KR20210084994A - Composition for enhancing skin cell regeneration containing Caragana sinica flower oil - Google Patents
Composition for enhancing skin cell regeneration containing Caragana sinica flower oil Download PDFInfo
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- KR20210084994A KR20210084994A KR1020190177564A KR20190177564A KR20210084994A KR 20210084994 A KR20210084994 A KR 20210084994A KR 1020190177564 A KR1020190177564 A KR 1020190177564A KR 20190177564 A KR20190177564 A KR 20190177564A KR 20210084994 A KR20210084994 A KR 20210084994A
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- composition
- skin
- oil
- wound healing
- lotion
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Abstract
Description
본 발명은 골담초 꽃 오일의 피부 재생 또는 주름 개선용 조성물에 대한 것으로서, 더욱 상세하게는 골담초(Caragana sinica)의 꽃에서 유기용매 추출법(solvent extraction)으로 추출한 오일의 피부 재생, 상처 치유 또는 주름개선용 조성물에 관한 것이다.The present invention relates to a composition for skin regeneration or wrinkle improvement of dandelion flower oil, and more particularly, for skin regeneration, wound healing or wrinkle improvement of oil extracted from the flower of Caragana sinica by solvent extraction to the composition.
피부(skin)는 인체의 가장 외부를 덮고 있는 조직으로 인체를 보호하고 장벽기능을 하여 외부의 화학적 또는 물리적 충격으로부터 몸을 보호하는 일차적인 방어기능을 한다. 피부는 표피(epidermis), 진피(dermis) 및 피하지방층(subcutis)의 3개의 층으로 구성되어 있다.The skin is the tissue that covers the outermost part of the human body and has a primary defense function that protects the human body and acts as a barrier to protect the body from external chemical or physical shocks. The skin is composed of three layers: the epidermis, the dermis, and the subcutis.
피부의 가장 바깥층에 위치하고 있는 표피층은 각질층, 투명층, 과립층, 유극층, 기저층으로 구성되어 있으며, 표피층의 95%가 각질형성세포(keratinocyte)로 구성되어있다. 각질형성세포는 피부에 침투하는 세균, 화학 물질 등에 효율적으로 방어역할을 하고, 다양한 사이토카인을 분비하여 피부의 염증반응과 면역반응에 관여한다. 또한, 각질형성세포의 이동과 증식은 상처치유와 피부 재생, 그리고 콜라겐(collagen type Ⅰ, collagen type Ⅳ) 합성을 유도하여 피부의 탄력 증진과 기저막 형성에 중요한 역할을 한다. 콜라겐은 알파체인에 따라 20종 이상의 콜라겐이 있으며, 사람의 피부에는 type I, III, IV, V, VI 및 VII 형 콜라겐이 분포하고 있다. 주로 type I 콜라겐은 피부 진피층의 85% 이상을 구성하고, type IV 콜라겐은 기저막(basement membrane)의 주요 구조 단백질이다. type I 및 IV 콜라겐의 합성은 피부의 탄력을 증진시키고, 기저막 형성을 향상시켜 피부 세포들의 부착과 증식이 조절되는 것으로 알려져 있다. 이로 인해, 피부 표피 각질형성세포의 이동과 증식, 그리고 콜라겐 합성의 유도는 피부 재생과 상처치유 뿐만 아니라 피부 탄력을 증진시키고 건강한 피부가 유지되도록 도와준다. The epidermal layer, located at the outermost layer of the skin, is composed of the stratum corneum, transparent layer, granular layer, stratum corneum, and basal layer, and 95% of the epidermal layer is composed of keratinocytes. Keratinocytes effectively defend against bacteria and chemicals that penetrate the skin, and secrete various cytokines to participate in the skin's inflammatory and immune responses. In addition, the migration and proliferation of keratinocytes plays an important role in improving skin elasticity and forming the basement membrane by inducing wound healing, skin regeneration, and collagen (collagen type I, collagen type IV) synthesis. There are more than 20 types of collagen according to the alpha chain, and types I, III, IV, V, VI and VII collagen are distributed in human skin. Mainly, type I collagen constitutes more than 85% of the dermal layer of the skin, and type IV collagen is a major structural protein of the basement membrane. It is known that the synthesis of type I and IV collagen enhances the elasticity of the skin and improves the formation of the basement membrane, thereby regulating the adhesion and proliferation of skin cells. For this reason, the migration and proliferation of epidermal keratinocytes, and induction of collagen synthesis, not only regenerate and heal wounds, but also enhance skin elasticity and help maintain healthy skin.
골담초(Caragana sinica)는 골담초 또는 기타 동속 근연 식물의 뿌리를 기원으로 하고, 한국 및 중국 산지에서자라는 콩과 식물이다. 높이는 약 2m이상으로, 가지가 사방으로 퍼지고, 줄기는 회갈색으로 가시가 뭉쳐나고 5개의 능선이 있다. 잎은 어긋나고, 홀수 1회 깃꼴겹잎이며, 작은 잎은 4개로 타원형인 특징이 있다. 작은 잎의 길이가 8~17 cm 인 것을 반용골담초(var. megalantha), 작은 잎이 12~18 cm 인 것을 좀골담초(C.microphylla)라고 한다. 잎과 가지에 플라보노이드, 열매 껍질에 알칼로이드, 여물지 않은 열매에는 이노시톨 등이 있으며, carpahenol A (resveratrol trimer), (+)-isoampelopson F (resveratrol dimer), caraphenol B(resveratrol dimer), caraphenol C (resveratrol dimer) 등의 올리고스틸벤(oligostilbene) 류를 함유하고 있는 것으로 보고되어 있다. 주요 효능으로는 강장, 이뇨, 류마티즘, 관절염, 대하증, 요통, 급성유선염, 이명, 뼈, 신경통에 효능이 있는 것으로 알려진 식물이다.Golddamcho ( Caragana sinica ) is a leguminous plant that originates from the root of Golddamcho or other closely related plants, and grows in Korea and China. The height is about 2m or more, the branches are spread out in all directions, the stem is grayish-brown, and there are 5 ridges. The leaves are alternate phyllotaxis and have an odd number of pinnate compound leaves, and there are 4 small leaves, which are oval. Those with small leaves of 8-17 cm in length are called var. megalantha, and those with small leaves of 12-18 cm are called C. microphylla. Flavonoids on leaves and branches, alkaloids on fruit skin, inositol in unripe fruit, carpahenol A (resveratrol trimer), (+)-isoampelopson F (resveratrol dimer), caraphenol B (resveratrol dimer), caraphenol C (resveratrol) dimer) and the like are reported to contain oligostilbene. It is a plant known to be effective for tonicity, diuresis, rheumatism, arthritis, hypothyroidism, back pain, acute mastitis, tinnitus, bone, and neuralgia.
천연물 유래 에센셜 오일은 식물의 2차 대사 산물 중 인체에 약리효과를 나타내는 휘발성 화학물질로, 주로 알코올, 알데하이드, 에스테르, 케톤 등의 여러가지 유기화합물로 구성되어 있다. 에센셜 오일의 독특한 향기 성분과 치료 성분은 피부, 순환계, 후각계를 통해 스트레스 해소, 피로 회복, 근육 이완 등의 효과를 나타낼 뿐만 아니라 항균작용, 면역 증진, 항알러지 작용이 있어 오래전부터 질병의 치료와 예방에 활용되었으며 노화 예방과 같은 미용에도 효과가 있다. Essential oils derived from natural products are volatile chemicals that have pharmacological effects on the human body among secondary metabolites of plants, and are mainly composed of various organic compounds such as alcohols, aldehydes, esters, and ketones. The unique fragrance and therapeutic ingredients of essential oils not only show effects such as relieving stress, recovering from fatigue, and relaxing muscles through the skin, circulation and olfactory systems, but also have antibacterial, immune-enhancing and anti-allergic effects, so they have been used in the treatment of diseases and diseases for a long time. It has been used for prevention and is also effective for beauty such as preventing aging.
