KR20180083835A - COMPOSITION COMPRISING THE EXTRACT OF ACER MONO Max. LEAF FOR ANTIOXIDANT EFFECT - Google Patents
COMPOSITION COMPRISING THE EXTRACT OF ACER MONO Max. LEAF FOR ANTIOXIDANT EFFECT Download PDFInfo
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- KR20180083835A KR20180083835A KR1020180081438A KR20180081438A KR20180083835A KR 20180083835 A KR20180083835 A KR 20180083835A KR 1020180081438 A KR1020180081438 A KR 1020180081438A KR 20180081438 A KR20180081438 A KR 20180081438A KR 20180083835 A KR20180083835 A KR 20180083835A
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Abstract
Description
본 발명은 고로쇠 잎 추출물을 유효성분으로 함유하는 항산화 활성을 갖는 조성물 및 그 제조방법에 관한 것이다.The present invention relates to a composition having an antioxidative activity containing a gonorrhea leaf extract as an active ingredient and a method for producing the same.
산소는 생명체를 유지하는데 없어선 안 될 가장 중요한 분자다. 예를 들어, 뇌세포는 30초만 산소가 공급되지 않으면 파괴되기 시작한다. 세포 안에서 에너지를 만드는 핵심 공장 역할을 하는 기관이 ‘미토콘드리아’ 이다. 미토콘드리아에 운반된 산소는 사람이 영양분(탄수화물, 단백질, 지방)을 섭취하여 만들어진 포도당을 분해시켜서 에너지를 생산한다. 에너지 생산과정에서 물과 이산화탄소 및 산소가 발생하는데, 이 산소를 활성산소종(Reactive oxygen species, ROS)라 한다. Oxygen is the most important molecule that can not exist to sustain life. For example, brain cells begin to break down if oxygen is not supplied for 30 seconds. Mitochondria is the organ that plays a key role in creating energy within cells. Oxygen transported to the mitochondria produces energy by digesting glucose produced by a person ingesting nutrients (carbohydrates, proteins, fats). During the energy production process, water, carbon dioxide and oxygen are generated, which is called reactive oxygen species (ROS).
활성산소종은 정상적인 세포 내 활성작용 과정에서 생성되며 세포분화, 유전자의 발현, 사이토카인에 대한 반응 정도를 포함한 다양한 생물학적 과정에 연관되어있다. 산화스트레스는 ROS의 생성과 이를 제거하는 항산화 반응 간의 불균형으로 인해 세포 내의 ROS가 증가하여 DNA나 단백질, 지질(lipid)과 반응하여 손상시키는 현상이며 이는 노화나 심장과 관련된 질병들의 핵심 원인으로 알려져 있다.Active oxygen species are produced during normal intracellular activity and are involved in a variety of biological processes including cell differentiation, expression of genes, and response to cytokines. Oxidative stress is an imbalance between the production of ROS and the antioxidant reaction that causes it to increase the intracellular ROS and react with DNA, protein, and lipid, which is known to be a major cause of aging and heart-related diseases .
대부분의 활성산소 종은 미토콘드리아의 전자 전달 사이에서 부산물로서 생성된다. 또한, ROS는 금속 촉매 산화 반응에 필요한 중간체를 형성하고 있다. 산소원자는 바깥 전자 껍질의 다른 궤도에 2개의 부대 전자를 가지고 있다. 이 전자 구조는 라디칼 형성의 영향을 받기 쉬운 산소를 만든다. 전자의 첨가에 의한 산소의 순차적 퇴화는 superoxide, hydrogen peroxide, hydroxyl radical, hydroxyl ion, nitric oxide 같은 ROS의 형성을 만들어 낸다.Most reactive oxygen species are produced as a by-product between the electron transport of the mitochondria. ROS also forms an intermediate for the metal catalytic oxidation reaction. The oxygen atom has two sub electrons in another orbit of the outer electron shell. This electronic structure creates oxygen that is susceptible to radical formation. Sequential degeneration of oxygen by the addition of electrons produces the formation of ROS such as superoxide, hydrogen peroxide, hydroxyl radical, hydroxyl ion, and nitric oxide.
활성 산소 종의 해독은 모든 호기성 생명체의 생존에 가장 중요하다. 보통 말하는 방어메커니즘의 상당수는 ROS의 제거와 생성 사이에 균형이 제공되고 이러한 필요성을 충족하도록 진화 되었다. ROS의 생성과 제거의 불균형은 산화적 스트레스라고 지칭한다. 세포는 ROS의 유해한 영향을 개선하기위한 방어 메커니즘의 다양성을 가지고 있다. Superoxide dismutase (SOD)는 과산화수소 (H2O2)와 산소(O2) 분자에 삽입 된 두 슈퍼 옥사이드 음이온의 변환을 촉매한다. Detoxification of reactive oxygen species is most important for the survival of all aerobic organisms. Many of the commonly mentioned defense mechanisms have evolved to meet this need by providing a balance between elimination and generation of ROS. The imbalance in the generation and elimination of ROS is referred to as oxidative stress. Cells have a variety of defense mechanisms to improve the harmful effects of ROS. Superoxide dismutase (SOD) catalyzes the conversion of two superoxide anions embedded in hydrogen peroxide (H 2 O 2 ) and oxygen (O 2 ) molecules.
2O2+ 2H H2O2+ O2 2O 2 + 2H H 2 O 2 + O 2
진핵 세포의 과산화소체에서, 카탈라아제 효소는 H2O2를 물과 산소로 변환 시키고, SOD로부터 시작된 활성산소의 해독을 완료한다.In the peroxidase of eukaryotic cells, the catalase enzyme converts H 2 O 2 to water and oxygen and completes the detoxification of active oxygen originating from SOD.
2H2O2 2 H2O + O2 2H 2 O 2 2 H 2 O + O 2
이러한 해독에 중요한 역할을 하는 비효소 소분자 항산화제들이 많이 있다. Glutathione은 활성산소종의 유해 효과에 대해 가장 중요한 세포 내 방어 역할을 수행한다. ROS 분자와의 반응은 Glutathione을 산화하지만, 환원 된 형태는 NADPH 의존성 환원 효소에 의해 산화 환원에서 재생된다. glutathione (GSSG)의 산화 형의 비율로 환원 형태인 (GSH)은 생물의 산화 스트레스의 동적 지표이다.There are many nonenzymatic small molecule antioxidants that play an important role in this detoxification. Glutathione plays an important role in intracellular defense against the harmful effects of reactive oxygen species. The reaction with the ROS molecule oxidizes Glutathione, but the reduced form is regenerated by redox reaction by NADPH-dependent reductase. Reduced form (GSH) is a dynamic index of the oxidative stress of the organism at the ratio of oxidative form of glutathione (GSSG).
