KR20180102415A - A compostion comprising Torreya nucifera seed extract or its fraction having anti-oxidation or anti-inflammation activity - Google Patents
A compostion comprising Torreya nucifera seed extract or its fraction having anti-oxidation or anti-inflammation activity Download PDFInfo
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- KR20180102415A KR20180102415A KR1020170029023A KR20170029023A KR20180102415A KR 20180102415 A KR20180102415 A KR 20180102415A KR 1020170029023 A KR1020170029023 A KR 1020170029023A KR 20170029023 A KR20170029023 A KR 20170029023A KR 20180102415 A KR20180102415 A KR 20180102415A
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Abstract
Description
본 발명은 비자 열매(Torreya nucifera seed) 추출물 또는 이의 분획물을 포함하는 항산화 또는 항염증용 조성물에 관한 것으로서, 보다 구체적으로 비자 열매 추출물 또는 이의 분획물을 유효성분으로 포함하는 항산화 또는 항염증용 화장료 조성물, 식품 조성물 및 약학 조성물에 관한 것이다.The present invention relates to an antioxidant or antiinflammatory composition comprising a Torreya nucifera seed extract or a fraction thereof, and more particularly, to a cosmetic composition for antioxidant or antiinflammation which comprises an extract of Visa or a fraction thereof as an active ingredient, Food compositions and pharmaceutical compositions.
인체는 산화촉진물질과 산화억제물질이 균형을 이루고 있으나, 여러 가지 요인들로 인하여 이러한 균형 상태를 잃고 산화를 촉진하는 방향으로 기울게 되면, 생체 내에 산화적 스트레스(oxidative stress)가 유발되어 세포손상 및 병리적 질환을 유발하게 된다. 이러한 산화적 스트레스의 직접적 원인이 되는 활성 산소종 (reactive oxygen species, ROS)은 화학적으로 불안정하고 반응성이 높아 DNA, 단백질, 지질 및 탄수화물과 같은 여러 생체물질과 쉽게 반응할 수 있으며, 생체 내 고분자들을 공격하여 세포와 조직에 비가역적인 손상을 일으키거나 돌연변이, 세포독성 및 암 등을 초래하게 된다.Although the human body has a balance of the oxidation promoting substance and the oxidation inhibiting substance, when the balance state is lost due to various factors and is inclined to promote the oxidation, oxidative stress is induced in the living body, Leading to pathological disease. Reactive oxygen species (ROS), which is a direct cause of oxidative stress, are chemically unstable and highly reactive, and can easily react with various biomaterials such as DNA, protein, lipid and carbohydrate. Attacks, causing irreversible damage to cells and tissues, or causing mutations, cytotoxicity, and cancer.
한편, 인체 내에서 대식세포는 병원체에 반응하여 종양 괴사 인자-α(tumor necrotic factor-α, TNF-α), 인터루킨-6(IL-6), 인터루킨-1β(IL-1β) 등과 같은 염증유발인자를 생성하고, 유도성 일산화질소 합성효소(inducible nitric oxide synthase, iNOS)와 사이클로옥시게나제-2(cyclooxygenase-2, COX-2)를 합성하여 일산화질소(NO) 및 프로스타글란딘(prostaglandin E2, PGE2)을 생성한다. 생리학적으로 NO는 세균과 종양을 제거하고 혈압을 조절하거나 신경 전달을 매개하는 등 다양한 역할을 한다. 그러나 염증 반응이 일어나게 되면 관련 세포들에서 iNOS의 발현이 증가하여 많은 양의 NO가 생성되고, 과도하게 생성된 NO는 조직의 손상, 유전자 변이, 신경 손상 등을 유발하며, 혈관 투과성을 증가시켜 부종 등의 염증 반응을 촉진시킨다.In the human body, macrophages respond to pathogens and cause inflammation such as tumor necrotic factor-α, TNF-α, interleukin-6 (IL-6), and interleukin-1β (NO) and prostaglandins (prostaglandin E2, PGE2) by synthesizing inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) ). Physiologically, NO plays a variety of roles such as removing bacteria and tumors, regulating blood pressure, or mediating neurotransmission. However, when the inflammatory reaction occurs, the expression of iNOS is increased in related cells, and a large amount of NO is produced. Excessively produced NO causes tissue damage, gene mutation, nerve damage, etc., And promote the inflammatory response.
염증 반응시에는 다양한 염증유발인자와 더불어 다양한 자유 라디칼(free radical)이 생성된다. 정상적인 자유 라디칼은 세포의 항상성 유지에 관여하며, 세포의 분화, 성장, 생존, 노화에 영향을 준다. 이러한 자유 라디칼 중 활성 산소종(reactive oxygen species, ROS)은 세포 내의 미토콘드리아에서 호흡과 면역 반응에 의한 산소의 산화와 환원 과정을 통해 끊임없이 생성된다. 유해한 ROS는 일반적으로 슈퍼옥사이드 디스뮤타제(superoxide dismutase), 카탈라제(catalase), 글루타치온 퍼옥시다제(glutathione peroxidase), 글루타민 리덕타제(glutamine reductase), 비타민 C, 비타민 E, 요산, 빌리루빈(bilirubin)과 같은 항산화 체계의 작용으로 제거된다. 그러나 자유 라디칼의 생성과 소거의 균형이 무너지게 되면 산화적 스트레스(oxidative stress)가 발생하게 되어 염증, 노화 또는 암과 같은 다양한 병리학적인 변화를 일으킨다. 실제로 많은 임상 질환에서 산화적 스트레스가 증가되는 것이 확인되었으며, 이러한 산화적 스트레스를 줄이기 위해 새로운 항산화 물질에 대한 다양한 연구가 이루어지고 있다.In the inflammatory reaction, various free radicals are produced along with various inflammation inducers. Normal free radicals are involved in maintaining the homeostasis of cells and affect cell differentiation, growth, survival, and aging. This reactive oxygen species (ROS) in the free radicals is produced constantly through oxidation and reduction of oxygen by respiration and immune reaction in mitochondria within the cell. Harmful ROS are usually caused by superoxide dismutase, catalase, glutathione peroxidase, glutamine reductase, vitamin C, vitamin E, uric acid, bilirubin, It is eliminated by the action of the same antioxidant system. However, when the balance between the generation and elimination of free radicals collapses, oxidative stress occurs, leading to various pathological changes such as inflammation, aging, or cancer. In fact, it has been confirmed that oxidative stress is increased in many clinical diseases. Various studies on new antioxidants have been made to reduce such oxidative stress.
이에, 본 발명자들은 항산화 또는 항염증 활성을 가지는 동시에 부작용이 적은 천연물 유래 소재를 개발하고자 예의 연구 노력한 결과, 비자 열매 추출물 또는 이의 분획물에서 자유 라디칼 소거능 및 NO 생성 저해능을 확인하여 본 발명을 완성하였다.Accordingly, the inventors of the present invention have made efforts to develop natural material-derived materials having antioxidant or anti-inflammatory activity and low side effects, and as a result, they have confirmed free radical scavenging ability and NO production inhibitory ability in visa fruit extract or its fractions.
본 발명의 하나의 목적은 비자 열매(Torreya nucifera seed) 추출물 또는 이의 분획물을 유효성분으로 포함하는 항산화 또는 항염증용 조성물을 제공하는 것이다.It is an object of the present invention to provide an antioxidant or anti-inflammatory composition comprising an extract of Torreya nucifera seed or a fraction thereof as an active ingredient.
