을 제공하고, 유효성분인 갈산이 근원세포의 생존율을 현저하게 증가시킬 뿐만 아니라, 근원세포가 근 세포로 분화를 하는 것과, 근 분화 관련 단백질의 발현량 변화를 확인함으로써, 본 발명을 완성하였다.The present invention is derived from the above requirements, the present invention provides a composition for promoting muscle differentiation comprising gallic acid as an active ingredient, gallic acid as an active ingredient not only increases the survival rate of myoblasts, but also the source The present invention was completed by identifying differentiation of cells into muscle cells and changes in the expression level of muscle differentiation-related proteins.
상기 목적을 달성하기 위하여, 본 발명은 갈산(gallic acid) 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 근원세포(myoblast)의 분화 촉진제를 제공한다.In order to achieve the above object, the present invention provides a differentiation promoter of myoblast (galolic acid) or a pharmaceutically acceptable salt thereof as an active ingredient.
또한, 본 발명은 상기 근원 세포 분화 촉진제를 유효성분으로 함유하는 근육 질환의 예방 또는 치료용 약학 조성물을 제공한다.The present invention also provides a pharmaceutical composition for the prevention or treatment of muscle diseases containing the source cell differentiation promoter as an active ingredient.
또한, 본 발명은 상기 근원 세포 분화 촉진제를 유효성분으로 함유하는 근육증가 촉진 또는 노인성 근 손실의 예방 또는 개선용 건강기능성식품 조성물을 제공한다.In addition, the present invention provides a health functional food composition for preventing or improving muscle growth or senile muscle loss, containing the source cell differentiation promoter as an active ingredient.
본 발명은 갈산을 유효성분으로 포함하는 근 분화 촉진용 조성물에 관한 것으로, 본 발명의 유효성분인 갈산이 근원세포의 세포 생존률을 증진시킬 뿐만 아니라, 근 세포로의 분화를 유도 및 촉진할 수 있으며, 근세포 분화 관련 단백질의 발현량을 조절하는 효과가 있는 것이다.The present invention relates to a composition for promoting muscle differentiation comprising gallic acid as an active ingredient, gallic acid as an active ingredient of the present invention not only enhances cell survival rate of myoblasts, but also induces and promotes differentiation into muscle cells. The effect is to regulate the expression level of muscle cell differentiation-related proteins.
도 1은 본 발명에 따른 갈산을 농도별로 처리하여 확인한 세포생존율의 결과이다.1 is a result of the cell survival rate confirmed by treating the gallic acid according to the concentration by concentration.
도 2는 본 발명에 따른 갈산을 근원세포에 처리한 후, 면역 블롯 분석으로 근분화 촉진과 관련된 단백질의 발현량 변화를 확인한 결과이다.2 is a result of confirming the change in the expression level of the protein associated with the promotion of muscle differentiation by immunoblot analysis after treating the gallic acid according to the present invention to the myoblasts.
도 3은 본 발명에 따른 갈산을 근원세포에 처리하고 48시간 후의 근원세포의 증식 및 분화 상태를 확인한 결과이다.Figure 3 is the result of confirming the proliferation and differentiation state of myoblasts after treatment with the gallic acid according to the present invention to myoblasts 48 hours.
본 발명은 갈산(gallic acid) 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 근원세포(myoblast)의 분화 촉진제에 관한 것이다.The present invention relates to a differentiation promoter of myoblasts comprising gallic acid or a pharmaceutically acceptable salt thereof as an active ingredient.
상기 갈산은 하기 화학식 1로 기재되는 구조식으로 이루어진 화합물이다. The gallic acid is a compound consisting of a structural formula represented by the following formula (1).
또한, 본 발명은 상기 근세포 분화 촉진제를 유효성분으로 함유하는 근육 질환의 예방 또는 치료용 약학 조성물에 관한 것이다.The present invention also relates to a pharmaceutical composition for preventing or treating muscle diseases containing the myocyte differentiation promoter as an active ingredient.
상기 근육 질환은 긴장감퇴증(atony), 근위축증(muscular atrophy), 근이영양증(muscular dystrophy), 근육 퇴화, 근경직증, 근위축성 축삭경화증, 근무력증, 악액질(cachexia) 및 근육감소증(sarcopenia) 중에서 선택된 어느 하나인 것이 바람직하지만 이에 한정하는 것은 아니다.The muscle disease is any one selected from atony, muscular atrophy, muscular dystrophy, muscle degeneration, myopathy, muscular atrophy, ataxia, cachexia and sarcopenia Although it is preferable, it is not limited to this.
