KR20180060169A - Composition for Preventing or Improving Atopic dermatitis Comprising Extract of Galium Aparine as Effective Component - Google Patents
Composition for Preventing or Improving Atopic dermatitis Comprising Extract of Galium Aparine as Effective Component Download PDFInfo
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- KR20180060169A KR20180060169A KR1020160159344A KR20160159344A KR20180060169A KR 20180060169 A KR20180060169 A KR 20180060169A KR 1020160159344 A KR1020160159344 A KR 1020160159344A KR 20160159344 A KR20160159344 A KR 20160159344A KR 20180060169 A KR20180060169 A KR 20180060169A
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Abstract
Description
본 발명은 식물 추출물을 유효성분으로 함유하는 아토피 피부염 예방 또는 개선용 조성물에 관한 것으로, 더욱 상세하게는 갈퀴덩굴 추출물을 유효성분으로 함유하는 아토피 피부염 예방 또는 개선용 건강기능식품 조성물, 약학 조성물 및 화장료 조성물에 관한 것이다.The present invention relates to a composition for preventing or ameliorating atopic dermatitis comprising an extract of plant as an active ingredient, and more particularly to a composition for preventing or ameliorating atopic dermatitis, ≪ / RTI >
일반적으로, 피부는 외부환경과 항상 접하고 있는 기관으로서, 수분손실을 막는 보호장벽으로서의 역할을 수행한다. 표피의 각질층 역시 피부에 가장 외측에 존재하면서 피부 밖으로 수분과 전해질이 소실되는 것을 억제함으로써, 피부의 건조를 막아 피부가 정상적인 대사과정을 수행할 수 있도록 환경을 제공한다. 또한, 외부의 물리적 손상과 화학물질로부터 인체를 보호하거나 세균, 바이러스 등이 피부로 침입하는 것을 막아주는 중요한 역할을 한다.In general, the skin is an organ that is always in contact with the external environment, and acts as a protective barrier to prevent moisture loss. The horny layer of the epidermis also exists in the outermost part of the skin and inhibits the loss of moisture and electrolytes outside the skin, thereby preventing the skin from drying and providing an environment in which the skin can perform a normal metabolic process. It also plays an important role in protecting the body from external physical damage, chemical substances, and preventing invasion of bacteria, viruses, etc. into the skin.
이러한 피부에 환경적, 유전적인 문제로 인하여 기능이 저하되어 생기는 질환이 아토피 피부염이다. 아토피성 피부염은 피부 장벽기능장애와 알레르기원과 미생물과 같은 환경 인자 사이의 복잡한 상호작용과 관련된 유일한 질병이다(Incovaria et al., Clin. Exp. Immunol., 153: 27-29, 2003). 멜라닌세포(melanocyte)와 각질세포(keratinocyte)를 포함하는 모든 피부세포는 활성산소종(reactive oxygen species, ROS)와 활성질소종(reactive nitrogen species, RNS)을 생산한다(Pelle et al., J. Invest. Dermatol., 124: 793-797, 2005; Kornina and Pastore, Front. Biosci. 1: 123-141, 2005). 덧붙여, 피부세포로부터 생성된 ROS는 또한 지질 분자 또는 리폭시게나제(lipoxigenase)와 시클로옥시게나제(cyclooxygenase)와 같은 산화환원-민감성 지질대사 효소와 반응함으로써 지질 라디칼(lipid radical), 과산화물(peroxide), 하이드로과산화물(hydroperoxide)와 같은 활성지질종(reactive lipid species) 또는 알데히드를 생산할 수 있다. 만성 염증질환인 아토피성 피부염은 그러한 산화 환원균형을 붕괴시켜서 ROS, RNS의 과잉생산과 지질 과산화물의 높은 수준을 야기하는데, 이는 결국 질병상태를 악화시키고 Th2-편향된 면역반응 쪽으로 치우키게 한다(Briganti and Picardo, J. Eur. Acad. Dermatol., 17: 663-669, 2003; Wu et al., Clin. Exp. Immunol., 135: 194-199, 2004).Atopic dermatitis is a disease caused by environmental and genetic problems in these skin. Atopic dermatitis is the only disease associated with complex interactions between skin barrier dysfunctions and environmental factors such as allergens and microorganisms (Incovaria et al., Clin. Exp. Immunol., 153: 27-29, 2003). All skin cells, including melanocytes and keratinocytes, produce reactive oxygen species (ROS) and reactive nitrogen species (RNS) (Pelle et al., J. Mol. Invest. Dermatol., 124: 793-797, 2005; Kornina and Pastore, Front. Biosci., 1: 123-141, 2005). In addition, ROS produced from skin cells also react with lipid radicals, peroxides, and lipid radicals by reaction with lipid molecules or redox-sensitive lipid metabolism enzymes such as lipoxigenase and cyclooxygenase. , Reactive lipid species such as hydroperoxide, or aldehydes. Atopic dermatitis, a chronic inflammatory disease, disrupts such redox balance, leading to overproduction of ROS, RNS and high levels of lipid peroxides, which eventually worsens the disease state and displaces it towards the Th2-biased immune response (Briganti and Picardo, J. Eur. Acad. Dermatol., 17: 663-669, 2003; Wu et al., Clin. Exp. Immunol., 135: 194-199, 2004).
피부에 나타나는 염증 반응은 붉어짐, 따끔거림, 화끈거림, 팽윤, 조직의 변화와 같은 5가지 현상으로 나타나는데, 이는 해로운 주위 환경, 즉 세균과 같은 외부 물질의 침입과 기계적 손상으로부터 생체를 보호하려는 생리적인 반응이다. 염증과 관계되는 대표적인 피부 트러블의 형태로는 아토피와 여드름 등이 있다. 특히, 아토피는 심한 가려움증이 특징적인데 피부에 자극을 가하면 가려움증이 더욱 심해지고, 자꾸 더 긁다보면 상처가 나게 된다. 이로 인해 세균감염(황색포도상구균, 연쇄상구균 등)이 일어나게 되어 염증이 발생한다. 아토피의 문제 중에 중요한 인자의 하나는 이때 일어나는 2차 감염 및 초항원 독소이다. 아토피 환자의 80~90%는 피부에 황색포도상구균(Staphylococcus aureus)과 연쇄상구균(Stereptococcus pyogenes)의 감염이 있는데, 이때 anti- Staphylococcus, IgE가 만들어지며, 이는 T-cell반응을 자극하는 초항원 독소에 의해 유발된다. 보다 구체적으로, 이러한 황색포도상구균의 초항원 독소는 랑게르한스 세포내 항원제시세포를 통하여 T임파구를 선택적으로 자극하여 인터루킨-1과 TNF-alpha를 생산하게 하고, 이들에 의해 자극된 T-cell에서 여러 가지세포내 사이토카인이라는 독소를 분비하여 염증을 일으키게 된다. 즉, 초항원 독소는 인터루킨-4와 인터루틴-5의 분비를 촉진하여 IFN-gamma의 분비는 저해하고, CLA의 수를 증가시켜 결국 IgE의 생성을 촉진시킨다. 이러한 IgE에 의하여 호염구에서 히스타민이라는 물질이 분비되어 아토피 피부염이 악화된다.The inflammatory reactions that appear on the skin appear as redness, tingling, burning, swelling, and changes in tissue, which are physiological and physiologic factors that protect the body from harmful environmental conditions such as intrusion of external substances such as bacteria and mechanical damage Reaction. Typical skin troubles associated with inflammation include atopy and acne. In particular, atopy is characterized by severe itching, irritation to the skin becomes more severe itching, more scratches and scratches. This causes bacterial infections (Staphylococcus aureus, Streptococcus, etc.) and inflammation occurs. One of the important factors in the problem of atopy is the secondary infections and superantigen toxins that occur at this time. Staphylococcus aureus and Stereptococcus pyogenes infect skin with at least 80-90% of atopic patients, where anti-Staphylococcus and IgE are produced, It is caused by toxins. More specifically, the superantigen toxin of Staphylococcus aureus selectively stimulates T lymphocytes through antigen presenting cells in Langerhans cells to produce interleukin-1 and TNF-alpha, and stimulates T-cell stimulation It secretes toxin called cytokine in branch cell and causes inflammation. That is, the superantigen toxin promotes the secretion of interleukin-4 and interleukin-5, thereby inhibiting the secretion of IFN-gamma and increasing the number of CLA, thereby promoting the production of IgE. Such IgE causes secretion of histamine from the basophil, and atopic dermatitis is exacerbated.