본 발명의 다른 목적은 상기 비자 열매 추출물 또는 이의 분획물을 유효성분으로 포함하는 항산화 또는 항염증용 화장료 조성물을 제공하는 것이다.Another object of the present invention is to provide a cosmetic composition for antioxidant or anti-inflammation comprising the above visa-fruit extract or a fraction thereof as an active ingredient.
본 발명의 또 다른 목적은 상기 비자 열매 추출물 또는 이의 분획물을 유효성분으로 포함하는 항산화 또는 항염증용 식품 조성물을 제공하는 것이다.Yet another object of the present invention is to provide a food composition for antioxidant or antiinflammation comprising the above visa-fruit extract or a fraction thereof as an active ingredient.
본 발명의 또 다른 목적은 상기 비자 열매 추출물 또는 이의 분획물을 유효성분으로 포함하는 항산화 또는 항염증용 약학 조성물을 제공하는 것이다.It is still another object of the present invention to provide a pharmaceutical composition for antioxidant or antiinflammation which comprises the above-mentioned Viscumin fruit extract or a fraction thereof as an active ingredient.
상기 목적을 달성하기 위한 하나의 양태로서, 본 발명은 비자 열매(Torreya nucifera seed) 추출물 또는 이의 분획물을 유효성분으로 포함하는, 항산화 또는 항염증용 조성물을 제공한다. In one aspect, the present invention provides an antioxidative or anti-inflammatory composition comprising an extract of Torreya nucifera seed or a fraction thereof as an active ingredient.
상기 조성물은 화장료 조성물, 식품 조성물, 약학 조성물일 수 있으나 이에 제한되지 않는다.The composition may be a cosmetic composition, a food composition, or a pharmaceutical composition, but is not limited thereto.
본 발명의 용어, “비자 열매(Torreya nucifera seed)”는 구과목 주목과의 비자나무의 열매로서, 기생충을 구제하고 대변이 잘 나오게 하며 기침을 멎게 하는 것으로 알려져 있다.The term " Torreya nucifera seed " of the present invention is known as a fruit of visa trees with a focus on old subjects, which rescues parasites, makes stools come out well, and stops coughing.
본 발명의 용어 "추출물"은 생약을 적절한 침출액으로 짜내고 침출액을 증발시켜 농축한 제제를 의미하는 것으로, 이에 제한되지는 않으나, 추출처리에 의해 얻어지는 추출액, 추출액의 희석액 또는 농축액, 추출액을 건조하여 얻어지는 건조물, 이들의 조정제물 또는 정제물일 수 있다. 본 발명의 추출물은 추출용매로 추출하거나 추출용매로 추출하여 제조한 추출물에 분획용매를 가하여 분획함으로써 제조할 수 있다. 상기 추출용매는 이에 제한되지는 않으나, 물, 유기용매 또는 이들의 혼합용매 등을 사용할 수 있으며, 상기 유기용매는 탄소수 1 내지 4의 알코올이나, 에틸아세테이트 또는 아세톤 등의 극성용매, 헥산 또는 디클로로메탄의 비극성 용매 또는 이들의 혼합용매를 사용할 수 있다. 또한, 구체적으로 물, 탄소수 1 내지 4의 알코올 또는 이들의 혼합용매를 사용할 수 있다. The term "extract " of the present invention means a preparation which is obtained by squeezing a herbal medicine with an appropriate leaching solution and concentrating the liquid by evaporating the leaching solution, and is not limited thereto. The extract is obtained by drying the extract, diluted solution, Dried products, controlled products thereof or purified products thereof. The extract of the present invention can be prepared by extracting with an extraction solvent or extracting with an extraction solvent and fractionating the extract with a fraction solvent. The extraction solvent may be, but is not limited to, water, an organic solvent or a mixed solvent thereof. The organic solvent may be an alcohol having 1 to 4 carbon atoms, a polar solvent such as ethyl acetate or acetone, hexane or dichloromethane Non-polar solvents or mixed solvents thereof may be used. Specifically, water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof may be used.
상기 비자 열매 추출물은 당업계에 공지된 일반적인 추출방법, 분리 및 정제방법을 이용하여 제조할 수 있다. 상기 추출방법으로는, 이에 제한되지는 않으나, 구체적으로 열탕 추출, 열수 추출, 냉침 추출, 환류 냉각 추출 또는 초음파 추출 등의 방법을 사용할 수 있다.The visceral fruit extract can be produced by a common extraction method, separation and purification method known in the art. Examples of the extraction method include, but are not limited to, hot-water extraction, hot water extraction, cold extraction, reflux cooling extraction, or ultrasonic extraction.
본 발명의 용어 "항산화"는 산화를 억제하는 작용을 의미하는 것으로, 인체는 산화촉진물질(prooxidant)과 산화억제물질(antioxidant)이 균형을 이루고 있으나 여러 가지 요인들에 의하여 이런 균형상태가 불균형을 이루게 되고 산화촉진 쪽으로 기울게 되면, 산화적 스트레스(oxidative stress)가 유발되어 잠재적인 세포손상 및 병리적 질환을 일으키게 된다. 이러한 산화적 스트레스의 직접적 원인이 되는 활성 산소종(reactive oxygen species, ROS)은 불안정하고 반응성이 높아 여러 생체물질과 쉽게 반응하고, 체내 고분자들을 공격하여 세포와 조직에 비가역적인 손상을 일으키거나 돌연변이, 세포독성 및 발암 등을 초래하게 된다. NO, HNO2, ONOO-와 같은 활성 질소종(reactive nitrogen species, RNS)은 염증 반응 시 대식세포 호중구 및 다른 면역 세포 들의 면역반응으로 인해 다량 생성되며, 이때 ROS도 같이 생성된다. 상기와 같은 활성산소는 체내에서 세포를 산화시켜 파괴시키며, 그에 따라 각종 질환에 노출되게 된다. 따라서, 본 발명의 비자 열매 추출물 또는 이의 분획물을 화장품에 포함시키면 항산화 효과를 달성함으로써, 건강증진에 기여할 수 있다.The term "antioxidant" of the present invention means an antioxidant action. The human body has a balance between prooxidant and antioxidant, but due to various factors, And leaning towards oxidative stimulation leads to oxidative stress leading to potential cellular damage and pathological disease. Reactive oxygen species (ROS), which is a direct cause of oxidative stress, is unstable and highly reactive. It reacts easily with various bio-materials, attacks irreversible damage to cells and tissues, Cytotoxicity and carcinogenesis. Active nitrogen species (RNS) such as NO, HNO 2 , and ONOO - are produced by the immune response of macrophage neutrophils and other immune cells during inflammatory reaction, and ROS is also produced. Such active oxygen oxidizes and destroys cells in the body, thereby exposing them to various diseases. Therefore, when the visa fruit extract or the fraction thereof of the present invention is contained in cosmetics, the antioxidative effect can be attained, thereby contributing to health promotion.
본 발명의 일실시예에서는 비자 열매 추출물 또는 이의 분획물의 DPPH 라디칼 소거활성(표 2 및 표 3)을 분석하여 강한 항산화능을 갖는 것을 확인하여, 본 발명의 항산화 효과는 자유 라디칼 소거능에 의해 달성되는 것일 수 있다.In one embodiment of the present invention, the DPPH radical scavenging activity (Tables 2 and 3) of the non-viscous fruit extract or its fractions was analyzed to confirm that it has a strong antioxidant ability. Thus, the antioxidative effect of the present invention is attained by free radical scavenging ability Lt; / RTI >
특히, 비자열매 메탄올 추출물의 에틸아세테이트 분획물이 다른 분획물에 비해, 현저히 우수한 항산화능을 갖는 것을 확인하였다.In particular, it was confirmed that the ethyl acetate fraction of the non-viscous methanol extract had remarkably excellent antioxidant ability as compared with the other fractions.
