KR101945864B1 - Cosmetic Composition Including Dendranthema zawadskii Extract for young face by Anti-aging, Skin-lifting and Anti-wrinkle - Google Patents
Cosmetic Composition Including Dendranthema zawadskii Extract for young face by Anti-aging, Skin-lifting and Anti-wrinkle Download PDFInfo
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- KR101945864B1 KR101945864B1 KR1020160114091A KR20160114091A KR101945864B1 KR 101945864 B1 KR101945864 B1 KR 101945864B1 KR 1020160114091 A KR1020160114091 A KR 1020160114091A KR 20160114091 A KR20160114091 A KR 20160114091A KR 101945864 B1 KR101945864 B1 KR 101945864B1
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- South Korea
- Prior art keywords
- cosmetic composition
- extract
- ulleung chrysanthemum
- aging
- ulleung
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Abstract
본 발명은 울릉국화 추출물을 유효성분으로 포함하는 세포노화 억제용 조성물 및 세포노화 억제와 주름개선 및 피부 탄력 개선을 통한 동안용 화장료 조성물에 관한 것이다. 본 발명의 울릉국화 추출물은 세포노화 관련 질환 및 염증성 질환을 치료하거나 예방할 수 있다. 특히 본 발명의 화장료 조성물은 주름 및 탄력 개선에 효과적이다.The present invention relates to a composition for inhibiting cell senescence comprising Ulleung chrysanthemum extract as an active ingredient, and a cosmetic composition for inhibiting cell senescence, improving wrinkles and improving skin elasticity. The Ulreung chrysanthemum extract of the present invention can treat or prevent cell senescence-related diseases and inflammatory diseases. In particular, the cosmetic composition of the present invention is effective for improving wrinkles and elasticity.
Description
본 발명은 울릉국화 추출물을 유효성분으로 포함하는 화장료 조성물에 관한 것으로, 더욱 상세하게는 노화 관련 질환에 유용하게 사용될 수 있는 울릉국화 추출물을 포함하는 세포노화 억제와 주름개선 및 피부 탄력 개선을 통한 동안(童顔)용 화장료 조성물에 관한 것이다.The present invention relates to a cosmetic composition comprising Ulleung chrysanthemum extract as an active ingredient, and more particularly to a cosmetic composition comprising Ulleung chrysanthemum extract which can be usefully used for aging-related diseases, The present invention relates to a cosmetic composition for a child's face.
노화를 지연시켜 퇴행성질환이 발병하지 않고 건강하게 질병 없이 오래 사는 것은 모든 사람의 바람으로 노화를 억제시키는 물질을 찾는 노력은 과거부터 현재까지도 계속해서 진행되고 있다. 노화를 촉진시키는 원인의 하나로 슈퍼옥사이드, 과산화수소, 수산화기 및 일중항산소와 같은 활성산소종들에 의한 산화 작용을 들 수 있다. 활성산소종들의 대사산물로 생성된 자유기(free radical)들이 생체 내에서 단백질, 생체막, DNA 등에 작용하여 산화적 손상을 주는데, 이들 작용을 억제시키기 위해서 항산화 성분에 대해 많은 연구가 이루어졌고, 항산화 작용을 갖는 새로운 소재를 탐색하기 위한 연구가 국내외에서 활발하게 이루어지고 있다. 그러나 노화현상은 산화에 의해서만 이루어지는 것이 아니고 텔로미어, 세포복제능력, 유전자손상 및 회복능력 등 수많은 요인에 의해서 영향을 받는 것으로 알려져 있기 때문에 천연물 및 약제의 항산화 효과가 아닌 노화억제능력을 평가를 위해서는 세포의 노화를 이용하는 방법이 필요하다.
The delay in the aging process, without degenerative disease, healthy and long-lasting without disease is an ongoing effort to find a substance that inhibits aging by the wind of all people. Oxidation by active oxygen species such as superoxide, hydrogen peroxide, hydroxyl group and singlet oxygen is one of the factors promoting aging. Free radicals generated by the metabolites of reactive oxygen species act on proteins, biomembranes, and DNA in vivo and cause oxidative damage. Many studies have been conducted on antioxidant components to suppress these effects, Researches are being actively conducted at home and abroad to search for new materials having action. However, since the aging phenomenon is not only caused by oxidation but is known to be influenced by numerous factors such as telomerase, cell replication ability, gene damage and recovery ability, it is necessary to evaluate the antioxidative effect of natural products and drugs, A method of using aging is needed.
세포노화는 정상 체세포가 일정 횟수 분열한 후 더 이상 분열하지 못하는 현상으로, 개체 및 조직의 노화에 기여하며, 세포의 비정상적인 증식과 암 형성을 억제하는 중요한 기전이다. 세포노화는 반복되는 체세포의 분열로 인해 염색체의 끝 부분인 텔로미어가 짧아져 생기거나, 암유전자 또는 암억제 유전자의 활성 증가, 과다한 산화스트레스, 자외선이나 방사선 조사, 항암제와 같은 세포독성 물질, 염증반응 등에 의해서도 생긴다.Cell senescence is a phenomenon in which normal somatic cells can no longer divide after a certain number of divisions, contributing to the aging of individuals and tissues, and is an important mechanism for inhibiting abnormal cell proliferation and cancer formation. Cell senescence is caused by the shortening of the telomere at the end of the chromosome due to repeated somatic cell division, the increase of the activity of the cancer gene or the cancer suppressor gene, the excessive oxidative stress, the cytotoxic substance such as ultraviolet ray or radiation, And the like.
노화된 세포는 모양이 커지고, 편평해지며, 핵에 이질염색질이 증가하고, 세포질에 공포가 많아지는 형태학적 특징과 함께, SA-β-gal (senescence-associated β-galatosidase) 활성이 증가하며, p53, p16INK4, p21과 같은 세포성장을 억제하는 단백질들의 양이 많아지고, 인슐린양성장인자 결합단백질 (insulin-like growth factor binding proteins; IGFBPs), 인터루킨-6 (interleukin-6), 전환성장인자-β (transforming growth-β;TGF-β), 인터페론 (interferon)과 같은 여러 가지 염증 단백질들을 분비한다.The senescence-associated β-galactosidase (SA-β-gal) activity increases with the morphological features that aged cells become larger, flattened, increase in heterochromatin in the nucleus, (IGFBPs), interleukin-6 (interleukin-6), and transgene-like growth factor-binding proteins (IGFBPs) (TGF-beta), interferon, and other inflammatory proteins.
세포노화는 단순히 개체나 조직의 노화에 기여할 뿐만 아니라, 여러 가지 다양한 질병의 병인에 중요한 역할을 한다. 노화세포는 류마티스성 관절염, 골관절염, 간염, 만성 피부손상 조직, 동맥경화 혈관조직 등과 같은 염증성 병변 조직에서 많이 관찰된다. 또한, 전립샘 증식증과 간염, 간암 등에서도 세포노화 현상이 관찰된다. Cell senescence not only contributes to the aging of individuals and tissues, but also plays an important role in the pathogenesis of various diseases. Aging cells are frequently observed in inflammatory lesions such as rheumatoid arthritis, osteoarthritis, hepatitis, chronic skin injured tissue, and arteriosclerotic vascular tissue. In addition, cell senescence is observed in proliferative hyperplasia, hepatitis, and liver cancer.
이와 같이 노화세포가 축적되면 노화세포는 잘 분열하지 못하므로 손상된 조직이 적절히 복구되지 못할 뿐만 아니라, 주위 조직을 분해하는 효소나 염증성 싸이토카인 등을 분비하므로 조직의 손상을 가속시키므로, 노화와 관련된 질병의 병인에 기여한다.As the aging cells accumulate, the aging cells do not divide well. Therefore, not only the damaged tissues are properly restored but also the enzymes that decompose the surrounding tissues or the inflammatory cytokines are secreted, accelerating the damage of the tissues. Contributes to the pathogenesis.
세포성 노화가 노화의 필수 원인 요소인 것으로 여겨지기 때문에, 세포성 노화를 지연시키는 방법을 개발하려는 노력이 있어왔다. 그 중에서 세포노화를 조절할 수 있는 물질을 탐색하여, 이를 질병의 예방과 치료에 활용하고자 하는 연구가 일부 보고되고 있다.Since cellular aging is considered to be an essential cause of aging, efforts have been made to develop methods for delaying cellular aging. Among them, some researches have been conducted to search for substances capable of regulating cell senescence and to utilize them for the prevention and treatment of diseases.
세포노화를 조절하는 것으로 알려진 물질로는 SIRT1 활성을 증가시키는 것으로 알려진 붉은 포도주에 많이 들어 있는 레스베라트롤(resveratrol), 각질세포 (keratinocyte) 에서 세포성 노화를 지연시키기 위한 레티노산의 용도 등이 대표적이다. 아울러 식물 추출물로부터 세포노화를 조절할 수 있는 물질을 동정하고 분리하여 화장품과 건강 기능성 식품 등에 활용하고자 하는 연구가 보고 되고 있다. Some of the substances known to regulate cell senescence include resveratrol, which is known to increase SIRT1 activity, and retinoic acid, which is used to delay cellular aging in keratinocytes. In addition, studies have been reported on the identification of substances capable of regulating cell senescence from plant extracts and their use in cosmetics and health functional foods.