본 발명의 약학 조성물에 포함되는 약학적으로 허용되는 담체는 제제 시에 통상적으로 이용되는 것으로서, 식염수, 멸균수, 링거액, 완충 식염수, 덱스트로즈 용액, 말토덱스트린 용액, 글리세롤, 에탄올, 락토스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산 칼슘, 알기네이트, 젤라틴, 규산 칼슘, 미세결정성 셀룰로스, 폴리비닐피롤리돈, 셀룰로스, 시럽, 메틸 셀룰로스, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 활석, 스테아르산 마그네슘 및 미네랄 오일 등을 포함하나, 이에 한정되는 것은 아니다. 상기 성분들 이외에 항산화제, 완충액, 정균제, 희석제, 계면활성제, 결합제, 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제 또는 보존제 등을 추가로 포함할 수 있다. 본 발명의 약학 조성물의 적합한 투여량은 제제화 방법, 투여 방식, 환자의 연령, 체중, 성, 병적 상태, 음식, 투여 시간, 투여 경로, 배설 속도 및 반응 감응성과 같은 요인들에 의해 다양하게 처방될 수 있다. 본 발명의 근육질환의 예방 또는 치료용 약학 조성물의 투여 경로는 목적 조직에 도달할 수 있는 한 일반적으로 허용되는 어떠한 경로를 통하여도 투여될 수 있다. 본 발명의 약학 조성물은 특별히 이에 제한되지 않으나, 목적하는 바에 따라 근육 내 투여, 점안 투여, 복강 내 투여, 정맥 내 투여, 피하 투여, 피내 투여, 경피 패치 투여, 경구 투여, 비내 투여, 폐내 투여, 직장 내 투여 등의 경로를 통해 투여될 수 있고, 구체적으로 근육 내 투여의 경로를 통해 투여될 수 있다.Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are conventionally used in the preparation, and include saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, lactose, water Cross, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzo 8, talc, magnesium stearate and mineral oil, and the like. In addition to the above components, antioxidants, buffers, bacteriostatic agents, diluents, surfactants, binders, lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents or preservatives may be further included. Suitable dosages of the pharmaceutical compositions of the present invention can be variously prescribed by such factors as the formulation method, mode of administration, age, weight, sex, morbidity, condition of food, time of administration, route of administration, rate of excretion and response to response of the patient. Can be. The route of administration of the pharmaceutical composition for the prophylaxis or treatment of the muscular diseases of the present invention may be administered via any route generally accepted as long as the target tissue can be reached. The pharmaceutical composition of the present invention is not particularly limited, but as desired, intramuscular administration, eye drop administration, intraperitoneal administration, intravenous administration, subcutaneous administration, intradermal administration, transdermal patch administration, oral administration, intranasal administration, pulmonary administration, It may be administered via a route such as rectal administration, and specifically may be administered through a route of intramuscular administration.
또한, 본 발명은 상기 근세포 분화 촉진제를 유효성분으로 함유하는 근육증가 촉진 또는 노인성 근 손실의 예방 또는 개선용 건강기능성식품 조성물에 관한 것이다.The present invention also relates to a health functional food composition for preventing or improving muscle growth or senile muscle loss, containing the muscle cell differentiation promoter as an active ingredient.
