KR20180081352A - Composition comprising Ocimene, Eugenol or as active ingredients for Preventing or treating muscle disease - Google Patents
Composition comprising Ocimene, Eugenol or as active ingredients for Preventing or treating muscle disease Download PDFInfo
- Publication number
- KR20180081352A KR20180081352A KR1020170002487A KR20170002487A KR20180081352A KR 20180081352 A KR20180081352 A KR 20180081352A KR 1020170002487 A KR1020170002487 A KR 1020170002487A KR 20170002487 A KR20170002487 A KR 20170002487A KR 20180081352 A KR20180081352 A KR 20180081352A
- Authority
- KR
- South Korea
- Prior art keywords
- muscle
- eugenol
- ocimene
- composition
- active ingredient
- Prior art date
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Abstract
Description
본 발명은 오시멘, 유게놀 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는 근육 질환 예방 또는 치료용 조성물에 관한 것이다.The present invention relates to a composition for preventing or treating muscle diseases comprising ocimen, eugenol or a pharmaceutically acceptable salt thereof as an active ingredient.
대한민국은 2000년 노인인구가 전체인구의 7.2%를 차지하여 고령화 사회에 진입하였으며, 2050년에는 초고령화사회(20% 이상)에 진입할 것으로 예측된다(2013년 고령자 통계, 통계청). 사람의 근육양은 나이가 들면서 감소하고(50~70세에 10~15% 정도, 그리고 70세~80세에서 30% 이상 감소), 이에 따라 근력과 근기능도 약화되는데, 이를 노인성 근감소증(sarcopenia)이라 한다. 노인성 근감소증은 활동장애와 보행장애를 유발하여 노인들의 독립적인 생활을 제한하는 주요 원인이 된다. 또한, 근감소증은 기초대사율을 저하시켜 인슐린 저항성을 높이고 2형 당뇨병 발생을 촉진하며, 고혈압 및 심혈관계 질환 발생위험을 3-5배 증가시킨다. 현재 근감소증 치료용도로 승인된 의약품은 전무한 실정이며, myostatin 억제물질 또는 기존 FDA 승인을 받은 타질환 치료제를 근감소증에 적용하는 약물재배치(drug repositioning) 기술이 개발 중에 있다.In Korea, the elderly population accounted for 7.2% of the total population in 2000 and entered the aging society. By 2050, it is expected to enter the aging society (more than 20%). The muscle mass of a person decreases with age (10-15% at 50-70 years of age, and 30% at 70-80 years of age), resulting in a weakening of muscle strength and myofunction. This is followed by sarcopenia, Quot; Elderly myopenia leads to dysfunctional and gait disturbances and is a major cause of restricting the independent life of the elderly. In addition, myopenia reduces basal metabolic rate, increases insulin resistance, promotes the development of type 2 diabetes, and increases the risk of hypertension and cardiovascular disease by 3-5 times. Currently, there are no approved drugs for the treatment of myopinia. Drug repositioning techniques are being developed to apply myostatin inhibitors or other existing FDA-approved drugs to myopenia.
근육은 크게 골격근(skeletal muscle), 심장근(cardiac muscle), 평활근(visceral muscle)으로 구분되고, 이 중 골격근은 인체에서 가장 많은 양으로 존재하는 조직으로, 체중의 40-45%를 차지한다. 골격근은 건(tendon)을 통해 뼈(bone)에 붙어서 뼈의 움직임 또는 힘을 만들어 내는 역할을 한다. 하나의 근육은 수많은 근섬유로 구성되어 있으며, 다시 근섬유는 액틴과 미오신으로 구성된 수많은 근원섬유로 만들어진다. 액틴과 미오신이 서로 겹쳐서 움직이면 근육의 길이가 짧아지거나 길어지면서 전체적인 근육의 수축과 이완을 유발하게 된다. 근원섬유 크기의 증가는 근섬유 두께의 증가를 의미하고, 그 결과 근육의 증가가 일어나게 된다.Muscles are divided into skeletal muscle, cardiac muscle, and visceral muscle, of which skeletal muscle is the most abundant in the human body, accounting for 40-45% of body weight. The skeletal muscle attaches to the bone through the tendon and acts to create movement or force of the bone. One muscle is made up of a number of muscle fibers, and the muscle fibers are made of numerous myofibers composed of actin and myosin. When actin and myosin overlap each other, muscle length is shortened or lengthened, causing overall muscle contraction and relaxation. An increase in myofibrillar size means an increase in muscle fiber thickness, resulting in an increase in muscle mass.
근육을 구성하는 근섬유의 유형은 ATP를 발생시키는 대사과정과 수축속도에 의해 주로 TypeⅠ, Type ⅡA 그리고 Type ⅡB로 구분된다. ‘Type Ⅰ 근섬유’는 수축속도가 느리고 많은 수의 미오글로빈과 미토콘드리아를 함유하고 있어 지속적이면서 낮은 강도의 유산소 활동을 하는데 적절하다. 타입Ⅰ 근섬유는 적색을 띄고 있어서 적색근이라고도 일컬어지며 대표적으로 가자미근(soleus)이 이에 속한다. 반면, ‘Type ⅡB 근섬유’는 수축속도가 빨라 매우 짧지만 높은 강도의 무산소 운동을 하는데 쓰이며, 미오글로빈의 함량이 적어 백색을 띄고 있다. ‘Type ⅡA 근섬유’는 앞서 언급한 두 가지 근섬유의 중간적인 특성을 띈다. 나이가 듦에 따라 근육의 부위별 Type Ⅰ, Ⅱ 근섬유의 조성이 달라질 뿐 아니라 모든 타입의 근섬유가 감소하게 된다.The types of muscle fibers constituting the muscles are mainly classified into Type I, Type IIA and Type IIB depending on the metabolic process that causes ATP and the rate of contraction. Type I muscle fiber is slow and has a large number of myoglobin and mitochondria and is suitable for sustained and low intensity aerobic activity. Type I muscle fibers are reddish, so they are also called red muscles, and the soleus is a typical example. On the other hand, 'Type IIB muscle fiber' has a very short contraction rate and is very short, but it is used for high intensity anaerobic exercise and has a low content of myoglobin and a white color. 'Type IIA muscle fiber' has the intermediate characteristics of the two muscle fibers mentioned above. As the age increases, not only does the composition of Type I and II muscle fibers vary, but also all types of muscle fibers decrease.
골격근은 환경에 따라 재생되어 유지되는 특징을 가지고 있으나, 이러한 특징은 나이가 듦에 따라 소실되고 결과적으로 노화가 진행되면서 근육양이 감소될 뿐 아니라 근력 역시 상실된다. 근육의 성장 및 재생에 관여하는 신호전달체계로는 insulin like growth factor 1(IGF-1)/AKT에 의해 매개되어 단백질 합성을 조절하는 신호전달이 있다. 근육세포막에 존재하는 IGF-1 receptor(IGF-1R)가 활성화되면 IRS1 및 PI3K 인산화를 통해 AKT 인산화가 증가되고 후자는 mTORC 인산화를 활성화시킨다. mTORC의 활성화는 ribosomal protein S6 kinase beta-1(p70S6K1)의 인산화를 증가시켜 mRNA 번역(translation)을 증가시키는 동시에 eukaryotic translation initiation factor 4 G(eIF4G)의 활성을 증가시키고, eukaryotic translation initiation factor 4E binding protein 1(4E-BP1) 단백질을 인산화시킨다. eIF4G와 4E-BP1은 eIF4F 복합체를 형성하는데 관여하는데 즉, eIF4G는 eIF4A 그리고 eIF4E와 결합하여 eIF4F 복합체를 형성하는 한편, 4E-BP1은 인산화되면 eIF4E와의 결합능이 저해되어 유리상태의 eIF4E를 증가시키게 된다. 후자는 다른 translation initiation factor들(eIF4G 및 eIF4A)와 결합하여 eIF4F 복합체를 형성하고, 이렇게 형성된 eIF4F 복합체는 리보솜 구조를 안정화시킴으로써 번역개시(translation initiation)를 촉진하여 궁극적으로 단백질 합성을 증가시키게 된다(Bodine et al., Akt/mTOR pathway is a crucial regulator of skeletal muscle hypertrophy and can prevent muscle atrophy in vivo. Nature cell biology, 3, 1014-1019, 2001).The skeletal muscle has characteristics that are regenerated and maintained according to the environment, but these characteristics disappear with aging, and as a result, as the aging progresses, the muscular volume decreases and the muscular strength is also lost. The signaling pathway involved in muscle growth and regeneration is signaling to regulate protein synthesis mediated by insulin like growth factor 1 (IGF-1) / AKT. Activation of IGF-1 receptor (IGF-1R) in muscle cell membranes increases AKT phosphorylation through IRS1 and PI3K phosphorylation and activates mTORC phosphorylation. Activation of mTORC increases phosphorylation of ribosomal protein S6 kinase beta-1 (p70S6K1), thereby increasing mRNA translation, increasing eukaryotic translation initiation factor 4 (eIF4G) activity, and inducing eukaryotic
또한 AKT 인산화는 glycogen synthase kinase 3 (GSK3)를 통해 eIF2B발현을 증가시켜 근섬유 성장을 촉진시키는 한편 단백질 분해 관련 전사인자인 forkhead box O(FOXO)의 발현을 억제함으로써 근손실을 억제하기도 한다. 근손실은 myostatin, transforming growth factor beta(TGF-β), 그리고 activin을 포함하는 TGF-β family의 receptor에 의해 매개되는 신호전달에 의해 조절된다. TGF-β type II receptor에 리간드가 결합하면 type I receptor를 인산화시키고, 후자는 smad 2/3 complex를 인산화시켜 결국 FOXO를 활성화시킨다. 후자는 muscle-specific ubiquitin-ligase인 muscle RING-finger protein-1(MURF1), Muscle Atrophy F-Box(MAFbx)/atrogin-1의 유전자 발현을 증가시키고, 이는 ubiquitin을 표적단백질의 lysine 부위에 부착시켜 단백질 분해를 촉진시키고, 결국 근육의 감소를 유도한다. (Gumucio et al., Atrogin-1, MuRF-1, and sarcopenia. Endocrine, 43, 12-21, 2013). In addition, AKT phosphorylation enhances eIF2B expression through glycogen synthase kinase 3 (GSK3), which promotes muscle fiber growth and suppresses muscle loss by suppressing expression of forkhead box O (FOXO), a proteolytic transcription factor. Muscle relaxation is regulated by signaling mediated by TGF-β family receptors including myostatin, transforming growth factor beta (TGF-β), and activin. The binding of the ligand to the TGF-β type II receptor phosphorylates the type I receptor, while the latter phosphorylates the smad 2/3 complex and ultimately activates FOXO. The latter increases the gene expression of muscle-specific ubiquitin-ligase, muscle RING-finger protein-1 (MURF1), Muscle Atrophy F-Box (MAFbx) / atrogin-1, which attaches ubiquitin to the lysine site of the target protein Promotes protein degradation, and ultimately leads to muscle loss. (Gumucio et al., Atrogin-1, MuRF-1, and sarcopenia, Endocrine, 43, 12-21, 2013).
따라서 본 발명자는 부작용이 적은 천연물에서 근육 단백질의 분해작용을 억제하고 합성을 촉진시킴으로써 근육의 증강 및 근손실 개선에 효과가 있는 식품 소재를 개발하는 연구를 진행하였다. Therefore, the present inventors have conducted research to develop a food material which is effective for enhancing muscles and muscle loss by inhibiting the degradation of muscle proteins and promoting their synthesis in natural products with low side effects.
본 발명은 오시멘(ocimene), 유게놀(eugenol) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는 근육 질환 예방 또는 치료용 약학적 조성물 등을 제공하고자 한다. The present invention is intended to provide a pharmaceutical composition for preventing or treating muscle diseases comprising ocimene, eugenol or a pharmaceutically acceptable salt thereof as an active ingredient.
그러나, 본 발명이 이루고자 하는 기술적 과제는 이상에서 언급한 과제에 제한되지 않으며, 언급되지 않은 또 다른 과제들은 아래의 기재로부터 당업자에게 명확하게 이해될 수 있을 것이다.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.
본 발명은 오시멘(ocimene), 유게놀(Eugenol) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는 근육 질환 예방 또는 치료용 약학적 조성물을 제공한다. The present invention provides a pharmaceutical composition for preventing or treating muscle diseases comprising ocimene, Eugenol or a pharmaceutically acceptable salt thereof as an active ingredient.
상기 조성물은 p-4E-BP1 또는 p-p70S6K1 단백질의 발현을 증가시킬 수 있다.The composition may increase the expression of p-4E-BP1 or p-p70S6K1 protein.
상기 조성물은 MuRF1(Muscle Ring-Finger Protein) 또는 MaFbx(Muscle atrophy F-box)의 발현을 감소시킬 수 있다.The composition may reduce the expression of MuRF1 (Muscle Ring-Finger Protein) or MaFbx (Muscle atrophy F-box).
상기 근육 질환은 근 기능 저하, 근육 감소, 근육 소모 또는 근육 퇴화로 인한 근육 질환일 수 있다.The muscle disorder may be a muscle disorder caused by muscle weakness, muscle weakness, muscle wasting or muscle regression.
