KR20170137546A - Composition for improving skin condition comprising herb extracts mixture - Google Patents
Composition for improving skin condition comprising herb extracts mixture Download PDFInfo
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- KR20170137546A KR20170137546A KR1020160069850A KR20160069850A KR20170137546A KR 20170137546 A KR20170137546 A KR 20170137546A KR 1020160069850 A KR1020160069850 A KR 1020160069850A KR 20160069850 A KR20160069850 A KR 20160069850A KR 20170137546 A KR20170137546 A KR 20170137546A
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- extract
- skin
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
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- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
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- A—HUMAN NECESSITIES
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- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/318—Foods, ingredients or supplements having a functional effect on health having an effect on skin health and hair or coat
Landscapes
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Abstract
Description
본 발명은 피부 적용시 안전하고, 피부 개선 효과가 우수한 복합 생약 추출물을 유효성분으로 포함하는 조성물에 관한 것이다.The present invention relates to a composition containing as an active ingredient a complex herbal extract which is safe when applied to skin and has excellent skin improving effect.
피부는 외부 환경으로부터 인체를 보호하고, 내부의 수분 및 유용 성분이 밖으로 유출되는 것을 막아주는 장벽 기능, 체온조절 기능, 배설 기능 등 다양한 생리적 기능을 담당하고 있는 중요한 기관이다. 그러나 다음과 같은 이유 등에 의해 피부 세포의 활성이 저하되고 피부 상태가 악화되는 현상이 일어날 수 있다.The skin is an important body that is responsible for various physiological functions such as barrier function, body temperature control function, excretion function, which protects the human body from the external environment and prevents the internal moisture and useful components from flowing out. However, due to the following reasons, the activity of skin cells may deteriorate and the skin condition may deteriorate.
피부의 진피층에 존재하는 콜라겐과 엘라스틴은 피부의 기계적 견고성, 결합조직의 저항력, 조직의 결합력 유지, 및 세포접착의 지탱 등의 기능을 수행하여 피부의 구성성분에 많은 영향을 미친다. 이러한 콜라겐은 스트레스, 고령화 또는 자외선 조사에 의한 광 노화에 의하여 감소되며, 한편, 엘라스틴은 자외선에 노출된 후 활성이 증가된 분해효소 엘라스테이즈에 의해 3차원 구조가 뒤틀려진다고 알려져있다. 이에 따라 피부 조직이 느슨해지고 탄력을 잃게 된다.Collagen and elastin present in the dermis of the skin perform functions such as mechanical rigidity of the skin, resistance of the connective tissue, maintenance of the cohesion of the tissue, and support of the cell adhesion, and have a great influence on the constituents of the skin. It is known that such collagen is reduced by photoaging due to stress, aging, or ultraviolet irradiation, while elastin is known to be distorted in three-dimensional structure by the enzymatic degradation enzyme Elastase after exposure to ultraviolet light. As a result, the skin tissue is loosened and the elasticity is lost.
표피의 세포 기능이 저하됨으로써 신진대사 작용이 원활하지 못하게 되어, 각질 탈락이 잘 일어나지 않아 각질이 과하게 축적된 상태에서 자외선을 받으면 탄력이 저하되어 주름 발생을 초래하게 된다. 또는 표피에 피지막이 형성되지 않을 경우 수분과 피지 분비가 감소하고 피부가 건조해지면서 주름이 형성되기도 한다.When the cell function of the epidermis is lowered, the metabolism is not smooth and the exfoliation does not occur well, and when the ultraviolet rays are received in a state where the keratin is overly accumulated, the elasticity is lowered and wrinkles are generated. If the skin is not formed on the epidermis, moisture and sebum secretion are reduced, and the skin becomes dry and wrinkles are formed.
피부가 자외선을 받으면 피부를 보호하기 위해 멜라닌을 합성한다. 합성된 멜라닌은 멜라노좀을 통해 피부의 각질세포로 옮겨진다. 이 각질 세포는 28일을 주기로 턴-오버(turn over) 현상이 일어난다. 따라서 생성된 멜라닌은 각질세포에 의해 28일을 주기로 소실되는 것이 일반적이나, 스트레스, 피부 노화 등에 의해 피부 세포 주기가 잘 조절되지 않게 되면 각질세포가 제시기에 탈락하지 않아 기미, 주근깨, 검버섯 등의 색소 침착이 일어나게 된다.When the skin receives ultraviolet rays, melanin is synthesized to protect the skin. The synthesized melanin is transferred to the keratinocytes of the skin through the melanoma. This keratinocyte turns over over a period of 28 days. Therefore, the melanin produced is generally lost by the keratinocyte periodically for 28 days. However, when the skin cell cycle is not properly controlled by the stress, skin aging, etc., keratinocytes do not fall out in the presentation period and the pigment such as spots, freckles, Calm occurs.
피부에서는 피지, 땀 성분 및 화장품 성분들이 피부 상재균에 의해 독성이 강한 물질로 분해되어 피부에 자극을 일으키고 염증을 유발시킬 수 있다. 또한 자외선으로 인한 자극으로 피부에 염증 매개 물질인 일산화질소(NO)의 생성을 증가시켜 피부 트러블이 유발될 수 있다. 일산화질소의 생성은 대부분 iNOS에 의한 것인데, iNOS는 LPS, 사이토카인 같은 자극에 의해 급격하게 유도되어 과량의 NO를 생성하는 것으로 알려져있다.In the skin, sebum, sweat and cosmetic ingredients are decomposed into substances that are toxic to the skin by the fungus, which can cause skin irritation and inflammation. In addition, skin irritation caused by ultraviolet rays increases the production of nitrogen monoxide (NO), which is an inflammation mediator, to cause skin troubles. The production of nitrogen monoxide is mostly caused by iNOS, and it is known that iNOS is rapidly induced by stimulation such as LPS and cytokine to produce excessive NO.
유해산소(toxic oxygen species)라고도 하는 활성산소종(reactive oxygen species, ROS)은 호흡 등과 같은 생리작용에 의해 세포에서 생성되는 독성물질로 끊임없이 생산되고 소멸하며, 정상적인 상태에서는 3-5% 정도로 존재한다. 이런 활성산소종은 수퍼옥시드 라디칼(Superoxide radical, O2-), 하이드록시라디칼 (hydroxyl radical, HO+)과 같은 자유라디칼(free radical: 화학적으로 최외각 전자궤도에 쌍을 이루고 있지 않은 원자나 분자로 매우 불안정하여 높은 반응성을 가지는 형태)로 존재하거나 혹은 과산화수소(hydorgen peroxide, H2O2)나 일중항산소(Singlet radical)와 같이 쌍을 이룬 전자를 가진 화합물의 형태로 존재한다. 활성산소종은 생리계 내에서 세균을 살균하는 생체 방어 작용을 하는 장점도 있지만, 일반적으로 생체 내에서 산화를 일으켜 질병의 원인이 되는 유해한 작용을 한다. 이러한 활성산소종은 생물분자를 공격하여 세포나 조직에 피해를 주며, 노화나 각종 성인병 질환에 관여하는 여러 종류의 질병을 야기한다는 보고가 있다.A reactive oxygen species (ROS), also known as a toxic oxygen species, is a cell-generated toxic substance produced by physiological actions such as respiration and is constantly produced and extinguished, with 3-5% in normal conditions . These reactive oxygen species are free radicals such as superoxide radicals (O2-) and hydroxyl radicals (HO +), which are chemically bound to atoms or molecules that are not paired with the outermost electron orbits Highly unstable and highly reactive) or in the form of compounds with paired electrons such as hydrogen peroxide (H 2 O 2 ) or singlet radicals. Active oxygen species have the advantage of biologically protecting bacteria in the physiological system, but generally cause oxidation in vivo, causing harmful effects that cause disease. It has been reported that these reactive oxygen species attack biological molecules and damage cells and tissues and cause various diseases related to aging and various diseases.