본 발명의 용어 "항염증"은 염증을 억제하는 작용을 의미하는 것으로, 염증반응의 조절은 대단히 복잡한 것으로 알려져 있는데, 이는 생체 내 복구체계의 증강 및 손상을 감소시키기 위한 것으로 알려져 있다. 그러나 반복되는 조직의 손상이나 재생에 의해 염증반응이 지속되면, 염증관련 세포에서 ROS와 RNS가 과다 생성되고 그 결과로 영구적인 유전자의 변형이 야기된다. 이처럼 ROS와 RNS는 생체 내 여러 가지 세포의 작용을 조절하는 염증 반응과 깊이 관련되어 있다. 염증 과정 중에는 많은 양의 염증유도 사이토카인(proinflammatory cytokines), 아질산(nitric oxide, NO) 그리고 프로스타글란딘(prostaglandin E2, PGE2)이 유도성 일산화질소 합성효소(inducible nitric oxide synthase, iNOS)와 사이클로옥시게나아제(cyclooxygenase-2, COX-2)에 의해 생성된다. 염증은 다양한 염증성 질환을 유발하는 원인으로써, 본 발명의 비자 열매 추출물 또는 이의 분획물을 포함하는 조성물은 항염증 작용을 통해 다양한 염증성 질환에 대한 예방 및 개선 효과를 가질 수 있다.The term "anti-inflammation" of the present invention means an action to suppress inflammation. It is known that the control of the inflammatory reaction is extremely complicated, which is known to reduce the enhancement and damage of the in vivo restoration system. However, if the inflammatory response is sustained by repeated tissue damage or regeneration, ROS and RNS are overproduced in inflammation-related cells, resulting in permanent gene deformation. Thus, ROS and RNS are deeply involved in the inflammatory response that regulates the action of various cells in vivo. During inflammation, large amounts of proinflammatory cytokines, nitric oxide (NO), and prostaglandin (prostaglandin E2, PGE2) are induced by inducible nitric oxide synthase (iNOS) and cyclooxygenase (cyclooxygenase-2, COX-2). The inflammation is a cause of various inflammatory diseases. The composition comprising the visa fruit extract or the fractions thereof according to the present invention may have anti-inflammatory action to prevent and improve various inflammatory diseases.
본 발명의 일실시예에서는 비자 열매 추출물 또는 이의 분획물의 NO 생성 저해능(도 2)을 분석한 결과, 유의적인 NO 생성 저해능을 확인함으로써, 항염증 효과를 갖는 것을 확인하였다. 따라서, 본 발명의 항염증 효과는 NO 생성 저해능에 의해 달성되는 것일 수 있다. In one embodiment of the present invention, the inhibitory effect on the NO production (FIG. 2) of the non-visceral fat extract or its fractions was analyzed. Therefore, the anti-inflammatory effect of the present invention may be achieved by inhibiting NO production.
특히, 비자열매 메탄올 추출물의 디클로로메탄 분획물이 다른 분획물에 비해 현저히 우수한 항염증 효과를 갖는 것을 확인하였다.In particular, it was confirmed that the dichloromethane fraction of non-viscous methanol extract had remarkably excellent anti-inflammatory effect as compared with other fractions.
다른 하나의 양태로서, 본 발명은 비자 열매 추출물 또는 이의 분획물을 유효성분으로 포함하는 항산화 또는 항염증용 화장료 조성물을 제공한다.In another aspect, the present invention provides a cosmetic composition for antioxidant or anti-inflammation comprising visa-fruit extract or a fraction thereof as an active ingredient.
본 발명의 용어 "화장료 조성물"은 일반적인 유화 제형 및 가용화 제형의 형태로 제조할 수 있다. 상기 유화 제형으로는 영양화장수, 크림, 에센스 등이 있으며, 상기 가용화 제형으로는 유연화장수 등이 있다. 적합한 제형은 이에 제한되지는 않으나, 예를 들어 용액, 겔, 고체 또는 반죽 무수 생성물, 수상에 유상을 분산시켜 얻은 에멀젼, 현탁액, 마이크로에멀젼, 마이크로캡슐, 미세과립구 또는 이온형(리포좀), 바이온형의 소낭 분산제의 형태, 크림, 스킨, 로션, 파우더, 연고, 스프레이 또는 콘실 스틱의 형태일 수 있다. 또한, 포말(foam)의 형태 또는 압축된 추진제를 더 함유한 에어로졸 조성물의 형태일 수 있다.The term "cosmetic composition" of the present invention can be prepared in the form of a general emulsified formulation and a solubilized formulation. Examples of the emulsified formulations include nutritive lotions, creams, essences, and the like, and the solubilization formulations include softening longevity. Suitable formulations include, but are not limited to, solutions, gels, solid or paste anhydrous products, emulsions obtained by dispersing the oil phase in water, suspensions, microemulsions, microcapsules, microgranules or ionic (liposomes) A cream, a skin, a lotion, a powder, an ointment, a spray, or a conical stick. It may also be in the form of a foam or in the form of an aerosol composition further containing a compressed propellant.
상기 화장료 조성물은 추가적으로 지방 물질, 유기 용매, 용해제, 농축제 및 겔화제, 연화제, 항산화제, 현탁화제, 안정화제, 발포제, 방향제, 계면활성제, 물, 이온형 또는 비이온형 유화제, 충전제, 금속이온봉쇄제, 킬레이트화제, 보존제, 비타민, 차단제, 습윤화제, 필수 오일, 염료, 안료, 친수성 또는 친유성 활성제, 지질 소낭 또는 화장료 조성물에 통상적으로 사용되는 임의의 다른 성분과 같은 통상적으로 사용되는 보조제를 함유할 수 있다.The cosmetic composition may further contain at least one selected from the group consisting of fatty substances, organic solvents, solubilizers, thickening and gelling agents, softening agents, antioxidants, suspending agents, stabilizers, foaming agents, perfumes, surfactants, water, ionic or nonionic emulsifiers, Auxiliaries such as ionic sequestering agents, chelating agents, preservatives, vitamins, blocking agents, moisturizers, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or any other ingredients conventionally used in cosmetic compositions ≪ / RTI >
또 다른 하나의 양태로서, 본 발명은 비자 열매 추출물 또는 이의 분획물을 유효성분으로 포함하는 항산화 또는 항염증용 식품 조성물을 제공한다.In another aspect, the present invention provides a food composition for antioxidant or antiinflammation comprising visa fruit extract or a fraction thereof as an active ingredient.
본 발명의 조성물을 식품 첨가물로 사용할 경우, 상기 비자 열매 추출물 또는 이의 분획물을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용할 수 있고, 통상의 방법에 따라 적절하게 사용할 수 있다. 유효 성분의 혼합양은 사용 목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있으며, 식품학적으로 허용가능한 식품 보조 첨가제를 추가로 포함할 수 있다. 본 발명의 조성물은 천연물로부터 유래한 추출물을 유효성분으로 하므로 안정성 면에서 문제가 없기 때문에 혼합량에 큰 제한은 없다.When the composition of the present invention is used as a food additive, the visa fruit extract or its fraction can be used as it is or can be used together with other foods or food ingredients, and can be suitably used according to ordinary methods. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment), and may further include a food-acceptable food-aid additive. Since the composition of the present invention contains an extract derived from a natural substance as an active ingredient, there is no problem in terms of stability, so there is no great limitation on the amount of the mixture.