설사 완벽한 항산화제가 만들어진다고 해도, 활성산소를 완벽하게 막아서도 안 된다. 생존에 꼭 필요한 물질이기 때문이다. 세포의 성장에 필수적이고 인간을 비롯한 동식물의 체내에 세균, 바이러스, 곰팡이 등의 이물질이 침입했을 경우, 이것을 녹여 없앰으로써 생체를 지키는 아주 중요한 역할을 한다. Even if a perfect antioxidant is made, it should not completely block active oxygen. It is a substance that is essential for survival. It is essential for cell growth and plays a very important role in protecting living organisms by dissolving foreign matter such as bacteria, viruses and fungi into the body of animals and plants including humans.
간에서는 활성산소가 해독 작용을 하기도 하고 어떤 활성산소는 암 세포를 죽이기도 한다. 또 활성산소가 인체의 세포 성장과 세포 자살에 관련된 다양한 생체 신호 전달 과정에서 매우 중요한 역할을 한다는 것이 밝혀지기도 했다. 초파리의 경우 활성산소를 만드는 효소의 유전자를 없애면 번식하지 못한다. 사람도 마찬가지다. 정자는 활성산소를 뿜어내는 관을 통과하지 않으면 성숙하지 않는다. 심지어 활성산소가 오히려 수명을 증가시키기도 한다. In the liver, active oxygen is detoxifying, and some reactive oxygen species kill cancer cells. It has also been shown that reactive oxygen species plays a very important role in various biological signal transduction processes related to human cell growth and apoptosis. In the case of Drosophila, it is impossible to reproduce by removing the gene of the enzyme that makes active oxygen. People are the same. Sperm are not mature unless they pass through a tube that emits oxygen. Even active oxygen increases life span.
2010년 캐나다 맥길 대학의 지그프리드 헤키미 박사는 꼬마선충을 활성산소를 많이 생산하도록 유전자 조작을 한 결과 수명이 단축되기는커녕 오히려 수명이 더 연장되었다고 밝혔다. 활성산소의 과다 생산이 체내의 보호와 수리라는 메커니즘을 작동시켜 오히려 수명이 연장된 것이다. 사람의 몸에도 활성산소가 적당량 있는 상태에서 최고의 성과를 낸다. 독과 약은 한 몸인 것이다. 적당한 정도가 좋은 것이지 완벽히 위험을 없애려다가는 오히려 위험해질 수 있는 것이다.In 2010, Dr. Siegfried Hekimi of McGill University in Canada said that genetic manipulation of small nematodes to produce more active oxygen resulted in a longer life span rather than a shorter life span. Excessive production of active oxygen has extended the life span by activating the mechanism of protection and repair in the body. It also gives the best results in a state where the amount of active oxygen is appropriate for a human body. Poison and medicine are one body. It is good enough, but it can be rather dangerous if you try to completely eliminate the risk.
고로쇠는 고로쇠나무, 오각풍, 수색수, 색목 이라고도 한다. 고로쇠나무는 단풍나무과에 속하는 낙엽교목으로 표고 100~1,800m에 자생하며 한국, 일본, 중국, 만주에까지 분포한다. 우리나라의 고로쇠나무는 내한성이 강하여 지리산, 백운산, 조계산 및 강원도 일대에서 주로 자생하고, 고로쇠나무(Acer pictum subsp. mono), 붉은고로쇠나무(A. mono for. rubrieps), 우산고로쇠나무(A. okamotoanum), 만주고로쇠(A.truncatum), 긴고로쇠나무(A. mono for. dissectum), 왕고로쇠나무(A. mono var. savatieri), 산 고로쇠나무(A.mono var. horizontale), 집게고로쇠나무(A. mono for. connivens), 털고로쇠나무(A. mono var. ambiguum) 등 9종의 품종과 변종이 생육하고 있는 것으로 알려져 있다.It is also known as a nori tree, a pentagonal tree, a searing number, and a tree. It is a deciduous arboreous tree belonging to the maple family. It grows at 100-1800m above sea level and is distributed in Korea, Japan, China, and Manchuria. Acer mono Korea is a strong and hardy and grows mainly in Jiri, White Clouds Mountain, Jogyesan and Gangwon one, Acer mono (Acer pictum subsp. mono), Red Acer mono (A. mono for. rubrieps), an umbrella Acer mono (A. okamotoanum), Acer mono ten thousand shares (A.truncatum), Long Acer mono (A. mono for. dissectum), king of Acer mono (A. mono var. savatieri ), A. mono var horizontale , A. mono for. connivens , A. mono var ambiguum , and so on . .
고로쇠라는 이름은 뼈에 이롭다는 듯의 한자어 골리수에서 유래하였다. 한방에서는 나무에 상처를 내어 흘러내린 즙을 풍당이라 하여 위장병, 폐병, 신경통, 관절염 환자들에게 약수로 마시게 하였다. 이처럼 고로쇠나무의 수액에 대한 음용의 역사가 깊고, 최근에는 수액 소비량도 증가하여 수액에 대한 연구도 진행되고 있으나 고로쇠 나무의 잎에 대한 연구는 거의 이루어지지 않고 있다.The name "gorozu" comes from the Korean word "gollyu" which seems to be beneficial to the bones. In one room, the juice that had been cut down by the wood was called zuchuan, and it was given to the patients with gastrointestinal disease, lung disease, neuralgia, and arthritis by drinking water. In recent years, the amount of consumption of sap has been increased, and studies on sap have been carried out. However, studies on the leaves of A. falciparum have been rarely carried out.