상기 건강기능성식품 조성물은 분말, 과립, 환, 정제, 캡슐, 캔디, 시럽 및 음료 중에서 선택된 어느 하나의 제형으로 제조되는 것이 바람직하지만 이에 한정하는 것은 아니다. 본 발명의 건강기능성식품 조성물은 갈산을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 혼합하여 제조될 수 있고, 통상적인 방법에 따라 적절하게 제조될 수 있다. 상기 가자 추출물을 첨가할 수 있는 식품의 예로는 카라멜, 육류, 소시지, 빵, 초콜릿, 캔디류, 스낵류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 수프, 음료수, 차, 드링크제, 알코올 음료 및 비타민 복합제 중에서 선택된 어느 하나의 형태일 수 있으며, 통상적인 의미에서의 건강기능성식품을 모두 포함한다. 즉, 상기 식품의 종류에는 특별한 제한은 없다. 상기 건강기능성식품 조성물은 여러 가지 영양제, 비타민, 광물(전해질), 합성 및 천연 풍미제, 착색제 및 증진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 또한, 천연 과일 주스 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 상기의 성분은 독립적으로 또는 조합하여 사용할 수 있다. 또한, 본 발명의 건강기능성식품 조성물은 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있으며, 상기 천연 탄수화물은 포도당, 과당과 같은 단당류, 말토스, 슈크로스와 같은 이당류, 덱스트린, 사이클로 덱스트린과 같은 다당류, 자일리톨, 소르비톨, 에리트리톨 등의 당 알코올이다. 상기 천연 탄수화물의 비율은 크게 중요하지 않지만, 본 발명의 조성물 100g에 대하여, 0.01~0.04g인 것이 바람직하고, 더욱 바람직하게는 0.02~0.03g을 포함하는 것이지만 이에 한정하지 않는다. 감미제로서는 타우마틴, 스테비아 추출물과 같은 천연 감미제, 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다. The health functional food composition is preferably made of any one selected from powder, granules, pills, tablets, capsules, candy, syrups and beverages, but is not limited thereto. The health functional food composition of the present invention may be prepared by adding gallic acid as it is or mixing with other food or food ingredients, and may be appropriately prepared according to a conventional method. Examples of foods to which the Gaza extract may be added include dairy products including caramel, meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, various soups, beverages, tea , Drink, alcoholic beverages and vitamin complexes may be in any one form selected, and includes all of the health functional foods in the conventional sense. That is, there is no particular limitation on the kind of food. The health functional food composition includes various nutrients, vitamins, minerals (electrolytes), synthetic and natural flavors, colorants and enhancers (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners. , pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. It may also contain pulp for the production of natural fruit juices and vegetable drinks. The above components can be used independently or in combination. In addition, the health functional food composition of the present invention may contain a variety of flavors or natural carbohydrates, etc. as additional components, the natural carbohydrates are monosaccharides such as glucose, fructose, maltose, sucrose, disaccharides, dextrin, cyclo Polysaccharides such as dextrin, sugar alcohols such as xylitol, sorbitol, and erythritol. Although the ratio of the said natural carbohydrate is not important, it is preferable that it is 0.01-0.04g with respect to 100g of the composition of this invention, More preferably, it contains 0.02-0.03g, but it is not limited to this. As the sweetener, natural sweeteners such as tautin, stevia extract, synthetic sweeteners such as saccharin, aspartame, and the like can be used.
이하, 실시예를 이용하여 본 발명을 더욱 상세하게 설명하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로 본 발명의 범위가 이들에 의해 제한되지 않는다는 것은 당해 기술분야에서 통상의 지식을 가진 자에게 있어 자명한 것이다. Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for explaining the present invention more specifically, it is obvious to those skilled in the art that the scope of the present invention is not limited by them.
실시예 1. 세포 배양Example 1. Cell Culture
본 발명의 실시예에서 사용된 세포주는 마우스 일차 근원세포(mouse primary myoblast)로 생후 3일 된 마우스의 뒷다리로부터 근육 조직을 분리하여 얻었다. 상기 세포주는 Ham's F-10 영양 혼합(F-10) 배지에 10% 코스믹 송아지 혈청(cosmic calf serum), 10ng/㎖의 염기성 섬유모세포생장인자(basic fibroblast growth factor), 1% 항생제를 첨가하여 표준 세포 배양법인 37℃, 5% CO
2 조건에서 폴리-L-오르니틴(poly-L-ornithin)과 콜라겐(collagen)으로 코팅한 배양 디쉬(culture dish)에서 배양하였다.The cell line used in the examples of the present invention was obtained by separating muscle tissue from the hind limbs of mice 3 days old as mouse primary myoblasts. The cell line was added with 10% cosmic calf serum, 10ng / ml basic fibroblast growth factor, and 1% antibiotic in Ham's F-10 nutrient mixture (F-10) medium. The cells were cultured in a culture dish coated with poly-L-ornithin and collagen at 37 ° C., 5% CO 2 .
실시예 2. 세포 생존율 분석Example 2. Cell Viability Assay
갈산(gallic acid)을 농도별로 24시간 동안 근원세포(myoblast)에 처리한 후. MTS(3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt) 어세이를 실시하였다. After gallic acid was treated with myoblasts for 24 hours at different concentrations. MTS (3- (4,5-dimethylthiazol-2-yl) -5- (3-carboxymethoxyphenyl) -2- (4-sulfophenyl) -2H-tetrazolium, inner salt) assay was performed.