상술한 여러 요인들에 의한 피부 노화를 억제하고, 미백, 주름, 탄력 또는 트러블을 개선하고, 항산화 효과가 있는 물질을 개발하고자 하는 연구가 계속되어 왔다.Research has been continued to suppress the skin aging caused by various factors as described above, to improve whitening, wrinkles, elasticity or trouble, and to develop a substance having an antioxidative effect.
상술한 효과가 있는 물질은 자연계에 널리 분포되어 있는데, 주로 식물로부터 유래된 물질을 사용해 식품, 화장품, 의약품 등의 원료로서 사용해 왔다. 그러나, 이 같은 자연계에서 유래된 물질은 효과가 크지 않아 유의미한 효과를 얻기 위해서는 다량으로 사용해야 하고, 이로 인한 독성 및 가격 상승 문제가 대두되었다.Materials having the above-mentioned effects are widely distributed in the natural world, and they have been mainly used as raw materials for foods, cosmetics, medicines and the like using materials derived from plants. However, the substances derived from such natural substances are not effective enough to be used in large quantities in order to obtain a meaningful effect, resulting in toxicity and price increase.
이러한 천연 물질의 문제점을 해결하기 위해, 화학적으로 합성된 물질의 개발이 이어졌다. 이들은 천연 물질에 비해 소량 사용으로도 월등히 우수한 효과를 발휘한다는 장점이 있으나, 반면 인체에 크고 작은 부작용을 유발시킬 수 있다는 치명적인 문제점이 있어 사용이 제한된다.In order to solve the problems of such natural materials, the development of chemically synthesized materials has continued. Although they have a merit that they exert a superior effect even when used in a small amount compared to a natural substance, their use is limited due to a fatal problem that can cause large and small side effects to the human body.
이에, 피부 개선에 우수한 효과를 발휘하면서도 천연물로부터 유래되어 안정성이 확보되는 물질의 개발이 절실한 실정이다.Accordingly, it is inevitable to develop a substance derived from a natural product and having stability, while exhibiting an excellent effect on skin improvement.
본 발명이 해결하고자 하는 과제는 피부 개선, 특히 미백, 주름, 탄력, 트러블 개선, 항산화에 우수한 효과를 발휘하면서도 천연물로부터 유래되어 안정성이 확보되는 복합 생약 추출물을 제공하는데 있다.A problem to be solved by the present invention is to provide a complex herbal medicine extract which is derived from natural materials and has stability, while exhibiting excellent effects on skin, in particular whitening, wrinkle, elasticity, trouble improvement and antioxidation.
상기 과제를 해결하기 위하여, 본 발명은 목단피 추출물, 당귀 추출물, 황기 추출물, 천궁 추출물, 생지황 추출물, 지부자 추출물, 백선피 추출물, 황정 추출물 및 노근 추출물을 유효성분으로 포함하는 피부 미백, 주름 개선, 탄력 개선, 트러블 개선 또는 항산화용 조성물(예를 들어, 화장료 조성물)을 제공한다.In order to solve the above-mentioned problems, the present invention provides skin whitening, wrinkle improvement, elasticity, and the like, which comprise, as an active ingredient, a herbal extract, a angelica edulis extract, an angelica edulis extract, an angelica edulis extract, (E. G., A cosmetic composition) for improving, improving the trouble or antioxidation.
[추출물][extract]
목단피(牧丹皮)는 작약과(Paeoniaceae) 목단(Paeonia suffruticosa Andrews)의 뿌리껍질이다.Mokdanpi (牧丹皮) is the peony (Paeoniaceae) Peony (Paeonia suffruticosa Andrews).
당귀(當歸, Angelica Gigas)는 미나리과에 속하며, 참당귀(Angelica Gigas Nakai)의 뿌리를 포함하고, 오장육부를 건강하게 하고 몸을 따스하게 하는 성질이 있어, 당귀의 몸통은 피를 조절하며 당귀의 윗부분은 보혈을 하는 효능이 있다. Angelica Gigas belongs to the persimmon, and contains the roots of Angelica Gigas Nakai . It has the property to make the body and the body healthy. The body of Angelica Gigas regulates the blood and the upper part of the Angelica gigas Has the effect of making blood.
황기(黃耆)는 콩과(Leguminosae)의 황기(Astragalus membranaceus Bunge) 또는 몽골황기(蒙古黃耆, Astragalus membranaceus Bunge var. mongholicus Hsiao)의 뿌리로서 그대로 또는 주피를 제거한 것이다. 황기는 약성이 온화하고 맛이 달다.Hwanggi (黄耆) of leguminosae (Leguminosae) ( Astragalus membranaceus Bunge) or Mongolian shaggy (蒙古 黄耆, Astragalus There are membranaceus Bunge. mongholicus Hsiao), or as a root. Hwanggi is mild and mild.
천궁(川芎)은 산형과(Umbelliferae)에 속하는 천궁(Cnidium officinale Makino) 또는 중국천궁(中國川芎, Ligusticum chuanxiong Hort)의 뿌리줄기로서 그대로 또는 끓는 물에 데친 것이다.The canton (川芎) belongs to the Umbelliferae ( Cnidium officinale Makino) or Chinese astronomy (Chinese 川芎, Ligusticum chuanxiong Hort), or poached in boiling water.
생지황(生地黃, Rehmanniae Radix Recens)은 현삼과(Scrophulariaceae)이고, 지황(Rehmannia glutinosa Liboschitz ex Steudel)의 신선한 뿌리로, 맛은 쓰고 약간 달며 성질은 몹시 차다.Rehmanniae Radix Recens is a scrophulariaceae and is a fresh root of Rehmannia glutinosa Liboschitz ex Steudel. Its taste is slightly worn, its quality is very cold.
지부자(地膚子,Kochiae Fructus)는 명아주과(Chenopodiaceae)에 속하는 댑싸리(Kochia scoparia Schrader)의 열매이다.Kochiae Fructus is a fruit of Kochia scoparia Schrader belonging to Chenopodiaceae.
백선피(白鮮皮)는 운향과(Rutaceae)의 여러해살이풀인 백선(Dictamnus dasycarpus Turczaininov)의 뿌리껍질이다.White pine bark is the roots of white pine ( Dictamnus dasycarpus Turczaininov), a perennial plant of rutaceae .
황정(黃精)은 백합과(Liliaceae)에 속하는 층층갈고리둥굴레(Polygonatum sibiricum Redoute), 진황정(Polygonatum falcatum A. Gray), 전황정(Polygonatum kingianum Coll. et Hemsley) 또는 다화황정(多花黃精, Polygonatum cyrtonema Hua)의 뿌리줄기이다.Hwangjeong (黃精) is tiered hook Polygonatum (Polygonatum sibiricum Redoute) belonging to Liliaceae (Liliaceae), binary hwangjeong (Polygonatum falcatum A. Gray, Polygonatum kingianum Coll. et Hemsley or Polygonatum polygonatum cyrtonema Hua).