본 발명의 식품 조성물은 통상적인 의미의 식품을 모두 포함할 수 있으며, 기능성 식품, 건강기능식품 등 당업계에 알려진 용어와 혼용 가능하다.The food composition of the present invention may include all foods having a conventional meaning, and may be mixed with terms known in the art such as functional foods, health functional foods, and the like.
본 발명의 용어 "기능성 식품"은 건강기능식품에 관한 법률 제6727호에 따른 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 제조 및 가공한 식품을 의미하며, "기능성"이라 함은 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건 용도에 유용한 효과를 얻을 목적으로 섭취하는 것을 의미한다.The term "functional food" in the present invention means a food prepared and processed by using a raw material or ingredient having functionality useful to the human body according to Law No. 6727 on health functional foods, and " And function of the nutrient for the purpose of obtaining a beneficial effect in health use such as controlling the nutrient or physiological action.
또한, 본 발명의 용어 "건강기능식품"은 건강보조의 목적으로 특정성분을 원료로 하거나 식품 원료에 들어있는 특정성분을 추출, 농축, 정제, 혼합 등의 방법으로 제조, 가공한 식품을 말하며, 상기 성분에 의해 생체방어, 생체리듬의 조절, 질병의 방지와 회복 등 생체조절기능을 생체에 대하여 충분히 발휘할 수 있도록 설계되고 가공된 식품을 말하는 것으로서, 상기 건강식품용 조성물은 질병의 예방 및 질병의 회복 등과 관련된 기능을 수행할 수 있다.The term "health functional food" of the present invention refers to a food prepared by processing a specific ingredient as a raw material for the purpose of health assisting or by extracting, concentrating, refining, mixing, or the like a specific ingredient contained in a food raw material, Refers to a food which is designed and processed so that the body control function such as bio-defense, regulation of biorhythm, prevention and recovery of disease and the like can be sufficiently exhibited to the living body by the above components. Recovery, and so on.
본 발명의 조성물이 사용될 수 있는 식품의 종류에는 제한이 없다. 아울러 본 발명의 비자 열매 추출물 또는 이의 분획물을 활성성분으로 포함하는 조성물은 당업자의 선택에 따라 식품에 함유될 수 있는 적절한 기타 보조 성분과 공지의 첨가제를 혼합하여 제조할 수 있다. 첨가할 수 있는 식품의 예로는 육류, 소세지, 빵, 쵸코렛, 캔디류, 스낵류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림 류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알콜 음료 및 비타민 복합제 등이 있으며, 본 발명에 따른 추출물을 주성분으로 하여 제조한 즙, 차, 젤리 및 주스 등에 첨가하여 제조할 수 있다.There is no limitation on the kind of food in which the composition of the present invention can be used. In addition, the composition containing the visa fruit extract or the fraction thereof as an active ingredient of the present invention can be prepared by mixing other suitable auxiliary ingredients that may be contained in foods and known additives, according to the selection of a person skilled in the art. Examples of foods that can be added include dairy products, such as meat, sausage, bread, chocolates, candies, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, Vitamin complex, and the like, and can be prepared by adding to the juice, tea, jelly, and juice prepared from the extract of the present invention as a main component.
또한, 본 발명에 적용될 수 있는 식품에는 예컨대, 특수영양식품(예: 조제유류, 영,유아식 등), 식육가공품, 어육제품, 두부류, 묵류, 면류(예: 라면류, 국수류 등), 건강보조식품, 조미식품(예: 간장, 된장, 고추장, 혼합장 등), 소스류, 과자류(예:스낵류), 유가공품(예: 발효유, 치즈 등), 기타 가공식품, 김치, 절임식품(각종 김치류, 장아찌 등), 음료(예: 과실, 채소류 음료, 두유류, 발효음료류 등), 천연조미료(예, 라면스프 등) 등 모든 식품을 포함할 수 있다.Examples of foods that can be used in the present invention include special nutritional foods such as crude oil, milk, baby food, meat products, fish meat products, tofu, mushrooms, noodles such as ramen noodles, (Such as soy sauce, doenjang, kochujang, mixed potatoes), sauces, confectionery (eg snacks), dairy products (eg fermented milk, cheese), other processed foods, kimchi, pickles ), Beverages (such as fruit, vegetable beverages, beverages, fermented beverages, etc.), and natural seasonings (eg, ramen soup, etc.).
본 발명의 건강기능식품 조성물이 음료의 형태로 사용될 경우에는 통상의 음료와 같이 여러 가지 감미제, 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상기 외에 본 발명의 건강기능식품 조성물은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 그밖에 천연 과일쥬스, 과일쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다.When the health functional food composition of the present invention is used in the form of a drink, it may contain various sweeteners, flavors, or natural carbohydrates as an additional ingredient such as ordinary beverages. In addition to the above, the health functional food composition of 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, pH adjusting agents, stabilizers, , Alcohols, carbonating agents used in carbonated drinks, and the like. It may also contain flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks.
또 다른 하나의 양태로서, 본 발명은 비자 열매 추출물 또는 이의 분획물을 유효성분으로 포함하는 항산화 또는 항염증용 약학 조성물을 제공한다. 상기 비자 열매 추출물 또는 이의 분획물에 관해서는 전술한 바와 같으며, 본 발명의 추출물은 활성산소를 제거하는 항산화 효과 및 항염증 효과를 갖기 때문에 약학 조성물로 활용하여 활성 산소로 인해 발생하는 질환들 및 염증성 질환들을 예방, 개선 및 치료하기 위한 의약품에 포함시킬 수 있다.In another aspect, the present invention provides a pharmaceutical composition for antioxidant or antiinflammation, which comprises a visa-derived fruit extract or a fraction thereof as an active ingredient. Since the extract of the present invention has antioxidative and antiinflammatory effects for removing active oxygen, it can be used as a pharmaceutical composition to treat diseases caused by active oxygen and inflammatory And can be included in medicines for preventing, improving and treating diseases.
상기 항산화에 대해서는 전술한 바와 같으며, 그에 따라 항산화를 목적으로 하는 약학 조성물에 포함될 수 있으며, 활성산소로 인해 발생하는 질환의 예방 또는 치료 효과를 가질 수 있다. 상기 활성산소로 인해 발생하는 질환들은 이에 제한되지는 않으나, 동맥경화증, 루게릭병, 파킨슨병, 알츠하이머, 근위축색경화증 및 헌팅톤병을 포함하는 퇴행성 신경질환, 심근경색, 협심증, 관상동맥질환, 허혈성 심장질환을 포함하는 심혈관 질환, 뇌졸중을 포함하는 허혈성 뇌질환, 당뇨병, 위염 및 위암을 포함하는 소화기계 질환, 암, 백혈병, 노화, 류마티스 관절염, 간염, 아토피성 피부염 등 다양한 질환을 포함할 수 있으며, 구체적으로 활성산소에 의해 발생되는 노화일 수 있다.The above-mentioned antioxidation is as described above, and thus can be included in a pharmaceutical composition for the purpose of antioxidation, and can have a preventive or therapeutic effect on a disease caused by active oxygen. The diseases caused by the active oxygen include, but are not limited to, degenerative neurological diseases including arteriosclerosis, Lou Gehrig's disease, Parkinson's disease, Alzheimer's disease, proximal axillary sclerosis and Huntington's disease, myocardial infarction, angina pectoris, coronary artery disease, May include various diseases such as cardiovascular diseases including diseases, ischemic brain diseases including stroke, digestive system diseases including diabetes, gastritis and gastric cancer, cancer, leukemia, aging, rheumatoid arthritis, hepatitis, atopic dermatitis, Specifically aging caused by active oxygen.
