KR102669920B1 - A composition for preventing or improving muscular disorders comprising extracts of undaria pinnatifida, pomelo, and wheat germ - Google Patents
A composition for preventing or improving muscular disorders comprising extracts of undaria pinnatifida, pomelo, and wheat germ Download PDFInfo
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- KR102669920B1 KR102669920B1 KR1020210022658A KR20210022658A KR102669920B1 KR 102669920 B1 KR102669920 B1 KR 102669920B1 KR 1020210022658 A KR1020210022658 A KR 1020210022658A KR 20210022658 A KR20210022658 A KR 20210022658A KR 102669920 B1 KR102669920 B1 KR 102669920B1
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- Prior art keywords
- pomelo
- seaweed
- wheat germ
- extract
- muscle
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- A—HUMAN NECESSITIES
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- 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
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- 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
- A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
- A23L17/60—Edible seaweed
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- 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
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- 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
- A23L7/00—Cereal-derived products; Malt products; Preparation or treatment thereof
- A23L7/10—Cereal-derived products
- A23L7/152—Cereal germ products
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A61K2236/30—Extraction of the material
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
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Abstract
본 발명은 미역, 포멜로 및 밀배아의 추출물을 포함하는 근육 질환의 예방 또는 개선용 조성물에 관한 것이다. 구체적으로 본 발명은 미역, 포멜로 또는 밀배아의 단일 추출물과 비교하여 3성분 복합 추출물이 월등하게 우수한 근육 질환의 개선 효과를 갖는 것을 특징으로 한다.The present invention relates to a composition for preventing or improving muscle disease comprising extracts of seaweed, pomelo, and wheat germ. Specifically, the present invention is characterized in that the three-component complex extract has a significantly better effect on improving muscle diseases compared to single extracts of seaweed, pomelo, or wheat germ.
Description
본 발명은 미역, 포멜로 및 밀배아의 추출물을 포함하는 근육 질환의 예방 또는 개선용 조성물에 관한 것이다. 구체적으로 본 발명은 미역, 포멜로 또는 밀배아의 단일 추출물과 비교하여 3성분 복합 추출물이 월등하게 우수한 근육 질환의 개선 효과를 갖는 것을 특징으로 한다.The present invention relates to a composition for preventing or improving muscle disease comprising extracts of seaweed, pomelo, and wheat germ. Specifically, the present invention is characterized in that the three-component complex extract has a significantly better effect on improving muscle diseases compared to single extracts of seaweed, pomelo, or wheat germ.
현대 과학기술 및 의학의 발달로 인간의 평균 수명이 연장됨에 따라 건강에 관심을 가지는 수요층이 점차 늘어나고 있다. 최근에는 젊은층에서 '근손실'이라는 용어가 새롭게 주목받으면서 근육에 대한 관심이 많아졌고, 이에 비례하여 관련 건강기능식품 및 의약품도 늘어나고 있다. 그 중에서도 특히 안전성에 대한 의구심이 없는 천연 물질을 이용한 조성물에 대한 연구가 활발하다. 그 예로 연잎추출물을 포함하는 식품조성물 (한국 등록특허 제10-2057226호), 굴을 이용한 기능성 발효물을 포함하는 근 기능 개선용 조성물 (한국 등록특허 제10- 2136886호) 등이 알려져 있다. As the average human lifespan is extended due to the development of modern science and technology and medicine, the number of consumers interested in health is gradually increasing. Recently, as the term 'muscle loss' has gained new attention among young people, interest in muscles has increased, and the number of related health functional foods and medicines is increasing in proportion. Among them, research is particularly active on compositions using natural substances with no doubts about their safety. Known examples include a food composition containing lotus leaf extract (Korean Patent No. 10-2057226) and a composition for improving muscle function containing functional fermentation using oysters (Korean Patent No. 10-2136886).
한편, 미역 (Undaria pinnatifida)은 일년생 해초이며 지방에 따라 차가 있으나 대체로 가을에서 경루 동안에 자라고 봄에서 초여름 동안에 유주자를 내어서 번식한다. 미역에는 칼슘의 함량이 많을 뿐 아니라 흡수율이 높아서 칼슘이 많이 요구되는 산모에게 좋고, 갑상선 호르몬의 주성분인 요오드의 함량도 높다. 또한, 혈압 강하작용을 하는 라미닌 (laminine)이라는 아미노산이 함유되어 있으며, 혈액 내의 콜레스테롤의 양을 감소시키는 효과도 알려져 있다. 이외에도, 섬유질 함량이 많아서 장운동을 촉진시킴으로써 변비 예방에도 좋고, 자극성 또한 적은 음식이다. 현재까지 보고된 미역의 생리 활성으로는 항혈액응고 활성과 항염증, 항산화, 항세균 작용 등이 있다. Meanwhile, seaweed (Undaria pinnatifida) is an annual seaweed that varies depending on the region, but generally grows from fall to mid-morning and reproduces by sending out zoospores during spring to early summer. Seaweed not only contains a lot of calcium, but also has a high absorption rate, making it good for pregnant women who require a lot of calcium. It also has a high content of iodine, the main component of thyroid hormones. In addition, it contains an amino acid called laminine, which has the effect of lowering blood pressure, and is also known to have the effect of reducing the amount of cholesterol in the blood. In addition, it is high in fiber, so it is good for preventing constipation by promoting intestinal movement, and is also a less irritating food. The physiological activities of seaweed reported to date include anti-blood coagulation activity, anti-inflammatory, antioxidant, and antibacterial activities.