한편, 국화과 산국속(Dendranthema)은 우리나라를 포함한 유럽, 아메리카 등지에 분포하고, 특히, 울릉국화 (Dendranthema zawadskii var. lucidum (Nakai) J.H.Park)는 경상북도 울릉도 성인봉의 산지에 난다. 잎은 깊게 비교적 가늘게 갈라지고 갈래는 피침형으로 두껍고 광채가 나며 잎자루가 있는 특징이 있다. 울릉국화에 대한 종래의 연구결과로는 울릉국화의 꽃과 잎, 줄기에서 DPPH 라디칼 소거능 및 ABTS 라디칼 소거능이 우수하다는 내용의 울릉국화의 항산화 효과에 대한 연구가 개시되어 있다(우정향 외 2, 국화과 Dendranthema속 식물 2종 80% 에탄올 추출물의 항산화 효과, Korean J. Plant Res., 23(1):47, 2010). 하지만 울릉 국화 추출물의 세포노화 억제 효능에 대한 결과는 보고된 바 없었다.On the other hand, Dendranthema is distributed in Europe, America, and Korea including Korea. Especially, Dendranthema zawadskii var. Lucidum (Nakai) J.H.Park is located in the mountain area of Adult of Ulleungdo in Kyungsangbuk province. Leaves are deeply and relatively thinly split, with lenticular lanceolate, thick, luminous and petiole-like. Previous studies on Ulleung chrysanthemum have shown that the antioxidant effect of Ulleung chrysanthemum is superior to DPRKH radical scavenging ability and ABTS radical scavenging ability in flowers, leaves and stems of Ulleung chrysanthemum (Ureung chrysanthemum 2, Antioxidative effects of 80% ethanol extracts of two species of Dendranthema spp., Korean J. Plant Res., 23 (1): 47, 2010). However, the results of inhibiting the cell senescence of Ulleung chrysanthemum extract were not reported.
본 발명은 울릉국화(Dendranthema zawadskii var. lucidum) 추출물을 유효성분으로 포함하는 화장료 조성물을 제공하는데 있다.
The present invention relates to a method for the production of < RTI ID = I have zawadskii . lucidum ) as an active ingredient.
그러나, 본 발명이 이루고자 하는 기술적 과제는 이상에서 언급한 과제에 제한되지 않으며, 언급되지 않은 또 다른 과제들은 아래의 기재로부터 당업자에게 명확하게 이해될 수 있을 것이다. However, the technical problem to be solved by the present invention is not limited to the above-mentioned problems, and other matters not mentioned can be clearly understood by those skilled in the art from the following description.
따라서, 본 발명은 울릉국화(Dendranthema zawadskii var. lucidum) 추출물을 유효성분으로 포함하는 화장료 조성물을 제공한다.Accordingly, the present invention provides a cosmetic composition comprising an extract of Dendranthema zawadskii var. Lucidum as an active ingredient.
상기 본 발명의 바람직한 일실시예에 따르면, 상기 상기 울릉국화(Dendranthema zawadskii var. lucidum) 추출물은 세포노화를 억제하는 것일 수 있다. According to a preferred embodiment of the present invention, the extract of Dendranthema zawadskii var. Lucidum may inhibit cell senescence.
상기 본 발명의 바람직한 다른 일실시예에 따르면, 상기 울릉국화 추출물은 줄기, 잎 및 꽃으로 이루어지는 울릉국화의 지상부로부터 추출한 것일 수 있다. According to another preferred embodiment of the present invention, the Ulleung chrysanthemum extract may be extracted from the ground part of Ulleung chrysanthemum comprising stem, leaf and flower.
상기 본 발명의 바람직한 또 다른 일실시예에 따르면, 상기 울릉국화 추출물은 SA-β-갈락토시다아제 활성을 저해하여 주름개선 및 피부 탄력 개선 효과를 가지는 것일 수 있다. According to another preferred embodiment of the present invention, the Ulleung chrysanthemum extract may inhibit SA-β-galactosidase activity, thereby improving wrinkles and improving skin elasticity.
상기 본 발명의 바람직한 또 다른 일실시예에 따르면, 상기 울릉국화 추출물은 섬유아세포(human dermal fibroblasts, HDFs)의 노화를 저해하는 것일 수 있다. According to another preferred embodiment of the present invention, the Ulleung chrysanthemum extract may inhibit the aging of human dermal fibroblasts (HDFs).
상기 본 발명의 바람직한 또 다른 일실시예에 따르면, 상기 울릉국화 추출물은 물, 알코올, 에틸아세테이트, 클로로포름 및 헥산으로 이루어진 군에서 선택된 어느 하나 이상의 용매로 추출한 것일 수 있다. According to another preferred embodiment of the present invention, the Ulleung chrysanthemum extract may be extracted with at least one solvent selected from the group consisting of water, alcohol, ethyl acetate, chloroform and hexane.
상기 본 발명의 바람직한 또 다른 일실시예에 따르면, 상기 화장료 조성물은 울릉국화 추출물을 2 내지 10 μg/ml 농도로 포함하는 것일 수 있다.According to another preferred embodiment of the present invention, the cosmetic composition may contain Ulleung chrysanthemum extract at a concentration of 2 to 10 μg / ml.
상기 본 발명의 바람직한 또 다른 일실시예에 따르면, 상기 울릉국화 추출물은 조성물 총 중량에 대하여 0.001 내지 10중량%로 포함하는 것일 수 있다.According to another preferred embodiment of the present invention, the Ulleung chrysanthemum extract may be contained in an amount of 0.001 to 10% by weight based on the total weight of the composition.
상기 본 발명의 발명의 바람직한 또 다른 일실시예에 따르면, 상기 화장료 조성물은 크림, 로션, 파우더, 계면활성제-함유 클린싱오일, 분말 파운데이션, 유탁액 파운데이션, 왁스 파운데이션, 페이스트, 에센스, 아이크림 중에서 선택되는 어느 하나의 제형인 것일 수 있다. According to another preferred embodiment of the present invention, the cosmetic composition may be selected from a cream, a lotion, a powder, a surfactant-containing cleansing oil, a powder foundation, an emulsion foundation, a wax foundation, a paste, Or a pharmaceutically acceptable salt thereof.
상기 본 발명의 바람직한 또 다른 일실시예에 따르면, 상기 울릉국화의 지상부의 알코올 추출물을 포함하는, 피부노화방지, 주름개선 및 피부 탄력 개선을 통한 동안(童顔)용 화장료 조성물을 제공한다.According to another preferred embodiment of the present invention, there is provided a cosmetic composition for preventing aging of the skin, improving wrinkles and improving skin elasticity, comprising the alcohol extract of the above-mentioned Ulleung chrysanthemum.
본 발명은 상기와 같은 문제점을 해결하기 위해 안출된 것으로, 본 발명은 울릉국화 추출물을 유효성분으로 포함하는 화장용 조성물에 관한 것으로, 상기 화장료 조성물은 세포노화 관련 질환을 치료하거나 예방할 수 있다. 특히 상기 화장료 조성물은 주름 및 탄력 개선에 효과적이다.Disclosure of the Invention The present invention has been conceived to solve the above-mentioned problems. The present invention relates to a cosmetic composition containing Ulleung chrysanthemum extract as an active ingredient, and the cosmetic composition can treat or prevent a cell senescence-related disease. In particular, the cosmetic composition is effective for improving wrinkles and elasticity.
도 1은 울릉국화 사진이다.
도 2는 섬유아세포(HDF)에서 울릉국화 추출물에 대한 MTT 분석 결과를 나타낸 그래프이다(Doxo : doxorubicin 0.5 μM (27.18 μg/ml), 5 : 울릉국화 추출물 5 μg/ml, 10 : 울릉국화 추출물 10 μg/ml, 20 : 울릉국화 추출물 20 μg/ml).
도 3은 섬유아세포에서 울릉국화 추출물에 대한 MTT 분석 결과를 나타낸 그래프이다(Doxo : doxorubicin 0.5 μM (27.18 μg/ml), 2 : 울릉국화 추출물 2μg/ml, 4 : 울릉국화 추출물 4 μg/ml, 8 : 울릉국화 추출물 8 μg/ml, NAC(N-acetyl-L-cysteine) : 5 mM (815.97 μg/ml)).
도 4는 울릉국화 추출물의 SA-β-gal 활성 억제능을 섬유아세포(HDF)에서 평가한 그래프(A) 및 광학현미경 관찰사진(B)이다.
도 5는 저농도의 울릉국화 추출물에서의 SA-β-gal 활성 억제능을 섬유아세포(HDF)에서 평가한 그래프(A) 및 광학현미경 관찰사진(B)이다.
도 6은 울릉국화 추출물 및 다른 구절초 추출물의 세포노화저해효과를 확인한 그래프이다. Fig. 1 is a photograph of Ulleung chrysanthemum.
2 is a graph showing the results of MTT analysis of Ulleung chrysanthemum extract in fibroblast (HDF) (Doxo: doxorubicin 0.5 μM (27.18 μg / ml), 5:
3 is a graph showing the results of MTT analysis of Ulleung chrysanthemum extract in fibroblasts (Doxo: doxorubicin 0.5 μM (27.18 μg / ml), 2:
FIG. 4 is a graph (A) and an optical microscope photograph (B) showing the inhibitory activity of SA-β-gal activity of Ulleung chrysanthemum extract on fibroblasts (HDF).
FIG. 5 is a graph (A) and an optical microscope photograph (B) showing the inhibitory activity of SA-β-gal activity inhibition in low concentration of Ulleung chrysanthemum extract in fibroblasts (HDF).
FIG. 6 is a graph showing the inhibitory effects of Ulleung chrysanthemum extract and other herb extracts on cell senescence.
이하, 본 발명을 보다 상세하게 설명한다.Hereinafter, the present invention will be described in more detail.
상술한 바와 같이, 국화과 식물 6종의 항산화 활성 및 항염증 활성에 대한 연구(대한민국 공개특허 제2014-0088504호)는 알려져 있고, 울릉국화의 라디칼 소거능에 대한 연구(woo et al., Korean J.Plant Res., 23(1):47, 2010)는 이루어져 있으나, 울릉국화 추출물의 항노화 및 피부의 주름 및 탄력 개선에 관한 연구는 이루어진 바 없었다.As described above, studies on the antioxidative and anti-inflammatory activities of six Asteraceae plants (Korean Patent Laid-Open Publication No. 2014-0088504) are known, and a study on the radical scavenging ability of Ulleung chrysanthemum (Woo et al. Plant Res., 23 (1): 47, 2010). However, no research has been conducted on the anti-aging and skin wrinkles and elasticity of Ulleung chrysanthemum extract.