또한, 상기 항염증에 대해서는 전술한 바와 같으며, 그에 따라 항염증, 즉 염증성 질환의 예방 또는 치료를 위한 약학 조성물에 포함될 수 있다. 상기 염증성 질환은 염증을 주병변으로 하는 질병을 총칭하는 의미로서, 이에 제한되지는 않으나, 알러지성 천식, 알러지성 비염, 알러지성 점막염, 두드러기 및 아나필락스(anaphylax)를 포함하는 알러지성 질환, 경피증(systemic sclerosis), 피부근염(dermatomyositis) 및 포함체 근육염(inclusion body myositis)을 포함하는 근병증, 관절염, 아토피성 피부염, 건선, 천식, 다발성 경화증, ssRNA 및 dsRNA 바이러스 감염증, 패혈증, 다발성 연골염, 경피증, 습진, 통풍, 치주질환, 베체트 증후군, 부종, 맥관염, 가와사키병, 당뇨병성 망막염, 자가 면역 췌장염, 혈관염, 사구체 신염, 급성 및 만성 기관지염, 및 인플루엔자 감염증일 수 있다.In addition, the anti-inflammatory activity is as described above, and thus can be included in a pharmaceutical composition for the prophylaxis or treatment of anti-inflammatory, i.e., inflammatory diseases. The inflammatory diseases include allergic diseases including allergic asthma, allergic rhinitis, allergic mucositis, urticaria, and anaphylax, although it is not limited thereto. Arthritis, atopic dermatitis, psoriasis, asthma, multiple sclerosis, ssRNA and dsRNA viral infection, sepsis, multiple chondritis, scleroderma, including systemic sclerosis, dermatomyositis and inclusion body myositis , Eczema, gout, periodontal disease, Behcet's syndrome, edema, vasculitis, Kawasaki disease, diabetic retinitis, autoimmune pancreatitis, vasculitis, glomerulonephritis, acute and chronic bronchitis, and influenza infection.
상기 본 발명의 약학조성물은 약학적으로 허용가능한 담체를 추가로 포함할 수 있다. 본 발명의 용어 "약학적으로 허용가능한"이란 상기 조성물에 노출되는 세포나 인간에게 독성이 없는 특성을 나타내는 것을 의미한다. 상기 담체는 완충제, 보존제, 무통화제, 가용화제, 등장제, 안정화제, 기제, 부형제, 윤활제 등 당업계에 공지된 것이라면 제한없이 사용할 수 있다.The pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable" of the present invention means that it exhibits properties that are not toxic to the cells or humans exposed to the composition. Such carriers may be used without limitation as long as they are known in the art such as buffers, preservatives, wetting agents, solubilizers, isotonic agents, stabilizers, bases, excipients and lubricants.
또한 본 발명의 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. 나아가, 연고제, 로션제, 스프레이제, 패취제, 크림제, 산제, 현탁제, 겔제 또는 젤의 형태의 피부 외용제의 형태로 사용될 수 있다. 본 발명의 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.In addition, the pharmaceutical composition of the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterilized injection solutions according to a conventional method have. Furthermore, it can be used in the form of an external preparation for skin in the form of ointments, lotions, spray agents, patches, creams, powders, suspensions, gels or gels. Examples of carriers, excipients and diluents that can be included in the composition of the present invention 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.
경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 상기 비자 열매 추출물 또는 이의 분획물에 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘카보네이트(calcium carbonate), 수크로스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제된다. 또한 단순한 부형제 이외에 마그네슘 스티레이트, 탈크 같은 윤활제들도 사용된다. 경구를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는 데 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.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. Liquid preparations for oral use may include various excipients such as wetting agents, sweetening agents, fragrances, preservatives, etc. in addition to water and liquid paraffin, which are simple diluents commonly used in suspension, liquid solutions, emulsions and syrups have. 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.
한편, 본 발명의 약학조성물은 약학적으로 유효한 양으로 투여한다. 본 발명의 용어 "투여"란, 적절한 방법으로 개체에게 소정의 물질을 도입하는 것을 의미하며 상기 조성물의 투여 경로는 목적 조직에 도달할 수 있는 한 어떠한 일반적인 경로를 통하여 투여될 수 있다. 복강내 투여, 정맥내 투여, 근육내 투여, 피하 투여, 피내 투여, 경구 투여, 국소 투여, 비내 투여, 폐내 투여, 직장내 투여될 수 있으나, 이에 제한되지는 않는다.Meanwhile, the pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. The term "administering" of the present invention means introducing a predetermined substance into an individual by an appropriate method, and the administration route of the composition can be administered through any conventional route so long as it can reach the target tissue. But are not limited to, intraperitoneal, intravenous, intramuscular, subcutaneous, intradermal, oral, topical, intranasal, intrathecal, rectal.
상기 용어 "개체"란 는 인간을 포함한 쥐, 생쥐, 가축 등의 모든 동물을 의미한다. 구체적으로, 인간을 포함한 포유동물일 수 있다.The term "individual" refers to all animals, including humans, including mice, mice, and livestock. Specifically, it may be a mammal including a human.
상기 용어 "약학적으로 유효한 양"이란 의학적 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료하기에 충분하며 부작용을 일으키지 않을 정도의 양을 의미하며, 유효 용량 수준은 환자의 성별, 연령, 체중, 건강상태, 질병의 종류, 중증도, 약물의 활성, 약물에 대한 민감도, 투여 방법, 투여 시간, 투여 경로, 및 배출 비율, 치료 기간, 배합 또는 동시에 사용되는 약물을 포함한 요소 및 기타 의학 분야에 잘 알려진 요소에 따라 당업자에 의해 용이하게 결정될 수 있다. 투여는 상기 권장 투여량을 하루에 한번 투여할 수도 있고, 수회 나누어 투여할 수도 있다.The term "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment and not causing side effects, and the effective dose level is determined by the patient's sex, Including, but not limited to, medicaments and other medical fields that are used in combination, or in combination with, or in combination with, a pharmaceutically acceptable carrier, excipient, Can be readily determined by those skilled in the art according to known factors. The administration may be carried out once per day, or divided into several doses.
본 발명의 비자 열매 추출물 또는 이의 분획물은 천연물을 원료로 하므로 화장료 조성물, 식품 조성물, 또는 약학적 조성물로 사용할 경우에도 일반적인 합성 화합물에 비하여 부작용이 덜할 수 있으므로 안전하게 포함되어 유용하게 사용될 수 있다.Since the visa fruit extract or the fraction thereof of the present invention is a natural material as a raw material, it can be used safely and usefully when it is used as a cosmetic composition, a food composition, or a pharmaceutical composition because its side effects are less than general synthetic compounds.
본 발명의 비자 열매 추출물 또는 이의 분획물을 포함하는 조성물은 항산화 또는 항염증 활성을 나타내므로, 산화 작용 및 염증 작용에 의해 유발되는 질환의 예방, 개선 또는 치료를 위한 또는 화장품, 식품 및 의약품 등에 이용될 수 있다.Since the composition comprising the visa fruit extract or the fraction thereof of the present invention exhibits antioxidant or anti-inflammatory activity, it can be used for preventing, ameliorating or treating a disease caused by oxidative and inflammatory action or for use in cosmetics, foods and pharmaceuticals .
도 1은 비자 열매 추출물의 항염증 활성을 나타낸 그래프이다. a) 비자 열매 물 추출물의 세포독성을 나타낸 그래프. b) 비자 열매 물 추출물의 NO 생성 저해능을 나타낸 그래프. c) 비자 열매 메탄올 추출물의 세포독성을 나타낸 그래프. d) 비자 열매 메탄올 추출물의 NO 생성 저해능을 나타낸 그래프.