아토피성 피부염의 치료 및 관리를 위해, 대부분의 피부과 진료 시 피부 표면의 수분을 유지시켜주는 보습제와 함께 염증반응을 호전시키기 위하여 스테로이드 호르몬 즉, 국소 부신피질호르몬 제제를 동시에 처방하는 것이 일반적이다. 하지만, 국소 부신피질호르몬을 장기간 사용하는 경우, 피부 위축, 혈관 확장, 색소 탈실 및 팽창선조의 발생 등 다양한 피부 부작용을 야기 시킨다는 문제점이 있다.For the treatment and management of atopic dermatitis, it is common to prescribe steroid hormone, or topical corticosteroid, at the same time in order to improve the inflammatory reaction with the moisturizing agent which maintains the moisture of the skin surface in most dermatological treatment. However, the use of topical corticosteroids for a long period of time causes various skin side effects such as skin atrophy, vasodilation, pigment desalination, and swelling ancestry.
이에 따라 안전성이 입증된 천연물로부터 치료효과가 높고 부작용이 적은 치료제를 찾고자 하는 시도가 많이 이루어지고 있다. 현재까지 알려진 항아토피 관련 천연 소재로는 단삼, 다래, 참소리쟁이, 삼백초, 자운고, 황련해독탕 등이 있다.Therefore, many attempts have been made to find therapeutic agents with high therapeutic efficacy and low side effects from natural products proved to be safe. Until now, there have been known anti-atopy related natural materials such as Dansam, Dahlia, Jamsuri, Saururus, Zaunggo, and Hwangryeun.
한국공개특허 제2016-0123592호는 목방충, 딱지꽃, 소목통, 지황, 조릿대 풀, 백선꽃, 남가재, 감초, 형개의 추출물에서 얻은 효소억제제의 제재로 아토피성 피부염증 완화에 효과를 갖는 추출조성물을 개시하였고, 한국등록특허 제1661688호는 n-헥산으로 기름성분이 제거된 참깨 분말의 추출물, 감태 추출물, 줄기부분의 로버참나무껍질 추출물, 어성초 추출물, 유근피 추출물 및 노각 추출물을 유효성분으로 포함하는 조성물로서 상기 추출물들은 정제수에 용해 또는 분산되며, 상기 정제수 100중량부를 기준으로 상기 추출물들 각각 0.5 내지 5 중량부를 포함되는 것을 특징으로 하는 아토피 증상의 예방, 완화 또는 치료용 조성물을 개시하였다.Korean Patent Laid-Open Publication No. 2016-0123592 discloses an agent for inhibiting atopic skin inflammation by using an enzyme inhibitor obtained from extracts of thrush insects, scabs, jojoba, rhubarb, sage grass, white flowers, southern lobster, licorice, Korean Patent No. 1661688 discloses an extract composition comprising sesame powder extract, rosemary extract, rosemary oak bark extract, rhizome extract, rhizome extract and lobster extract, which are oil-removed with n-hexane as an active ingredient Wherein the extracts are dissolved or dispersed in purified water and 0.5 to 5 parts by weight of each of the extracts is contained based on 100 parts by weight of the purified water, thereby providing a composition for preventing, alleviating or treating atopic symptoms.
한편, 갈퀴덩굴은 60~90㎝의 높이로 자라는 두해살이 덩굴풀로서, 줄기에는 네 개의 모가 있고 그 모 위에 밑으로 꼬부라진 작은 가시털이 있어서 다른 물체에 붙어 올라간다. 또한, 마디마다 피침 꼴의 작은 잎이 6~8장씩 둥글게 배열되어 있고 잎 가장자리와 뒷면의 잎맥 위에는 작은 가시털이 나 있으며, 잎겨드랑이 마다 2~3개의 꽃대가 자라나서 1~2송이의 작은 꽃을 피운다.On the other hand, raccoon is a two-year-old vine that grows at a height of 60 to 90 cm. There are four hairs on the stem and a small spiny hairs curled downward on the stem to climb on other objects. In addition, each node has 6 to 8 small leaflets arranged in a round shape. On the edge of leaves and leaf veins on the back side, there are small thorn hairs. Two to three peduncles grow on each side of the leaf, and one or two small flowers I smoke.
갈퀴덩굴에는 주로 쿼세틴-갈락토사이드(quercetin-galactoside) 등의 플라보노이드(flavonoid) 배당체, 아스페루로시드(asperuloside), 탄닌(tannin) 등이 함유된 것으로 알려져 있다. It is known that the extracts contain mainly flavonoid glycosides such as quercetin-galactoside, asperuloside, and tannin.
사용부위는 씨를 포함한 모든 부분이 약재로 쓰이며 진통, 이뇨, 해독, 소종 등의 효능을 가지고 있어, 신경통, 임질, 혈뇨, 장염 등을 다스리는 약으로도 사용되며 기타 악성종기와 종양을 치료하기 위해서도 사용된다고 알려져 있다. 그러나 아직까지 아토피 피부염의 치료에 효능이 있다는 것은 보고된 바 없다.All parts including seeds are used as medicinal parts and they have the efficacy of analgesic, diuretic, detoxification, and elimination. They are also used to treat neuralgia, gonorrhea, hemorrhoids and enteritis, and also to treat other malignant tumors and tumors. . However, it has not been reported that it is effective for the treatment of atopic dermatitis.
이에 본 발명자들은 아토피성 피부염 치료능을 갖는 천연 식물 추출물을 개발하고자 노력한 결과, 갈퀴덩굴 추출물이 아토피 피부염 치료와 관련된 항산화 활성, 염증 싸이토카인 발현 억제능 및 황색포도상구균에 대한 항균활성이 우수하다는 것을 확인하고 본 발명을 완성하게 되었다.Accordingly, the present inventors have made efforts to develop a natural plant extract having an ability to treat atopic dermatitis, and as a result, it has been confirmed that the extract of Extract is superior in antioxidative activity, inhibition of the expression of inflammatory cytokines and antimicrobial activity against Staphylococcus aureus in the treatment of atopic dermatitis Thereby completing the present invention.