노근(蘆根)은 벼과(Gramineae)에 속하는 갈대(Phragmites communis Trinius)의 뿌리줄기이다.根 root (根 root) belongs to the Gramineae ( Phragmites communis Trinius).
본 발명자들은 상기 추출물의 모두가 피부에 유익한 생물학적 활성을 가지며, 상기 추출물 중 2 이상의 복합 추출물, 바람직하게 목단피 추출물, 당귀 추출물, 황기 추출물, 천궁 추출물, 생지황 추출물, 지부자 추출물, 백선피 추출물, 황정 추출물 및 노근 추출물의 복합 추출물은 시너지적 생물학적 활성을 가짐을 발견하였다.The present inventors have found that all of the above extracts have a biological activity beneficial to the skin, and that the extracts of two or more of the above extracts, preferably a herbal extract, Angelica keiskei, Angelica keiskei, And a complex extract of Angelicae Radix extract have synergistic biological activity.
본 발명에 있어서, 용어 "추출물"은 상기 추출처리에 의하여 얻어지는 추출액, 상기 추출액의 희석액이나 농축액, 상기 추출액을 건조하여 얻어지는 건조물, 상기 추출액의 조정제물이나 정제물, 또는 이들의 혼합물 등, 추출액 자체 및 추출액을 이용하여 형성 가능한 모든 제형의 추출물을 포함한다.In the present invention, the term "extract" is intended to mean an extract obtained by the above-mentioned extraction treatment, a diluted or concentrated liquid of the extracted liquid, a dried material obtained by drying the extracted liquid, a controlled preparation or a purified product of the extracted liquid, And extracts of all formulations which can be formed using extracts.
상기 추출물에는 각각의 식물을 설명하며 특정된 부위 외에도 그의 잎, 줄기, 나무껍질, 뿌리, 꽃 또는 꽃눈, 과실, 종자, 수액 및 전체 식물이 포함될 수 있다.Each of the above-mentioned extracts describes each plant and may include leaves, stems, bark, roots, flowers or flowers, fruit, seeds, sap, and whole plants in addition to the specified parts.
상기 추출물을 제조하기 위하여, 통상의 기술자는 당업계에 공지된 임의의 적합한 방법을 사용할 수 있다. 예를 들어, 용매 추출법에 의할 수 있다. 식물 전체 또는 임의의 부분을 분쇄한 다음(예를 들어, 블렌더), 추출 용매를 처리하여, 용매 추출물을 수득할 수 있다. 분쇄 전에 건조 과정(예를 들어, 40 내지 70℃에서 15 내지 50시간 동안의 건조)을 거친 다음 분쇄할 수도 있다. 또한, 용매 추출물은 감압 증류 및 동결건조 또는 분무 건조 등과 같은 추가적인 과정에 의해 분말 상태로 제조될 수 있다.In order to prepare the above extract, one of ordinary skill in the art can use any suitable method known in the art. For example, a solvent extraction method can be used. The entire plant or any portion thereof may be ground (e.g., a blender) and then the extraction solvent may be treated to obtain a solvent extract. It may be subjected to a drying process (for example, drying at 40 to 70 DEG C for 15 to 50 hours) before pulverization and then pulverization. In addition, the solvent extract may be prepared in powder form by an additional process such as vacuum distillation and freeze-drying or spray-drying.
상기 사용되는 추출 용매의 종류는 특별히 제한되지 아니하며, 당해 기술 분야에서 공지된 임의의 용매를 사용할 수 있다. 상기 추출 용매의 비제한적인 예로는 물; 메탄올, 에탄올, 프로필알코올, 부틸알코올 등의 C1 내지 C4의 저급 알코올; 글리세린, 부틸렌글라이콜, 프로필렌글라이콜 등의 다가 알코올; 및 메틸아세테이트, 에틸아세테이트, 아세톤, 벤젠, 헥산, 디에틸에테르, 디클로로메탄 등의 탄화수소계 용매; 또는 이들의 혼합물을 사용할 수 있으며, 바람직하게, 물, 저급알코올을 단독으로 사용하거나 2종 이상 혼합하여 사용할 수 있다. 상기 용매를 사용하여 1회 이상 추출하여 용매 추출물을 제조할 수 있으며, 상기 용매 추출물을 감압 증류한 후 동결건조 또는 분무 건조하여 얻은 건조 추출물을 제조할 수 있다.The kind of the extraction solvent used is not particularly limited, and any solvent known in the art can be used. Non-limiting examples of the extraction solvent include water; C1 to C4 lower alcohols such as methanol, ethanol, propyl alcohol and butyl alcohol; Polyhydric alcohols such as glycerin, butylene glycol and propylene glycol; And hydrocarbon solvents such as methyl acetate, ethyl acetate, acetone, benzene, hexane, diethyl ether, and dichloromethane; Or mixtures thereof. Water and lower alcohols may be used alone or in combination of two or more. The solvent extract may be prepared by extracting the extract at least one time using the solvent, and the dry extract obtained by vacuum distillation or spray drying the solvent extract may be prepared.
상기 추출 용매의 양은 이용되는 추출 용매의 종류에 따라 다양할 수 있으나, 예를 들어 대상 식물의 건조 중량에 대하여 1 내지 20배, 또는 5 내지 20배로 사용할 수 있다.The amount of the extraction solvent may vary depending on the kind of the extraction solvent used, but may be, for example, 1 to 20 times, or 5 to 20 times the dry weight of the target plant.
이외에도, 당업계 공지된 다양한 추출 공정, 예를 들어, 온침(maceration), 인퓨전(infusion), 퍼콜레이션(percolation), 소화, 전즙(decoction), 고온 연속 추출(hot continuous extraction), 수성-알코올성 추출, 역류 추출, 마이크로파 보조 추출, 초음파 추출, 초임계 유체 추출, 조직편 추출(phytonic extract) (예를 들어, 하이드로-플루오로-카본 용매) 등) 등을 선택하여 사용할 수 있으며, 이들은 단독으로 수행되거나 2 종 이상의 방법을 병용하여 수행될 수 있다.In addition, various extraction processes known in the art such as, for example, maceration, infusion, percolation, digestion, decoction, hot continuous extraction, aqueous- (For example, hydrofluoro-carbon solvent), etc.), which may be used alone or in combination with one another, may be used, for example, May be carried out by using two or more methods in combination.
[조성물][Composition]
본 발명에 따른 조성물은 목단피 추출물, 당귀 추출물, 황기 추출물, 천궁 추출물, 생지황 추출물, 지부자 추출물, 백선피 추출물, 황정 추출물 및 노근 추출물을 유효성분으로 포함한다.The composition according to the present invention contains as an active ingredient an extract of Momordicae japonica, Angelica keiskei, Extract of Angelica keiskei, Extract of Angelica keiskei, Extract of Cryptomeria japonica, Extract of Cryptomeria japonica,
본 발명에 있어서 "유효성분으로 포함된다"는 의미는 본 발명에 따른 조성물로부터 피부 개선 효과를 나타낼 수 있는 정도로, 추출물이 첨가되는 것을 의미하고, 피부 전달 및 안정화 등을 위하여 다양한 성분을 부성분으로 첨가하여 다양한 형태로 제형화(formulation) 가능함을 포함하는 의미이다.In the present invention, the term "included as an active ingredient" means that the extract is added to the skin composition of the present invention to such an extent that it can exhibit a skin improving effect, and various components are added as a sub ingredient And it is possible to formulate in various forms.