이에 본 발명에서는 울릉국화 추출물을 유효성분으로 포함하는 화장료 조성물을 제공한다.Accordingly, the present invention provides a cosmetic composition comprising Ulleung chrysanthemum extract as an active ingredient.
본 발명에서의 "동안(童顔)"이란 나이 든 사람이 지니고 있는 어린아이 같은 얼굴을 의미하는 것으로, 피부의 노화가 억제되어 피부가 탄력이 있고, 주름이 없거나 적은 것을 의미한다.
The term " child face "in the present invention means a child-like face of an aged person, which means that the aging of the skin is suppressed, the skin is elastic, and there is no or little wrinkles.
본 발명에서는 상기 울릉국화 추출물을 유효성분으로 포함하는 조성물에 대해 MTT 분석법을 실시하였으며, 이를 통하여 울릉국화 추출물이 세포독성이 낮으며, 울릉국화 추출물을 처리한 실험군에서 세포 생존력이 증가한 것을 확인하였으며, SA-β-gal 활성이 저해되는 것을 확인할 수 있었다. 이를 통해 울릉국화 추출물이 직접적으로 세포노화를 억제하는 효능, 특히 피부 주름 개선 및 피부 탄력 개선효과를 가진 것을 확인할 수 있었다.
In the present invention, MTT assay was performed on the composition containing the Ulleung chrysanthemum extract as an active ingredient, and it was confirmed that the Ulleung chrysanthemum extract had low cytotoxicity, and the cell viability was increased in the test group treated with Ulleung chrysanthemum extract. It was confirmed that SA-beta-gal activity was inhibited. Thus, it was confirmed that the extract of Ulreung chrysanthemum directly has an effect of inhibiting cell senescence, in particular, improving wrinkles of skin and improving skin elasticity.
본 발명에서의 울릉국화의 학명은 Dendranthema zawadskii var. lucidum (Nakai) J.H. Park 또는 Chrysanthemum zawadskii var. lucidum (Nakai) Y.Lee이며, 이명으로는 섬들국화, 울능국화 또는 울릉구절초로도 불린다(도 1 참조). The scientific name of Ulleung chrysanthemum in the present invention is Dendranthema I have zawadskii . lucidum (Nakai) JH Park or Chrysanthemum I have zawadskii . It is also called lucidum (Nakai) Y.Lee, and the tinnitus is also called island nationalization, mystic chrysanthemum, or Ulleung ryukcho (see Fig. 1).
상기 울릉국화 추출물은 울릉국화의 지상부로부터 추출하는 것이 바람직한데 지상부(地上部)란, 식물체에서 뿌리부분이 아닌 식물 상부의 경엽 부분을 의미하며, 자세하게는 줄기, 잎, 꽃일 수 있다.It is preferable that the Ulleung chrysanthemum extract is extracted from the upper part of Ulleung chrysanthemum. The above ground part refers to a part of the upper part of the upper part of the plant which is not the root part in the plant, and in detail, it may be stem, leaf and flower.
상기 울릉국화 추출물은 물, 알코올, 에틸아세테이트, 클로로포름 또는 헥산 등의 단독 또는 혼합 형태인 용매로 추출할 수 있으며, 보다 바람직하게는 C1~4의 저급 알코올을 사용할 수 있다. 보다 더 바람직하게는 에탄올일 수 있다. 상기 용매를 이용한 일반적인 용매 추출법을 적용하여 추출할 수 있으며, 또한 칼럼 크로마토그래피를 이용하여 정제한 분획물일 수도 있다. 본 발명에 따른 추출물의 제조방법은 본 기술분야에 속하는 통상의 지식을 가진 자에게 알려진 식물 추출방법을 모두 적용할 수 있으며, 예컨대 물, 알코올 또는 혼합용매에 의한 추출, 중탕이나 상온에 의한 추출법 등을 포함하나 이에 한정되지 않는다. The Ulleung chrysanthemum extract may be extracted with a solvent such as water, alcohol, ethyl acetate, chloroform or hexane, or more preferably C1-4 lower alcohol. More preferably, it may be ethanol. The extract may be applied by a general solvent extraction method using the above-mentioned solvent, or it may be a fraction purified by using column chromatography. The method for producing the extract according to the present invention can be applied to any plant extraction method known to those of ordinary skill in the art. For example, extraction with water, alcohol or mixed solvent, extraction with hot water or room temperature But is not limited thereto.
이와 같이 추출된 울릉국화 추출물을 유효성분으로 포함하는 조성물은 섬유아세포(HDF)에서 세포독성을 거의 나타내지 않고, 세포의 증식을 유의하게 증가시켰다(도 2 및 3 참조).
The composition containing the extracted Ulleung chrysanthemum extract as an active ingredient showed little cytotoxicity in fibroblasts (HDF) and significantly increased cell proliferation (see FIGS. 2 and 3).
상기 울릉국화 추출물은 세포노화를 억제하는 것일 수 있다. 본 발명의 일 실시예에서 울릉국화 추출물의 항노화 효과를 발견하였으며, 이를 증명하기 위해 콜라게네이즈(collagenase) 저해 활성 평가, 엘라테이즈(elastase) 저해 활성 평가, DPPH 활성 평가, 크산틴 옥시데이즈(Xanthine oxidase) 저해활성 평가 등 여러가지 방법으로 세포 노화 억제 효과를 확인할 수 있지만, 그 중 가장 일반적인 방법인 SA-β-gal 활성 평가 시험(Dimri et al., A biomarker that identifies senescent human cells in culture and aging skin in vivo, Proc Natl Acad Sci 92(1995): 9363-9367)을 실시하였다. 그 결과 SA-β-gal 활성을 저해시켜 세포노화를 억제하는 것을 확인하였다(도 4 및 5 참조). 특히, 상기 울릉국화 추출물은 섬μ유아세포에서 현저히 우수한 세포노화 억제 효능을 보이는 것을 확인할 수 있다. The Ulleung chrysanthemum extract may inhibit cell senescence. In one embodiment of the present invention, the anti-aging effect of Ulleung chrysanthemum extract was found. To prove this, the collagenase inhibitory activity evaluation, the elastase inhibitory activity evaluation, the DPPH activity evaluation, the xanthine oxidase (Xanthine oxidase) inhibitory activity, the most common method is the SA-β-gal activity evaluation test (Dimri et al., A biomarker that identifies senescent human cells in culture and aging skin in vivo , Proc Natl Acad Sci 92 (1995): 9363-9367). As a result, it was confirmed that SA-β-gal activity was inhibited and cell senescence was inhibited (see FIGS. 4 and 5). In particular, it can be confirmed that the Ulleung chrysanthemum extract has a remarkably superior cell aging inhibitory effect in islet infant cells.
본 발명의 세포노화 억제용 조성물은 상기 울릉국화 추출물을 2 내지 10 μg/ml의 농도로 포함할 수 있다. 상기 조성물의 용매로는 울릉국화 추출물의 세포노화 억제 효과를 방해하지 않는 것이라면 특별히 제한되지 않으나, 보다 바람직하게는 물, 에탄올, 프로필 알코올, DMSO 등일 수 있다. The composition for inhibiting cell senescence of the present invention may contain the Ulleung chrysanthemum extract at a concentration of 2 to 10 μg / ml. The solvent of the composition is not particularly limited as long as it does not interfere with the cell aging inhibitory effect of Ulreung chrysanthemum extract, and more preferably water, ethanol, propyl alcohol, DMSO, and the like.
도 4 및 도 5에 나타난 바와 같이, 울릉국화 추출물의 세포노화 억제능을 실험한 결과, 5 내지 20 μg/ml의 울릉국화 추출물이 섬유아세포에서 노화를 유의하게 감소시키는 효과가 있었으며, 2 내지 4 μg/ml의 울릉국화 추출물에서도 섬유아세포의 노화를 유의하게 감소시키는 효과를 발휘하였다. As shown in FIGS. 4 and 5, the extract of Ulleung chrysanthemum showed an inhibitory effect on cell senescence, and the extract of Ulreung chrysanthemum with 5-20 .mu.g / ml showed significant reduction of senescence in fibroblasts, / ml of Ulreung chrysanthemum extract significantly reduced the aging of fibroblasts.
세포 독성 시험의 경우, 울릉국화 추출물은 10 μg/ml 농도에서도 세포독성이 없었다. 따라서 세포독성효과가 없으면서 우수한 세포노화 억제 효능을 나타내는 울릉국화 추출물 조성물의 농도는 보다 바람직하게는 2 내지 10 μg/ml일 수 있다. In the cytotoxicity test, Ulleung chrysanthemum extract was not cytotoxic even at a concentration of 10 μg / ml. Therefore, the concentration of the Ulleung chrysanthemum extract composition having no cytotoxic effect and exhibiting excellent cell senescence inhibitory effect may be more preferably 2 to 10 μg / ml.
본 발명의 바람직한 일실시예에 따르면, 상기 화장료 조성물은 울릉국화 추출물을 0.001 내지 10 중량% 농도로 포함할 수 있다.According to a preferred embodiment of the present invention, the cosmetic composition may contain Ulleung chrysanthemum extract at a concentration of 0.001 to 10% by weight.