이에 본 발명자들은 독성이 없으면서 부작용이 적고, 항산화 효과가 있는 새로운 천연물 소재를 제공하고자 한다. 현재 천연물 소재에 대한 관심도가 높아짐에 따라 고로쇠 같은 천연물에 관한 연구가 진행 되고 있는데, (우산고로쇠나무 추출물을 함유하는 트롬빈 저해 혈전증 예방 및 치료용 약학 조성물, 고로쇠 수액을 함유하는 위염 및 위궤양의 예방 및 치료용 조성물) 등의 특허가 있으나, 고로쇠 잎만을 가지고 그 항산화 활성에 관한 연구는 전무한 실정이다.Therefore, the present inventors intend to provide a novel natural material having no adverse effect, less side effects, and antioxidative effect. As the interest in natural materials has increased, studies on natural products such as gonorrhea have been carried out. (A pharmaceutical composition for prevention and treatment of thrombin inhibition thrombosis containing an umbrella tree extract, prevention of gastritis and gastric ulcer containing gonorrhea sap, Therapeutic composition). However, there is no research on the antioxidant activity of only the leaves of the gonorrhea leaf.
이에 본 발명자들은 독성이 없으면서 부작용이 적고, 항산화 효과가 있는 새로운 천연물 소재를 찾던 중 고로쇠 잎 추출물에 항산화 활성이 우수하다는 사실을 확인하고 본 발명을 완성하였다. Therefore, the inventors of the present invention confirmed that the antioxidative activity of the extract of the gonorrhea leaves is excellent while searching for a new natural material having no antioxidative effect while having little toxicity, and completed the present invention.
따라서, 본 발명의 목적은 고로쇠 잎 추출물을 유효성분으로 포함하는 항산화용 약학적 조성물을 제공하는 것이다. Accordingly, an object of the present invention is to provide a pharmaceutical composition for antioxidant comprising an extract of ganoderma lucidum as an active ingredient.
본 발명의 다른 목적은 고로쇠 잎 추출물을 유효성분으로 포함하는 항산화용 건강기능성 식품을 제공하는 것이다. Another object of the present invention is to provide a health functional food for antioxidation comprising an extract of ganoderma lucidum as an active ingredient.
본 발명의 다른 목적은 고로쇠 잎 추출물을 유효성분으로 포함하는 항산화용 화장료 조성물을 제공하는 것이다. Another object of the present invention is to provide a cosmetic composition for antioxidation comprising an extract of ganoderma lucidum as an active ingredient.
나아가 본 발명은 (a) 고로쇠 잎을 증류수 또는 유기용매로 환류추출하여 추출물을 수득하는 단계; (b) 상기 추출물을 감압농축하는 단계; 및 (c) 상기 감압농축한 추출물을 동결건조하는 단계;를 포함하는 고로쇠 잎 추출물을 유효성분으로 포함하는 항산화용 조성물의 제조방법을 제공하는 것이다. Further, the present invention relates to a method for producing an extract of the present invention, comprising the steps of: (a) refluxing ganoderma leaves with distilled water or an organic solvent to obtain an extract; (b) concentrating the extract under reduced pressure; And (c) lyophilizing the extract obtained by concentration under reduced pressure. The present invention also provides a method for producing an antioxidative composition comprising the extract of the present invention as an active ingredient.
상기 목적을 달성하기 위하여, 본 발명은 고로쇠 잎 추출물을 유효성분으로 포함하는 항산화용 약학적 조성물을 제공한다. In order to attain the above object, the present invention provides a pharmaceutical composition for antioxidant comprising an extract of ganoderma lucidum as an active ingredient.
본 발명의 일실시예에 있어서, 상기 추출물은 물, 메탄올, 에탄올, 부탄올 또는 아세톤에 가용한 추출물일 수 있다. In one embodiment of the present invention, the extract may be an extract soluble in water, methanol, ethanol, butanol or acetone.
본 발명의 일실시예에 있어서, 상기 조성물은 활성산소종을 제거하거나, 환원력을 가질 수 있다. In one embodiment of the present invention, the composition may remove active oxygen species or have a reducing power.
또한, 본 발명은 고로쇠 잎 추출물을 유효성분으로 포함하는 항산화용 건강기능성 식품을 제공한다.The present invention also provides a health functional food for antioxidation comprising an extract of ganoderma lucidum as an active ingredient.
또한, 본 발명은 고로쇠 잎 추출물을 유효성분으로 포함하는 항산화용 화장료 조성물을 제공한다. The present invention also provides a cosmetic composition for antioxidation comprising an extract of ganoderma lucidum as an active ingredient.
나아가 본 발명은 (a) 고로쇠 잎을 증류수 또는 유기용매로 환류추출하여 추출물을 수득하는 단계; (b) 상기 추출물을 감압농축하는 단계; 및 (c) 상기 감압농축한 추출물을 동결건조하는 단계;를 포함하는 고로쇠 잎 추출물을 유효성분으로 포함하는 항산화용 조성물의 제조방법을 제공한다. Further, the present invention relates to a method for producing an extract of the present invention, comprising the steps of: (a) refluxing ganoderma leaves with distilled water or an organic solvent to obtain an extract; (b) concentrating the extract under reduced pressure; And (c) lyophilizing the extract which has been concentrated under reduced pressure. The present invention also provides a method for preparing an antioxidant composition comprising the extract of the present invention as an active ingredient.
본 발명의 일실시예에 있어서, 상기 (a)단계는 고로쇠 잎의 30배(wt/v%)에 해당하는 부피의 증류수 또는 유기용매로 환류추출할 수 있다. In one embodiment of the present invention, the step (a) may be carried out by refluxing with distilled water or an organic solvent having a volume corresponding to 30 times (wt / v%) of the leaves of the gonorrhea leaf.
본 발명의 일실시예에 있어서, 상기 (a)단계의 유기용매는 70% 에탄올일 수 있다. In one embodiment of the present invention, the organic solvent of step (a) may be 70% ethanol.
본 발명의 고로쇠 잎 추출물은 활성산소종 (DPPH, ABTS radical)을 직접 제거하고 환원력 (Reducing power) 효과를 가지고 활성산소를 제거할 수 있으며, 산화스트레스에 의한 DNA 손상을 억제 할 수 있을 뿐만 아니라 항산화 성분 (Polyphenol, Flavonoid)을 함유하고 있어 항산화용 조성물로 유용하게 사용될 수 있는 효과가 있다. The present invention provides a method for removing active oxygen species (DPPH, ABTS radical) and reducing reactive oxygen species by reducing the oxidative stress, (Polyphenol, Flavonoid), which is useful as an antioxidant composition.
도 1은 본 발명에 의한 추출물의 DPPH 라디칼 소거능 측정결과를 나타낸 것이다.
도 2는 본 발명에 의한 추출물의 ABTS 라디칼 소거능 측정결과를 나타낸 것이다.