상기 근육 질환은 긴장감퇴증(atony), 근위축증(muscular atrophy), 근이영양증(muscular dystrophy), 근무력증, 악액질(cachexia), 경직성 척추 증후군(rigid spinesyndrome), 근위축성 측삭경화증(루게릭병, amyotrophic lateral sclerosis), 경직성 척추 증후군(rigid spinsesyndrome), 샤르코-마리-투스병(Charcot-Marie-Tooth disease) 및 근육 감소증(sarcopenia)으로 이루어진 군으로부터 선택되는 어느 하나 이상일 수 있다.The muscular diseases include atony, muscular atrophy, muscular dystrophy, myasthenia gravis, cachexia, rigid spinesyndrome, amyotrophic lateral sclerosis, amyotrophic lateral sclerosis, , Rigid spinsesyndrome, Charcot-Marie-Tooth disease, and sarcopenia. ≪ RTI ID = 0.0 >
본 발명은 오시멘(ocimene), 유게놀(Eugenol) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는 근육 분화 촉진, 근육 재생 또는 근육 강화용 약학적 조성물을 제공한다.The present invention provides a pharmaceutical composition for promoting muscle differentiation, muscle regeneration or muscle strengthening comprising ocimene, eugenol or a pharmaceutically acceptable salt thereof as an active ingredient.
본 발명은 오시멘(ocimene), 유게놀(Eugenol) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는 근 기능 개선용 건강기능성 식품 조성물을 제공한다.The present invention provides a health functional food composition for improving muscle function comprising ocimene, Eugenol or a pharmaceutically acceptable salt thereof as an active ingredient.
본 발명은 오시멘(ocimene), 유게놀(Eugenol) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는 근육 분화 촉진, 근육 재생 또는 근육 강화용 건강기능성 식품 조성물을 제공한다.The present invention provides a health functional food composition for promoting muscle differentiation, muscle regeneration or muscle strengthening comprising ocimene, Eugenol or a pharmaceutically acceptable salt thereof as an active ingredient.
본 발명은 오시멘(ocimene), 유게놀(Eugenol) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는 근육 질환 예방 또는 치료용 가축 사료용 조성물을 제공한다.The present invention provides a composition for preventing or treating muscle diseases, which comprises ocimene, eugenol or a pharmaceutically acceptable salt thereof as an active ingredient.
본 발명은 오시멘(ocimene), 유게놀(Eugenol) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는 근육 분화 촉진, 근육 재생 또는 근육 강화용 가축 사료용 조성물을 제공한다.The present invention provides a composition for promoting muscle differentiation, muscle regeneration or muscle strengthening for animal feed comprising ocimene, Eugenol or a pharmaceutically acceptable salt thereof as an active ingredient.
본 발명은 오시멘(ocimene), 유게놀(Eugenol) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는 근 기능 개선용 화장료 조성물을 제공한다.The present invention provides a cosmetic composition for improving muscle function comprising ocimene, eugenol or a pharmaceutically acceptable salt thereof as an active ingredient.
본 발명은 오시멘(ocimene), 유게놀(Eugenol) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는 근육 질환 예방 또는 치료용, 또는 근 기능 개선용 조성물에 관한 것으로, 상기 오시멘 또는 유게놀은 근육 세포에서 근단백질 합성 및 근육량 증가와 관련된 단백질의 발현을 증가시킬 수 있고, 근 단백질 분해에 관여하는 효소의 발현은 mRNA 수준에서부터 억제할 수 있으므로 근기능 저하, 근육 소모 또는 근육 퇴화로 인한 근육 질환에 있어서 근육 분화, 근육 재생, 근육량 증가를 통해 근력 강화 효과를 나타낼 수 있으며, 근육 감소를 억제할 수 있는 바, 근육 질환 예방 또는 치료용, 근육 분화, 근육 재생 및 근육량 증가용 또는 근 기능 개선에 이용될 수 있다. The present invention relates to a composition for preventing or treating muscular diseases or improving muscular function comprising ocimene, Eugenol or a pharmaceutically acceptable salt thereof as an active ingredient, Nol can increase the expression of proteins involved in muscle protein synthesis and muscle mass increase in muscle cells and can inhibit the expression of enzymes involved in muscle protein degradation from the level of mRNA and thus inhibit muscular dysfunction caused by muscle weakness or muscle degeneration In diseases, muscle differentiation, muscle regeneration, and increase in muscle mass can be used to enhance muscle strength, and it is possible to inhibit muscle decline, for prevention or treatment of muscle diseases, for muscle differentiation, for muscle regeneration and for increasing muscle mass, Lt; / RTI >
도 1은 마우스 근아세포에서 myotube의 두께 변화를 나타낸 것이다. (A)는 giemsa-wright 염색한 근관세포를 현미경으로 촬영하여 시각화한 것이고, (B)는 근관세포의 직경을 측정한 결과이며 각 값들은 세 개의 독립된 well에서 세 번의 결정 값의 평균±표준오차이다. P <0.05는 통계적 유의성을 나타낸다
도 2는 오시멘을 처리한 마우스 근아세포에서 단백질 분해 및 합성 관련 분자들의 발현 변화를 나타낸 것이다. (A)는 p-4E-BP1, Total 4E-BP1, p-p70S6K1, 및 Total p70S6K1의 단백질 수준을 나타낸 것이고, (B)는 MaFbx 및 MuRF1의 유전자 발현 수준을 나타낸 것이다. 각 값들은 세 개의 독립된 well에서 세 번의 결정 값의 평균±표준오차이다. P <0.05는 통계적 유의성을 나타낸다.
도 3은 마우스 근아세포에서 myotube의 두께 변화를 나타낸 것이다. (A)는 giemsa-wright 염색한 근관세포를 현미경으로 촬영하여 시각화한 것이고, (B)는 근관세포의 직경을 측정한 결과이며 각 값들은 세 개의 독립된 well에서 세 번의 결정 값의 평균±표준오차이다. P <0.05는 통계적 유의성을 나타낸다
도 4는 유게놀을 처리한 마우스 근아세포에서 단백질 분해 및 합성 관련 분자들의 발현 변화를 나타낸 것이다. (A)는 p-4E-BP1, Total 4E-BP1, p-p70S6K1, 및 Total p70S6K1의 단백질 수준을 나타낸 것이고, (B)는 MaFbx 및 MuRF1의 유전자 발현 수준을 나타낸 것이다. 각 값들은 세 개의 독립된 well에서 세 번의 결정 값의 평균±표준오차이다. P <0.05는 통계적 유의성을 나타낸다.Fig. 1 shows changes in the thickness of myotubes in mouse myoblasts. (A) is the visualization of gyemsa-wright stained canaliculus by microscopic observation, (B) is the result of measuring the diameter of canaliculus cells, and each value is the mean of standard deviation of three determinations in three independent wells to be. P <0.05 indicates statistical significance
Fig. 2 shows changes in expression of molecules involved in proteolysis and synthesis in oocimen-treated mouse myoblasts. (A) shows the protein levels of p-4E-BP1, Total 4E-BP1, p-p70S6K1 and Total p70S6K1, and (B) shows the gene expression levels of MaFbx and MuRF1. Each value is the mean ± standard error of three determinations in three independent wells. P <0.05 indicates statistical significance.
3 shows the change in the thickness of myotube in mouse myoblasts. (A) is the visualization of gyemsa-wright stained canaliculus by microscopic observation, (B) is the result of measuring the diameter of canaliculus cells, and each value is the mean of standard deviation of three determinations in three independent wells to be. P <0.05 indicates statistical significance
Fig. 4 shows changes in expression of molecules involved in proteolytic and synthetic processes in mouse enucleated myoblasts treated with yugenol. (A) shows the protein levels of p-4E-BP1, Total 4E-BP1, p-p70S6K1 and Total p70S6K1, and (B) shows the gene expression levels of MaFbx and MuRF1. Each value is the mean ± standard error of three determinations in three independent wells. P <0.05 indicates statistical significance.
본 발명자들은 모노테르펜(monoterpene)계 화합물인 오시멘 또는 페닐프로파노이드(phenylpropanoid)계 화합물인 유게놀이 근육단백질의 분해작용을 억제하고, 합성을 촉진시킴으로써 근육 증강 및 근 손실 개선에 효과가 있음을 확인함으로써, 본 발명을 완성하였다. The present inventors have found that it is effective for inhibiting the degradation of ogenic or phenylpropanoid-based compound, a monoterpene-based compound, yuzunor muscle protein, and promoting the synthesis, thereby improving muscle build-up and muscle loss By confirming, the present invention has been completed.
본 발명에서 "근"은 심줄, 근육, 건을 포괄적으로 지칭하고, "근 기능"은 근육의 수축에 의해 힘을 발휘하는 능력을 의미하며, 근육이 저항을 이겨내기 위하여 최대한으로 수축력을 발휘할 수 있는 능력인 근력, 근육이 주어진 중량에 얼마나 오랫동안 또는 얼마나 여러 번 수축과 이완을 반복할 수 있는지를 나타내는 능력인 근지구력, 단시간 내에 강한 힘을 발휘하는 능력인 순발력을 포함한다. 이러한 근 기능은 간이 주관하며, 근육량에 비례하고, "근 기능 개선"은 근 기능을 더 좋게 향상시키는 것을 의미한다.In the present invention, "muscle" refers to the tendons, muscles, and tendons comprehensively, and "muscular function" refers to the ability to exert its force by contraction of muscles. In order to overcome resistance, Muscular endurance which is the ability to indicate how long or how many times the muscle can repeat contraction and relaxation on a given weight, and ability to exert a strong force in a short period of time. These muscle functions are hosted by the liver, proportional to muscle mass, and "muscle function improvement" means better improvement of muscle function.
이하, 본 발명을 상세히 설명한다. Hereinafter, the present invention will be described in detail.
근육 질환 예방 또는 치료용 약학적 조성물Pharmaceutical compositions for preventing or treating muscle disorders
본 발명은 오시멘(ocimene), 유게놀(eugenol) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는 근육 질환 예방 또는 치료용 약학적 조성물을 제공한다.The present invention provides a pharmaceutical composition for preventing or treating muscular diseases comprising ocimene, eugenol or a pharmaceutically acceptable salt thereof as an active ingredient.
구체적으로, 상기 오시멘은 무색 또는 담황색의 투명한 액체로서, 물에는 녹지 않고, 기름과 에탄올에 용해되며, 다양한 식물들의 꽃향기를 구성하는 성분으로 모노테르펜(monoterpene)계 화합물로서, 구조식은 C10H16, 분자량은 136.2 g/mol이다. 오시멘은 하기 화학식 1로 표시될 수 있으며, IUPAC name은 (3E)-3,7-dimethylocta-1,3,6-triene이고, (E)-beta-ocimene, trans-beta-ocimene, beta-ocimene, (3E)-3,7-dimethylocta-1,3,6-triene, beta-ocimene 등의 이명을 가진다. Specifically, the five cement is as a colorless, clear liquid or pale yellow, water is not melted, soluble in oil and ethanol, as monoterpenes (monoterpene) based compound as a component constituting the floral fragrance of many plants, the structural formula is C 10 H 16 , and the molecular weight is 136.2 g / mol. O-cymene can be represented by the following formula (I), IUPAC name is (3E) -3,7-dimethylocta-1,3,6-triene, and (E) -beta-ocimene, trans- ocimene, (3E) -3,7-dimethylocta-1,3,6-triene, and beta-ocimene.
[화학식 1][Chemical Formula 1]
또한, 상기 오시멘과 동일한 효능을 갖는 범위 내에서 오시멘 수화물, 오시멘 유도체 등을 포함할 수 있고, 이의 용매 화합물이나 입체 이성질체 또한 포함할 수 있다.In addition, it may contain oxigenic hydrate, ocimene derivative, etc. within the range having the same effect as that of ocimer, and may also include a solvent or a stereoisomer thereof.