에센셜 오일의 이러한 효능·효과는 현대에도 이어져 제약이나 식품뿐만 아니라 화장품 분야까지 다양하게 연구 및 활용되고 있다. 특히 화장품 분야에 있어 최근에는 합성 향료 대신 에센셜 오일을 선호하는 경향이 증가하고 있어 에센셜 오일의 수요가 점점 증가하고 있는 추세이다. These effects and effects of essential oils have continued to the present day and are being studied and utilized in various fields, not only in pharmaceuticals and foods, but also in cosmetics. In particular, in the cosmetic field, there is a tendency to prefer essential oils instead of synthetic fragrances in recent years, so the demand for essential oils is increasing.
따라서, 상기와 같은 문제점을 해결하기 위하여 안출된 본 발명의 목적은, 골담초 오일을 유효성분으로 포함하는 피부 재생, 상처 치유 또는 주름개선용 조성물을 제공하는 것이다.Accordingly, an object of the present invention, devised to solve the above problems, is to provide a composition for skin regeneration, wound healing, or wrinkle improvement, which includes bone marrow oil as an active ingredient.
또한, 본 발명의 다른 목적은 골담초 오일을 유효성분으로 포함하는 피부 재생, 상처 치유 또는 주름개선용 화장료 조성물 또는 피부외용제 조성물 또는 식품 조성물을 제공하는 것이다.In addition, another object of the present invention is to provide a cosmetic composition for skin regeneration, wound healing or wrinkle improvement, a composition for external application for skin, or a food composition, which includes bone marrow oil as an active ingredient.
이상에서의 본 발명에서 해결하고자 하는 다양한 과제들은 이에 한정하는 것이 아니라, 후술할 실시예 및 청구범위에 기재된 사항을 통하여 본 발명이 속하는 분야의 통상의 지식을 가진 자에 의하여 분명하게 이해될 수 있을 것이다.Various problems to be solved in the present invention in the above are not limited thereto, but can be clearly understood by those of ordinary skill in the art to which the present invention pertains through the examples and claims to be described later. will be.
상기 과제를 해결하기 위한 본 발명에 일 실시예에 따르면, 골담초 오일을 유효성분으로 포함하는 피부 재생, 상처 치유 또는 주름개선용 조성물을 제공한다.According to one embodiment of the present invention for solving the above problems, there is provided a composition for skin regeneration, wound healing, or wrinkle improvement comprising bone marrow oil as an active ingredient.
또한, 상기 골담초 오일은 골담초(Caragana sinica)의 꽃으로부터 추출한 것을 특징으로 한다.In addition, the Goldamcho oil is characterized in that it is extracted from the flower of Goldamcho ( Caragana sinica ).
또한, 상기 오일은 물, 탄소수 1 내지 4의 알코올, 1,3-부틸렌글리콜, 아세톤, 에탄올, 에틸아세테이트, 부틸아세테이트, 헥산, 클로로포름 또는 이들의 혼합물을 이용하여 유기용매법에 의해 추출된 추출물인 것을 특징으로 한다.In addition, the oil is an extract extracted by an organic solvent method using water, an alcohol having 1 to 4 carbon atoms, 1,3-butylene glycol, acetone, ethanol, ethyl acetate, butyl acetate, hexane, chloroform, or a mixture thereof. characterized by being.
또한, 상기 오일은 상기 추출물에서 알코올을 이용하여 불용성 왁스를 제거한 것을 특징으로 한다.In addition, the oil is characterized in that the insoluble wax is removed from the extract using alcohol.
또한, 상기 오일은 휘발성 유기 화합물을 포함하는 것을 특징으로 한다.In addition, the oil is characterized in that it contains a volatile organic compound.
또한, 상기 휘발성 유기화합물은 7,11,15-trimethyl-3-methylidenehexadec-1-ene, α-linolenic acid, ethyl linoleate로 이루어진 그룹에서 선택되는 어느 하나 이상을 포함하는 것을 특징으로 한다.In addition, the volatile organic compound is characterized in that it contains any one or more selected from the group consisting of 7,11,15-trimethyl-3-methylidenehexadec-1-ene, α-linolenic acid, and ethyl linoleate.
또한, 상기 오일의 농도가 0.01μg/mL 내지 100μg/mL인 것을 특징으로 한다.In addition, it is characterized in that the concentration of the oil is 0.01 μg/mL to 100 μg/mL.
본 발명의 다른 실시예에 따르면, 골담초 오일을 유효성분으로 포함하는 피부 재생, 상처 치유 또는 주름개선용 화장료 조성물을 제공한다.According to another embodiment of the present invention, there is provided a cosmetic composition for skin regeneration, wound healing, or wrinkle improvement comprising bone marrow oil as an active ingredient.
또한, 상기 조성물은 피부외용연고, 크림, 유연화장수, 영양화장수, 팩, 에센스, 헤어토닉, 샴푸, 린스, 헤어 컨디셔너, 헤어 트리트먼트, 젤, 스킨로션, 스킨소프너, 스킨토너, 아스트린젠트, 로션, 밀크 로션, 모이스처 로션, 영양로션, 마사지 크림, 영양크림, 모이스처 크림, 핸드크림, 파운데이션, 영양에센스, 선스크린, 비누, 클렌징폼, 클렌징로션, 클렌징크림, 바디 로션 및 바디 클렌저로 이루어진 그룹에서 선택되는 어느 하나의 제형으로 제조된 것을 특징으로 한다.In addition, the composition is an external skin ointment, cream, softening lotion, nourishing lotion, pack, essence, hair tonic, shampoo, conditioner, hair conditioner, hair treatment, gel, skin lotion, skin softener, skin toner, astringent, lotion, Milk Lotion, Moisture Lotion, Nutrient Lotion, Massage Cream, Nutrient Cream, Moisture Cream, Hand Cream, Foundation, Nutrient Essence, Sunscreen, Soap, Cleansing Foam, Cleansing Lotion, Cleansing Cream, Body Lotion and Body Cleanser It is characterized in that it is prepared in any one formulation.
이어서, 골담초 오일을 유효성분으로 포함하는 피부 재생, 상처 치유 또는 주름개선용 피부외용제 조성물 또는 식품 조성물을 제공한다.Then, there is provided a composition for external application for skin or a food composition for skin regeneration, wound healing, or wrinkle improvement, comprising bone marrow oil as an active ingredient.
본 발명은 골담초로부터 추출된 에센셜 오일의 성분을 규명하고 이의 피부 재생 및 상처치유, 주름개선 효과를 입증한 것으로, 본 발명에 따른 골담초 오일은 각질형성세포에 처리 시 각질형성세포의 증식과 이동을 효과적으로 유도하고, 표피 각질형성세포에서 콜라겐의 맹아(sprouting) 자연성장(out growth)를 증진시키고, 표피 각질형성세포의 콜라겐 type Ⅰ 및 Ⅳ 합성과 분비를 촉진시킨다는 것을 확인함으로써 피부를 재생하고 주름을 개선시키는 효과가 있어 화장품 및 식품분야에서 유용하게 활용될 수 있다.The present invention identifies the components of essential oil extracted from bone marrow and proves its effects on skin regeneration, wound healing, and wrinkle improvement. The bone marrow oil according to the present invention improves the proliferation and migration of keratinocytes when treated with keratinocytes. By confirming that it effectively induces and promotes collagen spouting and out growth in epidermal keratinocytes, and promotes collagen type I and IV synthesis and secretion of epidermal keratinocytes, it regenerates the skin and reduces wrinkles It has an improving effect and can be usefully used in cosmetics and food fields.
이상에서의 본 발명에 따른 효과는 상기에 한정되는 것은 아니며, 기타 본 발명의 효과들은 후술할 실시예 및 청구범위에 기재된 사항을 통하여 본 발명이 속하는 분야의 통상의 지식을 가진 자에 의하여 분명하게 이해될 수 있을 것이다.The effects according to the present invention in the above are not limited to the above, and other effects of the present invention can be clearly identified by those of ordinary skill in the art through the examples and claims to be described later. can be understood
도 1은 본 발명의 실시예에 따른 골담초 꽃으로부터 에센셜 오일을 추출하는 과정을 나타내는 흐름도이다.
도 2는 본 발명의 실시예에 따른 골담초의 꽃으로부터 추출한 에센셜 오일의 성분 분석 결과표이다.
도 3은 골담초 꽃 에센셜 오일의 표피 각질형성세포에 대한 독성을 나타내는 그래프이다.