포멜로 (pomelo)는 쌍떡잎식물 무환자나무목 운향과에 속하는 과수와 그 열매로 학명은 시트러스 막시마(Citrus maxima)이다. 원산지는 동남아시아와 말레이시아 군도로 추정되며 중국에서도 수천년동안 야생종으로 자라기도 하였다. 오늘날에는 미국, 중국 남부, 일본, 인도네시아, 타이티, 인도 남부, 이스라엘 등의 열대와 아열대의 해안지대에서 주로 재배되며, 최대 주산지는 미국과 중국이다. 서양에서는 과육의 알맹이가 장식용으로도 활용되며, 마멀레이드, 잼, 초콜릿, 고기요리의 조미료 등으로도 활용된다. 또한 원산지인 동남아시아에서는 주로 소금물을 뿌려 디저트로 먹거나 음료, 샐러드 등으로 활용하여 먹으며, 중국에서는 잎과 과일 껍질을 삶아 차처럼 마시기도 한다.Pomelo is a fruit tree and its fruit belonging to the Rutaceae family of dicotyledonous plants, and its scientific name is Citrus maxima. Its origin is believed to be Southeast Asia and the Malaysian archipelago, and it has also grown as a wild species in China for thousands of years. Today, it is mainly cultivated in tropical and subtropical coastal areas of the United States, southern China, Japan, Indonesia, Tahiti, southern India, and Israel, with the largest producing areas being the United States and China. In the West, the pulp is used for decoration and as a seasoning for marmalade, jam, chocolate, and meat dishes. Also, in Southeast Asia, where it originates, it is mainly sprinkled with salt water and eaten as a dessert or used as a drink or salad, and in China, the leaves and fruit peel are boiled and drunk like tea.
또한, 밀배아 (wheat germ)는 밀종자의 약 2%를 차지하고 있고, 제분시에 글루텐분도 혼입하고 있는 조배아로 분리된다. 배아는 단백질, 지방, 각종 효소, 비타민, 미네랄 등을 풍부히 포함하고 있고 특히 비타민 E의 급원으로 중요하다. 사료용에는 조배아가 그대로 쓰이지만 건강식품용에는 글루텐분을 분리하여 순도를 높인 배아를 효소의 실활과 건조를 위해 가열처리하여 사용한다다. 밀배아는 피부미용, 항산화 작용, 혈액순환에 도움이 된다고 알려져 있다. In addition, wheat germ accounts for about 2% of wheat seeds, and is separated into crude embryos that also contain gluten during milling. Embryos are rich in protein, fat, various enzymes, vitamins, and minerals, and are especially important as a source of vitamin E. Crude embryos are used as is for feed, but for health foods, embryos whose purity is increased by separating the gluten are used by heat treatment to deactivate enzymes and dry them. Wheat germ is known to be helpful for skin care, antioxidant activity, and blood circulation.
인간은 노화에 따라 근세포의 직경감소와 양적 질적 감소를 특징으로 하는 근량 및 근력의 감소 등이 나타난다. 노화에 따른 근손실의 발생원인은 명확하게 제시되고 있지 않으나 대표적으로 미토콘드리아 결핍과 단백질 합성의 변이, 근육위성세포의 능력상실 등이 제시되고 있다. 따라서 골격근의 미토콘드리아 기능을 유지 또는 향상시키고 단백질의 합성능력을 향상시키는 것은 근육의 기능을 증가시키는 가장 기본적인 방법으로 제시되고 있다.As humans age, there is a decrease in the diameter of muscle cells and a decrease in muscle mass and strength characterized by quantitative and qualitative decreases. The cause of muscle loss due to aging is not clearly presented, but representative examples include mitochondrial deficiency, mutations in protein synthesis, and loss of ability of muscle satellite cells. Therefore, maintaining or improving the mitochondrial function of skeletal muscle and improving protein synthesis ability are presented as the most basic ways to increase muscle function.
본 발명자들은 근기능을 개선하여 근육질환을 예방 또는 개선할 수 있는 방법을 개발하기 위하여 지속적인 연구를 거듭한 결과, 미역, 포멜로 및 밀배아의 복합 추출물을 사용하였을 때, 각각의 단일 추출물을 사용하는 것보다 월등하게 우수한 근 기능 증진 및 개선 효과가 있음을 발견하여 본 발명을 완성하였다.The present inventors have conducted continuous research to develop a method to prevent or improve muscle disease by improving muscle function, and as a result, when using complex extracts of seaweed, pomelo, and wheat germ, it is better to use each single extract. The present invention was completed by discovering that it had a significantly better effect of promoting and improving muscle function.
본 발명은 미역, 포멜로 및 밀배아의 추출물을 포함하는 근육 질환의 치료를 위한 조성물을 제공하고자 한다. 상기 조성물은 식품 조성물, 약학 조성물 사료용 조성물일 수 있다.The present invention seeks to provide a composition for the treatment of muscle diseases comprising extracts of seaweed, pomelo and wheat germ. The composition may be a food composition, pharmaceutical composition, or feed composition.
본 발명은 미역, 포멜로, 및 밀배아 추출물을 포함하는, 근육 질환의 예방 또는 개선용 식품 조성물에 관한 것이다. 상기 근육 질환은 근위축증 (muscular atrophy), 근이영양증(muscular dystrophy), 근육 퇴화, 근기능 저하, 근무력증, 악액질 (cachexia) 또는 근육 감소증 (sarcopenia)일 수 있다.The present invention relates to a food composition for preventing or improving muscle disease, comprising extracts of seaweed, pomelo, and wheat germ. The muscle disease may be muscular atrophy, muscular dystrophy, muscle degeneration, decreased muscle function, myasthenia gravis, cachexia, or sarcopenia.
구체적으로, 본 발명의 조성물은 MyoD, MyoG, Myf5, MHC (myosin heavy chain), sirt1 및 단백질로 이루어진 군에서 하나 이상의 발현 또는 합성을 증가시킬 수 있다.Specifically, the composition of the present invention can increase the expression or synthesis of one or more proteins from the group consisting of MyoD, MyoG, Myf5, MHC (myosin heavy chain), sirt1, and proteins.