본 발명의 울릉국화 추출물을 유효성분으로 포함하는 화장료 조성물에는 상기 울릉국화 추출물과 더불어 필요에 따라 통상 화장료에 배합되는 다른 성분을 배합할 수 있다. 이외에 첨가할 수 있는 배합 성분으로서는 유지 성분, 보습제, 에몰리엔트제, 계면 활성제, 유기 및 무기안료, 유기 분체, 자외선 흡수제, 방부제, 살균제, 산화 방지제, 식물 추출물, pH 조정제, 알콜, 색소, 향료, 혈행 촉진제, 냉감제, 제한(制汗)제, 정제수 등을 들 수 있다.The cosmetic composition comprising the Ulleung chrysanthemum extract of the present invention as an active ingredient may be blended with the Ulleung chrysanthemum extract as described above and other ingredients usually added to cosmetics as required. Examples of the compounding ingredients that can be added in addition to the above components include a preservative component, a moisturizer, an emollient, a surfactant, an organic and inorganic pigment, an organic powder, an ultraviolet absorber, an antiseptic, a bactericide, an antioxidant, a plant extract, a pH adjuster, , A blood circulation accelerator, a cold agent, an antiperspirant agent, and purified water.
유지 성분으로서는 에스테르계 유지, 탄화수소계 유지, 실리콘계 유지, 불소계 유지, 동물 유지, 식물 유지 등을 들 수 있다.Examples of the oil retaining component include ester-based oil retaining, hydrocarbon-based oil retaining, silicone-based oil retaining, fluoric oil retaining, animal retention and plant retention.
에스테르계 유지로서는 트리2-에틸헥산산글리세릴, 2-에틸헥산산세틸, 미리스틴산이소프로필, 미리스틴산부틸, 팔미틴산이소프로필, 스테아르산에틸, 팔미틴산옥틸, 이소스테아르산이소세틸, 스테아르산부틸, 리놀레산에틸, 리놀레산이소프로필, 올레인산에틸, 미리스틴산이소세틸, 미리스틴산이소스테아릴, 팔미틴산이소스테아릴, 미리스틴산옥틸도데실, 이소스테아르산이소세틸, 세바신산디에틸, 아디핀산디이소프로필, 네오펜탄산이소알킬, 트리(카프릴, 카프린산)글리세릴, 트리2-에틸헥산산트리메틸롤프로판, 트리이소스테아르산트리메틸롤프로판, 테트라 2-에틸헥산산펜타엘리슬리톨, 카프릴산세틸, 라우린산데실, 라우린산헥실, 미리스틴산데실, 미리스틴산미리스틸, 미리스틴산세틸, 스테아르산스테아릴, 올레인산데실, 리시노올레인산세틸, 라우린산이소스테아릴, 미리스틴산이소트리데실, 팔미틴산이소세틸, 스테아르산옥틸, 스테아르산이소세틸, 올레인산이소데실, 올레인산옥틸도데실, 리놀레산옥틸도데실, 이소스테아르산이소프로필, 2-에틸헥산산세토스테아릴, 2-에틸헥산산스테아릴, 이소스테아르산헥실, 디옥탄산에틸렌글리콜, 디올레인산에틸렌글리콜, 디카프린산프로필렌글리콜, 디(카프릴,카프린산)프로필렌글리콜, 디카프릴산프로필렌글리콜, 디카프린산네오펜틸글리콜, 디옥탄산네오펜틸글리콜, 트리카프릴산글리세릴, 트리운데실산글리세릴, 트리이소팔미틴산글리세릴, 트리이소스테아르산글리세릴, 네오펜탄산옥틸도데실, 옥탄산이소스테아릴, 이소노난산옥틸, 네오데칸산헥실데실, 네오데칸산옥틸도데실, 이소스테아르산이소세틸, 이소스테아르산이소스테아릴, 이소스테아르산옥틸데실, 폴리글리세린올레인산에스테르, 폴리글리세린이소스테아르산에스테르, 시트르산트리이소세틸, 시트르산트리이소알킬, 시트르산트리이소옥틸, 락트산라우릴, 락트산미리스틸, 락트산세틸, 락트산옥틸데실, 시트르산트리에틸, 시트르산아세틸트리에틸, 시트르산아세틸트리부틸, 시트르산트리옥틸, 말산디이소스테아릴, 히드록시스테아르산 2-에틸헥실, 숙신산디2-에틸헥실, 아디핀산디이소부틸, 세바신산디이소프로필, 세바신산디옥틸, 스테아르산콜레스테릴, 이소스테아르산콜레스테릴, 히드록시스테아르산콜레스테릴, 올레인산콜레스테릴, 올레인산디히드로콜레스테릴, 이소스테아르산피트스테릴, 올레인산피트스테릴, 12-스테알로일히드록시스테아르산이소세틸, 12-스테알로일히드록시스테아르산스테아릴, 12-스테알로일히드록시스테아르산이소스테아릴 등의 에스테르계 등을 들 수 있다.Examples of ester-based fats include glyceryl tri-2-ethylhexanoate, cetyl 2-ethylhexanoate, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isostearyl isostearate, Butyl isopropyl myristate, isopropyl myristate, isopropyl myristate, isopropyl myristate, isopropyl myristate, isopropyl myristate, butyl, ethyl linoleate, isopropyl linoleate, ethyl oleate, isosilyl myristate, isostearic acid isostearyl, isostearyl palmitate, octyldodecyl myristate, Trimethylol propane, triisostearic acid trimethylol propane, tetra 2-ethylhexanoic acid pentaerythritol tetra (2-ethylhexanoate) , Decyl caprylate, decyl laurate, hexyl laurate, myristate decyl, myristyl myristate, myristine monoethyl stearate, stearyl stearate, decyl oleate, ricinoleic acid tri Isopropyl palmitate, isostearyl stearate, isostearyl stearate, isodecyl oleate, octyldodecyl oleate, octyldodecyl linoleate, isopropyl stearate, isopropyl stearate, isopropyl stearate, Ethylhexanoate, stearyl 2-ethylhexanoate, stearyl 2-ethylhexanoate, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate, propylene glycol dicaprate, di (capryl, capric acid) propylene glycol, Propylene glycol dicaprylate, propylene glycol dicaprylate, neopentyl glycol dicaprate, neopentyl glycol dioctanoate, glyceryl tricarboxylate, glyceryl triunsaturated acid, glyceryl triisopalmitate, glyceryl triisostearate, octyl dodecanoate Silane, octanoic acid, octanoic acid, octanoic acid, octanoic acid octanoate, octanoic acid octanoate, octanoic acid octanoate, octanoic acid octanoate, octanoic acid octanoate, octanoic acid octanoate, Octyldecyl lactate, octyldecyl lactate, octyldecyl lactate, polyglycerin oleic acid ester, polyglycerin isostearic acid ester, triisocetyl citrate, triisobutyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactate, But are not limited to, ethyl, acetyltriethyl citrate, acetyltributyl citrate, trioctyl citrate, diisostearyl malate, 2-ethylhexyl hydroxystearate, di-2-ethylhexyl succinate, diisobutyl adipate, diisopropyl sebacate, But are not limited to, dioctyl sebacate, stearic acid cholesteryl, isostearic acid cholesteryl, hydroxystearic acid cholesteryl, oleic acid cholesteryl, oleic acid dihydrocholesteryl, isostearic acid pitostearyl, Stearyl hydroxystearic acid isostearyl, 12-stearoyl hydroxystearic acid stearyl, 12-stearoyl hydroxystearic acid stearyl, One hydroxy stearic acid and the like esters such as cetearyl source.
탄화 수소계 유지로서는 스쿠알렌, 유동 파라핀, 알파-올레핀올리고머, 이소파라핀, 세레신, 파라핀, 유동 이소파라핀, 폴리부덴, 마이크로크리스탈린왁스, 바셀린 등의 탄화 수소계 유지 등을 들 수 있다.Hydrocarbon hydrocarbon-based fats and oils such as squalene, liquid paraffin, alpha-olefin oligomer, isoparaffin, ceresin, paraffin, floating isoparaffin, polybutene, microcrystalline wax and petrolatum are examples of such fats.
실리콘계 유지로서는 폴리메틸실리콘, 메틸페닐실리콘, 메틸시클로폴리실록산, 옥타메틸폴리실록산, 데카메틸폴리실록산, 도데카메틸시클로실록산, 디메틸실록산ㆍ메틸세틸옥시실록산 공중합체, 디메틸실록산ㆍ메틸스테알록시록산 공중합체, 알킬 변성 실리콘유, 아미노 변성 실리콘유 등을 들 수 있다.Examples of silicone based oils include polymethyl silicone, methylphenyl silicone, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, dimethylsiloxane-methyl cetyloxysiloxane copolymer, dimethylsiloxane-methylstearoxyloxane copolymer, alkyl Modified silicone oils, and amino-modified silicone oils.
불소계 유지로서는 퍼플루오로폴리에테르 등을 들 수 있다.Examples of the fluorine-based oil include perfluoropolyether and the like.
동물 또는 식물 유지로서는 아보카도유, 아르몬드유, 올리브유, 참깨유, 쌀겨유, 새플라워유, 대두유, 옥수수유, 유채유, 행인(杏仁)유, 팜핵유, 팜유, 피마자유, 해바라기유, 포도종자유, 면실유, 야자유, 쿠쿠이너트유(코코넛유), 소맥배아유, 쌀 배아유, 쉐아버터, 월견초유, 마커데이미아너트유, 메도홈유, 난황유, 우지(牛脂), 마유, 밍크유, 오렌지라피유, 호호바유, 캔데리러왁스, 카르나바왁스, 액상 라놀린, 경화피마자유 등의 동물 또는 식물 유지를 들 수 있다.
Examples of animal or vegetable oils include avocado oil, almond oil, olive oil, sesame oil, rice bran oil, new flower oil, soybean oil, corn oil, rape oil, apricot kernel oil, palm kernel oil, palm oil, castor oil, , Corn oil, coconut oil, cucumber nut oil (coconut oil), wheat germ oil, rice germ oil, shea butter, coltsfoot colostrum, marker daisy nut oil, meadow home oil, egg oil, Animal or vegetable oils such as oil, jojoba oil, canderir wax, carnauba wax, liquid lanolin, and hardened castor oil.