본 발명에 따른 조성물에 각각의 식물 추출물은 다음과 같은 비율로 포함될 수 있다. 예를 들어, 목단피 추출물 : 당귀 추출물 : 황기 추출물 : 천궁 추출물 : 생지황 추출물 : 지부자 추출물 : 백선피 추출물 : 황정 추출물 : 노근 추출물의 중량비는 1 : 0.1 ~ 10 : 0.1 ~ 10 : 0.1 ~ 10 : 0.1 ~ 10 : 0.1 ~ 10 : 0.1 ~ 10 : 0.1 ~ 10 : 0.1 ~ 10로 포함될 수 있으며, 1 : 0.1 ~ 5 : 0.1 ~ 5 : 0.1 ~ 5 : 0.1 ~ 5 : 0.1 ~ 5 : 0.1 ~ 5 : 0.1 ~ 5 : 0.1 ~ 5, 1 : 0.1 ~ 3 : 0.1 ~ 3 : 0.1 ~ 3 : 0.1 ~ 3 : 0.1 ~ 3 : 0.1 ~ 3 : 0.1 ~ 3 : 0.1 ~ 3, 1 : 0.1 ~ 2 : 0.1 ~ 2 : 0.1 ~ 2 : 0.1 ~ 2 : 0.1 ~ 2 : 0.1 ~ 2 : 0.1 ~ 2 : 0.1 ~ 2, 1 : 0.5 ~ 1.5 : 0.5 ~ 1.5 : 0.5 ~ 1.5 : 0.5 ~ 1.5 : 0.5 ~ 1.5 : 0.5 ~ 1.5 : 0.5 ~ 1.5 : 0.5 ~ 1.5로 포함될 수 있다.Each plant extract may be included in the composition according to the present invention in the following proportions. For example, the weight ratio of extracts of Root Extract: Huang Gui Extract: Huang Gui Extract: Huang Guo Extract: Huang Gui Extract: 0.1 to 5: 0.1 to 5: 0.1 to 5: 0.1 to 5: 0.1 to 5: 0.1 to 5: 0.1 to 10: 0.1 to 10: 0.1 to 10: 0.1 to 10: 0.1 to 10, 1: 0.1 to 3: 0.1 to 3: 0.1 to 3: 0.1 to 3: 0.1 to 3: 0.1 to 3: 0.1 to 3, 1: 0.1 to 2: 0.1 to 2: 0.1 to 2: 0.1 to 2: 0.1 to 2: 0.1 to 2: 0.1 to 2: 0.1 to 2, 1: 0.5 to 1.5: 0.5 to 1.5: 0.5 to 1.5: 0.5 to 1.5: 0.5 to 1.5: 0.5 to 1.5: 0.5 to 1.5: 0.5 to 1.5.
본 발명에 따른 조성물은 이의 최종 형태 중에 적어도 약 0.0001%, 0.0002%, 0.0003%, 0.0004%, 0.0005%, 0.0006%, 0.0007%, 0.0008%, 0.0009%, 0.0010%, 0.0011%, 0.0012%, 0.0013%, 0.0014%, 0.0015%, 0.0016%, 0.0017%, 0.0018%, 0.0019%, 0.0020%, 0.0021%, 0.0022%, 0.0023%, 0.0024%, 0.0025%, 0.0026%, 0.0027%, 0.0028%, 0.0029%, 0.0030%, 0.0031%, 0.0032%, 0.0033%, 0.0034%, 0.0035%, 0.0036%, 0.0037%, 0.0038%, 0.0039%, 0.0040%, 0.0041%, 0.0042%, 0.0043%, 0.0044%, 0.0045%, 0.0046%, 0.0047%, 0.0048%, 0.0049%, 0.0050%, 0.0051%, 0.0052%, 0.0053%, 0.0054%, 0.0055%, 0.0056%, 0.0057%, 0.0058%, 0.0059%, 0.0060%, 0.0061%, 0.0062%, 0.0063%, 0.0064%, 0.0065%, 0.0066%, 0.0067%, 0.0068%, 0.0069%, 0.0070%, 0.0071%, 0.0072%, 0.0073%, 0.0074%, 0.0075%, 0.0076%, 0.0077%, 0.0078%, 0.0079%, 0.0080%, 0.0081%, 0.0082%, 0.0083%, 0.0084%, 0.0085%, 0.0086%, 0.0087%, 0.0088%, 0.0089%, 0.0090%, 0.0091%, 0.0092%, 0.0093%, 0.0094%, 0.0095%, 0.0096%, 0.0097%, 0.0098%, 0.0099%, 0.0100%, 0.0200%, 0.0250%, 0.0275%, 0.0300%, 0.0325%, 0.0350%, 0.0375%, 0.0400%, 0.0425%, 0.0450%, 0.0475%, 0.0500%, 0.0525%, 0.0550%, 0.0575%, 0.0600%, 0.0625%, 0.0650%, 0.0675%, 0.0700%, 0.0725%, 0.0750%, 0.0775%, 0.0800%, 0.0825%, 0.0850%, 0.0875%, 0.0900%, 0.0925%, 0.0950%, 0.0975%, 0.1000%, 0.1250%, 0.1500%, 0.1750%, 0.2000%, 0.2250%, 0.2500%, 0.2750%, 0.3000%, 0.3250%, 0.3500%, 0.3750%, 0.4000%, 0.4250%, 0.4500%, 0.4750%, 0.5000%, 0.5250%, 0.550%, 0.5750%, 0.6000%, 0.6250%, 0.6500%, 0.6750%, 0.7000%, 0.7250%, 0.7500%, 0.7750%, 0.8000%, 0.8250%, 0.8500%, 0.8750%, 0.9000%, 0.9250%, 0.9500%, 0.9750%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8.0%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9.0%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% 또는 99% 이상의 범위로 본 발명의 1 이상의 추출물을 포함할 수 있다. 상기 %는 조성물의 총 중량 대비 중량 또는 총 부피 대비 부피를 기준으로 계산할 수 있으며, 농도는 조성물의 목적하는 효과 또는 조성물이 혼입되는 제품에 따라 조정될 수 있다.The compositions according to the present invention may contain at least about 0.0001%, 0.0002%, 0.0003%, 0.0004%, 0.0005%, 0.0006%, 0.0007%, 0.0008%, 0.0009%, 0.0010%, 0.0011%, 0.0012% , 0.0014%, 0.0015%, 0.0016%, 0.0017%, 0.0018%, 0.0019%, 0.0020%, 0.0021%, 0.0022%, 0.0023%, 0.0024%, 0.0025%, 0.0026% %, 0.0031%, 0.0032%, 0.0033%, 0.0034%, 0.0035%, 0.0036%, 0.0037%, 0.0038%, 0.0039%, 0.0040%, 0.0041%, 0.0042% 0.005%, 0.0057%, 0.0058%, 0.0059%, 0.0060%, 0.0061%, 0.0062%, 0.0063%, 0.0058% , 0.0064%, 0.0065%, 0.0066%, 0.0067%, 0.0068%, 0.0069%, 0.0070%, 0.0071%, 0.0072%, 0.0073%, 0.0074%, 0.0075%, 0.0076%, 0.0077% %, 0.0081%, 0.0082%, 0.0083%, 0.0084%, 0.0085%, 0.0086%, 0.0087%, 0.0088%, 0.0089%, 0.0090%, 0.0091%, 0.0092%, 0.0093%, 0.0094%, 0.0095% 0.0097%, 0.0098%, 0.0099%, 0.0100%, 0.0200%, 0.0250%, 0.0275 %, 0.0300%, 0.0325%, 0.0350%, 0.0375%, 0.0400%, 0.0425%, 0.0450%, 0.0475%, 0.0500%, 0.0525%, 