또한, 본 발명의 추출물은 미역, 포멜로 및 밀배아 중 하나 이상의 추출물의 혼합물이거나, 미역, 포멜로 및 밀배아의 혼합물의 추출물일 수 있다.Additionally, the extract of the present invention may be a mixture of one or more extracts of seaweed, pomelo, and wheat germ, or may be an extract of a mixture of seaweed, pomelo, and wheat germ.
본 발명의 추출물은 물, C1 내지 C6의 알코올, 아세트산, 및 이들의 혼합 용매로 이루어지는 군으로부터 선택되는 용매로 추출된 것일 수 있으며, 이에 제한되지 않는다.The extract of the present invention may be extracted with a solvent selected from the group consisting of water, C1 to C6 alcohol, acetic acid, and mixed solvents thereof, but is not limited thereto.
본 발명의 추출물에서 미역, 포멜로, 및 밀배아는 동일한 중량비를 가질 수 있다.In the extract of the present invention, seaweed, pomelo, and wheat germ may have the same weight ratio.
본 발명은 미역, 포멜로, 및 밀배아 추출물을 포함하는, 근육 질환의 예방 또는 치료용 약학 조성물에 관한 것이다.The present invention relates to a pharmaceutical composition for preventing or treating muscle diseases, comprising extracts of seaweed, pomelo, and wheat germ.
또한, 본 발명은 미역, 포멜로, 및 밀배아 추출물을 포함하는, 근육 질환의 예방 또는 개선용 사료 조성물에 관한 것이다.Additionally, the present invention relates to a feed composition for preventing or improving muscle disease, comprising extracts of seaweed, pomelo, and wheat germ.
본 발명에 따른 미역, 포멜로 및 밀배아의 추출물은 근육세포의 형성, 성장, 분화를 촉진시키고, Sirt1의 발현을 증가시켜 근 기능에 관여하는 미토콘드리아의 활성을 증진시키고, 단백질 합성을 증진시킴으로써 근 기능 증진 및 개선에 우수한 효과를 갖는다. The extract of seaweed, pomelo, and wheat germ according to the present invention promotes the formation, growth, and differentiation of muscle cells, enhances the activity of mitochondria involved in muscle function by increasing the expression of Sirt1, and promotes muscle function by enhancing protein synthesis. It has excellent effects on promotion and improvement.
도 1은 각 추출물 처리시 MyoD 발현 촉진 효과를 관찰한 결과를 나타낸다.
도 2는 각 추출물 처리시 MyoG 발현 촉진 효과를 관찰한 결과를 나타낸다.
도 3은 각 추출물 처리시 Myf5 발현 촉진 효과를 관찰한 결과를 결과를 나타낸다.
도 4는 각 추출물 처리시 MHC 발현 촉진 효과를 관찰한 결과를 나타낸다.
도 5는 각 추출물 처리시 Sirt1 발현 촉진 효과를 관찰한 결과를 나타낸다.
도 6은 각 추출물 처리시 promycine 반응 단백질 합성 효과를 관찰한 결과를 나타낸다Figure 1 shows the results of observing the effect of promoting MyoD expression upon treatment with each extract.
Figure 2 shows the results of observing the effect of promoting MyoG expression upon treatment with each extract.
Figure 3 shows the results of observing the effect of promoting Myf5 expression upon treatment with each extract.
Figure 4 shows the results of observing the effect of promoting MHC expression upon treatment with each extract.
Figure 5 shows the results of observing the effect of promoting Sirt1 expression upon treatment with each extract.
Figure 6 shows the results of observing the effect of promycine response protein synthesis upon treatment of each extract.
이하, 첨부한 도면을 참조하여 본 발명이 속하는 기술 분야에서 통상의 지식을 가진 자가 용이하게 실시할 수 있도록 본원의 실시태양 및 실시예를 상세히 설명한다. 그러나 본원은 여러 가지 형태로 구현될 수 있으며 여기에서 설명하는 실시태양 및 실시예에 한정되지 않는다. Hereinafter, with reference to the attached drawings, embodiments and examples of the present invention will be described in detail so that those skilled in the art can easily implement the present invention. However, the present application may be implemented in various forms and is not limited to the embodiments and examples described herein.
본원 명세서 전체에서, 어떤 부분이 어떤 구성 요소를 "포함" 한다고 할 때, 이는 특별히 반대되는 기재가 없는 한 다른 구성 요소를 제외하는 것이 아니라 다른 구성 요소를 더 포함할 수 있는 것을 의미한다.Throughout the specification of the present application, when a part "includes" a certain component, this means that it may further include other components rather than excluding other components unless specifically stated to the contrary.
본 발명은 미역, 포멜로, 및 밀배아 추출물을 포함하는, 근육 질환의 예방 또는 개선용 식품 조성물에 관한 것이다.The present invention relates to a food composition for preventing or improving muscle disease, comprising extracts of seaweed, pomelo, and wheat germ.
본 발명에서 "근육 질환"은 근위축증 (muscular atrophy), 근이영양증(muscular dystrophy), 근육 퇴화, 근기능 저하, 근무력증, 악액질 (cachexia) 또는 근육 감소증 (sarcopenia)일 수 있으며, 이에 제한되지 않는다.In the present invention, “muscle disease” may include, but is not limited to, muscular atrophy, muscular dystrophy, muscle degeneration, decreased muscle function, myasthenia gravis, cachexia, or sarcopenia.
본 발명에서 "식품 조성물"은 건강기능식품, 유제품, 발효제품 또는 식품 첨가물일 수 있다. 상기 "건강기능식품"은 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 제조 및 가공한 식품을 의미하며, "기능성"은 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건 용도에 유용한 효과를 얻을 목적으로 섭취하는 것을 의미한다. In the present invention, “food composition” may be a health functional food, dairy product, fermented product, or food additive. The above “health functional food” refers to food manufactured and processed using raw materials or ingredients with functional properties useful to the human body, and “functionality” refers to health purposes such as regulating nutrients for the structure and function of the human body or physiological effects. It means taking it for the purpose of obtaining useful effects.