상기 화장료 조성물은 당업계에서 통상적으로 제조되는 어떠한 제형으로도 제조될 수 있으며, 바람직하게는 크림, 로션, 파우더, 계면활성제-함유 클린싱오일, 분말 파운데이션, 유탁액 파운데이션, 왁스 파운데이션, 페이스트, 에센스, 아이크림 등의 제형일 수 있다. The cosmetic composition may be prepared in any form conventionally produced in the art, and is preferably a cream, a lotion, a powder, a surfactant-containing cleansing oil, a powder foundation, an emulsion foundation, a wax foundation, a paste, Eye cream, and the like.
구체적으로, 본 발명의 조성물은 스킨로션, 스킨소프너, 스킨토너, 아스트린젠트, 로션, 밀크로션, 모이스쳐 로션, 영양로션, 맛사지크림, 영양크림, 모이스처크림, 핸드크림, 파운데이션, 에센스, 영양에센스, 팩, 비누, 클렌징폼, 클렌징로션, 클렌징크림, 바디로션 및 바디클린저의 제형로 구성된 그룹에서 선택된 하나 이상의 제형을 가짐을 특징으로 하는 화장료 조성물을 포함한다.Specifically, the composition of the present invention can be used as a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutrition lotion, a massage cream, a nutritional cream, a moisturizing cream, a hand cream, And at least one formulation selected from the group consisting of soaps, cleansing foams, cleansing lotions, cleansing creams, body lotions and body cleansers.
본 발명의 제형이 페이스트, 크림 또는 겔인 경우에는 담체 성분으로서 동물섬유, 식물섬유, 왁스, 파라핀, 전분, 트라가칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크 또는 산화아연 등이 이용될 수 있다.When the formulation of the present invention is a paste, cream or gel, animal fibers, plant fibers, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide .
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 에스테르가 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 sorbitan fatty acid esters.
본 발명의 제형이 현탁액인 경우에는 담체 성분으로서 물, 에탄올 또는 프로필렌 글리콜과 같은 액상 희석제,When the formulation of the present invention is a suspension, a liquid diluent such as water, ethanol or propylene glycol,
에톡실화 이소스테아릴 알코올, 폴리옥시에틸렌 소르비톨 에스테르 및 폴리옥시에틸렌 소르비탄 에스테르와 같은 현탁제, 미소결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가 또는 트라가칸트 등이 이용될 수 있다.A suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth can be used.
본 발명의 제형이 계면-활성제 함유 클린징인 경우에는 담체 성분으로서 지방족 알코올 설페이트, 지방족 알코올 에테르 설페이트, 설포숙신산 모노에스테르, 이세티오네이트, 이미다졸리늄 유도체, 메틸타우레이트, 사코시네이트, 지방산 아미드 에테르 설페이트, 알킬아미도베타인, 지방족 알코올, 지방산 글리세리드, 지방산 디에탄올아미드, 식물성 유, 라놀린 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 may include aliphatic alcohol sulfates, aliphatic alcohol ether sulfates, sulfosuccinic acid monoesters, isethionates, imidazolinium derivatives, methyltaurate, Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters.
본 발명의 세포노화 억제용 화장료 조성물은 총 중량에 대하여 0.001 내지 10 중량%로 포함하는 것이 우수한 세포노화 억제 효능을 나타내기에 좋다. 너무 0.001 미만의 중량%로 포함하는 경우, 세포노화 억제 효능이 떨어지며, 10 중량%를 초과할 경우 세포독성을 나타낼수 있다. 더욱 바람직한 화장료 조성물의 중량%는 3 내지 7 중량%일 수 있다.
The cosmetic composition for inhibiting cell senescence of the present invention is contained in an amount of 0.001 to 10% by weight based on the total weight of the cosmetic composition. If it is contained at a weight percentage of less than 0.001%, the cell deterioration inhibiting effect is poor, and if it exceeds 10% by weight, cytotoxicity may be exhibited. More preferably, the weight percentage of the cosmetic composition may be 3 to 7% by weight.
이하, 본 발명을 실시예에 의해 상세히 설명하기로 한다. 그러나 이들 실시예는 본 발명을 보다 구체적으로 설명하기 위한 것으로서, 본 발명의 범위가 이들 실시예에 한정되는 것은 아니다.
Hereinafter, the present invention will be described in detail with reference to examples. However, these examples are intended to further illustrate the present invention, and the scope of the present invention is not limited to these examples.
1. 울릉국화 추출물 조성물의 제조1. Preparation of Ulleung chrysanthemum extract composition
2012년 7월 30일, 충북 음성군 소재에서 채취된 울릉국화(Dendranthema zawadskii var. lucidum (Nakai) J.H.Park) 시료의 지상부를 사용하였다. 해당 시료를 건조, 분쇄한 후 상온에서 에탄올로 추출한 후 여과하였으며, 추출액 중의 용매는 감압농축장치를 이용해서 제거하고, 수분은 동결건조기에서 제거하여 실험에 사용하였다. 건조시료 100 g으로부터 얻어진 추출물은 7.52 g이었고, 수율은 7.52 %였다.On July 30, 2012, the Dendranthema I have zawadskii. lucidum (Nakai) JHPark) was used. The sample was dried and pulverized, extracted with ethanol at room temperature and then filtered. The solvent in the extract was removed using a vacuum concentrator, and water was removed from the freeze dryer and used in the experiment. The extract obtained from 100 g of the dried sample was 7.52 g, and the yield was 7.52%.
얻어진 상기 울릉국화 추출물은 용매 DMSO에 100 μg/ml 농도로 녹여 울릉국화 추출물 조성물을 제조하였다.
The obtained Ulleung chrysanthemum extract was dissolved in a solvent DMSO at a concentration of 100 μg / ml to prepare a Ulleung chrysanthemum extract composition.
2. 세포 배양2. Cell culture
사람 섬유아세포 (Human dermal fibroblasts, HDFs)는 1% 항생제가 포함된 섬유아세포 배양배지 (FGM-2, Lonza, 스위스)을 이용하여 직경 100 mm 배양접시에 세포를 1 x 105개로 분주한 후, 37의 5% 이산화탄소 배양기에서 배양하였다. 배양접시의 바닥에 80~90% 정도 세포가 자라면, 2.5 x 트립신-EDTA 용액(Lonza, 스위스)을 처리하여 세포를 분리한 후, 계대 배양하였다. Human dermal fibroblasts (HDFs) were plated at 1 × 10 5 cells in a 100 mm diameter culture dish using a fibroblast culture medium (FGM-2, Lonza, Switzerland) containing 1% antibiotic, 37 in a 5% carbon dioxide incubator. If 80-90% of the cells were grown on the bottom of the culture dish, the cells were treated with 2.5 x trypsin-EDTA solution (Lonza, Switzerland) and subcultured.
세포의 분열 횟수(population doubling, PD)는 다음 식으로 조사하였다.
The population doubling (PD) was examined by the following equation.
PD= log2F/log2I (F=마지막 세포수, I=처음 세포수)
PD = log 2 F / log 2 I (F = number of cells last, I = number of cells first)
실험에 사용한 섬유아세포는 분열횟수가 8회 이하의 것을 사용하였다.
The fibroblasts used in the experiment were those having a number of divisions of 8 or less.
1. 세포 노화 유도 및 울릉국화 추출물 처리1. Cell aging induction and Ulleung chrysanthemum extract treatment
독소루비신(Sigma, USA)은 높은 농도에서는 세포를 죽이지만, 낮은 농도에서는 세포노화를 유도하는 것으로 알려져 있다. 상기 실시예 1-2의 방법으로 배양된 세포의 노화를 유도하기 위하여, 직경 100mm 배양접시에 섬유아세포를 150,000개 분주하고, 3일간 37, 5% 이산화탄소배양기에서 배양한 후, 세포 배양액을 제거하고, 세포를 1% 항생제가 포함된 섬유세포배양액으로 2회 세척한 후, 0.5 μM 독소루비신을 4시간 처리하였다.Doxorubicin (Sigma, USA) is known to kill cells at high concentrations but induce cell senescence at low concentrations. To induce senescence of the cells cultured by the method of Example 1-2, 150,000 fibroblasts were dispensed into a 100 mm diameter culture dish and cultured in a 37, 5% carbon dioxide incubator for 3 days. Then, the cell culture medium was removed , Cells were washed twice with fibroblast culture medium containing 1% antibiotic and treated with 0.5 μM doxorubicin for 4 hours.
독소루비신 0.5 μM을 4시간 처리한 세포를 50ml tube에 1% 항생제가 포함된 섬유세포배양액에 섬유아세포는 5,000개/ml가 되도록 하고 96웰 배양용기의 각 웰에 100 μl씩 분주하였다. 최종적으로 각 웰당 섬유아세포를 500개씩 분주하고, 하루 동안 37, 5% 이산화탄소배양기에서 배양하였다. 각 웰에 1% 항생제가 포함된 섬유세포배양액을 100 μl씩 더 넣어 준 후, 상기 실시예의 울릉국화 추출물을 처리하였다. 울릉국화 추출물은 각각 2, 4, 5, 8 또는 20 μg/ml 처리하였다. 이후 3일간 배양하여 SA-β-gal 염색을 실시하였다.Cells treated with 0.5 μM doxorubicin for 4 hours were dispensed into fibroblast culture medium containing 1% antibiotic in a 50 ml tube at a rate of 5,000 cells / ml and 100 μl in each well of a 96-well culture container. Finally, 500 fibroblasts per well were dispensed and cultured in 37, 5% CO2 incubator for one day. 100 μl of fibroblast culture medium containing 1% antibiotic was further added to each well, and the Ulleung chrysanthemum extract of the above-mentioned Example was treated. Ulleung chrysanthemum extracts were treated at 2, 4, 5, 8 or 20 μg / ml, respectively. The cells were then cultured for 3 days and stained with SA-β-gal.