0.0550%, 0.0575%, 0.0600%, 0.0625%, 0.0650% 0.0725%, 0.0725%, 0.0750%, 0.0775%, 0.0800%, 0.0825%, 0.0850%, 0.0875%, 0.0900%, 0.0925%, 0.0950%, 0.0975%, 0.1000%, 0.1250%, 0.1500%, 0.1750%, 0.2000% , 0.2250, 0.2500, 0.2750, 0.3000, 0.3250, 0.3500, 0.3750, 0.4000, 0.4250, 0.4500, 0.4750, 0.5000, 0.5250, 0.550, 0.5750, 0.6000, 0.6250 %, 0.6500%, 0.6750%, 0.7000%, 0.7250%, 0.7500%, 0.7750%, 0.8000%, 0.8250%, 0.8500%, 0.8750%, 0.9000%, 0.9250%, 0.9500%, 0.9750%, 1.0% 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9% , 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2% 5.4%, 5.7%, 5.8%, 5.9%, 6.0%, 6.1%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2% 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7% , 7.9%, 8. 9.2%, 9.4%, 9.4%, 9.5%, 9.6%, 8.3%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9.0% , 9.7%, 9.8%, 9.9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21% %, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 60%, 65%, 70%, 75% 85%, 90%, 95%, or 99% or more of the extract of the present invention. The% can be calculated on the basis of the total weight of the composition or the volume relative to the total volume, and the concentration can be adjusted according to the desired effect of the composition or the product into which the composition is incorporated.
본 발명의 조성물은 모든 유형의 비히클 내로 제형화될 수 있다. 적합한 비히클의 예로는 에멀젼 (예를 들어, 수중유, 유중수, 수중실리콘, 실리콘중수, 수중유중수, 수중유, 유중수중유, 실리콘중수중유 등), 크림, 로션, 용액 (예컨대, 수성 및 하이드로-알코올 용액), 무수 베이스 (예를 들어, 립스틱 및 파우더), 젤, 연고, 페이스트, 밀크, 액체, 에어로졸, 고체 형태 또는 아이 젤리를 포함하나, 이에 제한되지 않는다.The compositions of the present invention may be formulated into any type of vehicle. Examples of suitable vehicles include, but are not limited to, emulsions (e.g., oil, water, water in silicone, water in silicone, water in heavy oil, water in oil, water in heavy water, But are not limited to, a hydro-alcoholic solution), a dry basis (e.g., lipstick and powder), a gel, ointment, paste, milk, liquid, aerosol, solid form or eye jelly.
또한, 본 발명의 조성물은 피부와 같은 표적 영역으로의 운반을 위해 캡슐화될 수도 있다. 캡슐화 기술은 예를 들어, 피부에 성분을 전달하는 운반 비히클로서 사용될 수 있는 리포좀, 소낭, 및/또는 나노입자 (예를 들어, 성분이 트랩핑(trap되고, 캡슐화되고/되거나 흡수되는 폴리머 물질을 포함하는 생분해성 및 비생분해성 콜로이드성 입자, 예를 들어 나노스피어(nanosphere) 및 나노캡슐을 포함)의 사용을 포함할 수 있으나, 이에 제한되지 않는다.The compositions of the present invention may also be encapsulated for delivery to a target area, such as the skin. Encapsulation techniques include, for example, the use of liposomes, follicles, and / or nanoparticles (e. G., Components that are trapped, encapsulated, and / or absorbed polymeric materials that can be used as a delivery vehicle But are not limited to, the use of biodegradable and non-biodegradable colloidal particles, including, for example, nanospheres and nanocapsules.
본 발명에 따른 조성물은 다양하게 제품화될 수 있다. 예를 들어, 화장료 조성물, 식품 조성물 등으로 제품화될 수 있다.The composition according to the present invention can be variously commercialized. For example, cosmetic compositions, food compositions, and the like.
상기 화장료 조성물은 예를 들어, 일반적인 유화 제형 및 가용화 제형의 형태로 제조된 것일 수 있다. 예를 들어, 유연 화장수 또는 영양 화장수 등과 같은 화장수, 훼이셜 로션, 바디로션 등과 같은 유액, 영양 크림, 수분 크림, 아이크림, 마사지 크림 등과 같은 크림, 에센스, 세럼, 화장연고, 스프레이, 오일젤, 젤, 팩, 선 스크린, 메이크업베이스, 액체 타입, 고체 타입 또는 스프레이 타입 등의 파운데이션, 파우더, 클렌징 크림, 클렌징 로션, 클렌징 오일과 같은 메이크업 제거제, 클렌징 폼, 비누, 바디 워쉬 등과 같은 세정제 등의 제형을 가질 수 있으나, 이에 제한되지 않고 당업계 공지된 다양한 형태로 제형화될 수 있다.The cosmetic composition may be prepared, for example, in the form of a general emulsified formulation and a solubilized formulation. For example, creams, essences, serums, cosmetic ointments, sprays, oil gels, gels such as lotions such as lotion, facial lotion, body lotion and the like such as flexible lotion or nutrition lotion, nutrition cream, A lotion, a cleansing lotion, a makeup remover such as a cleansing oil, a cleansing foam, a soap, a body wash and the like such as a foundation, a pack, a sunscreen, a makeup base, a liquid type, a solid type or a spray type, But may be formulated into various forms known in the art without limitation.