본 발명의 "치료"는, 본 발명의 조성물의 투여로 근육 질환의 진행이 지연 또는 멈춰서 증세를 호전시키거나, 근 기능이 강화되는 등의 근육 질환의 증세를 이롭게 변경하는 모든 행위를 의미한다.“Treatment” of the present invention refers to any action that beneficially changes the symptoms of a muscle disease, such as improving symptoms by delaying or stopping the progression of the muscle disease or strengthening muscle function, by administering the composition of the present invention.
본 발명의 "예방"은 본 발명에 따른 조성물의 투여에 의해 근육 질환의 발병을 억제 또는 지연시키는 모든 행위를 의미하고, "개선"은 본 발명의 추출물을 포함하는 조성물의 투여로 치료되는 상태와 관련된 파라미터, 예를 들면 증상의 정도를 적어도 감소시키는 모든 행위를 의미한다.“Prevention” in the present invention refers to any act of suppressing or delaying the onset of muscle disease by administration of the composition according to the present invention, and “improvement” refers to the condition treated by administration of the composition containing the extract of the present invention and It refers to any action that at least reduces the relevant parameters, such as the severity of symptoms.
본 발명의 "추출물"은 당업계에서 공지된 통상적인 추출용매를 이용하여 얻을 수 있다. 추출용매로는 극성 용매 또는 비극성 용매를 이용할 수 있다. 극성 용매로는 물, C1 내지 C6의 알코올 (예: 메탄올, 에탄올, 프로판올, 부탄올, 노말-프로판올, 이소-프로판올 및 노말-부탄올 등), 아세트산, 또는 상기 극성 용매들의 혼합물을 포함한다. 비극성 용매로는 아세톤, 아세토나이트릴, 에틸아세테이트, 메틸아세테이트, 부틸아세테이트, 플루오로알칸, 헥산, 에테르, 클로로포름, 디클로로메탄 또는 상기 비극성 용매들의 혼합물을 포함한다.The “extract” of the present invention can be obtained using a conventional extraction solvent known in the art. As the extraction solvent, a polar solvent or a non-polar solvent can be used. Polar solvents include water, C1 to C6 alcohols (e.g., methanol, ethanol, propanol, butanol, n-propanol, iso-propanol, and n-butanol, etc.), acetic acid, or mixtures of the above polar solvents. Non-polar solvents include acetone, acetonitrile, ethyl acetate, methyl acetate, butyl acetate, fluoroalkane, hexane, ether, chloroform, dichloromethane, or mixtures of the above non-polar solvents.
일 실시태양에서, 상기 추출물은 물, C1 내지 C6의 알코올, 아세트산, 및 이들의 혼합 용매로 이루어지는 군으로부터 선택되는 용매로 추출된 것일 수 있다. In one embodiment, the extract may be extracted with a solvent selected from the group consisting of water, C1 to C6 alcohol, acetic acid, and mixed solvents thereof.
본 발명의 추출물은 열수 추출, 냉침 추출, 환류 냉각 추출, 초음파 추출 또는 당업계에 알려진 통상적인 추출방법을 통해 추출한 것일 수 있다.The extract of the present invention may be extracted through hot water extraction, cold needle extraction, reflux cooling extraction, ultrasonic extraction, or conventional extraction methods known in the art.
본 발명에 사용된 용어 "추출물"은 당업계에서 조추출물 (crude extract)로 통용되는 것을 의미하지만, 광의적으로 추출물을 추가적으로 분획 (fractionation)한 분획물도 포함한다. 즉 추출물은 상술한 용매를 이용하여 얻은 것뿐만 아니라, 여기에 정제 과정을 추가적으로 적용하여 얻은 것을 포함한다. 예컨대, 상기 추출물을 일정한 분자량 컷-오프 값을 갖는 한외여과막을 통과시켜 얻은 분획, 다양한 크로마토그래프 (크기, 전하, 소수성 또는 친화성에 따른 분리를 위해 제작된 것)에 의한 분리 등, 추가적으로 실시된 다양한 정제 방법을 통해 얻어진 분획도 본 발명의 생약 추출물에 포함된다.The term “extract” used in the present invention refers to what is commonly used in the art as a crude extract, but in a broad sense also includes fractions obtained by additional fractionation of the extract. That is, the extract includes not only those obtained using the above-mentioned solvents, but also those obtained by additionally applying a purification process. For example, fractions obtained by passing the extract through an ultrafiltration membrane with a certain molecular weight cut-off value, separation by various chromatographs (designed for separation according to size, charge, hydrophobicity, or affinity), etc. Fractions obtained through purification methods are also included in the herbal extract of the present invention.
본 발명의 추출물은 미역, 포멜로 및 밀배아 중 하나 이상의 추출물의 혼합물이거나, 미역, 포멜로 및 밀배아의 혼합물의 추출물일 수 있다.The extract of the present invention may be a mixture of one or more extracts of seaweed, pomelo, and wheat germ, or may be an extract of a mixture of seaweed, pomelo, and wheat germ.
본 발명의 추출물에서, 미역, 포멜로, 및 밀배아는 동일한 중량비를 가질 수 있다.In the extract of the present invention, seaweed, pomelo, and wheat germ may have the same weight ratio.
또한, 본 발명은 미역, 포멜로, 및 밀배아 추출물을 포함하는, 근육 질환의 예방 또는 치료용 약학 조성물에 관한 것이다.Additionally, the present invention relates to a pharmaceutical composition for preventing or treating muscle diseases, comprising extracts of seaweed, pomelo, and wheat germ.