도 4는 골담초 꽃 에센셜 오일이 표피 각질형성세포의 증식에 미치는 효과를 나타내는 그래프이다.
도 5는 골담초 꽃 에센셜 오일이 표피 각질형성세포의 이동에 미치는 효과를 나타내는 그래프이다.
도 6은 골담초 꽃 에센셜 오일이 표피 각질형성세포에서 콜라겐 맹아(sprouting) 자연성장(out growth)에 미치는 영향을 나타낸 것이다.
도 7은 골담초 꽃 에센셜 오일이 콜라겐 type I과 Ⅳ 합성에 미치는 효과를 나타내는 그래프이다.1 is a flow chart showing a process of extracting essential oil from a flower according to an embodiment of the present invention.
2 is a table showing the results of component analysis of essential oils extracted from flowers of Goldamcho according to an embodiment of the present invention.
Figure 3 is a graph showing the toxicity of the flower essential oil of Goldamcho flower to epidermal keratinocytes.
FIG. 4 is a graph showing the effect of Goldamcho flower essential oil on the proliferation of epidermal keratinocytes.
FIG. 5 is a graph showing the effect of Goldamcho flower essential oil on the migration of epidermal keratinocytes.
6 is a graph showing the effect of Goldamcho flower essential oil on collagen sprouting and out growth in epidermal keratinocytes.
7 is a graph showing the effect of Goldamcho flower essential oil on collagen type I and IV synthesis.
이하, 본 발명에 대하여 보다 상세하게 설명하도록 한다. 이하의 설명에 있어, 당업자에게 주지 저명한 기술에 대해서는 그 상세한 설명을 생략할 수 있다. 또한, 본 발명을 설명함에 있어서, 관련된 공지 기능 또는 구성에 대한 구체적인 설명이 본 발명의 요지를 불필요하게 흐릴 수 있다고 판단되는 경우에는 그 상세한 설명을 생략할 수 있다. 또한, 본 명세서에서 사용되는 용어(terminology)들은 본 발명의 바람직한 실시예를 적절히 표현하기 위해 사용된 용어들로서, 이는 사용자, 운용자의 의도 또는 본 발명이 속하는 분야의 관례 등에 따라 달라질 수 있다.Hereinafter, the present invention will be described in more detail. In the following description, detailed descriptions of well-known techniques known to those skilled in the art may be omitted. In addition, in describing the present invention, when it is determined that a detailed description of a related known function or configuration may unnecessarily obscure the gist of the present invention, the detailed description may be omitted. In addition, the terms used in this specification are terms used to properly express the preferred embodiment of the present invention, which may vary according to the intention of a user or operator, or customs in the field to which the present invention belongs.
따라서 본 용어들에 대한 정의는 본 명세서 전반에 걸친 내용을 토대로 내려져야 할 것이다. 명세서 전체에서, 어떤 부분이 어떤 구성요소를 "포함"한다고 할 때, 이는 특별히 반대되는 기재가 없는 한 다른 구성요소를 제외 하는 것이 아니라 다른 구성 요소를 더 포함할 수 있는 것을 의미한다.Therefore, definitions of these terms should be made based on the content throughout this specification. Throughout the specification, when a part "includes" a certain component, it means that other components may be further included, rather than excluding other components, unless otherwise stated.
본 발명은 골담초 오일을 유효성분으로 포함하는 피부 재생 또는 주름개선용 조성물을 제공한다.The present invention provides a composition for skin regeneration or wrinkle improvement comprising bone marrow oil as an active ingredient.
또한, 상기 골담초 오일은 골담초(Caragana sinica)의 꽃에서 추출된 것일 수 있다.In addition, the Goldamcho oil may be extracted from the flowers of Goldamcho ( Caragana sinica ).
또한, 일 실시예에서, 골담초의 꽃은 꽃이 피기 직전의 꽃봉오리일 수 있으며, 건조되지 않은 꽃일 수 있다.Also, in one embodiment, the flower of Golddammcho may be a bud just before flowering, or may be a flower that has not been dried.
또한, 상기 골담초 오일은 수증기증류법, 직접증류법, 퍼코레이션(percolation)법, 압착법, 냉침법, 유기용제법 등의 공지의 다양한 추출법에 의해 추출될 수 있는데, 바람직하게는 휘발성 용제를 이용한 유기용매 추출법(solvent extraction)에 의해 추출하는 것이 골담초 꽃에 포함된 소량의 유용성분을 더욱 많이 추출할 수 있어서 더욱 적합하다.In addition, the gold leaf oil can be extracted by various known extraction methods such as steam distillation, direct distillation, percolation, compression, chilling, organic solvent, etc., preferably an organic solvent using a volatile solvent. Extraction by solvent extraction is more suitable because it is possible to extract more of the small amount of useful components contained in the flower of dandelion.
또한, 상기 오일은 물, 탄소수 1 내지 4의 알코올, 1,3-부틸렌글리콜, 아세톤, 에탄올, 에틸아세테이트, 부틸아세테이트, 헥산, 클로로포름 또는 이들의 혼합물을 이용하여 유기용매법에 의해 추출된 에센셜 오일일 수 있다.In addition, the oil is an essential oil extracted by an organic solvent method using water, an alcohol having 1 to 4 carbon atoms, 1,3-butylene glycol, acetone, ethanol, ethyl acetate, butyl acetate, hexane, chloroform, or a mixture thereof. It may be oil.
또한, 상기 오일은 상기 추출물에서 알코올을 이용하여 불용성 왁스를 제거한 오일일 수 있으며, 바람직하게는 상기 유기용매법으로 추출하여 얻은 침천물로부터 콘크리트(concrete)를 획득하여 에틸알코올로 용해시켜 불용성 왁스를 제거한 앱솔루트(absolute)이다.In addition, the oil may be an oil in which insoluble wax is removed from the extract using alcohol, and preferably, concrete is obtained from the precipitate obtained by extraction by the organic solvent method, and the insoluble wax is removed by dissolving it with ethyl alcohol. It is absolute.
구체적으로, 상기 앱솔루트는 골담초 꽃을 휘발성 용제를 이용한 유기용매 추출법으로 추출하여 콘크리트(concrete)를 얻는 단계; 및 상기 얻은 콘크리트를 알코올로 추출하여 불용성 왁스(wax)를 제거하고 앱솔루트(absolute)를 얻는 단계에 의해 추출될 수 있다.Specifically, the absolute step of obtaining concrete (concrete) by extracting the flower of the Golden ivy by an organic solvent extraction method using a volatile solvent; And extracting the obtained concrete with alcohol to remove insoluble wax (wax) can be extracted by the step of obtaining an absolute (absolute).
일 실시예로서, 상기 앱솔루트는 도 1의 흐름도와 같이 추출할 수 있다. 우선, 건조하지 않은 골담초(Caragana sinica)의 꽃을 헥산(n-hexane)에 침전시킨 뒤, 얻은 추출물을 회전식 증발기(rotary evaporation)를 이용하여 농축시켜서 콘크리트를 얻은 다음, 상기 얻은 콘크리트에 에틸알코올(ethyl alcohol)을 넣어 용해시킨 후, 진공 냉각 필터(vacuum cooling filtration) 및 회전식 증발기를 이용하여 분리 또는 정제함으로써, 본 발명에 따른 골담초 앱솔루트를 제조할 수 있다.As an embodiment, the absolute may be extracted as shown in the flowchart of FIG. 1 . First, after precipitating the flowers of undried gold leaf ( Caragana sinica ) in hexane (n-hexane), the obtained extract is concentrated using rotary evaporation to obtain concrete, and then ethyl alcohol ( ethyl alcohol) is added and dissolved, and then separated or purified using a vacuum cooling filtration and a rotary evaporator, thereby preparing the bone marrow absolute according to the present invention.
상기 유기용매 추출법과 추출물로부터 앱솔루트를 분리하는 방법은 이 기술 분야에서 보통의 지식을 가진 자가 변형할 수 있는 모든 방법을 포함하는 것이 가능하다.The organic solvent extraction method and the method of separating absolute from the extract may include all methods that can be modified by a person of ordinary skill in the art.
또한, 상기 오일은 휘발성 유기 화합물을 포함할 수 있다.In addition, the oil may contain a volatile organic compound.