상기 화장료 조성물에는 본 발명의 목적을 해하지 않는 이상, 본 발명에 따른 추출물 외에 당업계에서 화장료 조성물의 성분으로 공지된 통상의 성분을 임의로 선택하여 추가로 포함할 수 있다. 예를 들어, 방향제 (합성 및 천연), 염료 및 색 성분, 흡수제, 에멀젼화제, 안정화제, 윤활제, 용매, 보습제 (예: 연화제(emollient), 습윤제(humectant), 피막형성제, 폐색제(occlusive agent), 및 피부의 천연 보습 메커니즘에 영향을 주는 작용제(agent)를 포함함), 발수제(water-repellant), UV 흡수제 (물리적 및 화학적 흡수제, 예를 들어 파라아미노벤조산("PABA") 및 상응하는 PABA 유도체, 이산화티탄, 산화아연 등), 에센셜 오일(essential oil), 비타민 (예: A, B, C, D, E 및 K), 미량 금속 (예: 아연, 칼슘 및 셀레늄), 항-자극제(anti-irritant) (예: 스테로이드 및 비스테로이드성 항염증제), 항-미생물제(anti-microbial agent), 항산화제 (예: BHT 및 토코페롤), 킬레이트제 (예: 다이소듐 EDTA 및 테트라소듐 EDTA), 보존제 (예: 메틸파라벤 및 프로필파라벤), pH 조정제 (예: 수산화나트륨 및 시트르산), 흡수제(예를 들어, 알루미늄 전분 옥테닐석시네이트, 카올린, 옥수수 전분, 귀리 전분, 사이클로덱스트린, 탈크(talc), 및 제올라이트), 피부 블리칭(bleaching) 및 라이트닝(lightening) 작용제 (예를 들어, 하이드로퀴논 및 니아신아미드 락테이트), 습윤제 (예: 글리세린, 프로필렌 글리콜, 부틸렌 글리콜, 펜틸렌 글리콜, 소르비톨, 우레아 및 만니톨), 각질제거제(exfoliant) (예: 알파-하이드록시산 및 베타-하이드록시산, 예를 들어 락트산, 글리콜산 및 살리실산; 및 이들의 염), 발수제(waterproofing agent) (예: 마그네슘/알루미늄 하이드록시드 스테아레이트), 피부 컨디셔닝제(skin conditioning agent) (예: 알로에 추출물, 알란토인(allantoin), 비사볼롤(bisabolol), 세라마이드(ceramide), 다이메티콘, 히알루론산, 및 다이포타슘 글리시리제이트), 및 증점제(thickening agent) (예:조성물의 점도를 증가시킬 수 있는 물질, 예를 들어 카복실산 폴리머, 가교된 폴리아크릴레이트 폴리머, 폴리아크릴아미드 폴리머, 다당류 및 검(gum)) 및 실리콘 함유 화합물(예: 실리콘 오일 및 폴리오르가노실록산) 등이 포함될 수 있다.In addition to the extract according to the present invention, the above-mentioned cosmetic composition may further include optional ingredients commonly known as ingredients of the cosmetic composition in the art, so long as the object of the present invention is not impaired. Emulsifying agents, stabilizers, lubricants, solvents, moisturizers (e.g. emollients, humectants, film forming agents, occlusive agents, emulsifiers, water-repellant, a UV absorber (including physical and chemical absorbents such as para-aminobenzoic acid ("PABA") and corresponding (Eg, PABA derivatives, titanium dioxide, zinc oxide, etc.), essential oils, vitamins such as A, B, C, D, E and K, trace metals such as zinc, calcium and selenium, Antioxidants (such as BHT and tocopherol), chelating agents (such as disodium EDTA and tetrasodium EDTA), antioxidants (for example, antimicrobial agents) , Preservatives (such as methylparaben and propylparaben), pH adjusting agents (such as sodium hydroxide and citric acid) (E.g., aluminum starch octenyl succinate, kaolin, corn starch, oat starch, cyclodextrin, talc, and zeolites), skin bleaching and lightening agents (e.g., Hydroquinone and beta-hydroxynaphthoic acid (e.g., hydroquinone and niacinamide lactate), wetting agents such as glycerin, propylene glycol, butylene glycol, pentylene glycol, sorbitol, urea and mannitol, exfoliants (E.g., magnesium / aluminum hydroxide stearate), skin conditioning agents (e.g., aloe extracts), and the like, , Allantoin, bisabolol, ceramide, dimethicone, hyaluronic acid, and dipotassium glycyrrhizate), and thickening agents (e.g., (E.g., silicone oils and polyorganosiloxanes), and the like, which are capable of increasing viscosity, such as carboxylic acid polymers, crosslinked polyacrylate polymers, polyacrylamide polymers, polysaccharides and gums, .
상기 식품 조성물은 예를 들어, 음료, 강화수(fortified water), 에너지 드링크(energy drink), 영양 드링크(nutritional drink), 고형 식품, 비타민, 보조제(Supplement) 등) 등으로 제형화 될 수 있으나, 이에 제한되지 않는다. 상기 보조제에는 비타민, 무기질, 허브 또는 기타 식물, 아미노산, 효소 및 대사산물을 포함할 수 있다. 이러한 보조제는 경구 이용(oral consumption)에 적합하며, 경구로 투여될 수 있다.The food composition may be formulated into, for example, beverages, fortified water, energy drinks, nutritional drinks, solid foods, vitamins, supplements, etc., But is not limited thereto. Such adjuvants may include vitamins, minerals, herbs or other plants, amino acids, enzymes and metabolites. Such adjuvants are suitable for oral consumption and may be administered orally.
본 발명에 따른 조성물을 포함하는 키트로 제공될 수 있다. 본 발명에 따른 조성물은 용기에 포함되며, 용기는 병, 금속 튜브, 라미네이트 튜브, 플라스틱 튜브, 디스펜서, 압력 용기, 장벽 용기, 패키지, 컴파트먼트, 립스틱 용기, 컴팩트 용기, 화장용 조성물을 담을 수 있는 화장용 팬, 또는 다른 타입의 용기 예를 들어, 분산매체(dispersions) 또는 조성물 또는 유지되는 바람직한 병, 디스펜서, 또는 패키지 내로 주입 또는 블로우-몰드된 플라스틱 용기 등이 포함될 수 있으며, 이에 제한되지 않는다. 상기 키트에는 키트 또는 조성물을 사용하기 위한 지시서가 포함될 수 있으며, 지시서는 별개 용지에 기재될 수 있고, 또는 용기 표면, 용기 포장지의 표면 상에 기재될 수 있다. 지시사항은 글자, 어구, 약어, 그림 또는 기호 등이 포함되며, 이에 제한되지 않는다. 지시서에는 예를 들어, 키트 또는 조성물의 사용 방법, 적용 방법 및 유지 방법 등에 관한 지시사항이 포함될 수 있다. 용기에는 사전에 정해진 양에 따라 분배하여 담을 수 있다.May be provided as a kit comprising the composition according to the present invention. The composition according to the invention is contained in a container which can contain a bottle, a metal tube, a laminate tube, a plastic tube, a dispenser, a pressure vessel, a barrier container, a package, a compartment, a lipstick container, a compact container, A cosmetic pan, or other type of container, including, but not limited to, a plastic container that is injected or blow-molded into a suitable bottle, dispenser, or package to be maintained or dispersed, . The kit may include an instruction for using the kit or composition, the instruction sheet may be described on a separate sheet, or may be described on the surface of the container surface, the surface of the container wrapper. Instructions include, but are not limited to, letters, phrases, abbreviations, pictures or symbols. The instructions may include, for example, instructions on how to use, apply and maintain the kit or composition. The container can be dispensed according to a predetermined amount.
본 발명에 따른 조성물은 국소용 피부 조성물로 제공될 수 있다.The composition according to the present invention may be provided as a topical skin composition.
또한, 본 발명에 따른 조성물을 피부에 국소 적용하는 방법을 제공할 수 있다.In addition, a method of topically applying the composition according to the present invention to the skin can be provided.