본 발명의 약학 조성물은 목적하는 방법에 따라 비경구투여 또는 경구 투여할 수 있으며, 투여량은 환자의 체중, 연령, 성별, 건강상태, 식이, 투여시간, 투여방법, 배설률 및 질환의 중증도 등에 따라 그 범위가 다양하다. 또한 상기 조성물의 치료적으로 유효한 양은 투여방법, 목적부위, 환자의 상태에 따라 달라질 수 있으며, 인체에 사용시 투여량은 안전성 및 효율성을 함께 고려하여 적정량으로 결정되어야 한다.The pharmaceutical composition of the present invention can be administered parenterally or orally according to the desired method, and the dosage depends on the patient's weight, age, gender, health status, diet, administration time, administration method, excretion rate, and severity of the disease. The range is diverse. In addition, the therapeutically effective amount of the composition may vary depending on the administration method, target area, and patient's condition, and when used in the human body, the dosage should be determined as an appropriate amount considering safety and efficiency.
또한, 본 발명은 미역, 포멜로, 및 밀배아 추출물을 포함하는, 근육 질환의 예방 또는 개선용 사료 조성물에 관한 것이다.Additionally, the present invention relates to a feed composition for preventing or improving muscle disease, comprising extracts of seaweed, pomelo, and wheat germ.
사료 조성물은, 모든 비-인간 동물, 예를들어 비-인간 영장류, 양, 개, 소, 말 등에게 섭취시킬 수 있다.The feed composition can be fed to all non-human animals, such as non-human primates, sheep, dogs, cattle, horses, etc.
이하 실시예를 통하여 본 발명을 더욱 상세하게 설명하고자 하나, 하기의 실시예는 단지 설명의 목적을 위한 것이며 본원 발명의 범위를 한정하고자 하는 것은 아니다.The present invention will be described in more detail through examples below. However, the examples below are for illustrative purposes only and are not intended to limit the scope of the present invention.
[실시예 1][Example 1]
미역, 포멜로, 밀배아의 단일 추출물, 및 복합 추출물의 제조Preparation of single and complex extracts of seaweed, pomelo, and wheat germ
미역 (국내산)은 잘게 분쇄한 후 60% 에탄올을 사용하여 상온에서 24시간 추출, 여과, 농축 후 동결건조하여 시료로 사용하였다. Seaweed (domestic) was finely ground, extracted at room temperature for 24 hours using 60% ethanol, filtered, concentrated, and then freeze-dried to be used as a sample.
밀배아 추출물의 제조를 위하여 밀배아를 60% 에탄올을 사용하여 상온 18시간 추출, 여과 및 농축을 거쳐 분무건조하여 시료로 사용하였다. To prepare wheat germ extract, wheat germ was extracted with 60% ethanol at room temperature for 18 hours, filtered, concentrated, and spray dried to be used as a sample.
포멜로 추출물의 제조를 위하여 60% 에탄올을 사용하여 상온 18시간 추출, 여과 및 농축을 거쳐 분무건조하여 시료로 사용하였다. 또한 미역, 포멜로, 밀배아의 복합추출물은 각각의 단일추출물을 동량의 비율로 혼합하여 사용하였다.To prepare the pomelo extract, it was extracted with 60% ethanol at room temperature for 18 hours, filtered, concentrated, and spray-dried to be used as a sample. In addition, complex extracts of seaweed, pomelo, and wheat germ were used by mixing the respective single extracts in equal amounts.
하기 표 1과 같이 본 발명의 추출물을 제조하여 실험에 사용하였다.The extract of the present invention was prepared as shown in Table 1 below and used in the experiment.
[실시예 2][Example 2]
세포 배양 및 독성 평가 (MTT assay)Cell culture and toxicity evaluation (MTT assay)
실험에 사용한 마우스 근아세포 (mouse myoblast cell line, C2C12 cell)를 ATCC사 (Manassas, VA, USA)는 10% fetal bovine serum media (Gibco-BRL)를 이용하여 37℃, 5% CO2 인큐베이터에서 배양하여 실험에 사용하였다. 세포가 70% confluent 상태가 되면 2% HS (horse serum media, GibcoBRL)를 이용하여 근관세포(myotube)로 분화시켰다. 실시예 1에서 제조된 단일 추출물 및 복합 추출물을 대상으로, C2C12 세포에 대한 독성을 평가하기 위하여 MTT assay를 수행하였다. 세포는 1×104 cells/well로 시딩(seeding)하여 24시간 부착하였고 시료를 처리하여 24시간 후 독성평가를 진행하였다. The mouse myoblast cell line (C2C12 cell) used in the experiment was cultured by ATCC (Manassas, VA, USA) in an incubator at 37°C and 5% CO 2 using 10% fetal bovine serum media (Gibco-BRL). and was used in the experiment. When the cells reached 70% confluent, they were differentiated into myotube cells using 2% HS (horse serum media, GibcoBRL). MTT assay was performed on the single extract and complex extract prepared in Example 1 to evaluate toxicity to C2C12 cells. Cells were seeded at 1×10 4 cells/well and adhered for 24 hours. Samples were processed and toxicity assessed 24 hours later.
시료를 1, 2.5, 5, 10 μg/mL 농도로 연속희석하여 처리한 결과, 포멜로를 제외한 모든 농도에서 80% 이상의 생존율을 나타내어 독성이 확인되지 않았다. 포멜로 시료는 10 μg/mL 농도에서 약 70%의 생존율을 나타내어 독성이 확인되었고, 이에 따라 하기 실험은 5 μg/mL 이하의 농도에서 진행되었다.As a result of processing the samples by serial dilution at concentrations of 1, 2.5, 5, and 10 μg/mL, the survival rate was over 80% at all concentrations except pomelo, and no toxicity was confirmed. The pomelo sample showed a survival rate of about 70% at a concentration of 10 μg/mL, confirming its toxicity. Accordingly, the following experiment was conducted at a concentration of 5 μg/mL or less.