도 3은 본 발명에 의한 추출물의 환원력 (Reducing power) 측정결과를 나타낸 것이다.
도 4는 본 발명에 의한 추출물의 DNA 산화적 손상에 대한 보호효과 실험결과를 나타낸 것이다.Figure 1 shows the DPPH radical scavenging activity of the extract of the present invention.
FIG. 2 shows the ABTS radical scavenging activity of the extract of the present invention.
FIG. 3 shows the measurement results of the reducing power of the extract according to the present invention.
FIG. 4 shows the results of the protective effect test for DNA oxidative damage of the extract of the present invention.
이하, 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 발명의 고로쇠 잎 추출물을 유효성분으로 포함하는 항산화용 조성물은 하기와 같이 제조될 수 있다. The antioxidant composition comprising the gonorrhea leaf extract of the present invention as an active ingredient can be prepared as follows.
(a) 고로쇠 잎을 증류수 또는 유기용매로 환류추출하여 추출물을 수득하는 단계;(b) 상기 추출물을 감압농축하는 단계; 및 (c) 상기 감압농축한 추출물을 동결건조하는 단계;를 포함하는 고로쇠 잎 추출물을 유효성분으로 포함하는 항산화용 조성물의 제조방법을 제공한다. (a) refining the leaves of ganoderma lucidum with distilled water or an organic solvent to obtain an extract; (b) concentrating the extract at a reduced pressure; And (c) lyophilizing the extract which has been concentrated under reduced pressure. The present invention also provides a method for preparing an antioxidant composition comprising the extract of the present invention as an active ingredient.
본 발명에서 상기 (a)단계는 고로쇠 잎의 30배 (wt/v%)에 해당하는 부피의 열수 및 70% 에탄올로 환류추출을 수행할 수 있으며, 상기 (b) 단계는 50℃에서 감압농축을 진행 할 수 있고, 상기 (c) 단계의 동결건조는 -80 내지 -110℃에서 수행할 수 있다. In the present invention, the step (a) may be carried out by reflux extraction with hot water and 70% ethanol corresponding to 30 times (wt / v%) of the leaves of the gonorrhea leaf, and the step (b) And the lyophilization in the step (c) can be performed at -80 to -110 ° C.
본 발명은 상기의 제조공정으로 얻어진 고로쇠 잎 추출물을 유효성분으로 함유하는 항산화 조성물을 제공한다. The present invention provides an antioxidative composition containing the extract of A. ganoderma lucidum obtained by the above-mentioned manufacturing process as an active ingredient.
상기 조성물은 약학적으로 허용되는 담체 및 부형제를 포함하는 약학 조성물 및 화장품학적으로 허용가능한 기제를 포함하는 화장용 조성물을 포함한다.The composition comprises a cosmetic composition comprising a pharmaceutical composition comprising a pharmaceutically acceptable carrier and an excipient and a cosmetically acceptable base.
또한, 상기 화장료 조성물은 스킨로션, 스킨소프너, 스킨토너, 아스트린젠트, 로션, 밀크로션, 모이스쳐 로션, 영양로션, 맛사지크림, 영양크림, 모이스처크림, 핸드크림, 파운데이션, 에센스, 영양에센스, 팩, 비누, 클렌징폼, 클렌징로션, 클렌징 크림, 바디로션 및 바디클린저의 제형을 포함한다.The cosmetic composition may be at least one selected from the group consisting of a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutritional lotion, a massage cream, a nutrition cream, a moisturizing cream, a hand cream, a foundation, , Cleansing foams, cleansing lotions, cleansing creams, body lotions and body cleansers.
본 발명의 고로쇠 잎 추출물은 활성산소종 (DPPH, ABTS radical)을 직접 제거하고 환원력 (Reducing power) 효과를 가지고 활성산소를 제거할 수 있으며, 산화스트레스에 의한 DNA 손상을 억제 할 수 있을 뿐만 아니라 항산화 성분 (Polyphenol, Flavonoid)을 함유하고 있다. The present invention provides a method for removing active oxygen species (DPPH, ABTS radical) and reducing reactive oxygen species by reducing the oxidative stress, Contains a component (Polyphenol, Flavonoid).
또한, 고로쇠는 자연에서 얻어진 천연물질로서 이로부터 추출된 본 발명의 추출물들 역시 독성 및 부작용 등의 문제가 없다.In addition, the ganoderma is a natural substance obtained from nature, and the extracts of the present invention extracted therefrom are also free from toxicity and side effects.
본 발명의 항산화용 약학조성물은, 조성물 총 중량에 대하여 상기 추출물을 0.02 내지 50 중량%로 포함한다.The pharmaceutical composition for antioxidation of the present invention contains 0.02 to 50% by weight of the above extract relative to the total weight of the composition.
본 발명의 추출물을 포함하는 약학조성물은 약학적 조성물의 제조에 통상적으로 사용하는 적절한 담체, 부형제 및 희석제를 더 포함할 수 있다.The pharmaceutical compositions comprising the extract of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions.
본 발명의 추출물의 약학적 투여 형태는 이들의 약학적 허용가능한 염의 형태로도 사용될 수 있고, 또한 단독으로 또는 타 약학적 활성 화합물과 결합뿐만 아니라 적당한 집합으로 사용될 수 있다.The pharmaceutical dosage forms of the extract of the present invention may be used in the form of their pharmaceutically acceptable salts, and may be used alone or in combination with other pharmaceutically active compounds as well as in suitable aggregates.
본 발명에 따른 추출물을 포함하는 약학조성물은, 각각 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제형화하여 사용될 수 있다. 추출물을 포함하는 조성물에 포함될 수 있는 담체, 부형제 및 희석제로는 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 들 수 있다. 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 상기 추출물에 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘카보네이트(calcium carbonate), 수크로스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제된다. 또한 단순한 부형제 이외에 마그네슘 스테아레이트, 탈크 같은 윤활제들도 사용된다. 경구를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제, 좌제가 포함된다. 비수성용제, 현탁제로는 프로필렌글리콜(propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세로제라틴 등이 사용될 수 있다.The pharmaceutical composition containing the extract according to the present invention can be administered orally in the form of powders, granules, tablets, capsules, oral preparations such as suspensions, emulsions, syrups and aerosols, external preparations, suppositories and sterilized injection solutions And can be used as formulations. Examples of carriers, excipients and diluents that can be included in the composition containing the extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose ), Lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, 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, glycerogelatin and the like can be used.