본 발명의 목적은 보다 안전하고 친환경적인 천연물 유래 아토피 피부염 예방 또는 개선용 조성물을 제공하는데 있다.It is an object of the present invention to provide a composition for prevention or improvement of atopic dermatitis derived from natural materials which is more safe and environmentally friendly.
상기 목적을 달성하기 위하여, 본 발명은 갈퀴덩굴 추출물을 유효성분으로 함유하는 아토피 피부염 예방 또는 개선용 건강기능식품 조성물을 제공한다.In order to achieve the above object, the present invention provides a health functional food composition for preventing or ameliorating atopic dermatitis, which comprises extracts of Extracts as an active ingredient.
본 발명은 또한 갈퀴덩굴 추출물을 유효성분으로 함유하는 아토피 피부염 예방 또는 치료용 약학 조성물을 제공한다.The present invention also provides a pharmaceutical composition for the prevention or treatment of atopic dermatitis, which comprises extracts of P. vivax as an active ingredient.
본 발명은 또한 갈퀴덩굴 추출물을 유효성분으로 함유하는 아토피 피부염 개선용 화장료 조성물을 제공한다.The present invention also provides a cosmetic composition for improving atopic dermatitis, which contains an extract of Extract as an active ingredient.
본 발명은 또한 갈퀴덩굴 추출물을 유효성분으로 함유하는 아토피 피부염 개선용 물티슈를 제공한다.The present invention also provides a wet tissue for improving atopic dermatitis, which contains an extract of Extract of Rape as an active ingredient.
본 발명에 따른 갈퀴덩굴 추출물을 함유하는 조성물은 항산화 활성, 염증 싸이토카인 발현 억제능 및 황색포도상구균에 대한 항균활성이 우수하고, 세포독성이 거의 없으므로, 아토피 피부염 예방 및 치료를 위한 건강기능식품, 약학조성물, 화장료 조성물의 유효성분으로 유용하게 사용될 수 있다.The composition containing the extract of the extract according to the present invention has excellent antioxidative activity, inhibitory effect on inflammatory cytokine expression and antimicrobial activity against Staphylococcus aureus, and has almost no cytotoxicity. Therefore, the composition for health functional food and pharmaceutical composition for prevention and treatment of atopic dermatitis , And can be effectively used as an effective ingredient of a cosmetic composition.
도 1은 본 발명의 일 실시예에 따른 갈퀴덩굴 추출물의 제조 공정도이다.
도 2는 본 발명의 일 실시예에 따라 제조된 갈퀴덩굴 추출물(AJ1, AJ2, AJ3)의 황색포도상 구균에 대한 항균활성을 평가한 사진이다.
도 3은 본 발명의 일 실시예에 따라 제조된 갈퀴덩굴 추출물(AJ1, AJ2, AJ3)의 항산화능을 평가한 그래프이다.
도 4는 본 발명의 일 실시예에 따라 제조된 갈퀴덩굴 추출물(AJ1, AJ2, AJ3)의 세포생존율을 평가한 그래프이다.
도 5~7은 본 발명의 일 실시예에 따라 제조된 갈퀴덩굴 추출물(AJ1, AJ2, AJ3)의 염증 싸이토카인 발현 억제능을 평가한 그래프이다.FIG. 1 is a view illustrating a process for preparing extracts of Extracts according to an embodiment of the present invention.
FIG. 2 is a photograph showing the antimicrobial activity of Streptomyces sp. Extract (AJ1, AJ2, AJ3) prepared according to one embodiment of the present invention against Staphylococcus aureus.
FIG. 3 is a graph showing the antioxidant activity of the extracts AJ1, AJ2, and AJ3 produced according to one embodiment of the present invention.
FIG. 4 is a graph showing the cell survival rate of extracts AJ1, AJ2, and AJ3 produced according to one embodiment of the present invention.
FIGS. 5 to 7 are graphs for evaluating the inhibitory effect of inflammatory cytokines on the extracts of extracts (AJ1, AJ2, AJ3) prepared according to one embodiment of the present invention.
본 발명에서는 갈퀴덩굴 추출물의 아토피 피부염 예방 또는 치료효과를 확인하고자 하였다. In the present invention, the effect of preventing or treating atopic dermatitis of Extract of Extract was investigated.
본 발명에서는 갈퀴덩굴 추출물이 항산화 활성, 염증 싸이토카인 발현 억제능 및 황색포도상구균에 대한 항균활성이 우수하다는 것을 확인하였다.In the present invention, it was confirmed that the extracts of Phellinus linteus have excellent antioxidative activity, inhibitory effect on inflammatory cytokine expression, and antibacterial activity against Staphylococcus aureus.
즉, 본 발명의 일 실시예에서는 갈퀴덩굴 잎을 에탄올로 추출하고, 감압농축하여 갈퀴덩굴 에탄올 추출물을 획득하였다. 그리고 획득한 에탄올 추출물을 헥산과 물로 순차적으로 분획하여 헥산 분획물 및 물 분획물을 획득하고, 획득한 (a) 에탄올 추출물, (b) 물 분획물 및 (c) 에탄올 추출물과 물 분획물 혼합물이 항산화 활성, 염증 싸이토카인 발현 억제능 및 황색포도상구균에 대한 항균활성이 우수하다는 것을 확인할 수 있었다.That is, in one embodiment of the present invention, the extracts of the extracts of the extracts were obtained by extracting the leaves of the extracts with ethanol and concentrating them under reduced pressure. (A) an ethanol extract, (b) a water fraction, and (c) a mixture of ethanol extract and a water fraction, wherein the obtained ethanol fraction is sequentially fractionated with hexane and water to obtain a hexane fraction and a water fraction, It was confirmed that the anticancer activity against antitumor activity against Staphylococcus aureus was excellent.
따라서, 본 발명은 일 관점에서, 갈퀴덩굴 추출물을 유효성분으로 함유하는 아토피 피부염 예방 또는 개선용 건강기능식품 조성물에 관한 것이다.Accordingly, in one aspect, the present invention relates to a health functional food composition for preventing or ameliorating atopic dermatitis, which comprises extracts of L. japonica as an active ingredient.
상기 갈퀴덩굴은 쌍떡잎식물 용담목 꼭두서니과의 두해살이 덩굴식물로, 한국, 일본, 사할린 등지에서 서식하는데, 본 발명에서는 갈퀴덩굴이라면 특별한 제한없이 이용할 수 있다. 상기 갈퀴덩굴은 잎, 뿌리, 열매 등을 모두 이용할 수 있으나, 잎을 이용하는 것이 바람직하다.The extract is a biennial vine plant of the dicotyledonous species of the dicotyledonous plant. It can be used in Korea, Japan, Sakhalin, and the like. It is preferable to use leaves, roots, fruits and the like, but leaves are preferred.