도 2는 비자 열매 메탄올 추출물 및 이의 분획물의 항염증 활성을 나타낸 그래프이다. a) 비자 열매 메탄올 추출물 및 이의 분획물을 50μg/mL 농도로 처리한 세포독성 분석 그래프. b) 비자 열매 메탄올 추출물 및 이의 분획물을 50μg/mL 농도로 처리한 NO 생성 저해능 분석 그래프. c) 비자 열매 메탄올 추출물 및 이의 분획물을 100μg/mL 농도로 처리한 세포독성 분석 그래프. d) 비자 열매 메탄올 추출물 및 이의 분획물을 100μg/mL 농도로 처리한 NO 생성 저해능 분석 그래프. e) 비자 열매 메탄올 추출물 및 이의 분획물을 200μg/mL 농도로 처리한 세포독성 분석 그래프. f) 비자 열매 메탄올 추출물 및 이의 분획물을 200μg/mL 농도로 처리한 NO 생성 저해능 분석 그래프.FIG. 1 is a graph showing the anti-inflammatory activity of visa-fruit extract. FIG. a) A graph showing the cytotoxicity of visceral fruit water extract. b) Graph showing inhibitory effect on NO production of visa fruit water extract. c) A graph showing the cytotoxicity of methanol extract of visa fruit. d) Graph showing inhibitory effect of NO production on non-viscous methanol extracts.
2 is a graph showing the anti-inflammatory activity of the non-viscous methanol extract and fractions thereof. a) Cytotoxicity analysis graph of methanol extract of visa fruit and its fraction treated at a concentration of 50 μg / mL. b) Analysis of NO production inhibitory activity by treatment with methanol extract of visa and its fractions at a concentration of 50 μg / mL. c) Cytotoxicity analysis of methanol extract of visa and its fractions treated at 100 μg / mL concentration. d) Graph of NO production inhibition assay treated with 100 μg / mL of non-viscous methanol extract and its fractions. e) Cytotoxicity analysis of methanol extract of visa and its fractions treated at a concentration of 200 μg / mL. f) Graph of NO production inhibition assay treated with 200 μg / mL of non-viscous methanol extract and fractions thereof.
이하 본 발명을 하기 예에 의해 상세히 설명한다. 다만, 하기 예는 본 발명을 예시하기 위한 것일 뿐, 하기 예에 의해 본 발명의 범위가 제한되는 것은 아니다.Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are only for illustrating the present invention, and the scope of the present invention is not limited by the following examples.
실시예Example 1. 비자 열매 추출 및 분획 1. Visa fruit extraction and fractionation
실험에 사용한 비자 열매 (Torreya nucifera seed)는 2013년 서울에 소재한 약령시장에서 구입하여, 시료 50 g씩 각각 물과 메탄올로 추출하여 동결 건조한 것을 사용하였다. 또한 메탄올 추출물 및 그 분획물을 획득하기 위하여 비자 열매 10 kg을 메탄올 9 L로 70℃ 에서 3시간 동안 3회에 걸쳐 진탕하면서 추출한 후 여과, 감압 농축하여 총 1630 g의 메탄올 추출물을 얻었다. 이 메탄올 추출물을 물:메탄올 (9:1, v/v)의 혼합용매로 녹인 다음 동량의 헥산을 넣어 흔든 다음 평형화시켜 헥산 가용부를 모아 무수 황산 나트륨으로 처리한 다음 여과하여 농축하였다. 이와 같은 방법으로 2회 더 반복하여 헥산 층을 얻었다. 동일한 방법으로 디클로로메탄, 에틸아세테이트, n-부탄올 순서로 분획하였다.Torreya nucifera seeds were purchased from the Yakushin market in Seoul in 2013 and were extracted with water and methanol (50 g each) and lyophilized. In order to obtain the methanol extract and fractions thereof, 10 kg of visa fruit was extracted with 9 L of methanol at 70 ° C for 3 hours with shaking three times, filtered and concentrated under reduced pressure to obtain a total of 1630 g of methanol extract. This methanol extract was dissolved in a mixed solvent of water and methanol (9: 1, v / v), equilibrated with hexane and then equilibrated. The hexane-soluble portion was collected, treated with anhydrous sodium sulfate, filtered and concentrated. By repeating this procedure twice more, a hexane layer was obtained. In the same manner, dichloromethane, ethyl acetate and n-butanol were fractionated.
실험예Experimental Example 1. 비자 열매 추출물의 총 페놀성 함량 분석 1. Analysis of total phenolic content of visa fruit extract
일반적으로 추출물질의 1차적 생리활성 검정을 위해 다양한 화학적 산화제를 이용한 전자공여능 또는 항산화능 측정 등의 방법이 가장 빈번하게 사용되고 있으며 이와 같은 활성 검증 시스템을 기반으로 하여 발굴되는 물질 중에는 페놀성 화합물이 상당수를 점유하고 있다. 따라서 페놀성 물질의 함량 측정이 일반적으로 진행되고 있으며 이러한 페놀성 물질의 정량방법으로 Folin-Denis 방법이 가장 널리 이용되고 있다. 총 페놀성 함량은 증류수 또는 pH 완충용액에 녹인 시료 80 μL에 50% Folin-Ciocalteu시약 20 μL을 첨가하여 5분간 방치한 후, 2% Na2CO3용액 100 μL을 가하였다. 상온에서 30분간 또는 일정시간 반응시킨 후 마이크로 플레이트 리더를 이용하여 750 nm에서 흡광도를 측정하였다. 총 페놀성 함량은 타닌산을 정량하여 작성한 표준곡선으로부터 계산하였다.In general, for the first-order physiological activity test of extracts, methods of measuring electron donating ability or antioxidant ability using various chemical oxidizing agents are most frequently used. Among the substances found based on such activity verification system, there are many phenolic compounds It is occupied. Therefore, the content of phenolic substances is generally measured, and Folin-Denis method is most widely used as a method of quantifying phenolic substances. The total phenolic content was determined by adding 20 μL of 50% Folin-Ciocalteu reagent to 80 μL of the sample dissolved in distilled water or pH buffer solution, and keeping it for 5 minutes. Then, 100 μL of 2% Na 2 CO 3 solution was added. After incubation at room temperature for 30 minutes or for a period of time, the absorbance was measured at 750 nm using a microplate reader. The total phenolic content was calculated from the standard curve prepared by quantifying tannic acid.
그 결과, 표 1에 나타난 바와 같이 비자열매 물 추출물과 메탄올 추출물의 총 페놀성 함량은 비자열매추출물에서 100 g 당 각각 0.73 ± 0.01 g과 0.63 ± 0.01 g으로 나타났다.As a result, as shown in Table 1, the total phenolic contents of visa fruit water extract and methanol extract were 0.73 ± 0.01 g and 0.63 ± 0.01 g per 100 g of visa fruit extract, respectively.