또한, 상기 휘발성 유기화합물은 7,11,15-trimethyl-3-methylidenehexadec-1-ene, α-linolenic acid, ethyl linoleate로 이루어진 그룹에서 선택되는 어느 하나 이상을 포함할 수 있으나, 이에 제한되는 것은 아니다.In addition, the volatile organic compound may include at least one selected from the group consisting of 7,11,15-trimethyl-3-methylidenehexadec-1-ene, α-linolenic acid, and ethyl linoleate, but is not limited thereto. .
또한, 이들 성분의 함량은 구체적으로 α-linolenic acid 59.14 중량%, ethyl linoleate 29.73 중량%, 7,11,15-trimethyl-3-methylidenehexadec-1-ene 11.13 중량%의 범위로 앱솔루트 내에 포함될 수 있다.In addition, the content of these components is specifically α-linolenic acid 59.14% by weight, ethyl linoleate 29.73% by weight, 7,11,15-trimethyl-3-methylidenehexadec-1-ene 11.13% by weight It can be included in the absolute range.
이에 따라 본 발명에 따른 골담초 꽃에서 추출한 오일(앱솔루트)을 유효성분으로 포함하는 피부 재생, 상처 치유 또는 주름개선용 조성물로 사용될 수 있으며, 약학적으로 유효한 양의 골담초 꽃에서 추출한 오일(앱솔루트)을 단독으로 포함하거나 하나 이상의 약학적으로 허용되는 담체, 부형제 또는 희석제를 포함할 수 있다.Accordingly, it can be used as a composition for skin regeneration, wound healing, or wrinkle improvement comprising the oil (absolute) extracted from the flower according to the present invention (absolute). It may contain alone or one or more pharmaceutically acceptable carriers, excipients or diluents.
또한, 상기 담체, 부형제 및 희석제의 예로는, 락토즈, 덱스트로즈, 수크로즈, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 폴리비닐피롤리돈, 물, 메틸하이드록시벤조에이트, 프로필하이드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 들 수 있다. 또한, 충진제, 항응집제, 윤활제, 습윤제, 향료, 유화제 및 방부제 등을 추가로 포함할 수 있다.In addition, examples of the carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. In addition, fillers, anti-agglomeration agents, lubricants, wetting agents, fragrances, emulsifiers and preservatives may be further included.
또한, 상기 오일의 농도가 0.01μg/mL 내지 100μg/mL일 수 있으며, 바람직하게는 0.1μg/mL 내지 100μg/mL 이다.In addition, the concentration of the oil may be 0.01 μg/mL to 100 μg/mL, preferably 0.1 μg/mL to 100 μg/mL.
본 발명은 골담초 오일을 유효성분으로 포함하는 피부 재생, 상처 치유 또는 주름개선용 화장료, 피부외용제 또는 건강기능식품 조성물을 제공한다.The present invention provides a cosmetic, external preparation for skin, or health functional food composition for skin regeneration, wound healing, or wrinkle improvement, comprising Goldamcho oil as an active ingredient.
또한, 상기 조성물은 피부외용연고, 크림, 유연화장수, 영양화장수, 팩, 에센스, 헤어토닉, 샴푸, 린스, 헤어 컨디셔너, 헤어 트리트먼트, 젤, 스킨로션, 스킨소프너, 스킨토너, 아스트린젠트, 로션, 밀크 로션, 모이스처 로션, 영양로션, 마사지 크림, 영양크림, 모이스처 크림, 핸드 크림, 파운데이션, 영양에센스, 선스크린, 비누, 클렌징폼, 클렌징로션, 클렌징크림, 바디 로션 및 바디 클렌저로 이루어진 그룹에서 선택되는 어느 하나의 제형으로 제조된 것일 수 있다.In addition, the composition is an external skin ointment, cream, softening lotion, nourishing lotion, pack, essence, hair tonic, shampoo, conditioner, hair conditioner, hair treatment, gel, skin lotion, skin softener, skin toner, astringent, lotion, Milk Lotion, Moisture Lotion, Nutrient Lotion, Massage Cream, Nutrient Cream, Moisture Cream, Hand Cream, Foundation, Nutrient Essence, Sunscreen, Soap, Cleansing Foam, Cleansing Lotion, Cleansing Cream, Body Lotion and Body Cleanser It may be prepared in any one formulation.
또한, 본 발명에 따른 화장료 조성물에 있어서, 상기 화장료 조성물은 상기 골담초 오일을 상기 조성물 총 중량에 대하여 0.0001~90 중량부의 양으로 함유할 수 있으나, 바람직한 효과를 제공할 수 있는 유효량을 포함한다면 이에 제한되지 않는다.In addition, in the cosmetic composition according to the present invention, the cosmetic composition may contain the bone marrow oil in an amount of 0.0001 to 90 parts by weight based on the total weight of the composition, but if it contains an effective amount that can provide a desirable effect, it is limited thereto doesn't happen
또한, 본 발명의 조성물은 포유동물에 투여된 후 활성 성분의 신속, 지속 또는 지연된 방출을 제공할 수 있도록 당업계에 공지된 방법을 사용하여 제형화될 수 있으며, 경구 또는 비경구 투여를 위한 다양한 형태로 제형화 될 수 있다. 제형은 분말, 과립, 정제, 에멀젼, 시럽, 에어로졸, 연질 또는 경질 젤라틴 캡슐, 멸균 주사용액, 멸균 분말, 연고, 로션, 크림, 고약, 페이스트, 겔, 패치, 분무제의 형태일 수 있다.In addition, the compositions of the present invention may be formulated using methods known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal, and may be formulated in a variety of ways for oral or parenteral administration. It can be formulated in the form Formulations may be in the form of powders, granules, tablets, emulsions, syrups, aerosols, soft or hard gelatin capsules, sterile injectable solutions, sterile powders, ointments, lotions, creams, salves, pastes, gels, patches, sprays.
또한, 상기 다양한 제형의 조성물에 있어서, 상기한 필수 구성성분 이외에도 당해 기술 분야의 통상의 지식을 가진 자의 기술 수준에 비추어 특별한 제한 없이 외용제의 제형 또는 사용 목적 등에 따라 다양한 물질들을 선정하여 배합할 수 있다.In addition, in the composition of the various formulations, in addition to the above-mentioned essential components, in view of the technical level of those skilled in the art, various substances can be selected and formulated according to the formulation or purpose of use of the external preparation without special limitation. .
나아가 본 발명에 따른 조성물은 상기 기술한 바와 같이 약학적 조성물로 사용할 수 있을 뿐만 아니라, 건강기능식품으로도 사용할 수 있다. 예컨대, 식품의 주원료, 부원료, 식품 첨가제, 기능성 식품 또는 음료로 용이하게 활용할 수 있다.Furthermore, the composition according to the present invention can be used not only as a pharmaceutical composition as described above, but also as a health functional food. For example, it can be easily utilized as a main raw material of food, an auxiliary raw material, a food additive, a functional food or a beverage.
본 발명에서 사용되는 용어 “식품”이란, 영양소를 한 가지 또는 그 이상 함유하고 있는 천연물 또는 가공품을 의미하며, 바람직하게는 어느 정도의 가공 공정을 거쳐 직접 먹을 수 있는 상태가 된 것을 의미하며, 통상적인 의미로서, 식품, 식품첨가제, 기능성식품 및 음료를 모두 포함하는 것을 말한다.The term “food” used in the present invention means a natural product or processed product containing one or more nutrients, and preferably means a state that can be eaten directly through a certain processing process, and is usually In the meaning of phosphorus, it refers to all foods, food additives, functional foods and beverages.