본 발명에 있어서, 상기 갈퀴덩굴 추출물은 갈퀴덩굴을 유기용매 또는 물로 추출하여 얻을 수 있다. 상기 유기 용매로는 에탄올, 메탄올, 부탄올, 프로판올 등의 극성 용매; 헥산, 에테르, 벤젠, 톨루엔 등의 무극성 용매; 클로로포름, 에틸아세테이트, 아세톤, 메틸렌클로라이드 등의 중간극성 용매를 예시할 수 있으나, 이에 한정되는 것은 아니다. In the present invention, the extract of the Extract can be obtained by extracting the Extract of the Extract with an organic solvent or water. Examples of the organic solvent include polar solvents such as ethanol, methanol, butanol, and propanol; Non-polar solvents such as hexane, ether, benzene, and toluene; Chloroform, ethyl acetate, acetone, methylene chloride and the like, but are not limited thereto.
상기 갈퀴덩굴 추출물은 에탄올 등의 유기용매 또는 물에 갈퀴덩굴을 침지, 중탕, 증류시켜 추출하거나 초임계 추출 또는 마이크로파 추출법으로 추출할 수 있으며, 여과 및 농축과정을 통하여 획득할 수 있다.The extract can be obtained by immersing the extract in an organic solvent such as ethanol or water, extracting the extract by boiling or distilling, extracting the extract by supercritical extraction or microwave extraction, and filtering and concentrating the extracted extract.
상기 분획물은 유기용매의 극성에 따라 용해되는 화합물들을 분리하는 통상의 방법으로 획득할 수 있으며, 본 발명에서는 먼저 추출한 갈퀴덩굴 추출물에 각각 순차적으로 헥산 및 물로 분획하여 헥산 분획물 및 물 분획물을 획득하였다. The fractions can be obtained by a conventional method for separating the compounds to be dissolved according to the polarity of the organic solvent. In the present invention, the extracts of the extracts of the extracts of the extracts were sequentially fractionated with hexane and water to obtain hexane fractions and water fractions.
본 발명의 상기 건강기능식품은 아토피 피부염을 예방하거나 개선하기 위해 섭취할 수 있는 것이면 특별히 제한되지 않는다.The health functional food of the present invention is not particularly limited as long as it can be ingested to prevent or ameliorate atopic dermatitis.
본 발명의 갈퀴덩굴 추출물을 식품첨가물로 사용하는 경우, 갈퀴덩굴 추출물을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 유효 성분의 혼합양은 그의 사용목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있다. 일반적으로, 식품 또는 음료의 제조시에 본 발명의 추출물은 원료에 대하여 0.1 이상 15중량% 이하, 바람직하게는 0.1 이상 10중량% 이하의 양으로 첨가된다. 그러나, 건강 및 위생을 목적으로 하거나 또는 건강 조절을 목적으로 하는 장기간의 섭취의 경우에는 상기 양은 상기 범위 이하일 수 있으며, 안전성 면에서 아무런 문제가 없기 때문에 유효성분은 상기 범위 이상의 양으로도 사용될 수 있다.When the extract of the extract of the present invention is used as a food additive, it can be used as it is or in combination with other food or food ingredients, and can be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to its intended use (prevention, health or therapeutic treatment). Generally, the extract of the present invention is added in an amount of 0.1 to 15% by weight, preferably 0.1 to 10% by weight based on the raw material, when the food or drink is prepared. 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, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range .
상기 식품의 종류에는 특별한 제한은 없다. 상기 물질을 첨가할 수 있는 식품의 예로는 육류, 소시지, 빵, 초콜릿, 캔디류, 스낵류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알콜 음료 및 비타민 복합제 등이 있으며, 통상적인 의미에서의 건강식품을 모두 포함한다.There is no particular limitation on the kind of the food. Examples of foods to which the above substances can be added include dairy products including meats, sausages, breads, chocolates, candies, snacks, confectionery, pizza, ramen noodles, gums, ice cream, soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include healthy foods in a conventional sense.
본 발명의 건강 음료 조성물은 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상술한 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드, 말토스, 슈크로스와 같은 디사카라이드 및 덱스트린, 사이클로덱스트린과 같은 폴리사카라이드, 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 감미제로서는 타우마틴, 스테비아 추출물과 같은 천연 감미제나, 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다.The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. The above-mentioned natural carbohydrates include 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.
상기 외에 본 발명의 추출물은 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 탄산화제 등을 함유할 수 있다. 이러한 성분은 독립적으로 또는 혼합하여 사용할 수 있다. 이러한 첨가제의 비율은 크게 중요하진 않지만 본 발명의 조성물 100 중량부당 0.01~0.1 중량부의 범위에서 선택되는 것이 일반적이다.In addition to the above, the extract of the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and its salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, A carbonating agent used in a carbonated beverage, and the like. These components may be used independently or in combination. The proportion of such additives is not critical, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.
또한, 본 발명은 다른 관점에서 갈퀴덩굴 추출물을 유효성분으로 함유하는 아토피 피부염 예방 또는 치료용 약학 조성물에 관한 것이다.In addition, the present invention relates to a pharmaceutical composition for preventing or treating atopic dermatitis, which comprises, as an active ingredient, Extract of Extract from the other aspect.
본 발명의 약학 조성물에서, 갈퀴덩굴 추출물은 상기 건강식품 조성물에서 기재한 바와 같다.In the pharmaceutical composition of the present invention, the extract of Extract is as described in the above-mentioned health food composition.
본 발명의 아토피 피부염 예방 또는 치료용 약학 조성물은 약학 조성물 총 중량에 대하여 상기 추출물을 0.02 내지 80 중량%, 바람직하게는 0.02 내지 50 중량%로 포함할 수 있다.The pharmaceutical composition for preventing or treating atopic dermatitis of the present invention may contain 0.02 to 80% by weight, preferably 0.02 to 50% by weight of the extract, based on the total weight of the pharmaceutical composition.
본 발명의 추출물을 포함하는 약학 조성물은 약학적 조성물의 제조에 통상적으로 사용되는 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 preparation of pharmaceutical compositions.
본 발명의 추출물의 약학적 투여 형태는 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 alone or in combination with other pharmacologically active compounds as well as in suitable aggregates.
본 발명에 따른 추출물을 포함하는 약학 조성물은, 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 약학 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, 셀룰로오스, 메틸 셀룰로오스, 미정질 셀룰로오스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유 등을 포함한 다양한 화합물 혹은 혼합물을 예시할 수 있다. 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 경구 투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 상기 추출물에 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘카보네이트(calcium carbonate), 수크로오스(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 단순 희석제인 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, 글리세로젤라틴 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 pharmaceutical composition containing the extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium Mention may be made of various compounds or mixtures including silicates, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil and the like . 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, Or 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. Examples of the suppository base include witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like.