[표 1][Table 1]
실험예Experimental Example 2. 비자 열매 추출물 또는 이의 2. Visa fruit extract or object 분획물의Fraction 항산화 활성 분석 Antioxidant activity analysis
2-1. 비자 열매 추출물의 항산화 활성 분석2-1. Antioxidant activity analysis of visa fruit extract
전자공여능은 항산화 작용의 지표로 사용되고 있으며 식물 추출물의 항산화능 측정에 많이 사용되고 있다. 전자공여작용은 인체 내에서 생성되는 자유 라디칼의 전자를 공여하여 자유 라디칼에 의한 노화와 질병을 억제하는 작용으로 이용되고 있다. 전자공여능 측정은 DPPH 라디칼 소거법을 이용하여 측정하였다. DPPH는 비교적 안정한 자유 라디칼로써, 아스코르브산, 토코페롤, 폴리하이드록시 방향족 화합물, 방향족 아민류에 의해 환원되어 짙은 자색이 탈색되는 원리를 이용하여 항산화 활성을 간단히 측정할 수 있는 동시에 식물체의 항산화 활성과도 연관성이 매우 높기 때문에 많이 이용되고 있는 방법이다. 비자열매 추출물을 농도별 (64~1600 μg/mL)로 메탄올에 녹여 준비한 후 96 웰 플레이트에 메탄올에 용해된 1.5×10-4 M DPPH 40 μL와 각 시료 160 μL를 분주한 혼합액을 실온에서 30분간 반응시킨 후, 마이크로 플레이트 리더를 이용하여 520 nm에서 흡광도를 측정하였다. 시료를 첨가하지 않은 음성 대조군과 비교하여 자유 라디칼 소거활성을 백분율로 나타내고, 50% 저해 농도 (Inhibitory Concentration, IC50)를 계산하였다. 양성 대조군으로는 대표적인 항산화제인 아스코르브산 (=비타민 C)을 사용하였으며 측정값은 3회 반복 실험의 평균값으로 나타내었다. 억제능의 백분율 공식은 다음과 같다.The electron donating ability is used as an index of antioxidant activity and is widely used for measuring the antioxidant activity of plant extracts. The electron donating action is used to donate electrons of free radicals generated in the human body and to inhibit aging and disease by free radicals. The electron donating ability was measured by the DPPH radical scavenging method. DPPH is a relatively stable free radical, which can be easily measured by using ascorbic acid, tocopherol, polyhydroxy aromatic compounds, aromatic amines, and decolorizing of deep purple color, and it is also related to the antioxidant activity of plants Is a very popular method. Visible fruit extracts were prepared by dissolving in methanol (64 ~ 1600 μg / mL) by concentration and 40 μL of 1.5 × 10 -4 M DPPH dissolved in methanol and 160 μL of each sample were added to 96 well plates. After the reaction, the absorbance was measured at 520 nm using a microplate reader. Free radical scavenging activity was expressed as a percentage and a 50% inhibitory concentration (IC 50 ) was calculated as compared to the negative control to which no sample was added. As a positive control, ascorbic acid (= vitamin C), a representative antioxidant, was used. The percent inhibition formula is as follows.
DPPH 라디칼 소거 활성 (%)={1-(A-B)/C}×100DPPH radical scavenging activity (%) = {1- (A-B) / C} 100
A: 시료 흡광도 520nm A: sample absorbance 520 nm
B: 색 대조군 흡광도 520nm B: color control absorbance 520 nm
C: 대조군 흡광도 520nm C: Control absorbance 520 nm
표 2에 나타난 바와 같이, 50% 소거 농도를 나타내는 IC50 값이 물추출물의 경우, 103.93 ± 2.19 μg/mL, 메탄올추출물은 81.25 ± 6.32 μg/mL으로 각각 나타났다.As shown in Table 2, IC 50 values showing 50% eradication concentration were 103.93 ± 2.19 μg / mL for water extract and 81.25 ± 6.32 μg / mL for methanol extract, respectively.
[표 2][Table 2]
2-2. 비자 열매 메탄올 추출물 및 2-2. Visa fruit methanol extract and 분획물의Fraction 항산화 활성 분석 Antioxidant activity analysis
비자열매 메탄올 추출물이 물추출물보다 활성이 높음을 확인함에 따라 메탄올 추출물 및 5종 분획물(헥산, 디클로로메탄, 에틸아세테이트, n-부탄올, 물 분획물)의 항산화 활성을 64, 320, 1600 μg/mL 시료 처리 농도에서 비교 분석하였다. 비자열매의 DPPH 라디칼 소거 활성은 에틸아세테이트 층 (8.20 ± 0.45 μg/mL) > 메탄올 추출물 (81.25 ± 6.32 μg/mL) > n-부탄올 층 (125.12 ± 7.33 μg/mL) > 디클로로메탄 층 (203.53 ± 4.40 μg/mL) > 헥산 층 (407.91 ± 4.84 μg/mL) > 물 층 (450.47 ± 6.29 μg/mL)순으로 나타났으며, 농도 의존적으로 라디칼 소거 활성이 나타났다 (표 3).The antioxidative activities of methanol extracts and five fractions (hexane, dichloromethane, ethyl acetate, n-butanol, water fractions) were determined as 64, 320, and 1600 μg / mL, respectively, And compared at the treatment concentration. The DPPH radical scavenging activity of visaflower was determined by measuring the concentration of ethyl acetate layer (8.20 ± 0.45 μg / mL)> methanol extract (81.25 ± 6.32 μg / mL)> n-butanol layer (125.12 ± 7.33 μg / mL)> dichloromethane layer (203.53 ± 4.40 μg / mL), hexane layer (407.91 ± 4.84 μg / mL), water layer (450.47 ± 6.29 μg / mL), and concentration-dependent radical scavenging activity (Table 3).
[표 3][Table 3]
실험예Experimental Example 3. 비자 열매 추출물 또는 이의 3. Visa fruit extract or object 분획물의Fraction 항염증 활성 분석 Analysis of anti-inflammatory activity
3-1. 비자 열매 추출물의 항염증 활성 분석3-1. Analysis of anti-inflammatory activity of visa fruit extract
항염증 활성의 주요 세포 실험 모델계로 사용되고 있는 뮤린 매크로파지 세포주(murine macrophage cell line)인 RAW 264.7을 American Type Tissue Collection (ATCC)로부터 구입하여 10% 소 태아 혈청 (FBS) 및 페니실린/스트렙토마이신이 포함된 DMEM 배지에서 배양하였다. RAW 264.7 쥐 대식세포주를 24-웰 플레이트에 웰 당 1.0 X 105 세포를 분주하여 24시간 동안 부착시킨 후 비자열매 물추출물 및 메탄올 추출물을 각각 50, 100, 150, 200, 250, 300 μg/mL으로 전 처리하고, 1시간 후에 1 μg/mL의 지질다당류 (LPS)를 처리하여 20 시간 동안 NO 생성을 유도하면서 각 추출물의 NO 생성 저해능의 정도를 Griess reaction을 통해 분석하였다. MAIN CELLS OF ANTI-INFLAMMATORY ACTIVITY RAW 264.7, a murine macrophage cell line used as a model system, was purchased from the American Type Tissue Collection (ATCC) and cultured with 10% fetal bovine serum (FBS) and penicillin / streptomycin And cultured in DMEM medium. RAW 264.7 mouse macrophage cells were plated on a 24-well plate at a rate of 1.0 × 10 5 cells per well for 24 hours, and then the visia fruit water extract and the methanol extract were added at 50, 100, 150, 200, 250 and 300 μg / mL . After 1 hour, 1 μg / mL of lipopolysaccharide (LPS) was treated to induce NO production for 20 hours, and the degree of NO production inhibition of each extract was analyzed by Griess reaction.