본 발명에 있어서, 용어 "국소 적용"은 조성물을 각질 조직의 표면상에 적용하거나 도포하는 것을 의미하고, "국소용 피부 조성물"은 각질 조직상에 국소 적용 또는 도포하기에 적합한 조성물을 포함한다. 이러한 조성물은 피부에 적용되는 경우 과도한 독성, 부적합성(incompatibility), 불안정성, 알러지 반응 등을 갖지 않는다는 점에서, 전형적으로 피부과학적으로 허용된다. 본 발명의 국소 피부 케어 조성물은, 피부에 적용된 후 현저한 적하(dripping) 또는 풀링(pooling)을 회피하기 위해, 선택된 점도를 지닐 수 있다.In the present invention, the term "topical application" means applying or applying the composition on the surface of keratinous tissue, and "topical skin composition" includes compositions suitable for topical application or application on keratinocytes. Such compositions are typically dermatologically acceptable in that they do not have excessive toxicity, incompatibility, instability, allergic response, etc. when applied to the skin. The topical skin care compositions of the present invention may have a selected viscosity to avoid significant dripping or pooling after application to the skin.
[피부 개선 효과][Skin improvement effect]
본 발명에 따른 조성물은 생물학적 활성을 가지며, 피부 개선에 우수한 효과를 발휘한다.The composition according to the present invention has biological activity and exerts excellent effects on skin improvement.
보다 구체적으로, 본 발명에 따른 조성물은 피부 미백 효과가 있다.More specifically, the composition according to the present invention has a skin whitening effect.
상기 피부 미백은 멜라닌 형성 과정을 억제 시켜 멜라닌 생성을 억제하는 것을 말한다.The skin whitening refers to inhibiting melanin formation and inhibiting melanin formation.
본 발명자들은 본 발명에 따른 복합 추출물을 멜라노마 세포에 처리하여 생성된 멜라닌의 양을 측정한 결과, 각각의 추출물을 처리하는 경우에 비해 상승된 멜라닌 생성 억제 효과를 보임을 확인할 수 있었고, 피부 미백 효과가 공지되어 있는 알부틴에 상응하는 효과를 발휘함을 알 수 있었다. 본 발명에 따른 조성물은 멜라닌 생성 저해 효과가 뛰어나, 우수한 미백 효과를 나타냄을 확인하였다.As a result of measuring the amount of melanin produced by treating the melanoma cells according to the present invention, the inventors of the present invention confirmed that the melanin production inhibitory effect was enhanced compared with the case of treating each extract, It was found that the effect exerted an effect corresponding to known arbutin. It was confirmed that the composition according to the present invention was excellent in the effect of inhibiting melanin formation and exhibited excellent whitening effect.
또한, 본 발명에 따른 조성물은 주름 및 탄력 개선 효과가 있다.In addition, the composition according to the present invention has an effect of improving wrinkles and elasticity.
상기 피부 주름이란 피부가 쇠하여 생긴 잔줄을 의미하며, 유전자에 의한 원인, 피부 진피에 존재하는 콜라겐과 엘라스틴의 감소, 외부환경 등에 의해 유발될 수 있다.The skin wrinkles are caused by the skin of the skin and can be caused by the cause of the gene, collagen and elastin present in the skin dermis, and the external environment.
상기 주름 개선은 주름이 생성되는 것을 억제 또는 저해하거나, 이미 생성된 주름을 완화시키는 것을 말한다.The wrinkle improvement refers to suppressing or inhibiting the generation of wrinkles, or alleviating the already generated wrinkles.
상기 피부 탄력이란 진피층에 존재하는 엘라스틴(elastin)으로 구성된 탄력섬유 및 콜라겐(collagen)이라고 하는 교원섬유에 의해 나타나는 것으로, 상기 탄력 개선이란 탄력 감소를 억제 또는 저해하거나, 탄력을 유지 또는 향상시키는 것을 말한다.The skin elasticity refers to elastic fibers composed of elastin existing in the dermal layer and collagen fibers called collagen. The elasticity improvement refers to suppressing or inhibiting elasticity reduction or maintaining or improving elasticity .
본 발명자들은 본 발명에 따른 복합 추출물을 처리한 결과, 각각의 추출물을 처리하는 경우에 비해 상승된 콜라겐 합성 촉진 효과 및 엘라스테이즈 활성 저해 효과를 보임을 확인할 수 있었고, 피부 콜라겐 합성 촉진 효과가 공지되어 있는 비타민 C에 상응하는 효과를 보이며, 피부 엘라스테이즈 활성 저해 효과가 공지되어 있는 퀘세틴에 상응하는 효과를 발휘함을 알 수 있었다. 본 발명에 따른 조성물은 콜라겐 합성 촉진 효과 및 엘라스테이즈 활성 저해 효과가 뛰어나, 우수한 피부 주름 및 탄력 개선 효과를 나타냄을 확인하였다.As a result of treating the complex extract according to the present invention, the inventors of the present invention confirmed that they exhibited an enhanced collagen synthesis promoting effect and an elastase activity inhibiting effect, respectively, as compared with the case of treating each extract, And vitamin C, and the effect of quercetin, which is known to inhibit the activity of the skin elastase, is exerted. The composition according to the present invention is excellent in collagen synthesis promoting effect and elastase activity inhibiting effect and shows excellent wrinkle and elasticity improvement effect.
또한, 본 발명에 따른 조성물은 트러블 개선 효과가 있다.Further, the composition according to the present invention has an effect of improving the trouble.
상기 트러블은 피부자극 및 염증과 같은 증상을 말하며, 염증 반응에는 외부 자극에 의한 동통, 발열, 발적, 종창, 기능상실 등을 특징으로 하며, 조직학적으로는 소동맥, 모세혈관 및 소정맥의 투과성과 혈류증가를 동반한 확장, 혈장단백을 함유한 혈장의 삼출, 백혈구의 염증부위로의 이동 등을 포함한 복잡한 증상 등이 관찰되며, 피부자극 반응에는 홍반, 가려움, 발열 등을 특징으로 한다.The above-mentioned troubles are symptoms such as skin irritation and inflammation. The inflammatory reaction is characterized by pain, fever, redness, swelling and malfunction due to external stimuli. Histologically, the permeability of the arterioles, capillaries, , Complicated symptoms including enlargement accompanied by increase, excretion of plasma containing plasma protein, migration to inflammation site of leukocyte, and skin irritation reaction are characterized by erythema, itching, fever and the like.
상기 트러블 개선은 피부 자극 반응 및 염증 반응을 억제 또는 저해하거나, 이미 진행 중인 피부자극 반응 또는 염증 반응을 완화시키는 것을 말한다.The improvement of the trouble refers to inhibiting or inhibiting the skin irritation reaction and the inflammation reaction, or alleviating the skin irritation reaction or the inflammation reaction already in progress.
본 발명자들은 본 발명에 따른 복합 추출물을 처리하여 NO 생성 저해 효과를 측정한 결과, 각각의 추출물을 처리하는 경우에 비해 상승된 NO 생성 저해 효과를 보임을 확인할 수 있었고, NO 생성 저해 효과가 공지되어 있는 L-NMMA에 상응하는 효과를 발휘함을 알 수 있었다. 본 발명에 따른 조성물은 NO 생성 저해를 통해 우수한 피부 트러블 개선 효과를 나타냄을 확인하였다.As a result of measuring the inhibitory effect on NO production by treating the compound extract according to the present invention, the present inventors confirmed that the inhibitory effect on NO production was enhanced compared to the case of treating each extract, and NO inhibitory effect was known And exhibited an effect corresponding to that of L-NMMA. It was confirmed that the composition according to the present invention exhibits excellent skin trouble improving effect through inhibition of NO production.