상기 식품 조성물을 첨가할 수 있는 식품으로는 예를 들어, 각종 식품류, 음료, 껌, 차, 비타민 복합체, 기능성 식품 등이 있다. 추가로, 특수영양식품(예, 조제유류, 영, 유아식 등), 식육가공품, 어육제품, 두부류, 묵류, 면류(예, 라면류, 국수류 등), 빵류, 건강보조식품, 조미식품(예, 간장, 된장, 고추장, 혼합장 등), 소스류, 과자류(예, 스낵류), 캔디류, 초코렛류, 껌류, 아이스크림류, 유가공품(예, 발효유, 치즈 등), 기타 가공식품, 김치, 절임식품(각종 김치류, 장아찌 등), 음료(예, 과실음료, 채소류 음료, 두유류, 발효음료류 등), 천연조미료(예, 라면스프 등)를 포함하나 이에 한정되지 않는다. 상기 식품, 음료 또는 식품첨가제는 통상의 제조방법으로 제조될 수 있다.Foods to which the food composition can be added include, for example, various foods, beverages, gum, tea, vitamin complexes, functional foods, and the like. In addition, special nutritional foods (eg, formula milk, young, baby food, etc.), processed meat products, fish meat products, tofu, jelly, noodles (eg, ramen, noodles, etc.), breads, health supplements, seasoned foods (eg, soy sauce) , soybean paste, red pepper paste, mixed paste, etc.), sauces, sweets (eg snacks), candy, chocolate, gum, ice cream, dairy products (eg fermented milk, cheese, etc.), other processed foods, kimchi, pickled foods (various kimchi) , pickles, etc.), beverages (eg, fruit beverages, vegetable beverages, soy milk, fermented beverages, etc.), and natural seasonings (eg, ramen soup, etc.), but are not limited thereto. The food, beverage or food additive may be prepared by a conventional manufacturing method.
본 발명에서 사용되는 용어 “기능성 식품”또는 “건강기능성 식품”이란 식품에 물리적, 생화학적, 생물 공학적 수법 등을 이용하여 해당 식품의 기능을 특정 목적에 작용, 발현하도록 부가가치를 부여한 식품군이나 식품 조성이 갖는 생체방어리듬조절, 질병방지와 회복 등에 관한 체내조절기능을 생체에 대하여 충분히 발현하도록 설계하여 가공한 식품을 의미하며, 구체적으로는 건강 기능성 식품일 수 있다. 상기 기능성 식품에는 식품학적으로 허용 가능한 식품 보조 첨가제를 포함할 수 있으며, 기능성 식품의 제조에 통상적으로 사용되는 적절한 담체, 부형제 및 희석제를 더욱 포함할 수 있다.The term “functional food” or “health functional food” as used in the present invention refers to a food group or food composition in which added values are added to the food to act and express the function of the food for a specific purpose by using physical, biochemical, or biological engineering methods, etc. It refers to a food that is designed and processed to sufficiently express the body control functions related to biological defense rhythm control, disease prevention and recovery, etc. to the living body, and specifically, it may be a health functional food. The functional food may include a food supplementary additive that is pharmaceutically acceptable, and may further include an appropriate carrier, excipient and diluent commonly used in the manufacture of functional food.
또한, 본 발명에 따른 건강기능식품 조성물에 있어서, 상기 건강기능식품 조성물은 정제, 과립제, 분말제, 드링크제와 같은 액체, 캐러멜, 겔, 바 등으로 제형화될 수 있다. 각 제형의 건강기능식품 조성물은 골담초 오일 이외에 해당 분야에서 통상적으로 사용되는 성분들을 제형 또는 사용 목적에 따라 당업자가 어려움 없이 적의 선정하여 배합할 수 있으며, 다른 원료와 동시에 적용할 경우 상승효과가 일어날 수 있다.In addition, in the health functional food composition according to the present invention, the health functional food composition may be formulated in tablets, granules, powders, liquids such as drinks, caramel, gels, bars, and the like. The health functional food composition of each formulation can be formulated without difficulty by those skilled in the art without difficulty depending on the formulation or purpose of use of ingredients commonly used in the field in addition to Goldamcho oil, and when applied simultaneously with other ingredients, a synergistic effect may occur. have.
상기 외에 본 발명의 건강기능식품 조성물은 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 증진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율 또한 당업자에 의해 적절히 선택될 수 있다.In addition to the above, the health functional food composition of the present invention contains various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavoring agents, colorants and enhancers (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. These components may be used independently or in combination. The proportion of these additives may also be appropriately selected by those skilled in the art.
본 발명의 상기 조성물은 항산화, 항노화, 항고혈압, 항관절염, 미백, 피부 보습용 조성물을 포함한다.The composition of the present invention includes a composition for antioxidant, anti-aging, anti-hypertension, anti-arthritis, whitening, and skin moisturizing.
본 발명의 상기 조성물은 본 발명의 유효 활성을 저해하지 않는 범위에서 항산화, 피부 노화 방지 및/또는 피부 보습의 효과를 위한 공지의 천연물 추출물이나 기타 성분을 추가로 포함시켜 이용할 수 있다.The composition of the present invention can be used by additionally including a known natural extract or other ingredients for antioxidant, skin aging prevention and/or skin moisturizing effects in a range that does not inhibit the effective activity of the present invention.
본 발명에서 사용되는 용어 “에센셜 오일”은 방향유(식물의 잎, 줄기, 열매, 꽃, 뿌리 따위에서 채취한 향기로운 휘발성의 물질)을 통칭하는 용어로서, 본 발명은 골담초의 꽃으로부터 추출된 오일을 유효성분으로 포함한다. 이때, 본 발명의 골담초 꽃 오일은 건체 또는 생체 꽃으로부터 추출된 것이 바람직하고, 그 중에서도 건조되지 않은 골담초의 꽃망울로부터 추출된 것이 더욱 많은 유용성분을 얻을 수 있어서 더욱 바람직하다.The term “essential oil” used in the present invention is a generic term for aromatic oils (fragrant and volatile substances collected from leaves, stems, fruits, flowers, roots, etc.) included as an active ingredient. In this case, it is preferable that the flower oil of the sagebrush of the present invention is extracted from dried or living flowers, and among them, the extract from the flower buds of the flower buds that have not been dried is more preferable because more useful components can be obtained.
본 발명에서 사용되는 용어 “앱솔루트”는 상기 유기용매 추출법에 의해 추출된 콘크리트(concrete)를 알코올로 추출하여 불용성 왁스(wax)를 제거한 것을 뜻한다. 즉, 상기 유기용매 추출법에 의해 유용성분을 추출하는 경우 수증기증류법에 의하는 것보다 소량의 유용성분을 더욱 많이 추출할 수 있는 장점이 있지만, 상기 유기용매 추출법에 의해 추출된 콘크리트에는 휘발성분이 아닌 왁스성분까지 함께 포함될 수 있는 단점을 가지는바, 본 발명은 상기 콘크리트로부터 불용성인 왁스를 제거한 앱솔루트만을 이용하는 것이 더욱 바람직하다.The term “absolute” used in the present invention means that insoluble wax is removed by extracting concrete extracted by the organic solvent extraction method with alcohol. That is, when useful components are extracted by the organic solvent extraction method, there is an advantage that a small amount of useful components can be extracted more than by the steam distillation method. However, in the concrete extracted by the organic solvent extraction method, wax is not a volatile component. Since it has a disadvantage that even components may be included together, it is more preferable to use only absolute, in which insoluble wax is removed from the concrete, in the present invention.
본 발명에서 사용되는 용어 “캐리어 가스”는 기체 크로마토그래피에서 쓰는 기체로 수소, 헬륨, 질소, 아르곤을 쓴다.The term “carrier gas” used in the present invention is a gas used in gas chromatography, and hydrogen, helium, nitrogen, and argon are used.
본 발명에서 설명되는 “약학적으로 유효한 양”이란 상기 생리활성성분이 동물 또는 사람에게 투여되어 목적하는 생리학적 또는 약리학적 활성을 나타내기에 충분한 양을 말한다. 그러나 상기 약학적으로 유효한 양은 증상의 정도, 환자의 연령, 체중, 건강상태, 성별, 투여 경로 및 치료기간 등에 따라 적절히 변화될 수 있다.As used herein, the term "pharmaceutically effective amount" refers to an amount sufficient to exhibit the desired physiological or pharmacological activity when the physiologically active ingredient is administered to an animal or human. However, the pharmaceutically effective amount may be appropriately changed according to the severity of symptoms, the age, weight, health status, sex, administration route, and treatment period of the patient.
본 발명에서 설명되는 “약학적으로 허용되는”이란 생리학적으로 허용되고 인간에게 투여될 때, 통상적으로 위장 장애, 현기증과 같은 알레르기 반응 또는 이와 유사한 반응을 일으키지 않는 것을 말한다. "Pharmaceutically acceptable" as used in the present invention means that it is physiologically acceptable and does not normally cause gastrointestinal disorders, allergic reactions such as dizziness or similar reactions when administered to humans.