[실시예 3][Example 3]
근모세포 (Myoblast)의 초기분화 촉진 효과 1Effect of promoting early differentiation of myoblasts 1
C2C12 세포의 분화초기 marker 중 하나인 MyoD의 발현량을 확인하기 위한 실험을 수행하였다. MyoD (myoblast determination protein 1; 근 아세포 결정 단백질 1)은 근육 분화를 조절하는데 중요한 역할을 하는 동물의 단백질에 해당한다.An experiment was performed to check the expression level of MyoD, one of the early differentiation markers of C2C12 cells. MyoD (myoblast determination protein 1) is an animal protein that plays an important role in regulating muscle differentiation.
C2C12 세포는 2×105 cells/well로 시딩(seeding)하여 48시간동안 100% confluent 할 때까지 배양하였다. 이후 2% HS와 1% PS가 첨가된 DMEM (분화배지)로 교체와 동시에 실시예 1의 추출물을 처리하여, 2일차에 세포를 수확하였다. 이때 세포는 3회 반복하여 풀링(pooling)하여 실험에 사용하였다. C2C12 cells were seeded at 2×10 5 cells/well and cultured for 48 hours until 100% confluent. Afterwards, it was replaced with DMEM (differentiation medium) supplemented with 2% HS and 1% PS, and simultaneously treated with the extract of Example 1, and the cells were harvested on the second day. At this time, the cells were repeated three times, pooled, and used in the experiment.
도 1에 나타낸 바와 같이, 미역, 포멜로 또는 밀배아 단일 추출물은 대조군 대비 1.4배 이하의 수치를 나타내고, 미역 및 포멜로의 복합 추출물 또한 대조군 대비 1.4배 이하의 수치를 나타내었다. 반면, 미역, 포멜로 및 밀배아의 복합 추출물은 대조군 대비 약 1.8배 이상의 월등하게 우수한 효과를 나타내었다.As shown in Figure 1, the single extract of seaweed, pomelo or wheat germ showed a value of 1.4 times or less compared to the control group, and the combined extract of seaweed and pomelo also showed a value of 1.4 times or less compared to the control group. On the other hand, the complex extract of seaweed, pomelo, and wheat germ showed a significantly more effective effect of about 1.8 times more than the control group.
결국, 미역, 포멜로 또는 밀배아 단일 추출물과, 미역 및 포멜로의 복합 추출물은 유사한 효과를 나타내었으나, 미역, 포멜로 및 밀배아의 복합 추출물은 상기 단일 추출물 및 2성분 복합 추출물과 비교하여 월등하게 우수한 효과를 나타내는 것을 알 수 있다. In the end, the single extract of seaweed, pomelo or wheat germ and the complex extract of seaweed and pomelo showed similar effects, but the complex extract of seaweed, pomelo and wheat germ had a significantly superior effect compared to the single extract and the two-component complex extract. It can be seen that it represents .
[실시예 4][Example 4]
근모세포 (Myoblast)의 초기분화 촉진 효과 2Effect of promoting early differentiation of myoblasts 2
상기 실시예 3과 동일한 방법으로, C2C12 세포의 분화초기 marker 중 하나인 MyoG의 발현량을 확인하기 위한 실험을 수행하였다. MyoG (Myogenin, 미오게닌)은 골격근 발달, 근육 발생 및 회복의 조정에 관여하는 근육-특이적 전사 인자이다.In the same manner as in Example 3, an experiment was performed to confirm the expression level of MyoG, one of the early differentiation markers of C2C12 cells. Myogenin (MyoG) is a muscle-specific transcription factor involved in the coordination of skeletal muscle development, myogenesis and repair.
도 2에 나타낸 바와 같이, 미역, 포멜로 또는 밀배아 단일 추출물은 대조군 대비 1.4배 이하의 수치를 나타내고, 미역 및 포멜로의 복합 추출물 또한 대조군 대비 1.4배 이하의 수치를 나타내었다. 반면, 미역, 포멜로 및 밀배아의 복합 추출물은 대조군 대비 약 1.7배 이상의 월등하게 우수한 효과를 나타내었다.As shown in Figure 2, the single extract of seaweed, pomelo or wheat germ showed a value of 1.4 times or less compared to the control group, and the combined extract of seaweed and pomelo also showed a value of 1.4 times or less compared to the control group. On the other hand, the complex extract of seaweed, pomelo, and wheat germ showed a significantly more effective effect of about 1.7 times more than the control group.
결국, 미역, 포멜로 또는 밀배아 단일 추출물과, 미역 및 포멜로의 복합 추출물은 유사한 효과를 나타내었으나, 미역, 포멜로 및 밀배아의 복합 추출물은 상기 단일 추출물 및 2성분 복합 추출물과 비교하여 월등하게 우수한 효과를 나타내는 것을 알 수 있다. In the end, the single extract of seaweed, pomelo or wheat germ and the complex extract of seaweed and pomelo showed similar effects, but the complex extract of seaweed, pomelo and wheat germ had a significantly superior effect compared to the single extract and the two-component complex extract. It can be seen that it represents .
[실시예 5][Example 5]
근모세포 (Myoblast)의 초기분화 촉진 효과 3Effect of promoting early differentiation of myoblasts 3
상기 실시예 3과 동일한 방법으로, C2C12 세포의 분화초기 marker 중 하나인 Myf5의 발현량을 확인하기 위한 실험을 수행하였다. Myf5 (Myogenic factor 5)는 근육 분화 또는 근육 생성, 특히 골격근의 발달을 조절하는 데 중요한 역할을 하는 단백질이다.In the same manner as Example 3, an experiment was performed to confirm the expression level of Myf5, one of the early differentiation markers of C2C12 cells. Myf5 (Myogenic factor 5) is a protein that plays an important role in regulating muscle differentiation or muscle creation, especially the development of skeletal muscle.