본 발명의 조성물은 상기 제조방법에 따라 얻어진 고로쇠 잎 추출물을 포함하는 항산화용 화장료 조성물을 제공한다. The composition of the present invention provides a cosmetic composition for antioxidation comprising the extract of ganoderma lucidum obtained according to the above production method.
본 발명의 조성물은 유해한 활성산소를 억제를 위한 화장품, 세안제 및 샴푸 등에 다양하게 이용될 수있다. 본 조성물을 첨가할 수 있는 제품으로는, 예를 들어, 각종 크림, 로션, 스킨 등과 같은 화장품류와 샴푸, 린스, 클렌징, 세안제, 비누, 트리트먼트, 미용액 등이 있다.The composition of the present invention can be variously used for cosmetics, cleansers and shampoos for inhibiting harmful active oxygen. Examples of products to which the present composition can be added include cosmetics such as various creams, lotions, and skins, and shampoos, rinses, cleansers, cleansers, soaps, treatments, and essences.
본 발명의 화장료는 수용성 비타민, 유용성 비타민, 고분자 펩티드, 고분자 다당, 스핑고 지질 및 해초 엑기스로 이루어진 군에서 선택된 조성물을 포함한다. The cosmetic composition of the present invention comprises a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, high molecular weight peptides, polymeric polysaccharides, sphingolipids and seaweed extracts.
본 발명의 화장료 조성물에 포함되는 성분은 유효성분으로서 상기 추출물 이외에 화장료 조성물에 통상적으로 이용되는 성분들을 포함할 수 있으며, 예를 들면, 안정화제, 용해화제, 비타민, 안료 및 향료와 같은 통상적인 보조제 및 담체를 포함한다.The ingredients contained in the cosmetic composition of the present invention may contain, in addition to the above-mentioned extracts, the components conventionally used in cosmetic compositions, such as stabilizers, solubilizers, vitamins, pigments and fragrances, And a carrier.
본 발명의 항산화용 화장료 조성물은 당업계에서 통상적으로 제조되는 어떠한 제형으로도 제조될 수 있으며, 예를들어 유액, 크림, 화장수, 팩, 파운데이션, 로션, 미용액, 모발화장료 등을 들 수 있다.The antioxidant cosmetic composition of the present invention can be prepared in any formulations conventionally produced in the art, and examples thereof include emulsions, creams, lotions, packs, foundations, lotions, essences, and hair cosmetics.
본 발명의 추출물의 바람직한 투여량은 환자의 상태 및 체중, 질병의 정도, 약물형태, 투여경로 및 기간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다. 그러나, 바람직한 효과를 위해서, 본 발명의 추출물은 1일 0.0001 내지 100mg/kg으로, 바람직하게는 0.001 내지 100mg/kg으로 투여하는 것이 좋다. 투여는 하루에 한번 투여할 수도 있고, 수회 나누어 투여할 수도 있다. 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다.The preferred dosage of the extract of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the type of drug, the administration route and the period of time, but can be appropriately selected by those skilled in the art. However, for the desired effect, the extract of the present invention is preferably administered at 0.0001 to 100 mg / kg per day, preferably 0.001 to 100 mg / kg per day. The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way.
본 발명의 추출물은 쥐, 생쥐, 가축, 인간 등의 포유동물에 다양한 경로로 투여될 수 있다. 투여의 모든 방식은 예상될 수 있는데, 예를 들면, 경구, 직장 또는 정맥, 근육, 피하, 자궁내 경막 또는 뇌혈관내(intracerebroventricular) 주사에 의해 투여될 수 있다.The extract of the present invention can be administered to mammals such as rats, mice, livestock, humans and the like in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine or intracerebroventricular injections.
본 발명은 항산화 효과를 나타내는 상기 추출물 및 식품학적으로 허용 가능한 식품보조 첨가제를 포함하는 건강기능식품을 제공한다. 고로쇠 잎 추출물을 첨가할 수 있는 식품으로는, 예를 들어, 각종 식품류, 음료, 껌, 차, 비타민 복합제, 건강 기능성 식품류 등이 있다.The present invention provides a health functional food comprising the above extract and a pharmaceutically acceptable food-aid additive exhibiting an antioxidative effect. Foods to which the gonorrhea leaf extract can be added include, for example, various foods, beverages, gums, tea, vitamin complexes, and health functional foods.
또한, 항산화 효과를 목적으로 식품 또는 음료에 첨가될 수 있다. 이 때, 식품 또는 음료 중의 상기 추출물의 양은 전체 식품 중량의 0.01 내지 15 중량%로 가할 수 있으며, 건강 음료 조성물은 100 ㎖를 기준으로 0.02 내지 5g, 바람직하게는0.3 내지 1g의 비율로 가할 수 있다.It can also be added to foods or beverages for the purpose of antioxidant effect. At this time, the amount of the extract in the food or drink may be 0.01 to 15% by weight of the total food, and the health beverage composition may be added in a proportion of 0.02 to 5 g, preferably 0.3 to 1 g, based on 100 ml .
본 발명의 건강 기능성 음료 조성물은 지시된 비율로 필수 성분으로서 상기 추출물을 함유하는 외에는 다른 성분에는 특별한 제한이 없으며 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상술한 천연 탄수화물의 예는 모노사카라이드, 예를 들어, 포도당, 과당 등; 디사카라이드, 예를 들어 말토스, 슈크로스 등; 및 폴리사카라이드, 예를 들어 덱스트린, 시클로덱스트린 등과 같은 통상적인 당, 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 상술한 것 이외의 향미제로서 천연 향미제(타우마틴, 스테비아 추출물(예를 들어 레바우디오시드 A, 글리시르히진등) 및 합성 향미제(사카린, 아스파르탐 등)를 유리하게 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 조성물 100 ㎖당 일반적으로 약 1 내지 20g, 바람직하게는 약 5 내지 12g이다.The health functional beverage composition of the present invention has no particular limitation on the other ingredients other than the above-mentioned extract as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavors (tau martin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.