상기 오시멘의 수득방법은 특별히 한정되지 않으며, 상기 오시멘을 함유하고 있는 식물로부터 분리하거나, 공지된 제법을 사용하여 화학적으로 합성하거나, 시판되는 것을 사용할 수 있다. 구체적으로, 상기 오시멘을 함유하고 있는 식물은 꿀풀과에 속하는 Ocimum basilicum L. , 귤과에 속하는 Evodia rutaecarpa의 과실, Agathosma Betulina , Pimenta dioica (올스파이스), Mentha arvensis piperascens (박하), Psidium guajava(구아바), Lavandula latifolia(스파이크라벤더), Annona squamosa(커스터드애플), Tagetes minuta(만수국아재비), Lantana camara(란타나), Lavandula x intermedia(라반딘), Laurus nobilis(월계수), Pimenta acris(베이), Perilla frutescens (차조기;소엽), Citrus x aurantium (비터오렌지), Ribes nigrum(블랙커런트), Camellia sinensis(차나무), Agastache foeniculum(아니스히솝), Alpinia galangal(고량강), Mentha arvensis(박하), Cinnamomum camphora(녹나무), Carum carvi(캐러웨이), Nepeta cataria(개박하, 캐트닢), Apium graveolens(샐러리), Andropogon nardus(시트로넬라), Angelica sinensis(당귀), Salvia sclarea(클라리 세이지), Hyacinthus orientalis(히아신스), Myrtus communis(은매화, 머틀), Thymus capitatus , Coriandrum sativum(고수), Origanum onites, Hesperis matronalis , Foeniculum vulgare(회향), Boswellia sacra(유향), Ferula gummosa , Angelica archangelica(안젤리카), Agastache urticifolia , Satureja thymbra , Origanum vulgare hirtum(오레가노), Hyssopus officinalis(히솝풀), Mentha longifolia , Tagetes filifolia , Melissa officinalis(레몬밤), Thymus x citriodorus(레몬타임), Aloysia citrodora(레몬버베나), Levisticum officinale(러비지), Petroselinum crispum(파슬리), Pastinaca sativa, Trifolium pretense, Rosmarinus officinalis(로즈마리), Salvia officinalis , Pinus silvestris(유럽소나무), Glycine max(콩), Lllicium verum , Ocimum basilicum(바질), Artemisia dracunculus(타라곤), Cinnamomum verum , Mentha aquatic 등을 포함하나, 이에 한정되지 않는다.The method of obtaining the ocimene is not particularly limited, and may be separated from a plant containing the ocimene, chemically synthesized using a known production method, or commercially available. Specifically, the plants containing the ocimene are Ocimum basilicum L. , Evodia belonging to the mandarin family rutaecarpa of negligence, Agathosma Betulina, Pimenta dioica (Allspice), Mentha arvensis piperascens (mint), Psidium guajava (guava) , Lavandula latifolia (spike lavender) , Annona squamosa (custard apple) , Tagetes minuta , Lantana camara (Lantana) , Lavandula x intermedia (Laban Dean) , Laurus nobilis (laurel) , Pimenta acris (Bay) , Perilla frutescens (Lobster , lobules) , Citrus x aurantium (Bitter orange) , Ribes nigrum (Black currant) , Camellia sinensis (Tea tree) , Agastache foeniculum (anise hyssop) , Alpinia galangal (Ko Yang) , Mentha arvensis (mint) , Cinnamomum camphora (Camphor) , Carum carvi (caraway) , Nepeta cataria (catnip, cat) , Apium graveolens (celery) , Andropogon nardus , Angelica sinensis , Salvia sclarea , Hyacinthus, orientalis (hyacinth) , Myrtus communis (silver plum, myrtle) , Thymus capitatus , Coriandrum sativum (coriander) , Origanum onites, Hesperis matronalis , Foeniculum vulgare (fennel), Boswellia sacra (frankincense), Ferula gummosa, Angelica archangelica (angelica), Agastache urticifolia, Satureja thymbra, Origanum vulgare hirtum (oregano) , Hyssopus officinalis (hyssop) , Mentha longifolia , Tagetes filifolia , Melissa officinalis (lemon balm) , Thymus x citriodorus (lemon-time), Aloysia citrodora (lemon verbena) , Levisticum officinale ( lavage ) , Petroselinum crispum (Parsley) , Pastinaca sativa, Trifolium pretense, Rosmarinus officinalis (rosemary) , Salvia officinalis , Pinus silvestris (European pine) , Glycine max (beans) , Lllicium verum , Ocimum basilicum (basil) , Artemisia dracunculus (tarragon) , cinnamomum verum , Mentha aquatic , and the like.
또한, 상기 유게놀은 옅은 노란색의 액체로 강한 향을 지니며, Syzygium aromaticum(Clove, 정향나무), Cinnamomum zeylanicum(Cinnamon, 계피), Ocimum gratissimum(Basil, 바질), Anethum graveolens(Dill, 서양자초), Pimpinella anisum(Anise, 아니스), Laurus nobilis(Bay laurel, 월계수), Apium graveolens(Celery, 셀러리), Melissa officinalis(Lemon balm, 멜리사), Pimenta racemosa(Bay) 등의 식물에 주로 함유되어 있는 성분으로 페닐프로파노이드(phenylpropanoid)계 화합물이고, 구조식은 C10H12O2, 분자량은 164.20 g/mol이다. 유게놀은 하기 화학식 2로 표시될 수 있으며, aldehyde C-10, caprinaldehyde, decyl aldehyde, eugenoldehyde 등의 이명을 가진다. In addition, the yugenol is a pale yellow liquid with a strong aroma, including Syzygium aromaticum (Clove), Cinnamomum zeylanicum (cinnamon, cinnamon), Ocimum gratissimum (Basil, basil), Anethum graveolens (Dill), Pimpinella anisum ( Anise , Anice), Laurus It is a phenylpropanoid compound mainly contained in plants such as nobilis (Bay laurel, laurel), Apium graveolens (Celery, celery) , Melissa officinalis (Lemon balm, Melissa) and Pimenta racemosa (Bay) , the structural formula is C 10 H 12 O 2, molecular weight 164.20 g / mol. Yugenols may be represented by the following general formula (2), and have aliphatics such as aldehyde C-10, caprinaldehyde, decyl aldehyde, and eugenoldehyde.
[화학식 2](2)
또한, 상기 유게놀과 동일한 효능을 갖는 범위 내에서 유게놀 수화물, 유게놀 유도체 등을 포함할 수 있고, 이의 용매 화합물이나 입체 이성질체 또한 포함할 수 있다.In addition, it may contain a eugenol hydrate, a yugenol derivative and the like within the range having the same efficacy as the yugenol, and may also include a solvent or a stereoisomer thereof.
상기 유게놀의 수득방법은 특별히 한정되지 않으며, 상기 유게놀을 함유하고 있는 식물로부터 분리하거나, 공지된 제법을 사용하여 화학적으로 합성하거나, 시판되는 것을 사용할 수 있다.The method for obtaining the eugenol is not particularly limited, and the eugenol may be isolated from a plant containing the eugenol, chemically synthesized using a known production method, or commercially available.
본 발명에 따른 조성물은 p-4E-BP1 또는 p-p70S6K1 단백질의 발현을 증가시킬 수 있다. The composition according to the present invention can increase the expression of p-4E-BP1 or p-p70S6K1 protein.
또한, 본 발명에 따른 조성물은 MuRF1(Muscle Ring-Finger Protein) 또는 MaFbx(Muscle atrophy F-box)의 발현을 감소시킬 수 있다. 구체적으로, 단백질 합성과 관련이 있는 대표적인 분자로는 p70S6K1, 4E-BP1, 그리고 eIF members가 있고, 이 세 가지 분자들은 상위의 mTORC에 의해 활성이 조절된다. mTORc의 활성화는 p70S6K1를 인산화시키고, 활성화된 p70S6K1은 40S 리보솜단백질(ribosomal protein) S6를 인산화시켜서 mRNA 번역(translation)을 증가시키게 된다. 또한 mTORC의 활성화는 eIF4G의 활성을 증가시키는 동시에 4E-BP1을 인산화시키는데, 이 두 분자는 eIF4F 복합체를 형성하는데 관여한다. 즉, eIF4G는 eIF4A 그리고 eIF4E와 결합하여 eIF4F 복합체를 형성하는 한편, 4E-BP1은 인산화되면 eIF4E와의 결합능이 저해되어 유리상태의 eIF4E를 증가시키게 된다. 후자는 다른 translation initiation factor들(eIF4G 및 eIF4A)과 결합하여 eIF4F 복합체를 형성하고, 이렇게 형성된 eIF4F 복합체는 리보솜 구조를 안정화시킴으로써 번역개시(translation initiation)를 촉진하여 궁극적으로 단백질 합성을 증가시키게 된다. MAFbx/Atrogin-1과 MuRF1은 muscle-specific ubiquitin-ligase로, ubiquitin을 표적단백질의 lysine 부위에 부착시켜 단백질 분해를 촉진시키고, 근육의 감소를 유도하는 대표적인 단백질로서, 본 발명의 조성물은 MuRF1(Muscle Ring-Finger Protein) 또는 MaFbx(Muscle atrophy F-box)의 발현을 감소시킴으로써, 근육의 감소를 저해할 수 있다. In addition, the composition according to the present invention can reduce the expression of MuRF1 (Muscle Ring-Finger Protein) or MaFbx (Muscle atrophy F-box). Specifically, p70S6K1, 4E-BP1, and eIF members are representative molecules involved in protein synthesis, and these three molecules are regulated by higher mTORCs. Activation of mTORc phosphorylates p70S6K1 and activated p70S6K1 phosphorylates the 40S ribosomal protein S6 to increase mRNA translation. Activation of mTORC also increases the activity of eIF4G and phosphorylates 4E-BP1, both of which are involved in the formation of the eIF4F complex. In other words, eIF4G binds eIF4A and eIF4E to form an eIF4F complex, while 4E-BP1 is phosphorylated, which inhibits binding to eIF4E and increases free eIF4E. The latter combines with other translation initiation factors (eIF4G and eIF4A) to form the eIF4F complex, which stabilizes the ribosome structure, thereby facilitating translation initiation and ultimately increasing protein synthesis. MAFbx / Atrogin-1 and MuRF1 are muscle-specific ubiquitin-ligase, which is a typical protein that attaches ubiquitin to the lysine site of the target protein to promote protein degradation and induce muscle loss. The composition of the present invention is MuRF1 Ring-Finger Protein) or MaFbx (Muscle atrophy F-box), thereby reducing muscle loss.
본 발명에서 상기 "근육 질환"은 근 기능 저하, 근육 감소, 근육 소모 또는 근육 퇴화로 인한 근육 질환으로 당업계에 보고된 질병인 것이 바람직하며, 구체적으로 상기 근육 질환은 긴장감퇴증(atony), 근위축증(muscular atrophy), 근이영양증(muscular dystrophy), 근무력증, 악액질(cachexia), 경직성 척추 증후군(rigid spinesyndrome), 근위축성 측삭경화증(루게릭병, amyotrophic lateral sclerosis), 경직성 척추 증후군(rigid spinsesyndrome), 샤르코-마리-투스병(Charcot-Marie-Tooth disease) 및 근육 감소증(sarcopenia)으로 이루어진 군으로부터 선택되는 어느 하나 이상인 것일 수 있으나, 이에 한정되지 않는다. 또한, 상기 근육 소모 또는 퇴화는 전적 요인, 후천적 요인, 노화 등을 원인으로 발생하며, 근육 소모는 근육량의 점진적 손실, 근육, 특히 골격근 또는 수의근 및 심장근육의 약화 및 퇴행을 특징으로 한다.In the present invention, the term " muscle disorder "is a disease reported in the art as a muscle disorder caused by muscle weakness, muscle loss, muscle wasting or muscle degeneration. Specifically, the muscle disorder is atony, The present invention relates to a method for the treatment of muscular atrophy, muscular dystrophy, myasthenia, cachexia, rigid spinesyndrome, amyotrophic lateral sclerosis, rigid spinsesyndrome, But is not limited to, at least one selected from the group consisting of Charcot-Marie-Tooth disease and sarcopenia. In addition, the muscle wasting or degeneration is caused by total factors, acquired factors, aging, etc., and muscle wasting is characterized by gradual loss of muscle mass, weakness and degeneration of muscles, particularly skeletal muscle or veterinary muscle and heart muscle.
상기 오시멘 또는 유게놀의 용량은 0.1μM 내지 1000μM의 농도로 포함되는 것이 바람직하나, 이에 한정되지 않는다. 이때, 오시멘 또는 유게놀이 상기 농도 범위 미만인 경우, 근육세포에서 단백질 합성 및 분해 활성이 저하되어, 근육 질환 예방 또는 치료 효과를 발휘하기 어려운 문제점이 있고, 오시멘 또는 유게놀이 상기 농도 범위를 초과하는 경우, 세포독성을 포함한 독성의 우려사항이 있을 수 있다.Preferably, the dose of oocimen or vesicle is in the range of 0.1 μM to 1000 μM, but is not limited thereto. At this time, when the concentration of oscimene or yuzenol is less than the above range, there is a problem that the protein synthesis and degradation activity is lowered in muscle cells, and it is difficult to exhibit the effect of preventing or treating muscle diseases. There may be concern about toxicity, including cytotoxicity.
본 발명에 따른 근육 질환 예방 또는 치료용 약학 조성물은 각각 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구제 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제형화되어 사용할 수 있고, 제형화를 위하여 약학 조성물의 제조에 통상적으로 사용되는 적절한 담체, 부형제 또는 희석제를 포함할 수 있다.The pharmaceutical composition for preventing or treating muscle diseases according to the present invention may be in the form of oral, granule, tablet, capsule, suspension, emulsion, syrup, aerosol or other oral preparation, external preparation, And may contain suitable carriers, excipients or diluents conventionally used in the manufacture of pharmaceutical compositions for formulation.
상기 담체 또는, 부형제 또는 희석제로는 락토즈, 덱스트로즈, 수크로오스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리게이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로즈, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유 등을 포함한 다양한 화합물 혹은 혼합물을 들 수 있다.The carrier or the excipient or diluent includes lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, Polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like.
제제화할 경우에는 보통 사용하는 충진제, 중량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 제조할 수 있다.In the case of formulation, a diluent or excipient such as a commonly used filler, a weight agent, a binder, a wetting agent, a disintegrant or a surfactant may be used.
경구 투여를 위한 고형제제는 상기 오시멘 또는 유게놀에 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘보네이트, 수크로스 또는 락토오스, 젤라틴 등을 섞어 제조할 수 있다. 또한 단순한 부형제 이외에 마그네슘 스테아레이트, 탈크 같은 윤활제들도 사용할 수 있다.The solid preparation for oral administration may be prepared by mixing at least one excipient such as starch, calcium carbonate, sucrose or lactose, gelatin and the like in osimen or eugenol. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used.