도 3에 나타낸 바와 같이, 미역, 포멜로 또는 밀배아 단일 추출물은 대조군과 유사한 수치를 나타내었다. 또한, 미역 및 포멜로의 복합 추출물은 단일 추출물과 유사한 수치를 나타내었다. 반면, 미역, 포멜로 및 밀배아의 복합 추출물은 대조군 대비 약 1.3배의 월등하게 우수한 효과를 나타내었다.As shown in Figure 3, single extracts of seaweed, pomelo or wheat germ showed similar values to the control group. Additionally, the composite extract of seaweed and pomelo showed similar values to the single extract. On the other hand, the complex extract of seaweed, pomelo, and wheat germ showed a significantly superior effect of about 1.3 times compared to the control group.
결국, 단일 추출물, 및 미역 및 포멜로의 복합 추출물은 대조군과 유사한 수치를 나타내었으나, 미역, 포멜로 및 밀배아의 3성분 복합 추출물은 월등하게 우수한 효과를 나타내는 것을 알 수 있다. In the end, the single extract and the complex extract of seaweed and pomelo showed similar values to the control group, but the three-component complex extract of seaweed, pomelo, and wheat germ showed a significantly superior effect.
[실시예 6][Example 6]
근관 성장 (Myotube hypertrophy)의 촉진 효과Promoting effect on root canal growth (Myotube hypertrophy)
C2C12 세포의 근관성장 촉진효과를 확인하기 위하여 2×10^5 cell/swell로 seeding하여 48시간 동안 100% confluent 할 때까지 배양하였다. 이후 2% hs와 1% ps가 첨가된 DMEM (분화배지)로 교체하여 4일동안 배양하였고 4일차에 시료를 처리하여 24시간 후 cell harvest 하였다. 이 때 세포는 3반복하여 pooling하여 실험에 사용하였으며 MHC (myosin heavy chain)의 발현량을 확인하기 위한 실험을 수행하였다.To confirm the myotube growth promoting effect of C2C12 cells, they were seeded at 2×10^5 cells/swell and cultured for 48 hours until 100% confluent. Afterwards, it was replaced with DMEM (differentiation medium) supplemented with 2% hs and 1% ps and cultured for 4 days. Samples were processed on the 4th day, and cells were harvested 24 hours later. At this time, the cells were pooled three times and used in the experiment, and an experiment was performed to confirm the expression level of MHC (myosin heavy chain).
도 4에 나타낸 바와 같이, 미역, 포멜로 또는 밀배아 단일 추출물은 대조군과 유사한 수치를 나타내었다. 또한, 미역 및 포멜로의 복합 추출물은 단일 추출물과 유사한 수치를 나타내었다. 반면, 미역, 포멜로 및 밀배아의 복합 추출물은 대조군 대비 약 1.3배의 월등하게 우수한 효과를 나타내었다.As shown in Figure 4, single extracts of seaweed, pomelo or wheat germ showed similar values to the control group. Additionally, the composite extract of seaweed and pomelo showed similar values to the single extract. On the other hand, the complex extract of seaweed, pomelo, and wheat germ showed a significantly superior effect of about 1.3 times compared to the control group.
결국, 단일 추출물, 및 미역 및 포멜로의 복합 추출물은 대조군과 유사한 수치를 나타내었으나, 미역, 포멜로 및 밀배아의 3성분 복합 추출물은 월등하게 우수한 효과를 나타내는 것을 알 수 있다. In the end, the single extract and the complex extract of seaweed and pomelo showed similar values to the control group, but the three-component complex extract of seaweed, pomelo, and wheat germ showed a significantly superior effect.
[실시예 7] [Example 7]
근관세포의 Sirt1 발현 실험Sirt1 expression experiment in myotube cells
상기 실시예 6과 동일한 실험방법으로 C2C12 세포 분화후기 marker 중 하나로 sirt1 발현을 측정하였다.Sirt1 expression was measured as one of the late differentiation markers of C2C12 cells using the same experimental method as in Example 6 above.
도 5에 나타낸 바와 같이, 미역 또는 포멜로 단일 추출물, 및 이의 복합 추출물은 대조군과 유사한 수치를 나타내고, 미역, 포멜로 및 밀배아의 복합 추출물은 대조군 대비 약 1.7배 이상의 월등하게 우수한 효과를 나타내었다.As shown in Figure 5, the single extract of seaweed or pomelo, and its complex extract showed similar values to the control group, and the complex extract of seaweed, pomelo, and wheat germ showed a significantly superior effect of about 1.7 times or more compared to the control group.
결국, 미역 또는 포멜로의 단일 추출물 및 이의 복합 추출물은 대조군과 비교하여 sirt1의 발현이 거의 증가하지 않으나, 미역, 포멜로 및 밀배아의 3성분 복합 추출물은 sirt1의 발현이 월등하게 증가하는 것을 알 수 있다. In the end, the single extract of seaweed or pomelo and its complex extract barely increased the expression of sirt1 compared to the control group, but the three-component complex extract of seaweed, pomelo, and wheat germ significantly increased the expression of sirt1. .
[실시예 8] [Example 8]
단백질 합성 실험 (SUnSET, surface sencing of translation assay)Protein synthesis experiment (SUnSET, surface sencing of translation assay)
단백질 합성률은 흔히 방사능 동위원소 추적자 (e.g., 3H-phenyalanine, 35S-methionine, 15N-lysine, 13C-leucine 등)를 사용하여 표지된 아미노산이 얼마나 많이 근육 단백질에 통합되어 단백질로 변화하는가를 수량화하는 방법을 통해 측정되어 왔다. 이러한 기술은 매우 성공적이었지만 비용과 시간이 많이 소요되며 일반적으로 단일 세포 수준에서 단백질 합성을 측정할 수 없다. 그러나 최근에는 배양 세포에서 단백질 합성을 측정하기 위해 화합물 puromycin을 사용하는 SUnSET assay인 비동위원소 기술이 개발되었다. Protein synthesis rate is often measured using radioactive tracers (e.g., 3H-phenyalanine, 35S-methionine, 15N-lysine, 13C-leucine, etc.) to quantify how much of the labeled amino acid is incorporated into muscle protein and converted into protein. It has been measured using this method. Although these techniques have been very successful, they are expensive, time-consuming, and generally cannot measure protein synthesis at the single-cell level. However, recently a non-isotopic technique has been developed, the SUnSET assay, which uses the compound puromycin to measure protein synthesis in cultured cells.