한편, 음성대조군으로 용매인 DMSO를 처리하였으며, 양성대조군으로는 양성대조군 5 mM (815.97 μg/ml) NAC를 3일간 처리 한 후 MTT 분석을 통해 독성 평가를 실시하였다.
As a negative control, DMSO was used as a negative control. To the positive control group, 5 mM (815.97 μg / ml) NAC was treated for 3 days and then toxicity was evaluated by MTT analysis.
2. 울릉국화 추출물의 세포 독성 평가 - 2. Evaluation of cytotoxicity of Ulleung chrysanthemum extract- MTTMTT 분석 analysis
상기 처리 후 3일 동안 37, 5% 이산화탄소배양기에서 배양한 후, 세포의 성장 정도를 MTT 방법으로 확인하였다. 96 well 배양용기의 각 웰에 0.1% MTT(3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide) 용액(Sigma, USA)을 50 μl씩 넣고 3시간 동안 37, 5% 이산화탄소배양기에서 반응시키고, 배양액과 MTT용액을 제거한 후 DMSO 100 μl를 첨가하여 형성된 결정을 녹여 마이크로플레이트 리더를 이용하여 550 nm에서 흡광도를 측정하였으며, 시료를 처리하지 않은 대조실험결과에 대한 백분율 (%)로 표시한 결과를 표 1 또는 도2 및 도3에 나타내었다.
After the cells were cultured in a 37, 5% carbon dioxide incubator for 3 days after the treatment, the degree of cell growth was confirmed by MTT method. 50 μl of 0.1% MTT (3- (4,5-dimethylthiazol-2yl) -2,5-diphenyltetrazolium bromide) solution (Sigma, USA) was added to each well of a 96- After incubation, the culture solution and MTT solution were removed, and 100 μl of DMSO was added to dissolve the crystals. The absorbance was measured at 550 nm using a microplate reader. The percentages (% The results are shown in Table 1, FIG. 2, and FIG.
실험결과 표 1 및 도 2에서 보이는 바와 같이, 섬유아세포에서 상기 실시예에 따른 울릉국화 추출물이 5 μg/ml 에서는 94.8 ± 2.7%의 세포생존율을 보였으며, 10 μg/ml 에서는 80.7 ± 2.7%의 세포 생존율, 그리고 20 μg/ml 에서는 74.8 ± 3.1%의 세포 생존율을 보였다. As shown in Table 1 and FIG. 2, in the fibroblasts, the cell survival rate was 94.8 ± 2.7% at 5 μg / ml and 80.7 ± 2.7% at 10 μg / ml Cell viability and cell viability of 74.8 ± 3.1% at 20 μg / ml.
더욱 낮은 농도의 울릉국화 추출물로 실험한 결과는 표 2 및 도 3과 같다. 울릉국화 추출물이 2 μg/ml 에서는 98.32 ± 0.69 %의 세포생존율을 보였으며, 4 μg/ml 에서는 97.84 ± 2.91 %의 세포 생존율, 그리고 8 μg/ml 에서는 94.81 ± 0.28 %의 세포 생존율을 보였다.The results of the experiment with Ulleung chrysanthemum extract of lower concentration are shown in Table 2 and FIG. The cell viability was 98.32 ± 0.69% at 2 μg / ml, 97.84 ± 2.91% at 4 μg / ml, and 94.81 ± 0.28% at 8 μg / ml.
즉, 도 2 및 도 3에 보이는 바와 같이, 2 내지 10 μg/ml 농도에서 세포독성이 거의 없거나 미미하게 나타난 것을 확인할 수 있다.
That is, as shown in FIG. 2 and FIG. 3, it can be confirmed that the cytotoxicity was little or insignificant at a concentration of 2 to 10 μg / ml.
1. 세포 노화 유도 및 울릉국화 추출물 처리1. Cell aging induction and Ulleung chrysanthemum extract treatment
상기 실시예 2-1과 같은 방법으로, 직경 100 mm 배양접시에 섬유아세포를 150,000개 분주하고, 3일간 37, 5% 이산화탄소배양기에서 배양한 후, 세포 배양액을 제거하고, 세포를 1% 항생제가 포함된 섬유세포배양액으로 2회 세척한 후, 0.5μM 독소루비신을 4시간 처리하였다.In the same manner as in Example 2-1, 150,000 fibroblasts were dispensed into a 100 mm diameter culture dish, and the cells were cultured in a 37%, 5% carbon dioxide incubator for 3 days. Then, the cell culture was removed and the cells were treated with 1% After washing twice with the fibroblast culture medium, 0.5 μM doxorubicin was treated for 4 hours.
독소루비신 0.5 μM을 4시간 처리한 세포를 50ml 튜브에 1% 항생제가 포함된 섬유세포배양액에 섬유아세포 5,000개/ml이 되도록 하고 96웰 배양용기의 각 웰에 100 μl씩 분주하였다. 최종적으로 각 웰당 섬유아세포 500개를 분주하고, 하루 동안 37, 5% 이산화탄소배양기에서 배양하였다. 각 웰에 1% 항생제가 포함된 섬유세포배양액을 100 μl씩 더 넣어 준 후, 상기 실시예의 울릉국화 추출물을 처리하였다. 울릉국화 추출물은 각각 2, 4, 5, 8 또는 20 μg/ml를 처리하였다. 이후 3일간 배양하여 SA-β-gal 염색을 실시하였다.Cells treated with 0.5 μM doxorubicin for 4 hours were dispensed into a 50 ml tube in a fibroblast culture medium containing 1% antibiotic at 5,000 cells / ml and 100 μl in each well of a 96-well culture container. Finally, 500 fibroblasts per well were dispensed and cultured in 37, 5% CO2 incubator for one day. 100 μl of fibroblast culture medium containing 1% antibiotic was further added to each well, and the Ulleung chrysanthemum extract of the above-mentioned Example was treated. Ulleung chrysanthemum extracts were treated with 2, 4, 5, 8, or 20 μg / ml, respectively. The cells were then cultured for 3 days and stained with SA-β-gal.
한편, 음성대조군으로 용매인 DMSO를 처리하였으며, 양성대조물질로는 5 mM (= 815.97 ug/ml) NAC 를 처리하여 비교실험하였다.
Meanwhile, DMSO, a negative control, was treated with 5 mM (= 815.97 ug / ml) NAC as a positive control.
2. SA-β-갈락토시다아제 활성 염색2. SA-β-Galactosidase activity staining
상기 처리 후 3일 동안 37, 5% 이산화탄소배양기에서 배양하고, 세포노화 저해에 대한 울릉국화 추출물의 효과를 SA-β-gal 활성 염색으로 조사하였다. β-갈락토시데이스는 갈락토스와 유기물 사이에 형성된 β-글라이코시딕 결합을 가수분해시키는 엑소글라이코시데이즈이며, 노화가 진행될수록 세포의 리소좀에 다량으로 축적되고, 그 발현 정도를 5-bromo-4-chloro-3-indolyl-βD-galactopyranoside를 사용하여 식별하고 정량화시킬 수 있다. 이 때, SA-β-gal 염색법으로 노화된 세포는 세포질이 파란색으로 염색된다. The cells were cultured in 37, 5% CO2 incubator for 3 days after the treatment, and the effect of Ulleung chrysanthemum extract on cell senescence inhibition was investigated by SA-β-gal activity staining. β-galactosidase is exoglycosidase that hydrolyzes β-glycosidic bonds formed between galactose and organic matter. As aging progresses, a large amount of β-galactosidase accumulates in lysosomes of cells, bromo-4-chloro-3-indolyl- [beta] D-galactopyranoside. At this time, cells aged by SA-β-gal staining are stained cytoplasm in blue.
울릉국화 추출물을 3일 동안 처리한 후, 세포를 인산완충액으로 2번 세척하고, 3.7% 파라포름알데하이드로 세포를 1분간 고정한 후, 고정액을 제거하였다. SA-β-gal 염색 용액(40 mM citric acid/phosphate [pH 5.8], 5 mM potassium ferrocyanide, 5 mM potassium ferricyanide, 150 mM NaCl, 2 mM MgCl2, X-gal 1 mg/ml)을 24웰 배양용기에는 500μl를 넣어 준 후, 은박지로 싸서 37에서 18시간 동안 반응시켰다. 인산완충용액 (PBS)으로 2번 세척한 후 인산완충용액으로 2회 세척 한 후, 광학현미경으로 파란색으로 염색된 세포를 관찰하였다. SA-β-gal 활성 정도는 총 50개의 세포 중에서 세포질에 파란색으로 염색된 세포 수를 측정하여 DMSO 처리군을 100%로 하고, 울릉국화 추출물 처리군의 결과는 DMSO 처리군에 대한 백분율(%)로 표시하였다. 이러한 실험결과는 표2 또는 도3 및 도4에 평균 ± 편차로 나타내었으며, 통계적 유의성은 Newman-keuls 방법에 의해 결정하였다.
The Ulleung chrysanthemum extract was treated for 3 days, the cells were washed twice with phosphate buffer, the cells were fixed with 3.7% paraformaldehyde for 1 minute, and the fixative solution was removed. 5 mM potassium ferricyanide, 5 mM potassium ferricyanide, 150 mM NaCl, 2 mM MgCl2, X-gal 1 mg / ml) in a 24 well culture vessel Was added, and then wrapped in silver foil and reacted at 37 for 18 hours. The cells were washed twice with phosphate buffered saline (PBS), washed twice with phosphate buffered saline, and then stained blue with an optical microscope. The degree of SA-β-gal activity was determined by counting the number of cells stained blue in the cytoplasm among 50 cells, and the percentage of SA-β-gal activity in DMSO-treated group was 100% Respectively. These experimental results are shown as mean ± deviation in Table 2 or Figures 3 and 4, and statistical significance was determined by the Newman-keuls method.