경구를 위한 액상 제제로는 현탁액, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용하는 단순 희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등을 포함할 수 있다.Examples of the liquid preparation for oral administration include suspensions, solutions, emulsions, syrups and the like. In addition to water and liquid paraffin which are commonly used diluents, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included .
비경구 투여를 위한 제제에는 멸균된 수용액, 비수용성제, 현탁제, 유제, 동결건조 제제, 좌제가 포함된다. 비수성용제, 현탁제로는 프로필렌글리콜, 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등을 사용할 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세롤젤라틴 등을 사용할 수 있다.Formulations for parenteral administration include sterile aqueous solutions, non-aqueous agents, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As a base for suppositories, witepsol, macrogol, tween 61, cacao paper, laurin, glycerol gelatin and the like can be used.
본 발명에 따른 근육 질환 예방 또는 치료용 약학 조성물의 바람직한 투여량은 환자의 상태, 체중, 질병의 정도, 약물형태, 투여경로 및 기간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다. 그러나, 바람직한 효과를 위해서는 1일 0.0001 내지 2,000 mg/kg으로, 바람직하게는 0.001 내지 2,000 mg/kg으로 투여할 수 있다. 투여는 하루에 한 번 투여할 수도 있고, 수회 나누어서 투여할 수도 있다. 다만, 상기 투여량에 의해서 본 발명의 범위를 한정하는 것은 아니다.The preferred dosage of the pharmaceutical composition for preventing or treating muscle disorders according to the present invention varies depending on the condition of the patient, the body weight, the degree of the disease, the drug form, the administration route and the period, but can be appropriately selected by those skilled in the art. However, for a desired effect, the dose may be 0.0001 to 2,000 mg / kg, preferably 0.001 to 2,000 mg / kg per day. The administration may be carried out once a day or divided into several doses. However, the scope of the present invention is not limited by the dosage.
본 발명에 따른 근육 질환 예방 또는 치료용 약학 조성물은 쥐, 생쥐, 가축, 인간 등의 포유 동물에 다양한 경로로 투여할 수 있다. 투여의 모든 방식은 예를 들면, 경구, 직장 또는 정맥, 근육, 피하, 자궁 내 경막 또는 뇌혈관내(intracerebroventricular) 주사에 의해서 투여할 수 있다.The pharmaceutical composition for preventing or treating muscle disorders according to the present invention can be administered to mammals such as rats, mice, livestock, and humans in various routes. All modes of administration can be administered, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine or intracerebroventricular injections.
근육 분화 촉진, 근육 재생 또는 근육 강화용 약학적 조성물A pharmaceutical composition for promoting muscle differentiation, muscle regeneration or muscle strengthening
오시멘(ocimene), 유게놀(eugenol) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는 근육 분화 촉진, 근육 재생 또는 근육 강화용 약학적 조성물을 제공한다. 상기 오시멘 또는 유게놀의 구체적인 내용은 전술한 바와 같다. There is provided a pharmaceutical composition for promoting muscle differentiation, muscle regeneration or muscle strengthening comprising ocimene, eugenol or a pharmaceutically acceptable salt thereof as an active ingredient. The specific contents of ocimene or eugenol are as described above.
근육의 성장은 섬유크기(fiber size)의 증가에 의해 및/또는 섬유수의 증가에 의해 일어날 수 있다. 상기 근육의 성장은 A) 습윤중량(wet weight)의 증가, B) 단백질 함량의 증가, C) 근섬유 수의 증가, D) 근섬유 직경의 증가에 의해 측정될 수 있다. 근섬유 성장의 증가는 직경을 단면 타원체의 단축으로 정의할 때 직경의 증가로 정의될 수 있다. 유용한 치료제는 이전에 유사하게 처리된 대조군 동물(즉, 근육 성장 화합물로 처리되지 않은 퇴행된 근육조직을 갖는 동물)에 비해 적어도 10% 정도 근육이 퇴행된 동물에 있어서 습윤증량, 단백질 함량 및/또는 직경을 10% 이상, 더욱 바람직하게 50% 이상, 및 가장 바람직하게 100% 이상 증가시키는 것이다. 근섬유의 수를 증가시킴으로써 성장을 증가시키는 화합물은 그것이 질병에 걸린 조직에서 근섬유의 수를 적어도 1%, 더욱 바람직하게 적어도 20%, 그리고 가장 바람직하게 적어도 50% 증가시킬 때 치료제로 유용하다. 이러한 백분율값은 화합물이 투여되어 국부적으로 작용하는 경우에 비처리되고 질병에 걸리지 않은 비교 포유동물에 있어서 또는 대측성인 병에 걸리지 않은 근육에 있어서의 기초수준에 대하여 상대적으로 결정된 것이다. Muscle growth can be caused by an increase in fiber size and / or an increase in fiber number. The muscle growth can be measured by A) an increase in wet weight, B) an increase in protein content, C) an increase in the number of myofibers, and D) an increase in myofiber diameter. The increase in muscle fiber growth can be defined as the increase in diameter when the diameter is defined as the short axis of the section ellipsoid. A useful therapeutic agent is one that is at least 10% more muscle-degenerated than the previously similarly treated control animal (i. E., An animal having degenerated muscle tissue not treated with a muscle growth compound) The diameter is increased by 10% or more, more preferably by 50% or more, and most preferably by 100% or more. Compounds that increase growth by increasing the number of muscle fibers are useful as therapeutic agents when it increases the number of muscle fibers in the diseased tissue by at least 1%, more preferably by at least 20%, and most preferably by at least 50%. These percent values are determined relative to baseline levels in untreated, non-diseased, comparable mammals, or in non-adulterated muscles, when the compound is administered and locally acting.
근육 재생은 근육 모세포로부터 새로운 근섬유가 형성되는 과정을 의미한다. 재생을 위한 유용한 치료제는 상술한 바와 같이 적어도 약 1%, 더욱 바람직하게 적어도 20%, 및 가장 바람직하게 적어도 50% 새로운 섬유(new fiber)의 수를 증가시킨다.Muscle regeneration is the process by which new muscle fibers are formed from myoblasts. Useful therapeutic agents for regeneration increase the number of new fibers by at least about 1%, more preferably at least 20%, and most preferably at least 50%, as described above.
근세포의 분화는 수축기관(미오피브릴)과 같은 근섬유의 성분들을 특정하는 근육 발생 프로그램(muscle developmental program)의 유도를 의미한다. 분화를 위한 유용한 치료제는 유사하게 처리된 대조군 동물에 있는 동등한 조직에 비하여, 질병에 걸린 조직에 있는 모든 근섬유 성분의 양을 약 10%이상, 더욱 바람직하게 50%이상, 및 가장 바람직하게 100%이상 증가시킨다.Differentiation of muscle cells implies induction of a muscle developmental program that specifies the components of muscle fibers, such as contractile organs (myopirbil). A useful therapeutic agent for differentiation is an amount of at least about 10%, more preferably at least 50%, and most preferably at least 100%, of the amount of all muscle fiber components in the diseased tissue, as compared to an equivalent tissue in a similarly treated control animal .
구체적으로, 본 발명의 일 구현예에 따르면, dexamethasone에 의해 감소한 마우스 근아세포에 오시멘 또는 유게놀을 처리한 경우, 상기 마우스 근아세포의 myotube가 유의적으로 증가하였음을 확인할 수 있다. 즉, 본 발명의 오시멘 또는 유게놀은 마우스 근아세포에서 myotube의 두께를 증가시킴으로써 근손실을 억제하고, 근육의 성장을 촉진시킬 수 있다. In particular, according to one embodiment of the present invention, when mycobacteria reduced by dexamethasone were treated with ocimene or eugenol, myotubes of the mouse myoblasts were significantly increased. That is, the ocimene or eugenol of the present invention can suppress muscle loss by increasing the thickness of myotube in mouse myoblasts, and can promote muscle growth.
또한, dexamethasone에 의해 감소한 마우스 근아세포에 오시멘 또는 유게놀을 처리한 경우, 단백질 합성에 관련이 있는 p-4E-BP1 및 p-p70S6K 단백질의 발현을 유의적으로 증가시킬 뿐만 아니라, 근육 감소를 유도하는 단백질인 MuRF1 및 Mafbx/atrogin1의 발현을 유의적으로 감소시킴을 확인할 수 있다. 즉, 본 발명의 오시멘 또는 유게놀은 마우스 근아세포에서 4E-BP1 및 p70S6K 단백질의 인산화를 증가시키고, MuRF1 및 Mafbx/atrogin1 유전자 발현을 억제함으로써 근육의 양을 증가시킬 수 있다. In addition, treatment with ocimene or eugenol in mouse myoblasts reduced by dexamethasone not only significantly increased the expression of p-4E-BP1 and p-p70S6K proteins involved in protein synthesis, Induced expression of MuRF1 and Mafbx / atrogin1, respectively. That is, the oocimen or eugenol of the present invention can increase the amount of muscle by increasing the phosphorylation of 4E-BP1 and p70S6K proteins in mouse muscle myoblasts and inhibiting MuRF1 and Mafbx / atrogin1 gene expression.
근 기능 개선용 건강기능성 식품 조성물Health functional food composition for improving muscle function
본 발명은 오시멘(ocimene), 유게놀(eugenol) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는 근 기능 개선용 건강기능성 식품 조성물을 제공한다. 상기 오시멘 또는 유게놀의 구체적인 내용은 전술한 바와 같다. The present invention provides a health functional food composition for improving muscle function comprising ocimene, eugenol or a pharmaceutically acceptable salt thereof as an active ingredient. The specific contents of ocimene or eugenol are as described above.
본 발명에 따른 근 기능 개선용 건강기능식품에 있어서, 상기 오시멘 또는 유게놀을 건강기능식품의 첨가물로 사용하는 경우 이를 그대로 첨가하거나 다른 식품 또는 식품성분과 함께 사용할 수 있고, 통상적인 방법에 따라 적절하게 사용할 수 있다. 유효 성분의 혼합양은 예방, 건강 또는 치료 등의 각 사용 목적에 따라 적합하게 결정할 수 있다.In the health functional food for improving muscle function according to the present invention, when oscimene or eugenol is used as an additive for health functional food, it may be added as it is or may be used together with other foods or food ingredients, It can be used properly. The amount of the active ingredient to be mixed may be suitably determined according to each use purpose such as prevention, health, or treatment.
건강기능식품의 제형은 산제, 과립제, 환, 정제, 캡슐제의 형태뿐만 아니라 일반 식품 또는 음료의 형태 어느 것이나 가능하다.Formulations of health functional foods may be in the form of powders, granules, pills, tablets, capsules, as well as in the form of ordinary foods or beverages.
상기 식품의 종류에는 특별히 제한은 없고, 상기 물질을 첨가할 수 있는 식품의 예로는 육류, 소세지, 빵, 쵸콜렛, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알콜 음료 및 비타민 복합제 등이 있으며, 통상적인 의미에서의 식품을 모두 포함할 수 있다.There is no particular limitation on the type of the food, and examples of the food to which the above substance can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, , Various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and may include foods in a conventional sense.
일반적으로, 식품 또는 음료의 제조시에 상기 오시멘 또는 유게놀은 원료 100 중량부에 대하여 15 중량부 이하, 바람직하게는 10 중량부 이하의 양으로 첨가할 수 있다. 그러나, 건강 및 위생을 목적으로 하거나 또는 건강 조절을 목적으로 하는 장기간의 섭취의 경우에는 상기 양은 상기 범위 이하일 수 있으며, 또한 본 발명은 천연물로부터의 분획물을 이용하는 점에서 안전성 면에서 문제가 없으므로 상기 범위 이상의 양으로도 사용할 수 있다.Generally, the ocimene or the eugenol may be added in an amount of not more than 15 parts by weight, preferably not more than 10 parts by weight, based on 100 parts by weight of the raw material. However, in the case of long-term consumption intended for health and hygiene purposes or for health control purposes, the amount may be less than the above range. Further, since the present invention uses fractions from natural products, there is no problem in terms of safety, Or more.
본 발명에 따른 건강기능식품 중 음료는 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로 함유할 수 있다. 상술한 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드, 말토스, 슈크로스와 같은 디사카라이드 및 덱스트린, 사이클로덱스트린과 같은 폴리사카라이드, 자일리톨, 소르비톨, 에리트리톨 등의 당알콜일 수 있다. 감미제로서는 타우마틴, 스테비아 추출물과 같은 천연 감미제나, 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명에 따른 음료 100 mL당 약 0.01 ~ 0.04 g, 바람직하게는 약 0.02 ~ 0.03 g일 수 있다.The beverage in the health functional food according to the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. The above-mentioned natural carbohydrates may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as 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 may be about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 mL of the beverage according to the present invention.
상기 외에 본 발명에 따른 근 기능 개선용 건강기능식품은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산음료에 사용되는 탄산화제를 함유할 수 있다. 그 밖에 본 발명의 수면 개선용 조성물은 천연 과일쥬스, 과일쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 혼합하여 사용할 수 있다. 이러한 첨가제의 비율은 제한되지 않으나 본 발명의 건강기능식품 100 중량부 대비 0.01 ~ 0.1 중량부의 범위에서 선택되는 것이 일반적이다.In addition to the above, the functional food for improving muscle function according to the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickening agents, , Preservatives, glycerin, alcohols, and carbonated beverages. In addition, the composition for improving sleep of the present invention may contain flesh for the production of natural fruit juice, fruit juice drink and vegetable drink. These components may be used independently or in combination. The ratio of such additives is not limited, but is generally selected in the range of 0.01 to 0.1 parts by weight based on 100 parts by weight of the health functional food of the present invention.