Puromycin은 박테리아 Streptomyces alboniger가 생산하는 aminonucleoside 항생제인데 aminoacyl-transfer RNA (aminoacyl-tRNA; 특히 tyrosyl-tRNA)의 구조적 유사체이므로 펩티드 결합의 형성을 통해 긴 펩티드 사슬에 포함될 수 있다. 따라서 전체 번역속도를 억제하지 않는 매우 낮은 농도에서 puromycin 표지 펩타이드가 형성되는 속도는 전체 단백질 합성속도를 반영하게 된다.Puromycin is an aminonucleoside antibiotic produced by the bacterium Streptomyces alboniger. It is a structural analog of aminoacyl-transfer RNA (aminoacyl-tRNA; especially tyrosyl-tRNA), so it can be incorporated into long peptide chains through the formation of peptide bonds. Therefore, at very low concentrations that do not inhibit the overall translation rate, the rate at which puromycin-labeled peptides are formed reflects the overall protein synthesis rate.
SUnSET assay를 통해 C2C12 마우스 유래 근아세포(myobloast)에서 단백질 합성률을 확인하기 위하여 6 well plate에 95%의 성장치 (confluence)까지 도달시킨 세포를 2% 말 혈청 (horse serum)이 함유된 DMEM 배지로 옮겨 4일 동안 배양하여 근관세포 (myotube)로 분화시켰다. 그 후 23시간 동안 각 단일추출물 및 복합추출물을 처리하고 추가적으로 1시간 동안 puromycin (1 mM)과 물질을 동시에 처리한 후, Western blot을 통해 puromycin을 측정하여 단백질 합성률을 확인하였다.To check the protein synthesis rate in myoblasts derived from C2C12 mice through the SUnSET assay, cells that reached 95% confluence in a 6-well plate were cultured in DMEM medium containing 2% horse serum. were transferred to and cultured for 4 days to differentiate into myotube cells. Afterwards, each single extract and composite extract were treated for 23 hours, puromycin (1 mM) and the substance were simultaneously treated for an additional 1 hour, and puromycin was measured through Western blot to confirm the protein synthesis rate.
도 6에 나타낸 바와 같이, 미역 포멜로 또는 밀배아 단일 추출물은 대조군 대비 유사하거나 더 낮은 수치를 나타내고, 미역, 포멜로 및 밀배아의 복합 추출물은 대조군 대비 약 1.5배의 월등하게 우수한 효과를 나타내었다. 결국, 미역, 포멜로 또는 미역의 단일 추출물은 대조군과 비교하여 단백질 합성이 거의 증가하지 않거나 오히려 감소하였으나, 미역, 포멜로 및 밀배아의 3성분 복합 추출물은 단백질 합성이 월등하게 증가하는 것을 알 수 있다. As shown in Figure 6, the single extract of seaweed pomelo or wheat germ showed similar or lower values compared to the control group, and the combined extract of seaweed, pomelo and wheat germ showed a significantly superior effect of about 1.5 times compared to the control group. In the end, the single extract of seaweed, pomelo, or seaweed showed little or even a decrease in protein synthesis compared to the control group, but the three-component complex extract of seaweed, pomelo, and wheat germ significantly increased protein synthesis.
Claims (14)
A food composition for preventing or improving muscle disease comprising extracts of seaweed, pomelo, and wheat germ, wherein the seaweed, pomelo, and wheat germ have the same weight ratio.
The method of claim 1, wherein the muscle disease is at least one selected from the group consisting of muscular atrophy, muscular dystrophy, muscle degeneration, decreased muscle function, myasthenia gravis, cachexia, and sarcopenia. A food composition for preventing or improving muscle disease.
The method of claim 1, wherein the composition increases the expression of one or more selected from the group consisting of MyoD, MyoG, Myf5, MHC (myosin heavy chain), and sirt1 or increases the amount of protein in muscle cells. Food composition for prevention or improvement.
The food composition for preventing or improving muscle disease according to claim 1, wherein the extract is a mixture of one or more extracts of seaweed, pomelo, and wheat germ, or an extract of a mixture of seaweed, pomelo, and wheat germ. .
The food composition for preventing or improving muscle disease according to claim 1, wherein the extract is extracted with a solvent selected from the group consisting of water, C1 to C6 alcohol, acetic acid, and mixed solvents thereof.
A pharmaceutical composition for preventing or treating muscle disease, comprising extracts of seaweed, pomelo, and wheat germ, wherein the seaweed, pomelo, and wheat germ have the same weight ratio.
The method of claim 7, wherein the muscle disease is one or more selected from the group consisting of muscular atrophy, muscular dystrophy, muscle degeneration, decreased muscle function, myasthenia gravis, cachexia, and sarcopenia. A pharmaceutical composition for preventing or treating muscle diseases.
The pharmaceutical composition for preventing or treating muscle diseases according to claim 7, wherein the extract is extracted with a solvent selected from the group consisting of water, C1 to C6 alcohol, acetic acid, and mixed solvents thereof.
A feed composition for preventing or improving muscle disease, comprising extracts of seaweed, pomelo, and wheat germ, wherein the seaweed, pomelo, and wheat germ have the same weight ratio.
The method of claim 11, wherein the muscle disease is one or more selected from the group consisting of muscular atrophy, muscular dystrophy, muscle degeneration, decreased muscle function, myasthenia gravis, cachexia, and sarcopenia. A feed composition for preventing or improving muscle disease.
The feed composition for preventing or improving muscle disease according to claim 11, wherein the extract is extracted with a solvent selected from the group consisting of water, C1 to C6 alcohol, acetic acid, and mixed solvents thereof.
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