실험결과, 섬유아세포에서 상기 실시예에 따른 울릉국화 추출물이 5 μg/ml 에서는 29.99 ± 8.84%의 SA-β-gal 활성이 보였으며, 20 μg/ml 에서는 11.62 ± 0.60%의 SA-β-gal 활성을 보였다. As a result, SA-β-gal activity was 29.99 ± 8.84% at 5 μg / ml, and SA-β-gal was 11.62 ± 0.60% at 20 μg / ml in the fibroblasts Lt; / RTI >
더욱 낮은 농도의 울릉국화 추출물로 실험한 결과는 도 5와 같다. 울릉국화 추출물 2 μg/ml 에서는 61.19 ± 4.93 %의 SA-β-gal 활성을 보였으며, 4 μg/ml 에서는 54.81 ± 4.11 %의 SA-β-gal 활성, 그리고 8 μg/ml 에서는 39.35 ± 2.46 %의 SA-β-gal 활성을 보였다.The results of the experiment with lower concentration of Ulleung chrysanthemum extract are shown in FIG. Β-gal activity was 61.19 ± 4.93% at 4 μg / ml, 54.81 ± 4.11% at SA, and 39.35 ± 2.46% at 8 μg / ml at the concentration of 2 μg / Of SA-β-gal activity.
즉, 도 4 및 도 5에 보이는 바와 같이, 2 내지 20 μg/ml 농도에서 울릉국화 추출물이 우수한 SA-β-gal 활성 감소를 보여, 세포노화 억제 효능이 우수한 것으로 나타났다.
That is, as shown in FIG. 4 and FIG. 5, the extract of Ulreung chrysanthemum showed an excellent SA-β-gal activity reduction at a concentration of 2 to 20 μg / ml and showed excellent antioxidant activity.
1. 세포 배양1. Cell culture
사람 혈관내피세포 (Human umbilical vein endothelial cells, HUVECs)는 10% 우태아혈청 및 1% 항생제가 포함된 혈관내피세포 배양배지 (FGM-2, Lonza, 스위스)을 이용하여 직경 100 mm 배양접시에 세포를 1 x 105개로 분주한 후, 37의 5% 이산화탄소 배양기에서 배양하였다. 배양접시의 바닥에 80~90% 정도 세포가 자라면, 2.5 x 트립신-EDTA 용액(Lonza, 스위스)을 처리하여 세포를 분리한 후, 계대 배양하였다. 이 때 상기 배양은 EGM-2 배양액을 사용하였다. Human umbilical vein endothelial cells (HUVECs) were cultured in a 100 mm diameter culture dish using a vascular endothelial cell culture medium (FGM-2, Lonza, Switzerland) containing 10% fetal bovine serum and 1% Were dispensed at 1 × 10 5 , and then cultured in a 37% 5% carbon dioxide incubator. If 80-90% of the cells were grown on the bottom of the culture dish, the cells were treated with 2.5 x trypsin-EDTA solution (Lonza, Switzerland) and subcultured. At this time, EGM-2 culture medium was used for the culture.
세포의 분열 횟수(population doubling, PD)는 다음 식으로 조사하였다.
The population doubling (PD) was examined by the following equation.
PD= log2F/log2I (F=마지막 세포수, I=처음 세포수)
PD = log 2 F / log 2 I (F = number of cells last, I = number of cells first)
실험에 사용한 섬유아세포는 분열횟수가 8회 이하의 것을 사용하였다.
The fibroblasts used in the experiment were those having a number of divisions of 8 or less.
2. 세포 노화 유도 및 울릉국화 추출물 처리2. Cell aging induction and Ulleung chrysanthemum extract treatment
상기 실시예 2-1과 같은 방법으로, 직경 100 mm 배양접시에 HUVEC(Human umbilical vein endothelial cell)을 150,000개 분주하고, 3일간 37, 5% 이산화탄소배양기에서 배양한 후, 세포 배양액을 제거하고, 세포를 1% 항생제가 포함된 EMDM배양액으로 2회 세척한 후, 0.5 μM 독소루비신을 4시간 처리하였다.In the same manner as in Example 2-1, 150,000 HUVECs (Human Umbilical Vein Endothelial Cells) were dispensed into a 100 mm diameter culture dish and cultured for 3 days in a 37, 5% carbon dioxide incubator. Cells were washed twice with EMDM medium containing 1% antibiotic and treated with 0.5 μM doxorubicin for 4 hours.
독소루비신 0.5 μM을 4시간 처리한 세포를 50ml 튜브에 10% 우태아혈청 및 1% 항생제가 포함된 EGM-2 배양액에 HUVEC 5,000개/ml이 되도록 하고 96웰 배양용기의 각 웰에 100 μl씩 분주하였다. 최종적으로 각 웰당 HUVEC 500개를 분주하고, 하루 동안 37, 5% 이산화탄소배양기에서 배양하였다. 각 웰에 1% 항생제가 포함된 EGM-2 배양액을 100 μl씩 더 넣어 준 후, 상기 실시예의 울릉국화 추출물을 처리하였다. 울릉국화 추출물은 20 μ/ml를 처리하였다. 이후 3일간 배양하여 SA-β-gal 염색을 실시하였다.Cells treated with 0.5 μM doxorubicin for 4 hours were seeded in a 50 ml tube in an EGM-2 culture medium containing 10% fetal bovine serum and 1% antibiotic at a concentration of 5,000 cells / ml, and 100 μl of each cell was dispensed into each well of a 96- Respectively. Finally, 500 HUVECs per well were dispensed and cultured in 37, 5% CO2 incubator for one day. 100 μl of EGM-2 culture medium containing 1% antibiotic was further added to each well, and the Ulleung chrysanthemum extract of the above-mentioned Example was treated. Ullung chrysanthemum extract was treated with 20 μ / ml. The cells were then cultured for 3 days and stained with SA-β-gal.
한편, 음성대조군으로 용매인 DMSO를 처리하였으며, 양성대조물질로는 5 mM (= 815.97 ug/ml) NAC 를 처리하여 비교실험하였다.
Meanwhile, DMSO, a negative control, was treated with 5 mM (= 815.97 ug / ml) NAC as a positive control.
3. SA-β-갈락토시다아제 활성 염색3. Staining of SA-β-galactosidase activity
상기 처리 후 3일 동안 37, 5% 이산화탄소배양기에서 배양하고, 세포노화 저해에 대한 울릉국화 추출물의 효과를 SA-β-gal 활성 염색으로 조사하였다. β-갈락토시데이스는 갈락토스와 유기물 사이에 형성된 β-글라이코시딕 결합을 가수분해시키는 엑소글라이코시데이즈이며, 노화가 진행될수록 세포의 리소좀에 다량으로 축적되고, 그 발현 정도를 5-bromo-4-chloro-3-indolyl-βD-galactopyranoside를 사용하여 식별하고 정량화시킬 수 있다. 이 때, SA-β-gal 염색법으로 노화된 세포는 세포질이 파란색으로 염색된다. The cells were cultured in 37, 5% CO2 incubator for 3 days after the treatment, and the effect of Ulleung chrysanthemum extract on cell senescence inhibition was investigated by SA-β-gal activity staining. β-galactosidase is exoglycosidase that hydrolyzes β-glycosidic bonds formed between galactose and organic matter. As aging progresses, a large amount of β-galactosidase accumulates in lysosomes of cells, bromo-4-chloro-3-indolyl- [beta] D-galactopyranoside. At this time, cells aged by SA-β-gal staining are stained cytoplasm in blue.
울릉국화 추출물을 3일 동안 처리한 후, 세포를 인산완충액으로 2번 세척하고, 3.7% 파라포름알데하이드로 세포를 1분간 고정한 후, 고정액을 제거하였다. SA-β-gal 염색 용액(40 mM citric acid/phosphate [pH 5.8], 5 mM potassium ferrocyanide, 5 mM potassium ferricyanide, 150 mM NaCl, 2 mM MgCl2, X-gal 1 mg/ml)을 24웰 배양용기에는 500μl를 넣어 준 후, 은박지로 싸서 37에서 18시간 동안 반응시켰다. 인산완충용액 (PBS)으로 2번 세척한 후 인산완충용액으로 2회 세척 한 후, 광학현미경으로 파란색으로 염색된 세포를 관찰하였다. SA-β-gal 활성 정도는 총 50개의 세포 중에서 세포질에 파란색으로 염색된 세포 수를 측정하여 DMSO 처리군을 100%로 하고, 울릉국화 추출물 처리군의 결과는 DMSO 처리군에 대한 백분율(%)로 표시하였다. 이러한 실험결과는 표 5 또는 도 6에 평균 ± 편차로 나타내었으며, 통계적 유의성은 Student's t-test 방법에 의해 결정하였다.
The Ulleung chrysanthemum extract was treated for 3 days, the cells were washed twice with phosphate buffer, the cells were fixed with 3.7% paraformaldehyde for 1 minute, and the fixative solution was removed. 5 mM potassium ferricyanide, 5 mM potassium ferricyanide, 150 mM NaCl, 2 mM MgCl2, X-gal 1 mg / ml) in a 24 well culture vessel Was added, and then wrapped in silver foil and reacted at 37 for 18 hours. The cells were washed twice with phosphate buffered saline (PBS), washed twice with phosphate buffered saline, and then stained blue with an optical microscope. The degree of SA-β-gal activity was determined by counting the number of cells stained blue in the cytoplasm among 50 cells, and the percentage of SA-β-gal activity in DMSO-treated group was 100% Respectively. These experimental results are shown in Table 5 or FIG. 6 as mean ± deviation, and statistical significance was determined by Student's t-test method.
<비교예 1> 여러 구절초 추출물의 세포노화저해효과 확인≪ Comparative Example 1 > Confirmation of cell aging inhibitory effect of various herb extracts
산구절초, 서흥구절초, 포천구절초를 사용한 점을 제외하고는 상기 실시예 4와 동일한 방법으로 상기 구절초의 추출물을 제조 및 처리하여 실험하였다. The extract was prepared and treated in the same manner as in Example 4, except that Acanthopanax senticosus, Seocheong Rhizoma, and Pocheon Rhizoma were used.