상기 외에 본 발명의 추출물은 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 중진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산 음료에 사용되는 탄산화제 등을 함유할 수 있다. 그밖에 본 발명의 추출물들은 천연 과일 주스 및 과일 주스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율은 그렇게 중요하진 않지만 본 발명의 추출물 100 중량부 당 0 내지 약 20 중량부의 범위에서 선택되는 것이 일반적이다.In addition to the above, the extract of the present invention can also be used as a flavoring agent such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and aging agents (cheese, chocolate, etc.), pectic acid and its salts, Salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like. In addition, the extracts of the present invention may contain flesh for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components may be used independently or in combination. The proportion of such additives is not so critical, but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the extract of the present invention.
이하, 본 발명을 실시예에 의해 상세히 설명하기로 한다. 그러나 이들 실시예는 본 발명을 보다 구체적으로 설명하기 위한 것으로서, 본 발명의 범위가 이들 실시예에 한정되는 것은 아니다. Hereinafter, the present invention will be described in detail with reference to examples. However, these examples are intended to further illustrate the present invention, and the scope of the present invention is not limited to these examples.
<실시예 1> ≪ Example 1 >
고로쇠 잎 열수 추출물 및 에탄올 추출물의 제조방법Production method of hot-water extract and ethanol extract of gonorrhea leaf
건조된 고로쇠 잎의 30배 (wt/v%)에 해당하는 부피의 열수 및 70% 에탄올로 50℃에서 2시간씩 3번 환류추출하여 추출물을 얻었다. 각각의 열수 및 에탄올 추출물을 rotary evaporator (Ika, China)를 사용하여 60℃ 이하에서 감압농축하고, 이 농축액을 -80℃로 동결 건조하여 제조하였다. 최종 수율(yield)은 고로쇠 잎 열수 추출물이 17.93%, 고로쇠 잎 에탄올 추출물이 20.64%이다.Extracts were obtained by refluxing three times at 50 ° C for 2 hours with hot water and 70% ethanol corresponding to 30 times (wt / v%) of dried gonorrhea leaves. Each hot water and ethanol extract was concentrated under reduced pressure at 60 ° C or lower using a rotary evaporator (Ika, China), and the concentrate was lyophilized at -80 ° C. The final yield was 17.93% of hot water extract of ganoderma lucidum and 20.64% of ethanol extract of ganoderma lucidum.
<실시예 2> ≪ Example 2 >
고로쇠 잎 Leaf leaf 열수Heat number 추출물 및 에탄올 추출물의 페놀, 플라보노이드 및 아스코르빈산의 함량 확인 Determination of phenol, flavonoid and ascorbic acid content of extracts and ethanol extracts
상기 실시예 1에서 추출한 고로쇠 잎 열수 추출물 및 에탄올 추출물의 Phenolic, Flavonoid 함량 및 수율 (yield)은 하기 표 1에 나타낸 바와 같이, 최종 수율은 고로쇠 잎 열수 추출물이 17.93%, 고로쇠 잎 에탄올 추출물이 20.64%이다.The phenolic and flavonoid contents and yields of the hot-water extract and the ethanol extract of the ganoderma lucidum obtained in Example 1 were 17.93% and 20.64%, respectively, as shown in Table 1, to be.
페놀함량은 각각 1387.20 ± 23.62 mg, 546.10 ± 7.54 mg GAE/g of dry mass로 나타났고, 플라보노이드 함량은 각각 2258.32 ± 153.67 mg , 3064.48 ± 37.08 mg CE/g of dry mass로 매우 높은 함량을 나타났다. 고로쇠 잎 추출물의 수율은 낮은 편이지만 페놀, 플라보노이드, 아스코르빈산의 함량이 매우 높음을 확인할 수 있었다. Phenol contents were 1387.20 ± 23.62 mg and 546.10 ± 7.54 mg GAE / g of dry mass respectively. The content of flavonoid was 2258.32 ± 153.67 mg and 3064.48 ± 37.08 mg CE / g of dry mass respectively. The yield of phenolic compounds, phenolic compounds, flavonoids and ascorbic acid was very high.
<실시예 3>≪ Example 3 >
고로쇠 잎 열수 추출물 및 에탄올 추출물의 DPPH 라디칼 소거능 측정Determination of DPPH Radical Scavenging Ability of Hot Water Extract and Ethanol Extract of Panax ginseng CA Meyer
상기 실시예 1에서 추출한 고로쇠 잎 열수 추출물 및 에탄올 추출물을 10mg/ml의 농도로 만든 후 0.25, 0.5, 1, 2, 4 mg/ml의 농도로 희석하여 각각 1ml씩 제조하고, standard로 사용되는 BHT 10mg/ml를 메탄올(methanol) 1ml로 희석하여서 샘플과 같은 농도로 1ml씩 제조하였다.The hot-water extracts and the ethanol extracts of the hot pepper leaves obtained in Example 1 were diluted to a concentration of 10 mg / ml, and diluted to 0.25, 0.5, 1, 2 and 4 mg / ml, 10 mg / ml was diluted with 1 ml of methanol and 1 ml was prepared at the same concentration as the sample.
농도별로 만든 고로쇠 잎 추출물과 BHT를 96 well plate에 각각 80 ㎕씩 분주한 후 DW, methanol, DPPH 순서로 80 ㎕씩 분주한 후 빛과 반응하지 못하게 foil로 감싸준 뒤 실온에서 30분간 방치한 후 ELISA reader를 이용해 517nm 흡광도를 측정하였다. 80 ㎕ of each leaf extract and BHT were added to each well of a 96 well plate, and then 80 ㎕ of each of them was added to each well in the order of DW, methanol and DPPH. The plate was wrapped with foil so that it could not react with light, and then allowed to stand at room temperature for 30 minutes. absorbance at 517 nm was measured using a reader.
DPPH 라디칼 소거능 계산식은 아래와 같다. The calculation formula of DPPH radical scavenging ability is as follows.
Scavenging activity (%) = (1- Absorbance of sample / Absorbance of control) ×100Scavenging activity (%) = (1- Absorbance of sample / Absorbance of control) × 100
고로쇠 잎 열수 추출물 및 에탄올 추출물의 DPPH 라디칼 소거능을 측정한 결과, 고로쇠 잎 열수 및 70% 에탄올 추출물은 standard인 BHT와 비교하여 우수한 효과를 가짐을 알 수 있었다(도 1 참조).As a result of measuring the DPPH radical scavenging ability of the hot-water extract and the ethanol extract of the gonorrhea leaf, it was found that the hot-water and 70% ethanol extract of the gonorrhea leaf had an excellent effect as compared with the standard BHT (see FIG. 1).