근육 분화 촉진, 근육 재생 또는 근육 강화용 건강기능성 식품 조성물Health functional food composition for promoting muscle differentiation, muscle regeneration or muscle strengthening
오시멘(ocimene), 유게놀(eugenol) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는 근육 분화 촉진, 근육 재생 또는 근육 강화용 건강기능성 식품 조성물을 제공한다. 상기 오시멘 또는 유게놀의 구체적인 내용은 전술한 바와 같다.There is provided a health functional food composition for promoting muscle differentiation, muscle regeneration or muscle strengthening comprising ocimene, eugenol or a pharmaceutically acceptable salt thereof as an active ingredient. The specific contents of ocimene or eugenol are as described above.
근육 질환 예방 또는 치료용 가축 사료용 조성물Composition for livestock feed for prevention or treatment of muscle diseases
오시멘(ocimene), 유게놀(eugenol) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는 근육 질환 예방 또는 치료용 가축 사료용 조성물을 제공한다. 상기 오시멘 또는 유게놀의 구체적인 내용은 전술한 바와 같다.There is provided a composition for animal feed for preventing or treating muscle diseases comprising ocimene, eugenol or a pharmaceutically acceptable salt thereof as an active ingredient. The specific contents of ocimene or eugenol are as described above.
상기 가축은 소, 돼지, 닭, 오리, 염소, 양 및 말로 이루어진 군 중에서 선택된 1종의 가축인 것이 바람직하나, 이에 한정되지 않는다. The livestock is preferably one species selected from the group consisting of cattle, pigs, chickens, ducks, goats, sheep and horses, but is not limited thereto.
상기 사료용 조성물은 사료 첨가제를 포함할 수 있다. 본 발명의 사료첨가제는 사료관리법상의 보조사료에 해당한다.The feed composition may include a feed additive. The feed additive of the present invention corresponds to an auxiliary feed in the feed control method.
본 발명에서 용어, "사료"는 동물이 먹고, 섭취하며, 소화시키기 위한 또는 이에 적당한 임의의 천연 또는 인공 규정식, 한끼식 등 또는 상기 한끼식의 성분을 의미할 수 있다.The term "feed" as used herein in the context of the present invention may mean any natural or artificial diet, single meal, or the like ingredients for feeding, ingesting, digesting or suitable for the animal.
상기 사료의 종류는 특별히 제한되지 아니하며, 당해 기술 분야에서 통상적으로 사용되는 사료를 사용할 수 있다. 상기 사료의 비제한적인 예로는, 곡물류, 근과류, 식품 가공 부산물류, 조류, 섬유질류, 제약 부산물류, 유지류, 전분류, 박류 또는 곡물 부산물류 등과 같은 식물성 사료; 단백질류, 무기물류, 유지류, 광물성류, 유지류, 단세포 단백질류, 동물성 플랑크톤류 또는 음식물 등과 같은 동물성 사료를 들 수 있다. 이들은 단독으로 사용되거나 2 종 이상을 혼합하여 사용될 수 있다.The kind of the feed is not particularly limited, and feeds conventionally used in the art can be used. Non-limiting examples of such feeds include vegetable feeds such as cereals, muscle roots, food processing busines logistics, algae, fibers, pharmaceutical buses, oils, fats, pastes or grain by-products; Animal feeds such as proteins, inorganic substances, fats, oils, fats, oils, monocellular proteins, animal plankton or foods. These may be used alone or in combination of two or more.
또한 상기 사료첨가제는 추가적으로 단위동물에 허용되는 담체를 함유할 수 있다. 본 발명에 있어서는 상기 사료첨가제를 그대로 또는 공지의 담체, 안정제 등을 가할 수 있으며, 필요에 따라 비타민, 아미노산류, 미네랄 등의 각종 양분, 항산화제 및 기타의 첨가제 등을 가할 수도 있으며, 그 형상으로서는 분체, 과립, 펠릿, 현탁액 등의 적당한 상태일 수 있다. 본 발명의 사료첨가제를 공급하는 경우는 단위동물에 대하여 단독으로 또는 사료에 혼합하여 공급할 수 있다.The feed additive may additionally contain a carrier that is acceptable to the unit animal. In the present invention, the feed additive may be added as it is or a known carrier, stabilizer and the like may be added. Various nutrients such as vitamins, amino acids and minerals, antioxidants and other additives may be added as needed, Powders, granules, pellets, suspensions, and the like. When the feed additive of the present invention is supplied, it can be supplied to the unit animal singly or mixed with the feed.
근육 분화 촉진, 근육 재생 또는 근육 강화용 가축 사료용 조성물Composition for livestock feed for promoting muscle differentiation, muscle regeneration or muscle strengthening
오시멘(ocimene), 유게놀(eugenol) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는 근육 분화 촉진, 근육 재생 또는 근육 강화용 가축 사료용 조성물을 제공한다. 상기 오시멘 또는 유게놀의 구체적인 내용은 전술한 바와 같다. There is provided a composition for promoting muscle differentiation, muscle regeneration, or muscle strengthening for livestock feed comprising ocimene, eugenol or a pharmaceutically acceptable salt thereof as an active ingredient. The specific contents of ocimene or eugenol are as described above.
근 기능 개선용 For improving muscular function 화장료Cosmetics 조성물 Composition
본 발명은 오시멘(ocimene), 유게놀(eugenol) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는 근 기능 개선용 화장료 조성물을 제공한다. 상기 오시멘 또는 유게놀의 구체적인 내용은 전술한 바와 같다. The present invention provides a cosmetic composition for improving muscle function comprising ocimene, eugenol or a pharmaceutically acceptable salt thereof as an active ingredient. The specific contents of ocimene or eugenol are as described above.
본 발명의 화장료 조성물은 오시멘 또는 유게놀을 유효성분으로 함유하며 피부학적으로 허용 가능한 부형제와 함께 기초 화장품 조성물(화장수, 크림, 에센스, 클렌징 폼 및 클렌징 워터와 같은 세안제, 팩, 보디오일), 색조 화장품 조성물(화운데이션, 립스틱, 마스카라, 메이크업 베이스), 두발 제품 조성물(샴푸, 린스, 헤어컨디셔너, 헤어젤) 및 비누 등의 형태로 제조될 수 있다.The cosmetic composition of the present invention contains ocimene or eugenol as an active ingredient and contains basic cosmetic composition (cleanser, pack, body oil, such as lotion, cream, essence, cleansing foam and cleansing water) together with dermatologically acceptable excipients, (Shampoos, rinse, hair conditioner, hair gel) and soap, in the form of a color cosmetic composition (foundation, lipstick, mascara, makeup base)
상기 부형제로는 이에 한정되지는 않으나 예를 들어, 피부연화제, 피부 침투 증강제, 착색제, 방향제, 유화제, 농화제 및 용매를 포함할 수 있다. 또한, 향료, 색소, 살균제, 산화방지제, 방부제 및 보습제 등을 추가로 포함할 수 있으며, 물성개선을 목적으로 점증제, 무기염류, 합성 고분자 물질 등을 포함할 수 있다. 예를 들면, 본 발명의 화장료 조성물로 세안제 및 비누를 제조하는 경우에는 통상의 세안제 및 비누 베이스에 상기 오시멘 또는 유게놀을 첨가하여 용이하게 제조할 수 있다. 크림을 제조하는 경우에는 일반적인 수중유적형(O/W)의 크림베이스에 오시멘, 유게놀 또는 이의 염을 첨가하여 제조할 수 있다. 여기에 향료, 킬레이트제, 색소, 산화방지제, 방부제 등과 물성개선을 목적으로 한 단백질, 미네랄, 비타민 등 합성 또는 천연소재를 추가로 첨가할 수 있다. 본 발명의 화장료 조성물에 함유되는 오시멘 또는 유게놀의 함량은 이에 한정되지 않지만 전체 조성물 총중량에 대하여 0.001 내지 10 중량%인 것이 바람직하고, 0.01 내지 5중량%인 것이 더욱 바람직하다. 상기 함량이 0.001중량% 미만에서는 목적하는 항노화 또는 주름개선 효과를 기대할 수 없고, 10중량% 초과에서는 안전성 또는 제형상의 제조에 어려움이 있을 수 있다.Such excipients include, but are not limited to, emollients, skin penetration enhancers, colorants, perfumes, emulsifiers, thickeners and solvents. In addition, it may further contain flavors, pigments, bactericides, antioxidants, preservatives, moisturizers and the like, and may include thickeners, inorganic salts and synthetic polymeric substances for the purpose of improving physical properties. For example, when the cleanser and the soap are prepared with the cosmetic composition of the present invention, the ocimene or the eugenol can be easily added to the cleanser and the soap base. When a cream is prepared, it can be prepared by adding ocimene, eugenol or a salt thereof to a cream base of a typical underwater type (O / W). A synthetic or natural material such as a flavor, a chelating agent, a coloring matter, an antioxidant, an antiseptic, and a protein, a mineral, and a vitamin for the purpose of improving a physical property may be further added. The content of ocimer or eugenol contained in the cosmetic composition of the present invention is not limited thereto, but is preferably 0.001 to 10% by weight, more preferably 0.01 to 5% by weight, based on the total weight of the whole composition. If the content is less than 0.001% by weight, the desired anti-aging or wrinkle-reducing effect can not be expected. If the content is more than 10% by weight, safety or formability may be difficult.
상기한 바와 같이, 본 발명의 오시멘, 유게놀 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는 조성물은 근아세포에서 4E-BP1 및 p70S6K1 단백질 인산화를 증가시키고, MuRF1 및 MaFbx/atrogin1 유전자 발현을 억제함으로써 근기능 저하, 근육 소모 또는 근육 퇴화로 인한 근육 질환에 있어서 근육 분화, 근육 재생, 근육량 증가를 통해 근력 강화 효과를 나타낼 수 있으며, 근육 감소를 억제할 수 있는 바, 근육 질환 예방 또는 치료용, 근육 분화, 근육 재생 및 근육량 증가용 또는 근 기능 개선에 이용될 수 있다. As described above, the composition comprising ocimen, eugenol or a pharmaceutically acceptable salt thereof of the present invention as an active ingredient increases the phosphorylation of 4E-BP1 and p70S6K1 proteins in myobacterium, and the expression of MuRF1 and MaFbx / atrogin1 gene , Muscle fatigue, muscular exhaustion, or muscle degeneration can be inhibited to improve muscular strength, muscle regeneration, muscle mass increase, and muscle tone reduction, thereby preventing or treating muscle disorders , Muscle differentiation, muscle regeneration and muscle mass increase, or muscle function improvement.
이하, 본 발명의 이해를 돕기 위하여 바람직한 실시예를 제시한다. 그러나 하기의 실시예는 본 발명을 보다 쉽게 이해하기 위하여 제공되는 것일 뿐, 하기 실시예에 의해 본 발명의 내용이 한정되는 것은 아니다.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 following examples.
[[ 실시예Example ]]
준비예Preparation Example . 세포배양. Cell culture
마우스 근아세포(mouse myoblast cell line, C2C12 cell)를 ATCC사(Manassas, VA, USA)로부터 구매하였고, 구입한 세포를 10% fetal bovine serum media (Gibco-BRL)를 이용하여 37℃, 5% CO2 인큐베이터에서 배양하였다. 상기 배양된 세포가 80% confluent해지면 2% horse serum media (Gibco-BRL)를 이용하여 myotube로 분화시켰다.The mouse myoblast cell line (C2C12 cell) was purchased from ATCC (Manassas, Va., USA). The cells were cultured in 10% fetal bovine serum (Gibco-BRL) 2 incubator. When the cultured cells became 80% confluent, they were differentiated into myotubes using 2% horse serum media (Gibco-BRL).
실시예Example 1. One.
분화 4일 째 되는 날부터 이틀 간 dexamethasone (dexa; Sigma) 50 μM 및 오시멘 (ocimene; CAS Number 13877-91-3, Sigma) 100 μM을 함께 처리하였다. From the fourth day of differentiation, 50 μM dexamethasone (dexa; Sigma) and 100 μM ocimene (CAS Number 13877-91-3, Sigma) were treated together for two days.
비교예Comparative Example 1. One.
상기 실시예 1과 동일한 방법으로 오시멘 (ocimene; CAS Number 13877-91-3, Sigma) 100 μM을 처리하였다. 100 μM of ocimene (CAS Number 13877-91-3, Sigma) was treated in the same manner as in Example 1 above.
실시예Example 2. 2.
유게놀(eugenol; CAS Number 97-53-0, Sigma) 100 μM을 사용하였다는 점을 제외하고는 실시예 1과 동일한 방법으로 수행하였다. Was performed in the same manner as in Example 1, except that 100 μM of eugenol (CAS Number 97-53-0, Sigma) was used.
비교예Comparative Example 2. 2.
유게놀(eugenol; CAS Number 97-53-0, Sigma) 100 μM을 사용하였다는 점을 제외하고는 비교예 1과 동일한 방법으로 수행하였다. Was performed in the same manner as in Comparative Example 1, except that 100 μM of eugenol (CAS Number 97-53-0, Sigma) was used.