96웰 플레이트에 근원세포(myoblast)를 2×10
4 개의
세포/웰로 분주하고 24시간 동안 배양한 후, 각각의 웰을 시료가 농도별로 처리된 배지로 교환해 주었다. 24시간 후 MTS 시약을 첨가하고 마이크로 플레이트 리더를 이용하여 490nm에서 흡광도를 측정하였다. 시료를 처리하지 않은 대조군 대비 시료를 처리한 세포의 생존율을 백분율로 표시하여 상대적인 세포 독성을 측정하였다.2 x 10 4 myoblasts in 96-well plates After dispensing into cells / well and incubating for 24 hours, each well was exchanged with medium in which samples were treated by concentration. After 24 hours MTS reagent was added and the absorbance was measured at 490 nm using a microplate reader. The relative cytotoxicity was measured by expressing the survival rate of the cells treated with the samples as a percentage compared to the control without the samples.
식(2)Formula (2)
세포독성(%)=[(시료 첨가군의 흡광도-시료 자체의 흡광도)/대조군의 흡광도] × 100Cytotoxicity (%) = [(absorbance of the sample addition group-absorbance of the sample itself) / absorbance of the control group] × 100
그 결과, 도 1에 개시한 바와 같이 갈산은 근원세포(myoblast)의 세포 활성도를 증가시켰으며, 특히 갈산이 40㎍/㎖에서 178.32%의 세포 활성도를 나타내며 가장 큰 효과를 보였다. As a result, as shown in FIG. 1, gallic acid increased the cell activity of myoblasts, and especially gallic acid showed the greatest cell activity of 178.32% at 40 µg / ml.
실시예Example
3. 3.
갈산의Gallic
처리에 따른 근원세포에서의 In myoblasts following treatment
pax3pax3
/7과 / 7 lesson
myh3myh3
단백질 발현량 변화 확인 Confirmation of protein expression change
근원세포 분화에 미치는 영향을 알아보기 위하여, 20㎍/㎖의 갈산을 48시간 동안 근원세포에 처리한 후, 면역블롯팅 분석(immunoblotting analysis)를 통해 pax3/7과 myh3 단백질 변화를 확인하였다. 상세하게는 근원세포(myoblast)를 PBS로 세척 후 분해 완충용액(lysis buffer)[10mM Tris-HCl (pH 7.5), 100mM NaCl, 1mM EDTA, 10% 글리세롤(glycerol), 1% Triton X-100]를 이용해 분해물(lysate)을 추출하였다. 로딩버퍼(Laemmi sample buffer)에 같은 양의 단백질 분해물(protein lysate)를 혼합하고, SDS-PAGE를 통해 단백질 분해물을 크기별로 분리시킨 후, anti-pax3/7 항체, anti-myh3 항체 및 anti-GAPDH 항체를 이용하여 면역블롯을 실시하였다.In order to determine the effect on myoblast differentiation, 20 μg / ml gallic acid was treated in myoblasts for 48 hours, and then pax3 / 7 and myh3 protein changes were confirmed by immunoblotting analysis. Specifically, the myoblasts were washed with PBS, followed by lysis buffer [10 mM Tris-HCl (pH 7.5), 100 mM NaCl, 1 mM EDTA, 10% glycerol, 1% Triton X-100]. The lysate was extracted using. After mixing the same amount of protein lysate in the loading buffer (Laemmi sample buffer) and separating the protein lysates by size through SDS-PAGE, anti-pax3 / 7 antibody, anti-myh3 antibody and anti-GAPDH Immunoblot was performed using the antibody.
그 결과, 도 2에 개시한 바와 같이 갈산의 처리에 의해 myh3 및 pax3/7의 단백질 발현이 농도 의존적으로 증가하였다.As a result, as shown in FIG. 2, the protein expression of myh3 and pax3 / 7 was increased concentration-dependently by the treatment of gallic acid.
실시예 4. 갈산의 처리에 따른 근원세포의 증식과 분화 확인Example 4 Confirmation of Proliferation and Differentiation of Myoblasts Following Treatment of Gallic Acid
갈산에 의한 근원세포(myoblast)에서의 myh3 단백질 발현량 증가를 면역형광 어세이(immunofluorescence assay)를 통해 시각화하였다. Increased expression of myh3 protein in myoblasts by gallic acid was visualized by immunofluorescence assay.