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 100 mg/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. 또한, 본 발명에 의한 약학 조성물은 단독으로 투여되거나 다른 약제와 동등하게 또는 다른 약제를 보조하기 위해 함께 투여될 수 있다. 또한, 각 제형의 조성물에 있어서, 상기 필수 성분인 조성물 이외의 다른 성분들은 기타 외용제의 제형 또는 사용목적 등에 따라 당업자가 적의 선정하여 배합할 수 있으며, 이 경우 다른 원료와 동시에 적용할 경우 상승 효과가 일어날 수 있다.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 a daily dose of 0.0001 to 100 mg / kg, preferably 0.001 to 100 mg / kg. 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. In addition, the pharmaceutical composition according to the present invention may be administered alone, or may be administered together with other medicines in the same or in order to supplement other medicines. In addition, in the composition of each formulation, ingredients other than the above-mentioned essential ingredient may be mixed and selected by those skilled in the art according to the formulation or use purpose of the other external preparation. In this case, Can happen.
본 발명은 또 다른 관점에서, 갈퀴덩굴 추출물을 유효성분으로 함유하는 아토피 피부염 개선용 화장료 조성물에 관한 것이다.In another aspect, the present invention relates to a cosmetic composition for improving atopic dermatitis, which comprises an extract of Extract of Vinegar as an active ingredient.
본 발명의 화장료 조성물에서, 갈퀴덩굴 추출물은 상기 건강식품 조성물에서 기재한 바와 같다.In the cosmetic composition of the present invention, the extract of Extract is as described in the above-mentioned health food composition.
본 발명의 아토피 피부염 개선용 화장료 조성물은 피부외용연고, 크림, 유연화장수, 영양화장수, 팩, 에센스, 헤어토닉, 샴푸, 린스, 헤어 컨디셔너, 헤어 트리트먼트, 젤, 스킨로션, 스킨소프너, 스킨토너, 아스트린젠트, 로션, 밀크로션, 모이스처 로션, 영양로션, 마사지 크림, 영양크림, 아이크림, 모이스처 크림, 핸드 크림, 파운데이션, 영양에센스, 선스크린, 비누, 클렌징폼, 클렌징로션, 클렌징크림, 바디 로션 및 바디 클렌저로 이루어지는 군으로부터 선택된 제형을 가질 수 있으나, 이에 제한되지 않는다. 이들 각 제형의 조성물은 그 제형의 제제화에 필요하고 적절한 각종의 기제와 첨가물을 함유할 수 있으며, 이들 성분의 종류와 양은 당업자에 의해 용이하게 선정될 수 있다.The cosmetic composition for improving atopic dermatitis according to the present invention can be used as an ointment for external skin such as ointment for skin, cream, softening longevity, nutrition lotion, pack, essence, hair tonic, shampoo, rinse, hair conditioner, hair treatment, gel, skin lotion, , Lotion, milk lotion, moisturizing lotion, nutrition lotion, massage cream, nutritional cream, eye cream, moisturizing cream, hand cream, foundation, nutrition essence, sunscreen, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion And body cleansers, but are not limited thereto. The composition of each of these formulations may contain various kinds of bases and additives necessary for formulation of the formulation, and the kinds and amounts of these ingredients can be easily selected by those skilled in the art.
본 발명의 제형이 페이스트, 크림 또는 겔인 경우에는 담체 성분으로서 동물섬유, 식물섬유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크 또는 산아연 등이 이용될 수 있다.When the formulation of the present invention is a paste, cream or gel, animal fibers, plant fibers, wax, paraffin, starch, tracant, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide are used as carrier components .
본 발명의 제형이 파우더 또는 스프레이인 경우에는 담체 성분으로서 락토스, 탈크, 실리카, 알루미늄 히드록시드, 칼슘 실리케이트 또는 폴리아미드 파우더가 이용될 수 있고, 특히 스프레이인 경우에는 추가적으로 클로로플루오로히드로카본, 프로판/부탄 또는 디메틸 에테르와 같은 추진체를 포함할 수 있다.When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, / Propane or dimethyl ether.
본 발명의 제형이 용액 또는 유탁액의 경우에는 담체 성분으로서 용매, 용매화제 또는 유탁화제가 이용되고, 예컨대 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌글리콜, 1,3-부틸글리콜 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜 또는 소르비탄의 지방산 에스테르 등이 있다.In the case of the solution or emulsion of the present invention, a solvent, a solvent or an emulsifier is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid esters of sorbitan.
본 발명의 제형이 현탁액인 경우에는 담체 성분으로서 물, 에탄올 또는 프로필렌 글리콜과 같은 액상 희석제, 에톡실화 이소스테아릴 알코올, 폴리옥시에틸렌 소르비톨 에스테르 및 폴리옥시에틸렌 소르비탄 에스테르와 같은 현탁제, 미소결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가 또는 트라칸트 등이 이용될 수 있다.When the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.
본 발명의 제형이 계면-활성제 함유 클렌징인 경우에는 담체 성분으로서 지방족 알코올 설페이트, 지방족 알코올 에테르설페이트, 설포숙신산 모노에스테르, 이세티오네이트, 이미다졸리늄 유도체, 메틸타우레이트, 사르코시네이트, 지방산 아미드 에테르 설페이트, 알킬아미도베타인, 지방족 알코올, 지방산 글리세리드, 지방산 디에탄올아미드, 식물성 유, 리놀린 유도체 또는 에톡실화 글리세롤 지방산 에스테르 등이 이용될 수 있다.When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component is selected from aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolenic derivatives or ethoxylated glycerol fatty acid esters.
본 발명의 아토피 피부염 개선용 화장료 조성물에서, 상기 아토피 피부염 개선용 화장료 조성물은 상기 갈퀴덩굴 추출물을 상기 조성물의 총 중량을 기준으로 0.0001 내지 10% 함유되어 있을 수 있고, 바람직하게는 0.005 내지 1% 함유되어 있을 수 있으나, 바람직한 아토피 피부염의 개선 효과를 제공할 수 있는 유효량을 포함한다면 이에 제한되지 않는다.In the cosmetic composition for improving atopic dermatitis according to the present invention, the cosmetic composition for improving atopic dermatitis may contain 0.0001 to 10%, preferably 0.005 to 1% But it is not limited thereto, including an effective amount that can provide an improvement effect of the desired atopic dermatitis.
본 발명은 또 다른 관점에서, 갈퀴덩굴 추출물을 유효성분으로 함유하는 아토피 피부염 개선용 물티슈에 관한 것이다.In another aspect, the present invention relates to a wet tissue for improving atopic dermatitis, which contains an extract of Extract of Curcuma Asa as an active ingredient.
상기 물티슈는 갈퀴덩굴 추출물 함유 용액에 티슈재를 함침시켜 제조할 수 있다. 티슈재를 함침시키는 갈퀴 덩굴 추출물 함유 용액은 갈퀴덩굴 추출물을 상기 함유 용액의 총 중량을 기준으로 0.0001 내지 10% 함유되어 있을 수 있고, 바람직하게는 0.005 내지 1% 함유되어 있을 수 있다. 상기 티슈재는 물티슈의 제조에 일반적으로 사용하는 소재이면 되고 특별히 제한되는 것은 아니다. 예컨대, 본 발명의 티슈재는 천연 펄프, 가공 펄프, 부직포, 예컨대, 아크릴계 또는 폴리에스테르계와 같은 합성폴리머를 이용한 부직포 등이 될 수 있다. The wet tissue may be prepared by impregnating a tissue material with the extract solution of extract. The extract solution containing the extracts of the extracts for impregnating the tissue material may contain 0.0001 to 10%, preferably 0.005 to 1%, of the extract of Extract, based on the total weight of the solution. The above-mentioned tissue material is not particularly limited as long as it is a material generally used for producing a wet tissue. For example, the tissue material of the present invention may be a natural pulp, a processed pulp, a nonwoven fabric, a nonwoven fabric using a synthetic polymer such as acrylic or polyester, and the like.