실험예Experimental Example 1. 마우스 1. Mouse 근아세포를Myoblasts 이용한 Used 오시멘의Ocimene 근손실억제Muscle loss suppression 효능 efficacy
(1) (One) GiemsaGiemsa -wright staining-wright staining
상기 실시예 1에 따른 myotube를 Phosphate buffered saline(PBS)으로 2회 세척한 후 100% methanol으로 10분동안 고정하였다. 고정이 완료되면 상온에서 10분간 자연 건조시킨 후 myotube를 특이적으로 염색시키는 Giemsa-wright staining solution(아산제약, 서울)을 떨어뜨려 30분간 염색하였다.Myotube according to Example 1 was washed twice with phosphate buffered saline (PBS) and fixed with 100% methanol for 10 minutes. After completion of the fixation, the plate was allowed to stand for 10 minutes at room temperature, and Giemsa-wright staining solution (Asan Pharm, Seoul) was stained for 30 minutes.
(2) (2) MyotubeMyotube 두께 측정 Thickness measurement
상기 실험예 1에서 염색된 myotube를 형광현미경(IX 71, Olympus)을 이용하여 X20 배율로 촬영 후 image J software(USA)를 이용하여 분석하였다. 각 well에서 6 부분을 무작위로 선택하여 현미경 촬영하였으며, 각 well로부터 최소 100개의 myotube 두께를 분석하였다(3회 반복/Group).The myotubes stained in Experimental Example 1 were photographed at an X20 magnification using a fluorescence microscope (IX 71, Olympus) and analyzed using image J software (USA). Six sections were randomly selected in each well and examined microscopically. At least 100 myotube thicknesses were analyzed from each well (3 replicates / Group).
도 1은 마우스 근아세포에서 myotube의 두께 변화를 나타낸 그래프이다. 1 is a graph showing changes in the thickness of myotubes in mouse myoblasts.
도 1A에 나타난 바와 같이, 비교예 1의 경우, 정상세포(Basal)에 비해 myotube의 두께가 현저히 감소하였으며, 오시멘을 처리한 실시예 1의 경우, dexamethasone에 의해 감소한 myotube 두께를 다시 증가시키는 것을 시각적으로 확인할 수 있다. 또한, 도 1B에 나타난 바와 같이, 이를 image J software를 이용하여 수치화한 결과에서도 오시멘을 처리한 실시예 1의 경우, dexamethasone에 의해 감소한 myotube 두께를 36% 유의적으로 증가시킴을 확인할 수 있다. 즉, 오시멘은 마우스 근아세포에서 myotube의 두께를 증가시켜 근손실을 억제하고, 근성장을 촉진시키는 것을 알 수 있다.As shown in FIG. 1A, in Comparative Example 1, the thickness of myotubes was significantly reduced compared to that of normal cells. In Example 1 treated with oscimene, dexamethasone reduced myotube thickness again It can be confirmed visually. In addition, as shown in FIG. 1B, it was confirmed by numerical analysis using image J software that the oocimen treated Example 1 significantly increased myotube thickness reduced by dexamethasone by 36%. In other words, it can be seen that ocimer increases the thickness of myotube in mouse myoblasts, thereby suppressing muscle loss and promoting muscle growth.
실험예Experimental Example 2. 작용기작 규명 2. Identification of mechanism
(1) (One) TrizolTrizol method를 이용한 RNA 분리 및 RT- RNA isolation and RT- PCRPCR (reverse transcription-polymerase chain reaction) (reverse transcription-polymerase chain reaction)
마우스 근아세포 1*107 cells 당 Trizol 용액 334 ㎕을 첨가하여 갈아준 후, 4℃, 12,000 × g에서 10분간 원심분리 하였다. 상층액을 새 튜브로 옮긴 후 chloroform 67 ㎕을 첨가하고, vortex하였다. 다시 상층액을 새 튜브로 옮기고 상층액과 isopropanol의 비율이 1:1이 되도록 isopropanol을 첨가하였다. 10회 세게 흔든 다음 실온에서 15분 동안 방치하고, 12,000 × g, 4℃에서 10분간 원심분리 시킨 후 상층액을 제거하고, 남은 침전물에 70% ethanol 1 ml을 가한 후 7,500 × g, 4℃에서 5분 동안 원심분리 하였다. 에탄올을 제거한 후 RNA 침전물이 담긴 튜브를 실온에서 15분 동안 건조시키고, nuclease free water를 사용하여 RNA pellet을 용해시켰다. UV/VIS spectrophotometer(Beckman coulter, DU730)를 이용하여 260 nm 및 280 nm 파장에서 추출된 RNA 시료의 농도를 측정하고, agarose gel electrophoresis를 실시하여 RNA 시료의 integrity를 확인하였다.334 μl of Trizol solution was added per 1 × 10 7 cells of mouse myoblast, and the mixture was centrifuged at 12,000 × g for 10 minutes at 4 ° C. The supernatant was transferred to a new tube, and 67 μl of chloroform was added and vortexed. The supernatant was again transferred to a new tube and isopropanol was added so that the ratio of the supernatant and isopropanol was 1: 1. The mixture was centrifuged at 12,000 × g for 10 minutes at 4 ° C., and the supernatant was removed. To the remaining precipitate was added 1 ml of 70% ethanol and the mixture was centrifuged at 7,500 × g at 4 ° C. And centrifuged for 5 minutes. After removing the ethanol, the tube containing the RNA precipitate was dried at room temperature for 15 minutes, and the RNA pellet was dissolved using nuclease free water. The concentration of RNA extracted at 260 nm and 280 nm wavelength was measured using a UV / VIS spectrophotometer (Beckman coulter, DU730) and agarose gel electrophoresis was performed to confirm the integrity of the RNA sample.
마우스 근아세포에서 추출된 RNA시료를 대상으로 oligo dT primer와 superscript reverse transcriptase (GIBCO BRL, Gaithersburg, MD, USA)을 이용하여 reverse transcription을 수행함으로써 cDNA를 합성하였다. Reverse transcription을 통해 얻은 cDNA를 template로 하고 증폭하고자 하는 유전자 cDNA의 5‘과 3’ flanking sequence를 primer로 사용하여 PCR을 수행하였으며, 이때 사용된 primer sequence는 하기 표 1에 제시된 바와 같다. 증폭된 PCR 산물 1 ㎕를 1% agarose gel에 전기 영동하여 DNA band를 확인하였다. CDNA was synthesized by reverse transcription using oligo dT primer and superscript reverse transcriptase (GIBCO BRL, Gaithersburg, MD, USA) for RNA samples from mouse myoblast. PCR was carried out using 5 'and 3' flanking sequences of the cDNA to be amplified as a template, using the cDNA obtained from the reverse transcription as a template. The primer sequences used at this time are shown in Table 1 below. 1 μl of the amplified PCR product was electrophoresed on 1% agarose gel to confirm the DNA band.
temperature
(℃)Annealing
온도
(° C)
product
(bp)PCR
product
(bp)
(synonym: atrogin-1)MaFbx
(synonym: atrogin-1)
(synonym: TRAM63)MuRF1
(synonym: TRAM63)
(2) Western blotting(2) Western blotting
세포에서 western blotting을 수행하기 위해 media를 제거한 각 well에 500 μL의 100 mM Tris-HCl, pH 7.4, 5 mM EDTA, 50 mM sodium pyrophosphate, 50 mM NaF, 100 mM orthovanadate, 1% Triton X-100, 1 mM phenylmethanesulfonyl fluoride, 2 ㎍/mL aprotinin, 1 ㎍/mL pepstatin A, and 1 ㎍/mL leupeptin을 포함하는 lysis buffer를 넣고 harvest한 후 1,300 × g, 4℃에서 20분간 원심분리한 후 가운데 층을 취하고 Bradford 법에 따라 단백질을 정량 하였다(Bio-Rad). 단백질 40 ㎍을 SDS polyacrylamide gel에 전기영동시킨 후 nitrocellulose membranes (Amersham, Buckinghamshire, UK)으로 이동시켰다. Membrane을 tris-buffered saline과 tween 20 용액(TBS-T)을 이용하여 10분 동안 3회 반복하여 세척한 후 10% skim milk를 이용하여 60분간 차단하였다. Membrane을 1:1,000의 비율로 희석한 1차 항체에 넣어 4°C에서 부드럽게 흔들어 12시간 동안 배양한 후 TBS-T를 이용하여 세척하였고, membrane을 다시 1:2,000의 비율로 희석한 2차 항체와 함께 60분 동안 배양하고 세척하였다. 이 때, 1차 항체는 p70S6K1, phopho-p70S6K1(p-p70S6K1), 4E-BP1, phospho-4E-BP1(p-4E-BP1) 그리고 GAPDH (Cell Signaling Technology, Beverly, MA, USA)를 사용하였다. 최종적으로 단백질을 X-ray 필름에 ECL Western blot detection kit (RPN2106, Amersham, Arlington Heights, IL, USA)를 사용하여 시각화하였다. X-ray 필름에 시각화된 밴드를 스캔 후 Quantity One analysis software (Bio-Rad)를 이용하여 정량화하였다.To perform western blotting, 500 μL of 100 mM Tris-HCl, pH 7.4, 5 mM EDTA, 50 mM sodium pyrophosphate, 50 mM NaF, 100 mM orthovanadate, 1% Triton X-100, Lysis buffer containing 1 mM phenylmethanesulfonyl fluoride, 2 μg / mL aprotinin, 1 μg / mL pepstatin A, and 1 μg / mL leupeptin was harvested and centrifuged at 1,300 × g at 4 ° C. for 20 min. Proteins were quantified according to the Bradford method (Bio-Rad). 40 ㎍ of protein was electrophoresed on SDS polyacrylamide gel and transferred to nitrocellulose membranes (Amersham, Buckinghamshire, UK). The membranes were washed three times for 10 minutes using tris-buffered saline and
도 2는 오시멘을 처리한 마우스 근아세포에서 단백질 분해 및 합성 관련 분자들의 발현 변화를 나타낸 그래프이다. FIG. 2 is a graph showing changes in the expression of molecules involved in proteolysis and synthesis in oocimen-treated mouse myoblasts.
도 2에 나타난 바와 같이, 비교예 1의 경우, 정상세포(Basal)에 비해 단백질 합성과 관련이 있는 p-4E-BP1 및 p-p70S6K1 단백질 양이 유의적으로 감소하였고(도 2A), 단백질 분해 유전자인 MaFbx/atrogin1과 MuRF1 발현은 유의적으로 증가함을 확인할 수 있다(도 2B). 오시멘을 처리한 실시예 1의 경우, dexamethasone에 의해 감소한 p-4E-BP1 및 p-p70S6K 단백질 양을 다시 유의적으로 증가시키고(도 2A), MuRF1 및 MaFbx/atrogin1의 발현은 유의하게 감소시킴을 확인할 수 있다(도 2B). 즉, 오시멘은 마우스 근아세포에서 4E-BP1 및 p70S6K1 단백질 인산화를 증가시키고, MuRF1 및 MaFbx/atrogin1 유전자 발현을 억제함으로써 궁극적으로 근육의 양을 증가시키는데 관여하였을 것으로 사료된다.As shown in FIG. 2, in Comparative Example 1, the amount of p-4E-BP1 and p-p70S6K1 protein, which are involved in protein synthesis, was significantly decreased compared to that of normal cells (FIG. 2A) The expression of the genes MaFbx / atroginl and MuRF1 was significantly increased (Fig. 2B). In Example 1 treated with ocimene, the amount of p-4E-BP1 and p-p70S6K proteins reduced by dexamethasone was again significantly increased (Fig. 2A), and the expression of MuRF1 and MaFbx / atrogin1 was significantly decreased (Fig. 2B). In other words, ocimer may be involved in increasing the amount of 4E-BP1 and p70S6K1 protein phosphorylation in mouse myoblasts, ultimately increasing the amount of muscle by inhibiting MuRF1 and MaFbx / atrogin1 gene expression.
실험예Experimental Example 3. 마우스 3. Mouse 근아세포를Myoblasts 이용한 Used 유게놀의Eugenol 근손실억제Muscle loss suppression 효능 efficacy
(1) (One) GiemsaGiemsa -wright staining-wright staining
실시예 2에 따른 myotube를 사용하였다는 점을 제외하고는 실험예 3-(1)과 동일한 방법으로 수행하였다.The same procedure as in Experimental Example 3- (1) was carried out except that myotube according to Example 2 was used.
(2) (2) MyotubeMyotube 두께 측정 Thickness measurement
실시예 2에 따른 myotube를 사용하였다는 점을 제외하고는 실험예 3-(2)와 동일한 방법으로 수행하였다.The same procedure as in Example 3- (2) was carried out except that myotube according to Example 2 was used.
도 3은 마우스 근아세포에서 myotube의 두께 변화를 나타낸 그래프이다. 3 is a graph showing changes in the thickness of myotube in mouse myoblasts.