갈산을 처리하지 않은 대조군(control)과 비교하였을 때, 48시간 동안 분화 배지(differentiation media)로 분화를 유도한 근원세포(myoblast)인 대조군에서는 myh3 발현의 증가와 함께 세포융합(cell fusion) 현상이 일어나 myotube가 형성된 것을 관찰할 수 있었고, 20㎍/㎖의 갈산을 48시간 동안 처리해준 근원세포(myoblast)에서는 분화를 유도하지 않았음에도 myh3의 발현량의 증가와 함께 근세포(myocyte)의 형태로 세포의 형태가 변하며 분화가 유도된 것을 확인할 수 있었다(도 3).Compared with the control without gallic acid, cell fusion with myh3 expression was increased in the control group, which is a myoblast, which induced differentiation in differentiation media for 48 hours. It was observed that myotubes were formed, and in myoblasts treated with 20 ㎍ / ㎖ gallic acid for 48 hours, the cells in the form of myocytes with the increase of myh3 expression level were not induced differentiation. It was confirmed that the change in the differentiation was induced (Fig. 3).
Claims (4)
- 갈산(gallic acid) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는 근원세포(myoblast)의 분화 촉진제.A promoter for differentiation of myoblasts comprising gallic acid or a pharmaceutically acceptable salt thereof as an active ingredient.
- 제1항의 근원세포 분화 촉진제를 유효성분으로 함유하는 근육 질환의 예방 또는 치료용 약학 조성물. A pharmaceutical composition for the prevention or treatment of muscle diseases, comprising the source cell differentiation promoter of claim 1 as an active ingredient.
- 제2항에 있어서, 상기 근육 질환은 긴장감퇴증(atony), 근위축증(muscular atrophy), 근이영양증(muscular dystrophy), 근육 퇴화, 근경직증, 근위축성 축삭경화증, 근무력증, 악액질(cachexia) 및 근육감소증(sarcopenia) 중에서 선택된 어느 하나인 것을 특징으로 하는 근육 질환 예방 또는 치료용 약학 조성물.The method of claim 2, wherein the muscle disease is atony, muscular atrophy, muscular dystrophy, muscle degeneration, myopathy, muscular dystrophy, ataxia, cachexia, and sarcopenia sarcopenia) pharmaceutical composition for preventing or treating muscle diseases, characterized in that any one selected from.
- 제1항의 근원세포 분화 촉진제를 유효성분으로 함유하는 근육증가 촉진 또는 노인성 근 손실의 예방 또는 개선용 건강기능성식품 조성물. A health functional food composition for preventing or improving muscle growth or senile muscle loss, comprising the root cell differentiation promoter of claim 1 as an active ingredient.
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| KR100665315B1 (en) * | 2004-03-25 | 2007-01-09 | 한국원자력연구소 | New gallic acid-conjugated linoleic acid fatty ester, the preparation method thereof and composition containing the same |
| KR20090029735A (en) * | 2006-05-24 | 2009-03-23 | 디에스엠 아이피 어셋츠 비.브이. | Treating muscular disorders and improving muscular function |
| KR20170022427A (en) * | 2015-08-20 | 2017-03-02 | 숙명여자대학교산학협력단 | Pharmacentical compositions for Preventing or Treating Muscle Atrophy Diseases Comprising Bakuchiol |
| KR101842948B1 (en) * | 2016-10-13 | 2018-03-28 | 연세대학교 산학협력단 | Composition comprising Decanal or as active ingredients for Preventing or treating muscle disease |
| KR101923153B1 (en) * | 2018-08-02 | 2018-11-28 | 충남대학교산학협력단 | Composition for promoting differentiation of muscle cells containing gallic acid as effective component |
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| KR100665315B1 (en) * | 2004-03-25 | 2007-01-09 | 한국원자력연구소 | New gallic acid-conjugated linoleic acid fatty ester, the preparation method thereof and composition containing the same |
| KR20090029735A (en) * | 2006-05-24 | 2009-03-23 | 디에스엠 아이피 어셋츠 비.브이. | Treating muscular disorders and improving muscular function |
| KR20170022427A (en) * | 2015-08-20 | 2017-03-02 | 숙명여자대학교산학협력단 | Pharmacentical compositions for Preventing or Treating Muscle Atrophy Diseases Comprising Bakuchiol |
| KR101842948B1 (en) * | 2016-10-13 | 2018-03-28 | 연세대학교 산학협력단 | Composition comprising Decanal or as active ingredients for Preventing or treating muscle disease |
| KR101923153B1 (en) * | 2018-08-02 | 2018-11-28 | 충남대학교산학협력단 | Composition for promoting differentiation of muscle cells containing gallic acid as effective component |
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