[실시예][Example]
이하, 실시 예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것으로, 본 발명의 범위가 이들 실시예에 한정 되는 것은 아니다.Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are for illustrating the present invention only, and the scope of the present invention is not limited to these examples.
실시예 1: 갈퀴덩굴 추출물 및 분획물 분리Example 1: Extracting Extracts and Fractions from Extracts
도 1과 같이, 갈퀴덩굴 잎 50g에, 95% 에탄올 500g을 혼합하고 실온에서 방치한 후 여과하여 여액을 수집하였다. 수집된 여액을 진공농축기로 감압농축하여 갈퀴덩굴 에탄올 추출물(AJ1)을 획득하였다. As shown in Fig. 1, 500 g of 95% ethanol was mixed with 50 g of rape leaves, left at room temperature, and filtered to collect the filtrate. The collected filtrate was concentrated under reduced pressure using a vacuum concentrator to obtain a crude extract (AJ1).
다음으로, 갈퀴덩굴 에탄올 추출물을 순차적으로 헥산 및 물로 분획하여 각각 물 분획물 50g 및 헥산 분획물 50g을 얻은 후, 물 분획물을 동결건조시켜 물 분획물 동결건조분말(AJ2) 8.5g을 획득하고, 갈퀴덩굴 에탄올 추출물(AJ1)에 물 분획물 동결건조분말(AJ2)를 1:1 중량비로 혼합하여 시험시료(AJ3)를 제조하였다.Next, 50 mg of the water fraction and 50 g of the hexane fraction were fractionated successively with hexane and water, and the water fractions were lyophilized to obtain 8.5 g of the freeze-dried powder (AJ2) of the water fraction, The test sample (AJ3) was prepared by mixing the extract (AJ1) with the freeze-dried powder (AJ2) of the water fraction at a weight ratio of 1: 1.
실험예 1: 갈퀴덩굴 추출물의 황색포도상 구균에 대한 항균활성 평가Experimental Example 1: Evaluation of antimicrobial activity against Staphylococcus aureus extract
황색포도상 구균은 아토피 피부염 환자 피부병변의 90% 이상에서 발견되지만 정상인의 피부에서는 5% 미만만 발견된다. 또한 정상인에서는 균 집락을 이루더라도 그 수가 적으나, 아토피 피부염의 급성병변에서는 107 colony-forming units/cm까지 발견된다. 이러한 점들을 종합하여 볼 때 황색포도상 구균이 아토피 피부염 환자의 피부 염증 소견과 연관성이 있음을 알 수 있다(소아알레르기 호흡기, 제16권 제2호, 2006)Staphylococcus aureus is found in more than 90% of skin lesions in atopic dermatitis patients, but less than 5% in normal skin. In addition, the number is normal, but ever even made the bacteria colonies are found in acute lesions of atopic dermatitis up to 10 7 colony-forming units / cm . These findings suggest that Staphylococcus aureus is associated with skin inflammation in patients with atopic dermatitis (Pediatric Allergy Respiratory, Vol. 16, No. 2, 2006)
따라서, 본 실험예에서는 실시예 1에서 획득한 에탄올 추출물(AJ1)의 황색포도상 구균에 대한 항균활성을 하기와 같이 평가하고, 그 결과를 표 1 및 도 2에 나타내었다.Therefore, in this Experimental Example, the antimicrobial activity of the ethanol extract (AJ1) obtained in Example 1 against Staphylococcus aureus was evaluated as follows, and the results are shown in Table 1 and Fig.
황색포도상 구균(Staphylococcus aureus)(ATCC 6538)을 Brain heart infusion broth(DIFCO, USA)에 접종하여 35 ㅁ 1℃에서 18~24시간 동안 전 배양시켰다. 전 배양된 균 배양액을 생균수 1~9 ㅧ 107 CFU/㎖가 되도록 멸균생리식염수에 희석한 후 시험균액으로 사용하였다. Staphylococcus aureus (ATCC 6538) was inoculated into Brain heart infusion broth (DIFCO, USA) and preincubated at 35 ° C and 1 ° C for 18-24 hours. The pre-cultured bacterial culture was diluted in sterile physiological saline to a concentration of 1 ~ 9 ㅧ 10 7 CFU / ㎖ and used as a test strain.
실시예 1에서 획득한 갈퀴덩굴 에탄올 추출물 20㎎을 DMEM(w/o FBS) 20㎖에 용해시킨 시험용액 20㎖ 및 대조군(DMEM(w/o FBS)) 20㎖에 각각 시험균액 0.2㎖를 첨가하여 혼합한 후 상온에서 24시간 방치하였다. 최초 희석은 D/E neutralizing broth(DIFCO, USA)를 이용하였다. 중화된 시험액은 단계별로 희석하여 각 농도당 Petri dish 2매에 1㎖씩 분주하였다. 미리 준비된 45~50℃의 Tryptic soy agar(DIFCO, USA)를 Petri dish에 15~25㎖ 분주하고, 상온에서 응고시켰다. 응고된 Petri dish는 거꾸로 하여 35 ㅁ 1 ℃에서 24 ㅁ 2시간 동안 배양하였다. 초기 및 대조 세균수의 측정은 멸균생리식염수를 사용하여 실시하였다.0.2 ml of the test strain was added to 20 ml of the test solution in which 20 mg of the extract obtained from Example 1 was dissolved in 20 ml of DMEM (w / o FBS) and 20 ml of the control (DMEM (w / o FBS) The mixture was allowed to stand at room temperature for 24 hours. Initial dilution was performed with D / E neutralizing broth (DIFCO, USA). The neutralized test solution was diluted step by step and dispensed at 1 ml per 2 Petri dishes per concentration. Prepared Tryptic soy agar (DIFCO, USA) at 45 ~ 50 ℃ was dispensed in 15 ~ 25ml of Petri dish and solidified at room temperature. The solidified Petri dish was inverted and cultured at 35 ° C and 1 ° C for 24 hours and 2 hours. Initial and control bacterial counts were determined using sterile physiological saline.
(99.9 % 이상)<10
(99.9% or more)
( ) = 감소율(%) = [(A - B) / A] ㅧ 100() = Reduction rate (%) = [(A - B) / A] 100
A : 대조군의 일정시간 후 생균수 (평균)A: Number of viable cells (average) after a certain time in the control group
B : 시험군의 일정시간 후 생균수 (평균)B: Number of live cells (average) after a certain time in the test group
위 표에서 나타난 것과 같이, 시험결과 에탄올 추출물(AJ1)을 용해시킨 시험군에서는 황색포도상 구균이 99.9% 이상 감소한 것으로 나타나, 황색포도상 구균에 대한 항균활성이 우수한 것으로 나타났다.As shown in the above table, in the test group in which the ethanol extract (AJ1) was dissolved in the test, the amount of Staphylococcus aureus decreased by 99.9% or more, indicating that the antimicrobial activity against Staphylococcus aureus was excellent.