그 결과, 물추출물은 분석한 모든 농도에서 세포 독성을 보이지 않았고 (도 1a), 시료 처리 군에서 최소 12.9에서 최대 22.7%까지 NO 생성 저해능을 보였으나 농도의존성은 보이지 않았다 (도 1b). 반면 메탄올 추출물은 150 μg/mL 이상에서 농도의 증가에 따른 약한 세포 생존율의 저하가 나타났으나 300 μg/mL에서 81.4%로 그 정도는 크지 않았고 현미경 하에서 세포 독성은 관찰되지 않았다 (도 1c). 또한 모든 처리 농도에서 농도 의존적인 강한 NO 생성 저해능을 보여 LPS 처리군(44.5 μM) 대비 50, 100, 150, 200, 250, 300 μg/mL에서 각각 37.43, 30.88, 23.6, 16.45, 11.45, 5.31 μM의 NO 생성량을 나타내 15.9, 30.6, 47.0, 63.0, 74.3, 86.9%의 NO 생성 억제율을 보였다 (도 1d).As a result, the water extract showed no cytotoxicity at all the concentrations analyzed (FIG. 1A), and showed a minimum NO production inhibition of 12.9 to 22.7% in the sample treatment group but no concentration dependence (FIG. On the other hand, the methanol extract showed a decrease in cell viability with an increase in concentration above 150 μg / mL, but not so large as 81.4% at 300 μg / mL and no cytotoxicity under the microscope (FIG. In addition, there was a strong concentration-dependent inhibition of NO production at all treatment concentrations, which was 37.43, 30.88, 23.6, 16.45, 11.45, and 5.31 μM at 50, 100, 150, 200, 250 and 300 μg / (Fig. 1d), indicating 15.9, 30.6, 47.0, 63.0, 74.3, and 86.9% NO production, respectively.
3-2. 비자 열매 메탄올 추출물 및 3-2. Visa fruit methanol extract and 분획물의Fraction 항염증 활성 분석 Analysis of anti-inflammatory activity
비자열매 메탄올추출물이 농도의존적인 NO 생성 저해능을 보유함을 확인함에 따라 메탄올 추출물(M) 및 5종 분획물(헥산, 디클로로메탄, 에틸아세테이트, n-부탄올, 물 분획물)의 NO 생성 저해능을 50, 100, 200 μg/mL 시료 처리 농도에서 비교 분석하였다. (M) and five fractions (hexane, dichloromethane, ethyl acetate, n-butanol, water fractions) were 50, 50 and 100 ppm, respectively, as the visa-extract methanol extracts showed a concentration- 100, and 200 μg / mL, respectively.
그 결과, 도 2에서 나타난 바와 같이 활성의 정도는 디클로로메탄 층이 가장 높게 나타났으나 고농도 처리에서 세포증식억제능을 보였고 헥산 층과 에틸아세테이트 층은 메탄올 추출물과 유사한 정도의 활성을 나타내었으며 에틸아세테이트 층이 유사한 활성에 비해 세포증식 억제 정도는 메탄올 층보다 적게 나타났다. n-부탄올 층과 물 층 분획에서는 분석한 농도 범위에서는 활성을 나타내지 않았다.As a result, as shown in FIG. 2, the degree of activity was the highest in the dichloromethane layer, but the cell growth inhibition was shown in the high concentration treatment. The hexane layer and the ethyl acetate layer showed similar activity to the methanol extract, Compared with this similar activity, cell proliferation inhibition was lower than that of methanol layer. The n - butanol layer and the water layer fraction showed no activity at the concentration range analyzed.
이상의 설명으로부터, 본 발명이 속하는 기술분야의 당업자는 본 발명이 그 기술적 사상이나 필수적 특징을 변경하지 않고서 다른 구체적인 형태로 실시될 수 있다는 것을 이해할 수 있을 것이다. 이와 관련하여, 이상에서 기술한 실시 예들은 모든 면에서 예시적인 것이며 한정적인 것이 아닌 것으로서 이해해야만 한다. 본 발명의 범위는 상기 상세한 설명보다는 후술하는 특허 청구범위의 의미 및 범위 그리고 그 등가 개념으로부터 도출되는 모든 변경 또는 변형된 형태가 본 발명의 범위에 포함되는 것으로 해석되어야 한다.From the above description, it will be understood by those skilled in the art that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. In this regard, it should be understood that the above-described embodiments are to be considered in all respects as illustrative and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention without departing from the scope of the present invention as defined by the appended claims.
Claims (10)
A composition for antioxidant or antiinflammatory activity, comprising an extract of Torreya nucifera seed or a fraction thereof as an active ingredient.
상기 추출물은 물, 탄소수 1 내지 4의 알코올 또는 이들의 혼합용매로 추출한 것인, 항산화 또는 항염증용 조성물.
The method according to claim 1,
Wherein the extract is extracted with water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.
상기 분획물의 용매는 물, 탄소수 1 내지 4의 알코올, 에틸아세테이트, 아세톤, 헥산, 디클로로메탄 또는 이들의 혼합 용매인 것인, 항산화 또는 항염증용 조성물.
The method according to claim 1,
Wherein the solvent of the fraction is water, an alcohol having 1 to 4 carbon atoms, ethyl acetate, acetone, hexane, dichloromethane or a mixed solvent thereof.
상기 항산화용 조성물은 비자 열매 메탄올 추출물의 에틸아세테이트 분획물을 포함하는 것인, 항산화용 조성물.
The method according to claim 1,
Wherein the composition for antioxidation comprises an ethyl acetate fraction of a non-viscous methanol extract.
상기 항염증용 조성물은 비자 열매 메탄올 추출물의 디클로로메탄 분획물을 포함하는 것인, 항염증용 조성물.
The method according to claim 1,
Wherein the anti-inflammatory composition comprises a dichloromethane fraction of a non-viscous methanol extract.
상기 항산화 효과는 활성 산소종 제거에 의해 달성되는 것인, 항산화용 조성물.
5. The method of claim 4,
Wherein the antioxidant effect is achieved by removal of active oxygen species.
상기 항염증 효과는 NO 생성 저해능에 의해 달성되는 것인, 항염증용 조성물.
6. The method of claim 5,
Wherein the anti-inflammatory effect is achieved by inhibiting NO production.
A cosmetic composition for antioxidant or antiinflammatory comprising an extract of Torreya nucifera seed or a fraction thereof.
An antioxidant or antiinflammatory food composition comprising a Torreya nucifera seed extract or a fraction thereof.
A pharmaceutical composition for antioxidant or antiinflammatory, comprising a Torreya nucifera seed extract or a fraction thereof.
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| KR20050054112A (en) * | 2003-12-04 | 2005-06-10 | 주식회사 엘지생활건강 | The use of extracts from torreya nucifera, mucuna birdwoodiana and/or eriobotrya japonica as a anti-oxidant |
| KR20110054154A (en) * | 2009-11-17 | 2011-05-25 | (주)아모레퍼시픽 | Cosmetic composition containing immature fruit or seed extract of torreya nucifera |
| KR20130134480A (en) * | 2012-05-31 | 2013-12-10 | (주)아모레퍼시픽 | Extract of torreya sp.treated enzyme |
Non-Patent Citations (2)
| Title |
|---|
| Endo, Yasushi, et al. "Effects of Japanese torreya (Torreya nucifera) seed oil on lipid metabolism in rats." Nutrition 22.5 (2006): 553-558. |
| Journal of the Korean Society of Food Science and Nutrition, 2009, 38(1), pp. 1-8 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR102263888B1 (en) | 2021-01-29 | 2021-06-14 | (주)브이에스신비 | Preparation Methods for Fermentative Extract of Torreya Nucifera Seed from JEJU Having Anti-Inflammatory Activities |
Also Published As
| Publication number | Publication date |
|---|---|
| KR102154092B1 (en) | 2020-09-09 |
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