항산화 효과 비교를 위해 계산된 IC50 값을 하기 표2에 나타내었다. 고로쇠 잎 추출물의 DPPH 라디컬 소거 활성에 대해 계산된 IC50 값은 열수 추출물은 0.051 mg/ml, 70% 에탄올 추출물은 0.027 mg/ml로, standard로 사용한 BHT (0.021 mg/ml)과 비슷한 항산화 효과를 가진 것을 알 수 있었다(표 2 참조).The IC 50 values calculated for the comparison of antioxidant effects are shown in Table 2 below. The calculated IC 50 values for the DPPH radical scavenging activity of ganoderma lucidum extract were 0.051 mg / ml for the hot water extract and 0.027 mg / ml for the 70% ethanol extract. The antioxidative effect similar to that of the standard BHT (0.021 mg / ml) (See Table 2).
<실시예 4><Example 4>
고로쇠 잎 열수 추출물 및 에탄올 추출물의 ABTS 라디칼 소거능 측정Measurement of ABTS radical scavenging ability of hot-water extract and ethanol extract of gonorrhoeae leaf
상기 실시예 1에서 추출한 고로쇠 잎 열수 추출물 및 에탄올 추출물 10mg/ml의 농도로 만든 후 0.25, 0.5, 1, 2, 4 mg/ml의 농도로 희석하여 각각 1ml씩 제조하고, standard로 사용되는 아스코르브산(Ascorbic acid) 10mg/ml를 증류수 1ml로 희석하여서 샘플과 같은 농도로 1ml씩 제조하였다. The hot-water extract and the ethanol extract of the gonorrhoeae extract prepared in Example 1 were diluted to a concentration of 10 mg / ml, and diluted to a concentration of 0.25, 0.5, 1, 2, and 4 mg / ml to prepare 1 ml of ascorbic acid Ascorbic acid (10 mg / ml) was diluted with 1 ml of distilled water and 1 ml was prepared at the same concentration as the sample.
증류수에 용해한 7mM의 ABTS와 2.45mM의 Potassium persulphate을 혼합한 후, 안정화를 위해 암실상태에서 12-16시간 방치하였다. 이 용액은 상온에서 2일간 사용가능하다. 734nm에서 위의 혼합물이 0.70 ± 0.02이 되도록 PBS(0.01M, pH 7.4)로 희석하였다. 그리고 나서 고로쇠 잎 추출물과 ascorbic acid standard 를 각각 0.3 ㎖ 을 0.7 ㎖ of ABTS+ solution과 혼합 후 상온에서 5분 반응 시킨 뒤 blank에 대하여 734nm로 측정하였다. 대조군은 시료의 추출물 대신 용매가 사용되었으며, ABTS 라디칼의 소거 활성은 다음과 같이 계산 하였다.After mixing 7 mM ABTS and 2.45 mM Potassium persulphate dissolved in distilled water, they were allowed to stand for 12-16 hours in a dark room for stabilization. This solution can be used at room temperature for 2 days. At 734 nm, the above mixture was diluted with PBS (0.01 M, pH 7.4) to 0.70 + 0.02. Then, 0.3 ml of ascorbic acid standard and 0.70 ml of ABTS + solution were mixed with each other and incubated at room temperature for 5 minutes. Then, the blank was measured at 734 nm. In the control group, solvents were used instead of the extract of the sample, and the scavenging activity of the ABTS radical was calculated as follows.
Scavenging activity (%) = (1- Absorbance of sample / Absorbance of control) ×100Scavenging activity (%) = (1- Absorbance of sample / Absorbance of control) × 100
고로쇠 잎 열수 추출물 및 에탄올 추출물의 ABTS 라디칼 소거능을 측정한 결과, 고로쇠 잎 열수 및 70% 에탄올 추출물은 standard인 Ascorbic acid와 비교하여 우수한 효과를 가짐을 알 수 있었다(도 2 참조).As a result of measuring the ABTS radical scavenging ability of the hot-water extract and the ethanol extract of the gonorrhoeae leaf, it was found that the hot-water and 70% ethanol extract of gonorrhoeae had a superior effect compared with the standard Ascorbic acid (see FIG. 2).
또한, 표 2에서 IC50 값을 보면 열수 추출물은 0.018 mg/ml, 70% 에탄올 추출물은 0.004 mg/ml로, standard로 사용한 Ascorbic acid (0.015 mg/ml)와 비교했을 때 에탄올 추출물은 standard보다 더 항산화 효과가 좋음을 알 수 있었다. In addition, the IC 50 values in Table 2 are 0.018 mg / ml for the hot-water extract and 0.004 mg / ml for the 70% ethanol extract, compared to the standard Ascorbic acid (0.015 mg / ml) Antioxidant effect was good.
<실시예 5>≪ Example 5 >
고로쇠 잎 열수 추출물 및 에탄올 추출물의 환원력 측정Reducing power of hot-water extract and ethanol extract of gonorrhea leaf
환원력 측정은 Chung등에(Chung et al. 2005)의해 기술된 방법에 따라 약간 변형하여 실행하였다. 0.25, 0.5, 1, 2, 4 mg/ml 다양한 농도의 고로쇠 잎 추출물 또는 standard인 BHT, Ascorbic acid 0.5ml와 0.5ml의 Sodium Phosphate buffer(PBS)와 1% 페리시안화 칼륨(Potassium Ferricyanide)을 0.5ml를 각각 혼합 시킨 후 50℃에서 20분 동안 정치한 다음 반응이 끝난 혼합물을 각각 1ml씩 따서 새 tube로 옮긴 뒤 Trichloroacetic acid(TCA) 0.5ml를 첨가하여 centrifuge에서 3,000rpm으로 4℃에서 10분간 원심분리 하였다. Reduction power measurements were performed with slight modifications according to the method described by Chung et al. (Chung et al. 2005). BHT, 0.5 ml of Ascorbic acid, 0.5 ml of Sodium Phosphate buffer (PBS) and 1% Potassium Ferricyanide in 0.5 ml, 0.25, 0.5, 1, 2 and 4 mg / And then incubated at 50 ° C for 20 minutes.
원심분리 한 상층액 0.5ml에 DW 0.5ml와 염화제2철(Ferric chloride) 0.1ml를 첨가한 후 96 well plate에 0.15ml씩 분주 해준 뒤 ELISA reader를 이용해 700nm로 흡광도를 측정하였다. 측정한 값을 그래프로 나타내었다.0.5 ml of DW and 0.1 ml of ferric chloride were added to 0.5 ml of the centrifuged supernatant, and 0.15 ml of the solution was dispensed into a 96-well plate. The absorbance was measured at 700 nm using an ELISA reader. The measured values are shown in a graph.