본 발명에서 달리 명시하지 않는 한, 모든 화학 물질은 Sigma-Aldrich Corporation, Inc.(미국 미주리 주 세인트루이스)에서 구입하였다. Unless otherwise specified herein, all chemicals were purchased from Sigma-Aldrich Corporation, Inc. (St. Louis, MO, USA).
이하, 본 발명의 실시예를 첨부된 도면을 참고하여 보다 상세하게 설명하도록 한다. 그러나, 하기의 실시예는 본 발명의 내용을 구체화하기 위한 것일 뿐, 이에 의해 본 발명이 한정되는 것은 아닐 것이다.Hereinafter, embodiments of the present invention will be described in more detail with reference to the accompanying drawings. However, the following examples are only intended to embody the contents of the present invention, and the present invention will not be limited thereby.
<실시예 1> 골담초 꽃 앱솔루트의 제조<Example 1> Preparation of Golddammcho Flower Absolute
도 1에 도시된 공정을 참조하여 골담초 꽃으로부터 앱솔루트를 제조하였다. 골담초 꽃은 피기 직전의 꽃망울 전체를 채취(전라남도 해남, 하나로 농장)하여 세척 후, 건조하지 않은 상태에서 유기용매 추출법을 이용하여 추출하였다. 구체적으로, 골담초 꽃을 헥산(n-hexane)으로 추출하였고, 이렇게 얻은 침전물을 회전식증발기(rotary evaporation)에서 증발시켜서 농축시킴으로서 고형의 콘크리트(concrete)를 획득하였다. 그 다음, 상기 콘크리트를 에틸알코올(ethyl alcohol)을 넣어서 용해시킨 후, 진공 냉각 필터(vacuum cooling filtration)를 거쳐서 불용성 왁스를 제거하였으며, 다시 회전식 증발기(rotary evaporation)를 이용하여 용매를 제거하여 골담초 꽃 앱솔루트를 제조하였다. 그런 다음 상기 제조한 골담초 꽃 앱솔루트는 4℃의 저온고에서 보관하였다.With reference to the process shown in FIG. 1 , an absolute was prepared from the flowers of Golddammosa. Golddamcho flowers remove the entire bud just before blooming. It was collected (Haenam, Jeollanam-do, Hanaro Farm), washed, and extracted using an organic solvent extraction method in a non-dry state. In detail, the flower of dandelion flower was extracted with n-hexane, and the precipitate thus obtained was evaporated by rotary evaporation and concentrated to obtain solid concrete. Then, after dissolving the concrete by adding ethyl alcohol, the insoluble wax was removed through vacuum cooling filtration, and the solvent was removed using a rotary evaporation to remove the flower Absolute was prepared. Then, the prepared gold leaf flower absolute was stored at a low temperature of 4 ℃.
<실시예 2> 골담초 꽃 앱솔루트의 성분 분석<Example 2> Analysis of the components of Golddamel flower absolute
골담초 꽃 앱솔루트의 성분 분석은 KBSI(Korea Basic Science Institute, Seoul Center)에 의뢰하였다. 가스 크로마토그래피/질량분석계(6890N GC/5975i MS, Agilent, USA)를 사용하였고, GC/MS(Gas chromatography-mass spectrometry)의 컬럼(column)은 DB5-MS(30m×50μm, 0.25μm)를 사용하였으며, 캐리어 가스(carrier gas)는 헬륨을 사용하여 1mL/분으로 흐르게 하여 분석하였다. 오븐의 온도는 40℃-2℃/분-230℃-5℃/분-300℃(5분)으로 유지하였으며, 주입구(injection port)의 온도는 280℃에서 분석하였다. 또한 인터페이스(Interface)의 온도는 300℃, 이온소스(ion source)의 온도는 230℃, 분석관(analyzer)의 온도는 150℃ 에서 분석하였으며, 질량비정(Mass range)는 40 내지 800 m/z에서 분석하였다. 골담초 앱솔루트 성분의 확인은 휘발성분의 지체시간(retention indices, RI)과 표준물질의 지체시간과 일치하거나 휘발물질의 질량 스펙트럼과 Wiley 7Nist 05 library의 질량 스펙트럼을 비교하여 물질을 확인하였다. RI는 C6 내지 C24(n-alkane)를 이용하여 시료와 동일 조건에서 결정하였다. 그 결과, 총 16개의 성분으로 구성되어 있음을 확인할 수 있었으며, 주요 성분으로 α-linolenic acid, ethyl linoleate, 7,11,15-trimethyl-3-methylidenehexadec-1-ene 순으로 높은 함량을 나타내었다(도 2).The analysis of the components of Golddammcho Flower Absolute was commissioned by KBSI (Korea Basic Science Institute, Seoul Center). A gas chromatography/mass spectrometer (6890N GC/5975i MS, Agilent, USA) was used, and the column of GC/MS (Gas chromatography-mass spectrometry) was DB5-MS (30m×50μm, 0.25μm). and the carrier gas was analyzed by flowing at 1 mL/min using helium. The temperature of the oven was maintained at 40°C-2°C/min-230°C-5°C/min-300°C (5 minutes), and the temperature of the injection port was analyzed at 280°C. In addition, the temperature of the interface was analyzed at 300 °C, the temperature of the ion source at 230 °C, and the temperature of the analyzer at 150 °C, and the mass range was 40 to 800 m/z. analyzed. For the identification of Golddammcho absolute components, the retention indices (RI) of volatiles and retention times of standard substances were matched, or substances were identified by comparing the mass spectrum of volatiles with the mass spectrum of Wiley 7Nist 05 library. RI was determined under the same conditions as the sample using C 6 to C 24 (n-alkane). As a result, it was confirmed that it was composed of a total of 16 components, and the main components were α-linolenic acid, ethyl linoleate, and 7,11,15-trimethyl-3-methylidenehexadec-1-ene in the order of high content ( 2).