실험예 2: 갈퀴덩굴 추출물의 항산화능 평가Experimental Example 2: Evaluation of Antioxidative Ability of Extract of Extract
실시예 1에서 확보한, 갈퀴덩굴 에탄올 추출물(AJ1), 물 분획물 동결건조분말(AJ2) 및 갈퀴덩굴 에탄올 추출물 +물 분획물 동결건조분말(AJ3)의 항산화능을 평가하기 위해 DPPH assay법을 수행하고, 그 결과를 도 3에 나타내었다. The DPPH assay was performed to evaluate the antioxidative activity of the extracts of AJ1, AJ2, AJ2 and AJ3 obtained in Example 1 and extracts of freeze-dried and water-fraction freeze-dried (AJ3) , And the results are shown in Fig.
DPPH 라디칼 소거 활성은 Blois(Blois MS. Antioxidant determination by the use of a stable free radical. Nature 181: 1199-1200 (1958))의 방법으로 측정하였다. 시료(AJ1, AJ2, AJ3, ascorbic acid)를 DMEM(w/o FBS)로 녹여 5ng/㎖~10㎎/㎖이 되도록 정량하여 96 웰 플레이트에 각 시료를 50㎕를 주입하고, 동시에 0.3 mM DPPH 500㎕를 넣어 총량이 550㎕가 되도록 하였다. 실온에서 30분간 반응시킨 후 microplate reader로 517nm 파장에서 흡광도를 측정하고 그 결과를 도 3에 나타내었다.The DPPH radical scavenging activity was measured by the method of Blois (Blois MS. Antioxidant determination by the use of a stable free radical. Nature 181: 1199-1200 (1958)). The samples (AJ1, AJ2, AJ3, ascorbic acid) were dissolved in DMEM (w / o FBS) and quantitated to 5 ng / ml to 10 mg / ml. 50 ㎕ of each sample was injected into a 96- And the total amount was adjusted to 550 μl. After incubation at room temperature for 30 minutes, the absorbance at 517 nm was measured with a microplate reader. The results are shown in FIG.
DPPH 라디칼 소거 활성은 시료 용액의 첨가군과 무첨가군 사이의 흡광도의 차이를 백분율로 나타내었다.The DPPH radical scavenging activity was expressed as a percentage of absorbance difference between the addition group and the no addition group of the sample solution.
DPPH 라디칼 소거 활성(%) = {1 - (시료 첨가군 흡광도/시료 무첨가군 흡광도) } ㅧ 100DPPH radical scavenging activity (%) = {1- (absorbance of sample addition group / absorbance of sample addition group)} 100
도 3에 나타난 바와 같이, 갈퀴덩굴 에탄올 추출물(AJ1)은 positive control로 사용된 ascorbic acid보다는 상대적으로 낮지만 우수한 항산화능을 갖고 있으며, 물 분획물 동결건조분말(AJ2) 및 갈퀴덩굴 에탄올 추출물 +물 분획물 동결건조분말(AJ3)은 ascorbic acid와 유사 혹은 그 이상의 항산화능을 보유한다는 것을 확인할 수 있었다. As shown in FIG. 3, the extract of AJ1 was relatively lower than ascorbic acid used as a positive control, but had excellent antioxidant ability. The freeze-dried powder (AJ2) of water fraction and the ethanol extract + It was confirmed that freeze-dried powder (AJ3) possesses antioxidant ability similar to or higher than ascorbic acid.
실험예 3: 갈퀴덩굴 추출물의 세포생존율 평가Experimental Example 3: Evaluation of cell survival rate of Extract of Extracts
실시예 1에서 확보한, 갈퀴덩굴 에탄올 추출물(AJ1), 물 분획물 동결건조분말(AJ2) 및 갈퀴덩굴 에탄올 추출물 +물 분획물 동결건조분말(AJ3)의 독성을 확인하기 위하여, mouse macrophage인 RAW264.7 세포에서 세포생존율을 측정하였다. 즉, 96 well plate에 세포를 분주한 다음 페니실린 (100 IU/㎖), 스트렙토마이신 (100㎍/㎖), 10% FBS를 함유하는 DMEM배지를 넣고 37℃, 5% CO2를 포함하는 배양기에서 24시간 배양하였다. 배양 후, 배지를 제거하고 PBS로 세척한 다음, 시료(AJ1, AJ2, AJ3) 및 새로운 배지를 넣고 24시간 배양하였다. 24시간 이후, 세포의 생존율을 측정하기 위해 DMEM(w/o phenol red)에 1/10로 희석한 CCK8 solution을 접종하고 1시간 동안 배양기 내에서 배양한 다음 상등액을 microplate reader를 사용하여 450 nm 파장에서 흡광도를 측정하고, 세포 생존율을 계산하여, 그 결과를 도 4에 나타내었다. 세포 생존율은 세포에 아무 처리하지 않은 무처리군의 세포수를 기준(100%)으로 하여 상대적인 수치를 계산하였다.To confirm the toxicity of the extracts of the extracts of the extracts AJ1, AJ2 and AJ3 obtained in Example 1, the mouse macrophages RAW264.7 (AJ1) and the water extracts of freeze-dried (AJ3) Cell viability was measured in cells. That is, the busy cells in 96 well plate the following penicillin (100 IU / ㎖), streptomycin (100㎍ / ㎖), put into a DMEM medium containing 10% FBS in an incubator containing 37 ℃, 5% CO 2 And cultured for 24 hours. After culturing, the medium was removed, washed with PBS, and samples (AJ1, AJ2, AJ3) and fresh medium were added and cultured for 24 hours. After 24 hours, CCK8 solution diluted 1/10 in DMEM (w / o phenol red) was inoculated and incubated for 1 hour in an incubator to measure cell viability. The supernatant was transferred to a 450 nm wavelength , And the cell viability was calculated. The results are shown in Fig. The cell viability was calculated based on the number of cells in the untreated untreated group (100%).
도 4에 도시된 바와 같이, 갈퀴덩굴 에탄올 추출물(AJ1)은 1㎍/㎖의 농도에서 118.8% 이상의 세포 생존율을 보였고, 물 분획물 동결건조분말(AJ2)는 500㎍/㎖ 농도에서 147%의 생존율을 보였으며, 갈퀴덩굴 에탄올 추출물 +물 분획물 동결건조분말(AJ3)은 500ng/㎖의 농도에서 103%의 세포생존율을 보였다.As shown in FIG. 4, the survival rate of the extract of AJ1 was 118.8% or higher at a concentration of 1 μg / ml, and the survival rate of 147% at a concentration of 500 μg / ml of the lyophilized powder (AJ2) (AJ3) showed a cell viability of 103% at the concentration of 500ng / ㎖.