고로쇠 잎 열수 추출물 및 에탄올 추출물의 환원력 측정한 결과, 고로쇠 잎 열수 및 70% 에탄올 추출물은 standard인 Ascorbic acid와 BHT에 비교하여 우수한 효과를 가짐을 알 수 있었다(도 3 참조).As a result of the measurement of reducing power of hot pepper leaves and ethanol extracts of gonorrhea leaf, it was found that the hot water and 70% ethanol extract of gonorrhoeae had a superior effect compared with the standard Ascorbic acid and BHT (see FIG. 3).
<실시예 6>≪ Example 6 >
고로쇠 잎 열수 추출물 및 에탄올 추출물의 DNA 산화적 손상 정도 확인Identification of oxidative damage of DNA of hot-water extract and ethanol extract of gonorrhoeae leaf
H2O2에 의한 DNA의 산화적 손상에 대한 예방은 Tian &Hua (2005)의 실험방법을 약간 수정하여 실시하였다. 플라스미드 PBR 322 DNA (0.5 μg/μL) 1 μL은 FeSO4의 2 μL (0.08 ㎜), 30 % H2O2 3 μL (v / v)의 3 μL 증류수와 서로 다른 농도의 추출물 2 μL의 혼합물을 1시간 동안 37 ℃에서 반응시켰다. Prevention of oxidative damage of DNA by H 2 O 2 was carried out with slightly modified experimental method of Tian & Hua (2005). 1 μL of plasmid PBR 322 DNA (0.5 μg / μL) was mixed with 2 μL (0.08 ㎜) of FeSO 4 , 3 μL of 30 μL H 2 O 2 (v / v) distilled water and 2 μL of different concentrations of extract Was reacted at 37 ° C for 1 hour.
그 후 6X DNA loading dye 2 μL를 첨가한 뒤 실온에서 70V의 0.8% 아가로스겔전기영동(agarose gel electrophoresis) 시켰다. DNA 밴드 (supercoiled, linear and open circular)는 에티디움 브로마이드(ethidium bromide)로 염색시켰고, DNA 산화를 억제하는 효과는 control 값을 기준으로 한 supercoiled monomer의 증가와 감소로 평가하였다. Subsequently, 2 μL of 6X DNA loading dye was added, followed by 0.8% agarose gel electrophoresis at 70V at room temperature. DNA bands (supercoiled, linear and open circular) were stained with ethidium bromide. The effect of inhibiting DNA oxidation was evaluated by the increase and decrease of supercoiled monomer based on control values.
고로쇠 잎 추출물에 DNA 산화적 손상을 보호하는 효과가 있는지 여부를 확인한 결과, 대조군에 비하여 손상되었던 초나선 DNA가 선형형태로 변환되었음을 알 수 있었다(Lane 2). 또한, DNA는 열수 추출물과 70% 에탄올 추출물 처리되었을 때 Supercoiled의 DNA는 용량 의존적으로 복원되었음을 알 수 있었다(Lanes 3-5). As a result, it was found that the superhelical DNA, which was damaged compared with the control, was converted into a linear form (Lane 2). In addition, the DNA was recovered in a dose-dependent manner when the hot-water extract and the 70% ethanol extract were treated (Lanes 3-5).
따라서, 고로쇠 열수 추출물과 70% 에탄올 추출물 모두에서 DNA 손상에 대한 보호 효과가 있음을 확인할 수 있었고, 그 중 열수 추출물에서 DNA 손상에 대하여 더 유의적인 보호 효과를 보임을 알 수 있었다. Therefore, it was confirmed that both of the hot water extract and the 70% ethanol extract of the present invention had a protective effect against DNA damage, and the hot water extract showed a more significant protection effect against DNA damage.
<실시예 7>≪ Example 7 >
통계처리Statistical processing
모든 데이터는 평균± 표준 편차(SDM)로 제시하였다. 모든 실험은 3회 반복 실시하였다. 각 군 간의 측정치 비교는 one-way analysis of variance (ANOVA) test를 실시한 후 p < 0.05의 유의수준에서 차이를 검증하였다. GraphPad Prism 5 와 Microsoft Excel 2007은 그래프의 평가와 통계에 사용하였다. All data are presented as means ± SD (SDM). All experiments were repeated three times. The differences between the groups were analyzed by one-way analysis of variance (ANOVA).
이제까지 본 발명에 대하여 그 바람직한 실시예들을 중심으로 살펴보았다. 본 발명이 속하는 기술 분야에서 통상의 지식을 가진 자는 본 발명이 본 발명의 본질적인 특성에서 벗어나지 않는 범위에서 변형된 형태로 구현될 수 있음을 이해할 수 있을 것이다. 그러므로 개시된 실시예들은 한정적인 관점이 아니라 설명적인 관점에서 고려되어야 한다. 본 발명의 범위는 전술한 설명이 아니라 특허 청구범위에 나타나 있으며, 그와 동등한 범위 내에 있는 모든 차이점은 본 발명에 포함된 것으로 해석되어야 할 것이다.The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.
Claims (5)
상기 추출물은 물, 메탄올, 에탄올, 부탄올 또는 아세톤에 가용한 추출물인 것을 특징으로 하는 조성물.The method according to claim 1,
Wherein the extract is an extract soluble in water, methanol, ethanol, butanol or acetone.
상기 조성물은 활성산소종을 제거하거나, 환원력을 가지는 것을 특징으로 하는 조성물.The method according to claim 1,
Wherein the composition removes active oxygen species or has a reducing power.
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KR20120006779A (en) | 2010-07-13 | 2012-01-19 | 경북대학교 산학협력단 | Composition comprising the extract of acer mono max. for the prevention or treatment of gastritis or gastric ulcer |
KR20130049043A (en) | 2011-11-03 | 2013-05-13 | 안동대학교 산학협력단 | Pharmaceutical composition comprising the extract of betel nut of areca catechu l. for prevention and control of thrombosis |
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KR20130049043A (en) | 2011-11-03 | 2013-05-13 | 안동대학교 산학협력단 | Pharmaceutical composition comprising the extract of betel nut of areca catechu l. for prevention and control of thrombosis |
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