<실시예 3> 골담초 꽃 앱솔루트가 표피 각질형성세포에 미치는 독성조사<Example 3> Toxicity investigation of keratinocytes of Goldamcho flower Absolute on epidermal keratinocytes
사람 피부 유래 표피 각질형성세포(HaCat)를 10%의 우태아 혈청(fetal bovine serum)이 포함된 DMEM(Dulbeco’s Modified Eagle’s Medium)배지를 이용하여 배양하고 본 발명에 활용하였다. 표피 각질형성세포를 96 well microtiter plate에 각 well 당 세포수 5X103cells/100μL/well 이 되도록 주입하여 37℃, 5% CO2 배양기에서 부착시켰다. 골담초 꽃 앱솔루트 0.1 내지 100μg/mL 농도로 각 well에 100 μL씩 처리하여 배양하고, 대조군은 골담초 꽃 앱솔루트를 미처리하여 배양하였다. 이후, Ez-cytox와 DMEM(Dulbeco’s Modified Eagle’s Medium)를 1:2로 혼합하여 각 well 당 30 μL씩 처리한 후 배양기에서 30분간 반응시켰다. ELISA 플레이트 리더(Synerge2, BioTek, USA)를 이용하여 450 nm에서 흡광도 측정 후, 백분율로 환산하여 골담초 앱솔루트의 피부 표피각질형성세포에 대한 독성 결과를 확보하였다. 그 결과, 도 3에 나타난 바와 같이, 골담초 앱솔루트의 0.1 내지 100μg/mL의 농도에서는 유의한 세포독성이 관찰되지 않았다. 다만, 150μg/mL의 농도에서 세포 독성을 가지는 것으로 확인되었다(미도시). 따라서 본 발명에 따른 골담초 앱솔루트의 농도는 100μg/ml 이하의 농도가 바람직하다.Human skin-derived epidermal keratinocytes (HaCat) were cultured using DMEM (Dulbeco's Modified Eagle's Medium) medium containing 10% fetal bovine serum and used in the present invention. Epidermal keratinocytes were injected into a 96-well microtiter plate so that the number of cells per well was 5X10 3 cells/100 μL/well, and adhered in a 37° C., 5
<실시예 4> 골담초 꽃 앱솔루트가 표피 각질형성세포의 증식에 미치는 효과<Example 4> Effect of Goldamcho Flower Absolute on the proliferation of epidermal keratinocytes
사람 피부 유래 표피 각질형성세포(keratinocytes, HaCat)를 10%의 우태아 혈청(fetal bovine serum)이 포함된 DMEM(Dulbeco’s Modified Eagle’s Medium)배지를 이용하여 배양하고 본 발명에 활용하였다. 표피 각질형성세포를 96 well microtiter plate에 각 well 당 세포수 3 X 103 cells/100μL/well이 되도록 주입하여 37℃, 5% CO2 배양기에서 부착시켰다. 골담초 꽃 앱솔루트 0.1 내지 100μg/mL 농도로 각 well에 100μL씩 처리하여 배양한 후, Ez-cytox와 DMEM(Dulbeco’s Modified Eagle’s Medium)을 1:2로 혼합하여 각 well 당 30μL씩 처리한 후 배양기에서 30분간 반응시켰다. ELISA 플레이트 리더(Synerge2, BioTek, USA)를 이용하여 450nm에서 흡광도 측정 후, 백분율로 환산하였다. 그 결과, 본 발명에 따른 골담초 꽃 앱솔루트는 표피 각질형성세포의 증식을 농도 의존적으로 유도하는 것으로 나타났다(도 4). 여기서, 음성 대조군은 골담초 꽃 앱솔루트를 이용하지 않은 것이고, 양성 대조군은 종래에 상처치유 및/또는 피부 재생에 활용되고 있는 표피성장인자(epidermal growth factor, EGF 50ng/mL)를 이용한 것이다. 즉, 표피성장인자(50ng/mL)에 의한 표피 각질형성세포의 증식은 169.67±6.40% 유도가 되었으며, 양성 대조군과 비교하여 골담초 꽃 앱솔루트 10μg/mL 농도에서 121.00±9.81%의 증식 유도율을 나타내었고, 50μg/mL의 농도에서 160.56±12.36%의 최대 증식 유도율을 나타내었다. 이는 골담초 오일이 각질형성세포의 증식을 효과적으로 유도하여 피부를 재생시키거나 상처를 치유하는 효과를 가진다는 것을 나타낸다.Human skin-derived epidermal keratinocytes (HaCat) were cultured using DMEM (Dulbeco's Modified Eagle's Medium) medium containing 10% fetal bovine serum and used in the present invention. Epidermal keratinocytes were injected into a 96-well microtiter plate so that the number of cells per well was 3
<실시예 5> 골담초 꽃 앱솔루트가 표피 각질형성세포 이동에 미치는 효과<Example 5> Effect of Goldamcho flower absolute on epidermal keratinocyte migration
사람 피부 유래 표피 각질형성세포(HaCat)를 10%의 우태아 혈청(fetal bovine serum)이 포함된 DMEM(Dulbeco’s Modified Eagle’s Medium)배지를 이용하여 배양하였다. Boyden chamber assays를 사용하여 표피 각질형성세포의 이동률을 조사하였으며, 먼저 48 well Boyden microchemotaxis chamber의 lower chamber에 골담초 꽃 앱솔루트를 각각 0.1 내지 100μg/mL 의 농도를 처리하고, type I 콜라겐이 코팅된 8μm pore의 폴리카보네이트 막(polycarbonate membrane)을 올려 주었다. 이후, upper chamber에 표피 각질형성세포를 well당 1×106 cell/50μL씩 처리하였다. 48 well Boyden microchemotaxis chamber를 37℃ 배양기에서 3시간 30분 배양시킨 후, 폴리카보네이트 막(polycarbonate membrane)을 Diff-Quick solution(Dade Behring #B4132-11A)으로 염색 및 고정하였다. 그 다음, 폴리카보네이트 막(polycarbonate membrane)을 통과하여 이동한 세포의 수를 세었다. 그 결과, 골담초 꽃 앱솔루트는 표피 각질형성세포의 이동을 농도 의존적으로 유도하였으며, 특히, 골담초 꽃 앱솔루트 10μg/mL의 농도에서 260.91±19.50%, 50μg/mL의 농도에서 245.46±38.46%의 이동률을 나타내었다(도 5). 음성 대조군은 골담초 꽃 앱솔루트를 이용하지 않은 것이고, 양성 대조군은 종래에 상처치유 및/또는 피부 재생에 활용되는 있는 표피성장인자(epidermal growth factor, EGF 50ng/mL)를 이용한 것이다. 이러한 결과는 골담초 꽃 오일이 각질형성세포의 이동을 효과적으로 유도하여 피부를 재생시키거나 상처를 치유하는 효과를 가진다는 것을 나타낸다.Human skin-derived epidermal keratinocytes (HaCat) were cultured using DMEM (Dulbeco's Modified Eagle's Medium) medium containing 10% fetal bovine serum. The migration rate of epidermal keratinocytes was investigated using Boyden chamber assays, and first, each concentration of 0.1 to 100 μg/mL of alfalfa flower absolute was treated in the lower chamber of a 48 well Boyden microchemotaxis chamber, and 8 μm pores coated with type I collagen. of a polycarbonate membrane (polycarbonate membrane) was placed on it. Thereafter, epidermal keratinocytes were treated with 1×10 6 cells/50 μL per well in the upper chamber. After incubating a 48-well Boyden microchemotaxis chamber in a 37°C incubator for 3 hours and 30 minutes, a polycarbonate membrane was stained and fixed with Diff-Quick solution (Dade Behring #B4132-11A). Then, the number of cells that migrated through the polycarbonate membrane was counted. As a result, the bone marrow flower absolute induced the migration of epidermal keratinocytes in a concentration-dependent manner, and in particular, it showed a migration rate of 260.91±19.50% at a concentration of 10 μg/mL and 245.46±38.46% at a concentration of 50 μg/mL. was (Fig. 5). Negative control group did not use the flower Absolute of Gold dammosa, and positive control group used epidermal growth factor (EGF 50ng/mL) which is conventionally used for wound healing and/or skin regeneration. These results indicate that the flower oil of dandelion flower effectively induces the migration of keratinocytes and has the effect of regenerating the skin or healing wounds.
<실시예 6> 골담초 꽃 앱솔루트가 표피 각질형성세포 콜라겐 발아(sprouting)의 자연성장(out growth)에 미치는 영향<Example 6> Effect of Goldamcho Flower Absolute on the out growth of epidermal keratinocyte collagen sprouting
Type I 콜라겐과 표피 각질형성세포를 2.5×107 cells/mL이 되도록 섞은 후, NaOH(1N)용액을 처리하여 중화한 후, 24 well plate에 5μL씩 스포팅(spotting)하고, 10분동안 상온에서 건조하여 인공피부조직을 형성하였다. 샘플을 처리하고, 48시간 동안 37℃, 5% CO2 배양기에서 배양하였다. 배양액을 제거하고, Diff-Quik solution(Dade Behring #B4132-11A)을 이용하여 세포와 조직을 고정 및 염색하였으며, 염색된 세포와 조직을 촬영하여 결과를 확보하였다. 도 5는 본 발명의 실시예 1에 의해 추출된 골담초 꽃 앱솔루트의 표피 각질형성세포 콜라겐 발아(sprouting)의 자연성장(out growth)에 미치는 영향을 나타낸 결과이며, 음성 대조군은 골담초 꽃 앱솔루트를 이용하지 않은 것이고, 양성 대조군은 종래의 상처치유 및/또는 피부 재생에 활용되는 있는 표피성장인자(epidermal growth factor, EGF 50ng/mL)를 이용한 것이다. 그 결과, 골담초 꽃 앱솔루트는 표피 각질형성세포 콜라겐 발아(sprouting)의 자연성장(out growth)를 농도 의존적으로 유도하는 것으로 나타났다(도 6A). 그리고 골담초 꽃 앱솔루트 1μg/mL 농도에서 158.69±4.17%가 유도되었으며, 50μg/mL 농도에서 297.52±12.63%가 유도되었다(도 6B). 이러한 결과는 양성 대조군과 비교하여 골담초 오일이 각질형성세포 콜라겐 발아(sprouting)의 자연성장(out growth)를 효과적으로 유도하여 주름을 개선하는 효과를 가지는 것을 나타낸다.Type I collagen and epidermal keratinocytes 2.5×10 7 After mixing to make cells/mL, neutralized by treatment with NaOH (1N) solution, spotted by 5 μL in a 24-well plate, and dried at room temperature for 10 minutes to form artificial skin tissue. Samples were processed and incubated in a 37° C., 5% CO 2 incubator for 48 hours. The culture medium was removed, cells and tissues were fixed and stained using Diff-Quik solution (Dade Behring #B4132-11A), and the results were obtained by photographing the stained cells and tissues. 5 is a result showing the effect on the natural growth (out growth) of epidermal keratinocyte collagen spouting of the bone marrow flower absolute extracted according to Example 1 of the present invention, and the negative control group does not use the bone marrow flower absolute and the positive control group uses epidermal growth factor (EGF 50ng/mL), which is used for conventional wound healing and/or skin regeneration. As a result, it was shown that the flower Absolute of Goldamchoysis induces the out growth of epidermal keratinocyte collagen sprouting in a concentration-dependent manner (FIG. 6A). And 158.69±4.17% was induced at a concentration of 1 μg/mL of Goldamcho flower absolute, and 297.52±12.63% was induced at a concentration of 50 μg/mL ( FIG. 6B ). These results indicate that bone marrow oil has an effect of improving wrinkles by effectively inducing out growth of keratinocyte collagen sprouting as compared with the positive control.