이 때, 울릉국화 추출물, 산구절초, 서흥구절초 및 포천구절초는 각각 DZ, G2, G3 및 G4로 나타냈다. At this time, the extracts of Ulreung chrysanthemum, Mt., Mt. Seoheung, and Pocheon were DZ, G2, G3 and G4, respectively.
그 결과, 도 9에 나타난 바와 같이, 울릉국화 추출물은 포천구절초 추출물과 비교하여 약 5%p 내지 7%p 증가된 세포노화저해 효과를 나타냄을 확인할 수 있었다. 즉, 울릉 국화 추출물은 산구절초, 서흥구절초, 포천구절초 추출물에 비해 세포노화저해 효능이 우수한 것을 확인할 수 있다.
As a result, as shown in Fig. 9, it was confirmed that the extract of Ulreung chrysanthemum showed an inhibitory effect on cell senescence increased by about 5% ~ 7% p as compared with that of Pocheon Rhizoma extract. In other words, the extract of Ulreung chrysanthemum is superior to the extracts of Acanthopanax senticosus, Seoheung Kwangseo and Pocheon Rhizoma extracts.
이하, 본 발명의 울릉국화 추출물을 포함하는 화장료 조성물의 제제예를 설명하나, 본 발명은 이를 한정하고자 함이 아닌 단지 구체적으로 설명하고자 함이다.
Hereinafter, formulation examples of the cosmetic composition containing Ulrethrae chrysanthemum extract of the present invention will be described, but the present invention is not intended to be limited thereto but is specifically explained.
<제제예7> 에센스의 제조≪ Formulation Example 7 > Preparation of essence
에틸 알코올 4.00 중량%Ethyl alcohol 4.00 wt%
메틸 파라벤 0.10 중량%Methyl paraben 0.10 wt%
향 0.10 중량%Fragrance 0.10 wt%
수소처리된 캐스터 오일 0.60 중량%Hydrotreated castor oil 0.60 wt%
DL-α-토코페릴 아세테이트 0.02 중량%DL-a-tocopheryl acetate 0.02 wt%
글리세린 5.00 중량%Glycerin 5.00 wt%
1,3-부틸렌 글리콜 3.00 중량%1,3-butylene glycol 3.00 wt%
하이드록시 에틸 셀룰로오스 0.20 중량%0.20% by weight of hydroxyethylcellulose
잔탄검 0.40 중량%Xanthan gum 0.40 wt%
E.D.T.A-2Na 0.02 중량%0.02 wt% of E. D. T. A-2Na,
피쉬-콜라겐 4.00 중량%Fish-collagen 4.00 wt%
울릉국화 추출물 조성물 0.5~2.0 중량%Ulleung chrysanthemum extract composition 0.5 ~ 2.0 wt%
증류수 잔량
Distilled water balance
<제제예8> 클렌징폼의 제조≪ Formulation Example 8 > Preparation of Cleansing Foam
스테아린산 6.5 중량%6.5% by weight stearic acid
미리스틴산 28.0 중량%Myristic acid 28.0 wt%
자기유화형 모노스테아린산 글리세린 3.0 중량%3.0% by weight of self-emulsifiable glycerin monostearate
프로필렌 글리콜 5.0 중량%5.0% by weight of propylene glycol
농글리세린 10.0 중량%Concentrated glycerin 10.0 wt%
수산화나트륨 7.0 중량%Sodium hydroxide 7.0 wt%
에틸렌디아민테트라초산나트륨 0.1 중량%0.1% by weight of sodium ethylenediaminetetraacetate
울릉국화 추출물 조성물 7.0 중량%Ulleung chrysanthemum extract composition 7.0 weight%
향료 미량Fragrance
정제수 잔량
Purified water balance
Claims (10)
The present invention relates to an extract of Dendranthema zawadskii var. Lucidum, consisting of stem, leaf and flower, which is extracted from at least one solvent selected from the group consisting of water, alcohol, ethyl acetate, chloroform and hexane as doxorubicin ) For preventing skin aging.
The cosmetic composition for preventing skin aging according to claim 1, wherein the extract of Dendranthema zawadskii var. Lucidum inhibits cell senescence due to doxorubicin.
The cosmetic composition for preventing skin aging according to claim 1, wherein the Ulleung chrysanthemum extract has anti-SA-β-galactosidase activity and has an effect of improving wrinkles and skin elasticity by doxorubicin.
The cosmetic composition for preventing skin aging according to claim 1, wherein the Ulleung chrysanthemum extract inhibits aging of human dermal fibroblasts (HDFs) caused by doxorubicin.
The cosmetic composition for preventing skin aging according to claim 1, wherein the cosmetic composition comprises Ulleung chrysanthemum extract at a concentration of 2 to 10 μg / ml.
The cosmetic composition for preventing skin aging according to claim 1, wherein the Ulleung chrysanthemum extract comprises 0.001 to 10% by weight based on the total weight of the composition.
The cosmetic composition according to claim 1, wherein the cosmetic composition is any one of a cream, a lotion, a powder, a surfactant-containing cleansing oil, a powder foundation, an emulsion foundation, a wax foundation, a paste, an essence, (Doxorubicin) to prevent skin aging.
Preventing aging of skin and improving wrinkles caused by doxorubicin, including extracts of water from above ground Ulleung chrysanthemum comprising stem, leaf and flower, alcohol, ethyl acetate, chloroform and hexane And improving skin elasticity.
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| KR1020150124424 | 2015-09-02 | ||
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| KR1020160113493A Division KR101945862B1 (en) | 2015-09-02 | 2016-09-02 | Anti-aging Composition Including Dendranthema zawadskii Extract, and Cosmetic Composition for young face by Anti-aging, Skin-lifting and Anti-wrinkle |
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| KR20180026310A KR20180026310A (en) | 2018-03-12 |
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| KR1020160113493A KR101945862B1 (en) | 2015-09-02 | 2016-09-02 | Anti-aging Composition Including Dendranthema zawadskii Extract, and Cosmetic Composition for young face by Anti-aging, Skin-lifting and Anti-wrinkle |
| KR1020160114105A KR101945865B1 (en) | 2015-09-02 | 2016-09-05 | Anti-aging Composition Including Dendranthema zawadskii Extract |
| KR1020160114091A KR101945864B1 (en) | 2015-09-02 | 2016-09-05 | Cosmetic Composition Including Dendranthema zawadskii Extract for young face by Anti-aging, Skin-lifting and Anti-wrinkle |
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| KR1020160113493A KR101945862B1 (en) | 2015-09-02 | 2016-09-02 | Anti-aging Composition Including Dendranthema zawadskii Extract, and Cosmetic Composition for young face by Anti-aging, Skin-lifting and Anti-wrinkle |
| KR1020160114105A KR101945865B1 (en) | 2015-09-02 | 2016-09-05 | Anti-aging Composition Including Dendranthema zawadskii Extract |
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Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20170027594A (en) * | 2015-09-02 | 2017-03-10 | 대한민국(농촌진흥청장) | A composition comprising an extract of Dendranthema Zawadskii for preventing and treating inflammatory disease |
| KR102395725B1 (en) * | 2016-03-31 | 2022-05-10 | (주)아모레퍼시픽 | Skin external composition for anti-aging comprising chrysanthemum zawadskii herb. var. lucidum (nakai) t.lee |
| CN106890278A (en) * | 2017-04-17 | 2017-06-27 | 杨长安 | The health Chinese medicine composition of eliminating the phlegm dehumidifying anti-aging |
| KR102124698B1 (en) | 2018-08-20 | 2020-06-19 | 코스맥스 주식회사 | Cosmetic composition for improving skin wrinkle or skin aging comprising mixture extract of Chrysanthemum morfiolium, Aloe vera, and Hedera Helix |
| KR102117661B1 (en) * | 2018-11-26 | 2020-06-02 | 주식회사 예나 | Cosmetic composition for skin anti-wrinkle and anti-aging containing extracts of Chrysanthemum zawadskii subsp. lucidum (Nakai) Y.N.Lee, Solidago virgaurea and Anthriscus sylvestris |
| KR20210014489A (en) | 2019-07-30 | 2021-02-09 | (주)국심 | Anti-aging health food make use of regional product kyoungnam region and chrysanthemum |
| KR102350857B1 (en) | 2019-11-26 | 2022-01-14 | (주)에이텍 | Cosmetic composition containing lactic acid bacteria fermentation of chrysanthemum and magnolia extracted with YU SEONG hot spring water |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2013035776A (en) | 2011-08-08 | 2013-02-21 | Shiseido Co Ltd | Collagen production promoter |
| KR101244752B1 (en) * | 2010-07-27 | 2013-03-19 | 정연옥 | Cosmetic ingredient containing whitening, anti-aging, anti-oxidation and anti-microbial activities |
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2016
- 2016-09-02 KR KR1020160113493A patent/KR101945862B1/en active IP Right Grant
- 2016-09-05 KR KR1020160114105A patent/KR101945865B1/en active IP Right Grant
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101244752B1 (en) * | 2010-07-27 | 2013-03-19 | 정연옥 | Cosmetic ingredient containing whitening, anti-aging, anti-oxidation and anti-microbial activities |
| JP2013035776A (en) | 2011-08-08 | 2013-02-21 | Shiseido Co Ltd | Collagen production promoter |
Also Published As
| Publication number | Publication date |
|---|---|
| KR20170027685A (en) | 2017-03-10 |
| KR20180026311A (en) | 2018-03-12 |
| KR101945862B1 (en) | 2019-02-11 |
| KR20180026310A (en) | 2018-03-12 |
| KR101945865B1 (en) | 2019-02-11 |
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