도 3A에 나타난 바와 같이, 비교예 2의 경우, 정상세포(Basal)에 비해 myotube의 두께가 현저히 감소하였으며, 유게놀을 처리한 실시예 2의 경우, dexamethasone에 의해 감소한 myotube 두께를 다시 증가시키는 것을 시각적으로 확인할 수 있다. 또한, 도 3B에 나타난 바와 같이, 이를 image J software를 이용하여 수치화한 결과에서도 유게놀을 처리한 실시예 1의 경우, dexamethasone에 의해 감소한 myotube 두께를 28% 유의적으로 증가시킴을 확인할 수 있다. 즉, 유게놀은 마우스 근아세포에서 myotube의 두께를 증가시켜 근손실을 억제하고, 근성장을 촉진시키는 것을 알 수 있다.As shown in FIG. 3A, in Comparative Example 2, the thickness of myotubes was significantly decreased compared with that of normal cells. In Example 2, which was treated with eugenol, deoxamethasone reduced myotube thickness again It can be confirmed visually. In addition, as shown in FIG. 3B, it was confirmed by numerical analysis using image J software that Example 1 in which yugenol was treated significantly increased myotube thickness decreased by dexamethasone by 28%. In other words, it can be seen that yugenol increases myotube thickness in mouse myoblasts, thereby suppressing muscle loss and promoting muscle growth.
실험예Experimental Example 2. 작용기작 규명 2. Identification of mechanism
(1) (One) TrizolTrizol method를 이용한 RNA 분리 및 RT- RNA isolation and RT- PCRPCR (reverse transcription-polymerase chain reaction) (reverse transcription-polymerase chain reaction)
하기 표 2에 제시된 primer suquence를 사용하였다는 점을 제외하고는 실험예 2-(1)과 동일한 방법으로 수행하였다. (1) except that the primer suquence shown in Table 2 below was used.
temperature
(℃)Annealing
온도
(° C)
product
(bp)PCR
product
(bp)
(synonym: atrogin-1)MaFbx
(synonym: atrogin-1)
(synonym: TRAM63)MuRF1
(synonym: TRAM63)
(2) Western blotting(2) Western blotting
유게놀 처리한 마우스 근아세포를 사용하였다는 점을 제외하고는 실험예 1-(2)와 동일한 방법으로 수행하였다. The same procedure as in Experimental Example 1- (2) was performed except that the mouse muscle myoblasts treated with eugenol were used.
도 4는 유게놀을 처리한 마우스 근아세포에서 단백질 분해 및 합성 관련 분자들의 발현 변화를 나타낸 그래프이다. FIG. 4 is a graph showing changes in expression of molecules involved in proteolysis and synthesis in mouse enucleated ovalbumin-treated mouse myoblasts.
도 4에 나타난 바와 같이, 비교예 2의 경우, 정상세포(Basal)에 비해 단백질 합성과 관련이 있는 p-4E-BP1 및 p-p70S6K1 단백질 양이 유의적으로 감소하였고(도 4A), 단백질 분해 유전자인 MaFbx/atrogin1과 MuRF1 발현은 유의적으로 증가함을 확인할 수 있다(도 4B). 유게놀을 처리한 실시예 2의 경우, dexamethasone에 의해 감소한 p-4E-BP1 및 p-p70S6K 단백질 양을 다시 유의적으로 증가시키고(도 4A), MuRF1 및 MaFbx/atrogin1의 발현은 유의하게 감소시킴을 확인할 수 있다(도 4B). 즉, 유게놀은 마우스 근아세포에서 4E-BP1 및 p70S6K1 단백질 인산화를 증가시키고, MuRF1 및 MaFbx/atrogin1 유전자 발현을 억제함으로써 궁극적으로 근육의 양을 증가시키는데 관여하였을 것으로 사료된다.As shown in FIG. 4, in the case of Comparative Example 2, the amounts of p-4E-BP1 and p-p70S6K1 proteins, which are involved in protein synthesis, were significantly decreased as compared with normal cells (Basal) The expression of the genes MaFbx / atroginl and MuRF1 was significantly increased (Fig. 4B). In the case of eugenol-treated Example 2, the amount of p-4E-BP1 and p-p70S6K protein reduced by dexamethasone was again significantly increased (Fig. 4A), and the expression of MuRF1 and MaFbx / atrogin1 was significantly decreased (Fig. 4B). In other words, yugenol may be involved in increasing the amount of muscle 4F-BP1 and p70S6K1 protein phosphorylation in mouse myoblasts and ultimately increasing the amount of muscle by inhibiting MuRF1 and MaFbx / atrogin1 gene expression.
하기에 본 발명의 추출물을 함유하는 조성물의 제제예를 설명하나, 본 발명은 이를 한정하고자 함이 아닌 단지 구체적으로 설명하고자 함이다.Hereinafter, formulation examples of the composition containing the extract of the present invention will be described, but the present invention is not intended to be limited thereto but is specifically described.
제제예 1: 산제의 제조Formulation Example 1: Preparation of powder
오시멘 또는 유게놀 20 ㎎Ocimene or yugenol 20 mg
유당수화물 100 ㎎100 mg of lactose hydrate
탈크 10 ㎎10 mg of talc
상기의 성분들을 혼합하고 기밀포에 충진하여 산제를 제조하였다.The above ingredients were mixed and filled in an airtight container to prepare powders.
제제예 2: 정제의 제조Formulation Example 2: Preparation of tablets
오시멘 또는 유게놀 10 ㎎Ocimene or yugenol 10 mg
옥수수전분 100 ㎎
유당수화물 100 ㎎100 mg of lactose hydrate
스테아르산마그네슘 2 ㎎Magnesium stearate 2 mg
상기의 성분을 혼합한 후, 통상의 정제의 제조방법에 따라서 타정하여 정제를 제조하였다.After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.
제제예 3: 캅셀제의 제조Formulation Example 3: Preparation of capsules
오시멘 또는 유게놀 10 ㎎Ocimene or yugenol 10 mg
미결정셀룰로오스 3 ㎎ 3 mg of microcrystalline cellulose
유당수화물 14.8 ㎎Lactose hydrate 14.8 mg
스테아르산마그네슘 0.2 ㎎Magnesium stearate 0.2 mg
상기의 성분을 혼합한 후, 통상의 캅셀제의 제조방법에 다라서 젤라틴캡슐에 충전하여 캅셀제를 제조하였다.After mixing the above components, the capsules were filled in gelatin capsules in accordance with the usual methods for preparing capsules.
제제예 4: 주사제의 제조Formulation Example 4: Preparation of injection
오시멘 또는 유게놀 10 ㎎Ocimene or yugenol 10 mg
만니톨 180 ㎎180 mg mannitol
주사용 멸균 증류수 2974 ㎎2974 mg of sterile distilled water for injection
인산일수소나트퓸 26 ㎎26 mg of sodium hydrogen phosphate
상기의 성분을 혼합한 후, 통상의 주사제의 제조방법에 따라 1앰플당(2mL) 상기의 성분 함량으로 제조하였다.After the above components were mixed, they were prepared with the above ingredient contents per ampoule (2 mL) according to the usual injection preparation method.
제제예 5: 액제의 제조Formulation Example 5: Preparation of a liquid preparation
오시멘 또는 유게놀 10 ㎎Ocimene or yugenol 10 mg
이성화당 10 ㎎10 mg of isomerized sugar
만니톨 5 ㎎5 mg mannitol
정제수 적량Purified water quantity
레몬향 적량Lemon incense quantity
상기의 성분을 통상의 제조방법에 따라 정제수에 각각의 성분을 가하여 용해시키고 레몬향을 적량 가한 다음 정제수를 가하여 전체 100mL로 조절한 후 멸균시켜 갈색병에 충진하여 액제를 제조한다. The components are dissolved in purified water according to the usual preparation method, and the lemon flavor is added in an appropriate amount. Then, purified water is added to adjust the total volume to 100 mL, sterilized and filled in a brown bottle to prepare a liquid preparation.
제제예 6: 건강기능식품의 제조Formulation Example 6: Preparation of Health Functional Foods
오시멘 또는 유게놀 10 ㎎Ocimene or yugenol 10 mg
비타민 혼합물 적량Vitamin mixture quantity
비타민 A 아세테이드 70 ㎍Vitamin A Acetate 70 ㎍
비타민 E 1.0 ㎎Vitamin E 1.0 mg
비타민 B1 0.13 ㎎0.13 mg of vitamin B 1
비타민 B2 0.15 ㎎0.15 mg of vitamin B 2
비타민 B6 0.5 ㎎0.5 mg of vitamin B 6
비타민 B12 0.2 ㎍Vitamin B 12 0.2 g
비타민 C 10 ㎎10 mg vitamin C
비오틴 10 ㎍Biotin 10 μg
니코틴산아미드 1.7 ㎎Nicotinic acid amide 1.7 mg
엽산 50 ㎍50 ㎍ of folic acid
판토텐산 칼슘 0.5 ㎎Calcium pantothenate 0.5 mg
무기질 혼합물 적량Mineral mixture quantity
황산제1철 1.75 ㎎1.75 mg of ferrous sulfate
산화아연 0.82 ㎎0.82 mg of zinc oxide
탄산마그네슘 25.3 ㎎Magnesium carbonate 25.3 mg
제1인산칼륨 15 ㎎15 mg of potassium phosphate monobasic
제2인산칼슘 55 ㎎
구연산칼륨 30 ㎎Potassium citrate 30 mg
탄산칼슘 100 ㎎100 mg of calcium carbonate
염화마그네슘 24.8 ㎎24.8 mg of magnesium chloride
상기의 비타민 및 미네랄 혼합물의 조성비는 비교적 건강식품에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 그 배합비를 임의로 변형 실시하여도 무방하며, 통상의 건강식품 제조방법에 따라 상기의 성분을 혼합한 다음, 과립을 제조하고, 통상의 방법에 따라 건강식품 조성물 제조에 사용할 수 있다.Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.
제제예 7: 건강음료의 제조Formulation Example 7: Preparation of health drinks
오시멘 또는 유게놀 10 mgOcimene or
비타민 C 15 gVitamin C 15 g
비타민 E(분말) 100 gVitamin E (powder) 100 g
젖산철 19.75 g19.75 g of ferrous lactate
산화아연 3.5 g3.5 g of zinc oxide
니코틴산아미드 3.5 gNicotinic acid amide 3.5 g
비타민 A 0.2 gVitamin A 0.2 g
비타민 B1 0.25 gVitamin B 1 0.25 g
비타민 B2 0.3 g0.3 g of vitamin B 2
정제수 정량Purified water quantitation
통상의 건강음료 제조방법에 따라 상기의 성분을 혼합한 다음, 약 1 시간 동안 85℃에서 교반 가열한 후, 만들어진 용액을 여과하여 멸균된 2ℓ 용기에 취득하여 밀봉 멸균한 뒤 냉장 보관한 다음 본 발명의 건강음료 조성물 제조에 사용한다.The above components were mixed according to a conventional health drink manufacturing method, and the mixture was heated at 85 DEG C for about 1 hour with stirring, and the solution thus prepared was filtered to obtain a sterilized 2-liter container, which was sealed and sterilized, ≪ / RTI >
상기 조성비는 비교적 기호음료에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만 수요계층이나, 수요국가, 사용용도 등 지역적, 민족적 기호도에 따라서 그 배합비를 임의로 변형 실시하여도 무방하다.Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.
하기에 본 발명의 추출물을 함유하는 화장료 조성물의 제조예를 설명하나, 본 발명은 이를 한정하고자 함이 아닌 단지 구체적으로 설명하고자 함이다.Hereinafter, a preparation example of a cosmetic composition containing the extract of the present invention will be described, but the present invention is not intended to be limited thereto but is specifically explained.
제조예 1: 영양화장수(밀크로션)Production Example 1: Nutritional lotion (milk lotion)
본 발명의 오시멘 또는 유게놀 2.0 중량%2.0% by weight of the oicimene or eugenol of the present invention,
스쿠알란 5.0 중량%Squalane 5.0 wt%
밀납 4.0 중량%4.0 wt%
폴리솔베이트60 1.5 중량%Polysorbate 60 1.5 wt%
솔비탄세스퀴올레이트 1.5 중량%1.5% by weight of sorbitan sesquioleate
유동파라핀 0.5 중량%0.5% by weight liquid paraffin
카프릴릭/카프릭트리글리세라이드 5.0 중량%Caprylic / capric triglyceride 5.0 wt%
글리세린 3.0 중량%Glycerin 3.0 wt%
부틸렌글리콜 3.0 중량%3.0% by weight of butylene glycol
프로필렌글리콜 3.0 중량%3.0% by weight of propylene glycol
카르복시비닐폴리머 0.1 중량%Carboxyvinyl polymer 0.1 wt%
트리에탄올아민 0.2 중량%0.2% by weight triethanolamine
방부제, 색소, 향료 적량Preservative, pigment, perfume
정제수 to 100 중량%Purified water to 100%
상기의 배합비는 비교적 영양화장수에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 그 배합비를 임의로 변형 실시하여도 무방하며, 통상적인 화장품 분야에서의 제조방법에 따라 제조할 수 있다. Although the compounding ratio of the above-described ingredients is comparatively comparable to that of a nutritional lotion, the compounding ratio of the ingredients may be arbitrarily varied and can be prepared according to a conventional method in the field of cosmetics.