<실시예 7> 골담초 꽃 앱솔루트가 표피 각질형성세포의 type I 또는 IV 콜라겐 합성과 분비에 미치는 영향<Example 7> Effect of Goldenflower Absolute on the synthesis and secretion of type I or IV collagen in epidermal keratinocytes
96 well microtiter plate에 단클론 항-콜라겐(monoclonal anti-collagen) I 또는 IV Ab(2 μg/well/PBS)를 주입 후 12시간 동안 코팅하였다. 차단 완충액(Blocking buffer)로 1시간 동안 블로킹(blocking) 후, 각각의 샘플을 well당 100μL씩 처리하여 90분 동안 반응시켰다. 샘플은 대조군으로 표피 각질형성세포의 배양액(DMEM(Dulbeco’s Modified Eagle’s Media)배지와 CM(conditioned medium)배지)과 세포 용해물(Lysate)을 처리하였고, 실험군으로 골담초 꽃 앱솔루트가 각각 1μg/mL, 50μg/mL, 100μg/mL 농도로 처리된 표피 각질형성세포의 배양액(DMEM(Dulbeco’s Modified Eagle’s Media)배지와 CM(conditioned medium)배지)과 세포 용해물(Lysate)을 처리하였다. 그 후, 다클론 항-콜라겐(polyclonal anti-collagen) I 또는 IV Ab를 각 well에 처리하여 90분 동안 반응시킨 다음, 스트렙트아비딘-POD(streptavidin-POD)를 각 well에 처리하여 60분 동안 반응시켰다. ELISA pico chemiluminescent substrate를 각 well 당 100μL씩 주입하여 10분 동안 반응시킨 후, ELISA 플레이트 리더를 이용하여 루미네선스(luminescence) 정도를 측정하였고, 골담초 꽃 앱솔루트의 표피 각질형성세포의 type I 또는 IV 콜라겐 합성과 분비에 대한 결과를 확보하였다. 그 결과, 골담초 꽃 앱솔루트는 1μg/mL, 50μg/mL, 100μg/mL 농도에서 모두 표피 각질형성세포의 type I 과 Ⅳ의 콜라겐 합성과 분비를 유의하게 촉진시켰다. 특히, 골담초 꽃 앱솔루트가 50μg/mL 농도로 처리된 표피 각질형성세포의 배양액(DMEM(Dulbeco’s Modified Eagle’s Media)배지와 CM(conditioned medium)배지)의 콜라겐 type I과 Ⅳ의 합성과 분비가 각각 245.74±35.82%, 174.74±2.73% 유도되는 것으로 나타났다(도 7A, B). 이러한 결과는 골담초 오일이 콜라겐의 합성 또는 분비를 유도하여 주름개선 효과를 가진다는 것을 나타낸다.A 96-well microtiter plate was coated with monoclonal anti-collagen I or IV Ab (2 μg/well/PBS) for 12 hours after injection. After blocking for 1 hour with a blocking buffer, each sample was treated with 100 μL per well and reacted for 90 minutes. The samples were treated with epidermal keratinocyte culture medium (DMEM (Dulbeco's Modified Eagle's Media) medium and CM (conditioned medium) medium) and cell lysate as a control group, and as an experimental group, 1 μg/mL and 50 μg of B. A culture medium of epidermal keratinocytes (DMEM (Dulbeco's Modified Eagle's Media) and CM (conditioned medium)) and a cell lysate were treated at a concentration of /mL and 100μg/mL. Then, polyclonal anti-collagen (polyclonal anti-collagen) I or IV Ab was treated in each well and reacted for 90 minutes, and then streptavidin-POD (streptavidin-POD) was treated in each well for 60 minutes. reacted. After injecting 100 μL of ELISA pico chemiluminescent substrate into each well and reacting for 10 minutes, the degree of luminescence was measured using an ELISA plate reader. Results for synthesis and secretion were obtained. As a result, Blossom Root Flower Absolute significantly stimulated the synthesis and secretion of type I and IV collagen in epidermal keratinocytes at concentrations of 1 μg/mL, 50 μg/mL, and 100 μg/mL. In particular, the synthesis and secretion of collagen types I and IV in the culture medium of epidermal keratinocytes (DMEM (Dulbeco's Modified Eagle's Media) and CM (conditioned medium) medium) treated at a concentration of 50 μg/mL 35.82%, 174.74±2.73% were induced ( FIGS. 7A, B). These results indicate that bone marrow oil has a wrinkle-improving effect by inducing the synthesis or secretion of collagen.
이상에서 살펴본 바와 같이, 본 발명의 구체적인 실시예를 상세하게 설명되었으나, 본 발명의 사상을 이해하는 당업자는 동일한 사상의 범위 내에서 다른 구성요소를 추가, 변경, 삭제 등을 통하여, 퇴보적인 다른 발명이나 본 발명 사상의 범위 내에 포함되는 다른 실시예를 용이하게 제안할 수 있을 것이다. 그러므로 이상에서 기술한 실시예들은 모든 면에서 예시적인 것이며 한정적이 아닌 것으로 이해해야만 한다. 본 발명의 범위는 상술한 상세한 설명보다는 후술하는 특허청구의 범위에 의하여 나타내어지며, 특허청구의 범위의 의미 및 범위 그리고 그 균등 개념으로부터 도출되는 모든 변경 또는 변형된 형태가 본 발명의 범위에 포함되는 것으로 해석되어야 한다.As described above, although specific embodiments of the present invention have been described in detail, those skilled in the art who understand the spirit of the present invention may add, change, delete, etc. other components within the scope of the same spirit, and other degenerate inventions However, other embodiments included within the scope of the present invention may be easily proposed. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention is indicated by the following claims rather than the above detailed description, and all changes or modifications derived from the meaning and scope of the claims and their equivalents are included in the scope of the present invention. should be interpreted as
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| KR20140006139A (en) * | 2012-06-26 | 2014-01-16 | 주식회사 래디안 | Cosmetic composition with anti-uv and anti-wrinkle effect from caragana sinica rehder |
| KR20160135037A (en) * | 2015-05-15 | 2016-11-24 | 경북대학교 산학협력단 | Compositions for preventing or treating metabolic disorders comprising an extract of Caragana Sinica |
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| KR101890828B1 (en) * | 2018-04-10 | 2018-09-28 | 주식회사 디아이오 | Functional cosmetic composition and manufacturing method thereof |
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| KR20140006139A (en) * | 2012-06-26 | 2014-01-16 | 주식회사 래디안 | Cosmetic composition with anti-uv and anti-wrinkle effect from caragana sinica rehder |
| KR101413105B1 (en) | 2012-06-26 | 2014-07-02 | 주식회사 래디안 | Cosmetic composition with anti-UV and anti-wrinkle effect from Caragana sinica Rehder |
| KR20160135037A (en) * | 2015-05-15 | 2016-11-24 | 경북대학교 산학협력단 | Compositions for preventing or treating metabolic disorders comprising an extract of Caragana Sinica |
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