즉, 실시예 1에서 획득한 시료(AJ1, AJ2, AJ3)는 일정 농도에서 면역세포의 활성을 강화하고 생존율을 증대시키는 효과가 있음을 확인할 수 있었다. That is, it was confirmed that the samples (AJ1, AJ2, AJ3) obtained in Example 1 had the effect of enhancing the activity of immune cells and increasing the survival rate at a certain concentration.
실험예 4: 갈퀴덩굴 추출물의 염증 싸이토카인 발현억제능 평가 Experimental Example 4: Evaluation of the inhibitory effect of extracts of extracts of inflammation on the expression of cytokines
염증 싸이토카인은 아토피 피부염 등을 유발하는 중요 요인으로 염증 싸이토카인의 발현을 조절하는 소재는 피부 염증완화와 관련이 있다. 따라서 시료(AJ1, AJ2, AJ3)의 염증 싸이토카인 발현양을 확인하여 염증 억제 효능을 확인하였다.Inflammatory cytokines are important factors that cause atopic dermatitis and so on. Materials that regulate the expression of inflammatory cytokines are related to skin inflammation relief. Therefore, the inflammatory cytotoxic activity of the samples (AJ1, AJ2, AJ3) was confirmed by confirming the expression of inflammatory cytokines.
염증성 마크로파지는 lipopolysaccharide(LPS)와 인터페론에 의해 자극을 받으면 염증성 싸이토카인인 Interleukin류와 TNF의 발현을 증가시켜 염증반응을 유도한다. 따라서 세포의 염증반응을 유도하기 위하여 LPS와 INFγ를 처리하여 염증반응을 유도한 뒤 세포에 각각 시료(AJ1, AJ2, AJ3)를 처리하여 염증 싸이토카인 유전자의 발현을 확인하였다. Inflammatory macrophages induce inflammatory responses by stimulating the expression of interleukins and TNF, which are inflammatory cytokines, when stimulated by lipopolysaccharide (LPS) and interferon. Therefore, in order to induce the inflammatory response of the cells, the inflammatory reaction was induced by treatment with LPS and INFγ, and the cells were treated with the respective samples (AJ1, AJ2, AJ3) to confirm the expression of the inflammatory cytokine gene.
RAW 264.7 세포를 96 웰 플레이트에 최종농도가 2ㅧ105 cells/㎖가 되도록 분주한 뒤 37℃, 5% CO2 인큐베이터에서 24시간 배양한 후 시료(AJ1, AJ2, AJ3)를 농도별로 처리한 다음, 1시간 후 염증유도 물질인 LPS(1㎍/㎖)와 interferon-γ 를 처리하여 24시간 배양하였다. 배양 후 trizol을 사용하여 RNA를 추출하고 reverse transcription을 진행하였다. 역전사된 cDNA를 Taqman gene expression assay를 통해 realtime RT-PCR을 실시하여 면역 싸이토카인의 유전자 (IL1α, 6, TNFα) 발현을 분석하고, 그 결과를 도 5(AJ1), 도 6(AJ2) 및 도 7(AJ3)에 나타내었다.RAW 264.7 cells were seeded in 96-well plates to a final concentration of 2 × 10 5 cells / ㎖, cultured in a 5% CO 2 incubator at 37 ° C for 24 hours, treated with AJ1, AJ2, AJ3 Then, 1 hour later, LPS (1 μg / ml) and interferon-γ, which are inflammation inducers, were treated and cultured for 24 hours. After incubation, RNA was extracted using trizol and reverse transcription was performed. The reverse transcribed cDNAs were analyzed by real-time RT-PCR using the Taqman gene expression assay to analyze the expression of the genes of the immunocytokines (IL1 ?, 6, TNF?) And the results are shown in FIG. 5 (AJ1) (AJ3).
도 5에 나타난 바와 같이, 에탄올 추출물(AJ1)은 1㎍/㎖의 농도에서 IL6의 발현을 유의성 있게 감소시켰고, 도 6에 나타난 바와 같이, 물 분획물 동결건조분말(AJ2)는 5㎍/㎖, 25㎍/㎖ 및 50㎍/㎖ 농도에서 IL6의 발현을 유의성 있게 감소시켰고, 5㎍/㎖ 및 50㎍/㎖ 농도에서 TNFα의 발현을 유의성 있게 감소시켰음을 확인할 수 있었다.As shown in FIG. 5, the ethanol extract (AJ1) significantly decreased IL6 expression at a concentration of 1 μg / ml. As shown in FIG. 6, the freeze-dried powder (AJ2) IL6 expression was significantly decreased at the concentration of 25 / / ㎖ and 50 / / ㎖, and the expression of TNFα was significantly decreased at the concentration of 5 / / ㎖ and 50 / / ㎖.
또한, 도 7에 나타난 바와 같이, 갈퀴덩굴 에탄올 추출물 +물 분획물 동결건조분말(AJ3)은 500ng/㎖, 2.5㎍/㎖ 및 5㎍/㎖, 농도에서 각각 IL1α 및 IL6의 발현을 유의성 있게 감소시켰음을 확인할 수 있었다.In addition, as shown in Fig. 7, the expression of IL1 [alpha] and IL6 was significantly reduced at 500 ng / ml, 2.5 [mu] g / ml and 5 g / ml concentrations of ethanol extract of ragweed vinegar + lyophilized powder (AJ3) .
이상으로 본 발명 내용의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서 이러한 구체적 기술은 단지 바람직한 실시태양일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서, 본 발명의 실질적인 범위는 첨부된 청구 항들과 그것들의 등가물에 의하여 정의된다고 할 것이다.While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will readily appreciate that many modifications are possible in the exemplary embodiments without materially departing from the novel teachings and advantages of this invention. something to do. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.
Claims (8)
상기 갈퀴덩굴 추출물은 총 중량에 대하여 0.1 내지 15 중량%로 포함되는 것을 특징으로 하는 아토피 피부염 예방 또는 개선용 건강기능식품 조성물.The method according to claim 1,
The composition of claim 1, wherein the extract is present in an amount of 0.1 to 15% by weight based on the total weight of the composition.
상기 갈퀴덩굴 추출물은 총 중량에 대하여 0.02 내지 50 중량%로 포함되는 것을 특징으로 하는 아토피 피부염 예방 또는 치료용 약학 조성물.The method of claim 3,
The pharmaceutical composition for preventing or treating atopic dermatitis according to claim 1, wherein the extract is from 0.02 to 50% by weight based on the total weight of the extract.
상기 갈퀴덩굴 추출물은 총 중량에 대하여 0.0001 내지 10 중량%로 포함되는 것을 특징으로 하는 아토피 피부염 개선용 화장료 조성물.6. The method of claim 5,
The cosmetic composition for improving atopic dermatitis according to claim 1, wherein the extract is at a concentration of 0.0001 to 10% by weight based on the total weight of the cosmetic composition.
상기 갈퀴덩굴 추출물이 총 중량에 대하여 0.0001 내지 10 중량%로 포함되어 있는 함유 용액에 함침된 것을 특징으로 하는 아토피 피부염 개선용 물티슈.8. The method of claim 7,
Wherein the extract is impregnated with a solution containing 0.0001 to 10% by weight based on the total weight of the extract.
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