제조예 2: 유연화장수(스킨로션)Production Example 2: Flexible Longevity (Skin lotion)
발명의 오시멘 또는 유게놀 2.0 중량 %Ocimene or eugenol of the invention 2.0 wt%
글리세린 3.0 중량 %Glycerin 3.0 wt%
부틸렌글리콜 2.0 중량 %Butylene glycol 2.0 wt%
프로필렌글리콜 2.0 중량 %Propylene glycol 2.0 wt%
카르복시비닐폴리머 0.1 중량 %Carboxyvinyl polymer 0.1 wt%
PEG 12 노닐페닐에테르 0.2 중량 %PEG 12 nonyl phenyl ether 0.2 wt%
폴리솔베이트80 0.4 중량 %Polysorbate 80 0.4 wt%
에탄올 10.0 중량 %Ethanol 10.0 wt%
트리에탄올아민 0.1 중량 %0.1% by weight triethanolamine
방부제, 색소, 향료 적량Preservative, pigment, perfume
정제수 to 100 중량 %Purified water to 100%
상기의 배합비는 비교적 유연화장수에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 그 배합비를 임의로 변형 실시하여도 무방하며, 통상적인 화장품 분야에서의 제조방법에 따라 제조할 수 있다. Although the compounding ratio of the above-described components is a mixture of the components suitable for the relatively long softening time, it may be arbitrarily varied in its blending ratio, and it can be produced according to a conventional production method in the field of cosmetics.
제조예 3: 영양크림Production Example 3: Nourishing cream
본 발명의 오시멘 또는 유게놀 2.0 중량 % 2.0% by weight of the oicimene or eugenol of the present invention,
폴리솔베이트60 1.5 중량 %Polysorbate 60 1.5 wt%
솔비탄세스퀴올레이트 0.5 중량 %0.5% by weight of sorbitan sesquioleate
PEG60 경화피마자유 2.0 중량 %PEG 60 hardened castor oil 2.0 wt%
유동파라핀 10 중량 %10% by weight liquid paraffin
스쿠알란 5.0 중량 %Squalane 5.0 wt%
카프릴릭/카프릭트리글리세라이드 5.0 중량 %Caprylic / capric triglyceride 5.0 wt%
글리세린 5.0 중량 %Glycerin 5.0 wt%
부틸렌글리콜 3.0 중량 %3.0% by weight of butylene glycol
프로필렌글리콜 3.0 중량 %3.0% by weight of propylene glycol
트리에탄올아민 0.2 중량 %0.2% by weight triethanolamine
방부제 적량Antiseptic
색소 적량Pigment amount
향료 적량Fragrance quantity
정제수 to 100 중량 %Purified water to 100%
상기의 배합비는 비교적 영양크림에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 그 배합비를 임의로 변형 실시하여도 무방하며, 통상적인 화장품 분야에서의 제조방법에 따라 제조할 수 있다. Although the compounding ratio of the above-mentioned ingredients is comparatively comparable to that of the nutritional cream, the compounding ratio of the ingredient may be arbitrarily varied, and can be manufactured according to a conventional method in the field of cosmetics.
제조예 4: 마사지크림Production Example 4: Massage cream
본 발명의 오시멘 또는 유게놀 1.0 중량 %1.0% by weight of the oicimene or eugenol of the present invention,
밀납 10.0 중량 %Wax 10.0 wt%
폴리솔베이트60 1.5 중량 %Polysorbate 60 1.5 wt%
PEG 60 경화피마자유 2.0 중량 %PEG 60 hardened castor oil 2.0 wt%
솔비탄세스퀴올레이트 0.8 중량 %0.8% by weight of sorbitan sesquioleate
유동파라핀 40.0 중량 %Liquid paraffin 40.0 wt%
스쿠알란 5.0 중량 %Squalane 5.0 wt%
카프릴릭/카프릭트리글리세라이드 4.0 중량 %Caprylic / capric triglyceride 4.0 wt%
글리세린 5.0 중량 %Glycerin 5.0 wt%
부틸렌글리콜 3.0 중량 %3.0% by weight of butylene glycol
프로필렌글리콜 3.0 중량 %3.0% by weight of propylene glycol
트리에탄올아민 0.2 중량 %0.2% by weight triethanolamine
방부제, 색소, 향료 적량Preservative, pigment, perfume
정제수 to 100 중량 %Purified water to 100%
상기의 배합비는 비교적 마사지크림에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 그 배합비를 임의로 변형 실시하여도 무방하며, 통상적인 화장품 분야에서의 제조방법에 따라 제조할 수 있다. Although the compounding ratio of the above-mentioned ingredients is comparatively comparatively suitable for the massage cream, the compounding ratio thereof may be arbitrarily modified, and can be manufactured according to a conventional manufacturing method in the field of cosmetics.
제조예 4: 팩Production Example 4:
발명의 오시멘 또는 유게놀 1.0 중량 %1.0% by weight oscimene or eugenol of the invention,
폴리비닐알콜 13.0 중량 %Polyvinyl alcohol 13.0 wt%
소듐카르복시메틸셀룰로오스 0.2 중량 %0.2% by weight of sodium carboxymethylcellulose,
글리세린 5.0 중량 %Glycerin 5.0 wt%
알란토인 0.1 중량 %Allantoin 0.1 wt%
에탄올 6.0 중량 %6.0% by weight of ethanol
PEG 12 노닐페닐에테르 0.3 중량 %PEG 12 nonyl phenyl ether 0.3 wt%
폴리솔베이트60 0.3 중량 %Polysorbate 60 0.3 wt%
방부제, 색소, 향료 적량Preservative, pigment, perfume
정제수 to 100 중량 %Purified water to 100%
상기의 배합비는 비교적 팩에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 그 배합비를 임의로 변형 실시하여도 무방하며, 통상적인 화장품 분야에서의 제조방법에 따라 제조할 수 있다. Although the compounding ratio of the above components is comparatively comparatively compatible with the pack, the compounding ratio thereof may be arbitrarily varied and can be produced by a conventional method in the field of cosmetics.
제조예 6: 젤Production Example 6: Gel
본 발명의 오시멘 또는 유게놀 0.5 중량 %0.5% < RTI ID = 0.0 >% < / RTI >
에틸렌디아민초산나트륨 0.05 중량 %0.05% by weight of sodium ethylenediaminetetraacetate
글리세린 5.0 중량 %Glycerin 5.0 wt%
카르복시비닐폴리머 0.3 중량 %Carboxyvinyl polymer 0.3 wt%
에탄올 5.0 중량 %5.0% by weight of ethanol
PEG 60 경화피마자유 0.5 중량 %PEG 60 hardened castor oil 0.5 wt%
트리에탄올아민 0.3 중량 %Triethanolamine 0.3 wt%
방부제, 색소, 향료 적량Preservative, pigment, perfume
정제수 to 100 중량 %Purified water to 100%
상기의 배합비는 비교적 젤에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 그 배합비를 임의로 변형 실시하여도 무방하며, 통상적인 화장품 분야에서의 제조방법에 따라 제조할 수 있다. Although the compounding ratio of the above-mentioned ingredients is comparatively comparable to that of the gel, the blending ratio may be arbitrarily varied and can be produced by a conventional method in the field of cosmetics.
상기 배합비는 비교적 화장료 조성물에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 그외의 색채 화장품을 포함하는 다양한 용도의 화장품에 적용될 수 있는 것이고, 그 효능에 따라 인체에 얇게 도포하여 바를 수 있는 약제 즉, 연고로 제조에 이용될 수 있으며 수요계층이나, 수요국가, 사용용도 등 지역적, 민족적 기호도에 따라서 그 배합비를 임의로 변형 실시하여도 무방하다.Although the compounding ratio is comparatively comparatively suitable for the cosmetic composition, it can be applied to cosmetics for various uses including other color cosmetics. Depending on its effectiveness, it can be applied to a human body thinly, It can be used for manufacturing in ointment, and it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as demand level, demand country, use purpose, and the like.
하기에 본 발명의 추출물을 함유하는 가축 사료 조성물의 제조예를 설명하나, 본 발명은 이를 한정하고자 함이 아닌 단지 구체적으로 설명하고자 함이다.Hereinafter, a preparation example of a livestock feed composition containing the extract of the present invention will be described, but the present invention is not intended to be limited thereto but is specifically explained.
제조예 1: 사료첨가제의 제조Production Example 1: Preparation of Feed Additive
본 발명의 조성물 0.1~10%0.1 to 10% of the composition of the present invention,
제3 인산칼슘 1~20%Tribasic calcium phosphate 1 to 20%
비타민 E 0.01~0.1%0.01 to 0.1% of vitamin E
효소 분말 1~10%Enzyme powder 1 ~ 10%
유산균 0.1~10%Lactic acid bacteria 0.1 ~ 10%
포도당 20~90%Glucose 20-90%
제조예 2: 사료의 제조Production Example 2: Production of feed
상기 제조예 1의 사료첨가제를 유효성분으로 하여 하기와 같은 조성으로 사료를 제조하였다.Using the feed additive of Production Example 1 as an active ingredient, feeds were prepared in the following composition.
제조예 1의 사료 첨가제 0.1~10%Feed additives of Preparation Example 1 0.1 to 10%
밀기울 40~49.9%Bran 40 ~ 49.9%
마일로 21.20%Milo 21.20%
대두박 20.00%Soybean meal 20.00%
어분 3.00%Fish meal 3.00%
당밀 4.00%Molasses 4.00%
미네랄 1.53%Mineral 1.53%
비타민 0.27%Vitamin 0.27%
전술한 본 발명의 설명은 예시를 위한 것이며, 본 발명이 속하는 기술분야의 통상의 지식을 가진 자는 본 발명의 기술적 사상이나 필수적인 특징을 변경하지 않고서 다른 구체적인 형태로 쉽게 변형이 가능하다는 것을 이해할 수 있을 것이다. 그러므로 이상에서 기술한 실시예들은 모든 면에서 예시적인 것이며 한정적이 아닌 것으로 이해해야만 한다.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 (11)
A pharmaceutical composition for preventing or treating muscular diseases comprising ocimene, eugenol or a pharmaceutically acceptable salt thereof as an active ingredient.
상기 조성물은 p-4E-BP1 또는 p-p70S6K1 단백질의 발현을 증가시키는 것을 특징으로 하는
근육 질환 예방 또는 치료용 약학적 조성물.
The method according to claim 1,
Wherein said composition increases the expression of p-4E-BP1 or p-p70S6K1 protein
A pharmaceutical composition for preventing or treating a muscle disorder.
상기 조성물은 MuRF1(Muscle Ring-Finger Protein) 또는 MaFbx(Muscle atrophy F-box)의 발현을 감소시키는 것을 특징으로 하는
근육 질환 예방 또는 치료용 약학적 조성물.
The method according to claim 1,
The composition is characterized by reducing the expression of MuRF1 (Muscle Ring-Finger Protein) or MaFbx (Muscle atrophy F-box)
A pharmaceutical composition for preventing or treating a muscle disorder.
상기 근육 질환은 근 기능 저하, 근육 감소, 근육 소모 또는 근육 퇴화로 인한 근육 질환인 것을 특징으로 하는
근육 질환 예방 또는 치료용 약학적 조성물.
The method according to claim 1,
Wherein the muscle disease is a muscle disease caused by a decrease in muscular function, a decrease in muscle, a decrease in muscle wear or a muscle degeneration
A pharmaceutical composition for preventing or treating a muscle disorder.
상기 근육 질환은 긴장감퇴증(atony), 근위축증(muscular atrophy), 근이영양증(muscular dystrophy), 근무력증, 악액질(cachexia), 경직성 척추 증후군(rigid spinesyndrome), 근위축성 측삭경화증(루게릭병, amyotrophic lateral sclerosis), 경직성 척추 증후군(rigid spinsesyndrome), 샤르코-마리-투스병(Charcot-Marie-Tooth disease) 및 근육 감소증(sarcopenia)으로 이루어진 군으로부터 선택되는 어느 하나 이상인 것을 특징으로 하는
근육 질환 예방 또는 치료용 약학적 조성물.
5. The method according to any one of claims 1 to 4,
The muscular diseases include atony, muscular atrophy, muscular dystrophy, myasthenia gravis, cachexia, rigid spinesyndrome, amyotrophic lateral sclerosis, amyotrophic lateral sclerosis, , Rigid spinsesyndrome, Charcot-Marie-Tooth disease, and sarcopenia. In a preferred embodiment of the present invention,
A pharmaceutical composition for preventing or treating a muscle disorder.
A pharmaceutical composition for promoting muscle differentiation, muscle regeneration or muscle strengthening comprising ocimene, eugenol or a pharmaceutically acceptable salt thereof as an active ingredient.
A health functional food composition for improving muscle function comprising ocimene, eugenol or a pharmaceutically acceptable salt thereof as an active ingredient.
A health functional food composition for promoting muscle differentiation, muscle regeneration or muscle strengthening comprising ocimene, eugenol or a pharmaceutically acceptable salt thereof as an active ingredient.
A composition for the prevention or treatment of muscle diseases, comprising ocimene, eugenol or a pharmaceutically acceptable salt thereof as an active ingredient.
A composition for promoting muscle differentiation, muscle regeneration or muscle strengthening comprising ocimene, eugenol or a pharmaceutically acceptable salt thereof as an active ingredient.
A cosmetic composition for improving muscle function comprising ocimene, eugenol or a pharmaceutically acceptable salt thereof as an active ingredient.
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PCT/KR2018/000306 WO2018128483A1 (en) | 2017-01-06 | 2018-01-05 | Composition for preventing or treating muscular diseases, comprising ocimene, eugenol or pharmaceutically acceptable salt thereof as active ingredient |
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