또한, 본 발명에 따른 조성물은 항산화 효과가 있다.In addition, the composition according to the present invention has an antioxidative effect.
상기 항산화는 세포내 대사 또는 자외선의 영향으로 인한 산화적 스트레스에 따라 반응성이 높은 자유 라디칼(free radical) 또는 활성산소종(reactive oxygen species; ROS)에 의한 세포의 산화를 억제하는 것을 말하며, 자유 라디칼 또는 활성산소종을 제거하여 이로 인한 세포의 손상이 감소되는 것을 포함한다.The antioxidant refers to inhibition of cellular oxidation by free radicals or reactive oxygen species (ROS), which are highly reactive according to oxidative stress caused by intracellular metabolism or ultraviolet rays, and free radicals Or removal of reactive oxygen species, thereby reducing damage to the cells.
본 발명자들은 본 발명에 따른 복합 추출물을 처리하여 자유라디칼 소거능을 측정한 결과, 각각의 추출물을 처리하는 경우에 비해 상승된 자유라디칼 소거능을 보임을 확인할 수 있었고, 자유라디칼 소거능이 공지되어 있는 비타민 C에 상응하는 효과를 발휘함을 알 수 있었다. 이로써, 본 발명에 따른 조성물은 우수한 항산화 효과를 나타냄을 확인하였다.As a result of measuring the free radical scavenging ability of the complex extract according to the present invention, the inventors of the present invention confirmed that they exhibited an increased free radical scavenging ability as compared with the case of treating each extract, and found that the free radical scavenging ability of vitamin C And the effect is comparable to that of the conventional method. Thus, it was confirmed that the composition according to the present invention exhibits an excellent antioxidative effect.
또한, 본 발명에 따른 조성물은 항당화 효과가 있다.In addition, the composition according to the present invention has an antiherating effect.
상기 당화는 과도한 당의 존재로 인해 생기는 당과 단백질이 결합해 단백질이 파괴되는 현상을 말한다. 당화가 진행되면 콜라겐과 엘라스틴에 영향을 미쳐 피부 탄력 저하 외 각종 피부손상을 일으키게 된다.The glycation refers to a phenomenon in which a sugar and a protein are bound to each other due to the presence of excessive sugars and the protein is destroyed. As glycosylation progresses, it affects collagen and elastin, resulting in skin elasticity deterioration and other skin damage.
본 발명자들은 본 발명에 따른 복합 추출물을 처리하여 항당화 효과를 측정한 결과, 각각의 추출물을 처리하는 경우에 비해 상승된 항당화 효과를 보임을 확인할 수 있었고, 항당화 효과가 공지되어 있는 아미노구아니딘에 상응하는 효과를 발휘함을 알 수 있었다. 이로써, 본 발명에 따른 조성물은 우수한 항당화 효과를 나타냄을 확인하였다.As a result of measuring the antagonism effect by treating the complex extract according to the present invention, the inventors of the present invention have confirmed that they exhibit an elevated antihyperglycosylation effect as compared with the case of treating the respective extracts, and the aminoguanidine And the effect is comparable to that of the conventional method. Thus, it was confirmed that the composition according to the present invention exhibits excellent anticarcinogenic effect.
본 발명에 따른 복합 추출물은 피부에 안전하며, 항당화 효과, 피부 미백 효과, 피부 탄력 및 주름 개선 효과, 항산화 효과 및 피부 트러블 개선 효과를 포함한 피부 개선에 탁월한 효과를 갖는다.The complex extract according to the present invention is safe for the skin and has an excellent effect for improving the skin including anti-glycation effect, skin whitening effect, skin elasticity and wrinkle improving effect, antioxidative effect and skin trouble improving effect.
이하, 본 발명을 구체적으로 설명하기 위해 실시예 및 실험예를 들어 상세하게 설명하기로 한다. 그러나 본 발명에 따른 실시예 및 실험예는 여러 가지 다른 형태로 변형될 수 있으며, 본 발명의 범위가 아래에서 상술하는 실시예 및 실험예에 한정되는 것으로 해석 되어서는 안 된다. 본 발명의 실시예 및 실험예는 당업계에서 평균적인 지식을 가진 자에게 본 발명을 보다 완전하게 설명하기 위해서 제공되는 것이다.Hereinafter, the present invention will be described in detail with reference to Examples and Experimental Examples. However, the embodiments and experimental examples according to the present invention can be modified into various other forms, and the scope of the present invention should not be construed as being limited to the above-described embodiments and experiments. The embodiments and experimental examples of the present invention are provided to enable those skilled in the art to more fully understand the present invention.
<< 실시예Example 1> 목단피 추출물의 제조 1> Preparation of extract
목단피를 잘 건조하여 세절한 후, 건조 중량 100g을 플라스크에 넣고 추출 용매(증류수) 1000g으로 3일간 냉침하여 추출하였다. 냉침된 추출물을 0.2㎛의 기공 크기를 가진 필터로 여과하여 목단피 추출물을 제조하였다.After thoroughly drying the slices, the dried weight of 100 g was placed in a flask, and extracted with 1000 g of an extraction solvent (distilled water) for 3 days by cooling. The frozen extract was filtered with a filter having a pore size of 0.2 mu m to prepare a herringbone extract.
<< 실시예Example 2> 당귀 추출물의 제조 2> Preparation of Angelica gigantosa extract
당귀를 잘 건조하여 세절한 후, 건조 중량 100g을 플라스크에 넣고 추출 용매(증류수) 1000g으로 3일간 냉침하여 추출하였다. 냉침된 추출물을 0.2㎛의 기공 크기를 가진 필터로 여과하여 당귀 추출물을 제조하였다The Angelica gigas was dried well and cut into three pieces. 100 g of dry weight was put into a flask and extracted with 1000 g of an extraction solvent (distilled water) for 3 days by cooling. The frozen extract was filtered with a filter having a pore size of 0.2 mu m to prepare a Angelica gigantis extract
<< 실시예Example 3> 황기 추출물의 제조 3> Preparation of Hwanggi Extract
황기를 잘 건조하여 세절한 후, 건조 중량 100g을 플라스크에 넣고 추출 용매(증류수) 1000g으로 3일간 냉침하여 추출하였다. 냉침된 추출물을 0.2㎛의 기공 크기를 가진 필터로 여과하여 황기 추출물을 제조하였다.The dried yellow oak was thoroughly dried, and 100 g of dry weight was placed in a flask and extracted with 1000 g of an extraction solvent (distilled water) for 3 days by cooling. The frozen extract was filtered through a filter having a pore size of 0.2 mu m to prepare an extract of hwanggi.
<< 실시예Example 4> 천궁 추출물의 제조 4> Preparation of extract
천궁을 잘 건조하여 세절한 후, 건조 중량 100g을 플라스크에 넣고 추출 용매(증류수) 1000g으로 3일간 냉침하여 추출하였다. 냉침된 추출물을 0.2㎛의 기공 크기를 가진 필터로 여과하여 천궁 추출물을 제조하였다.The celestial ghouls were well dried and cut into small pieces. 100 g of dry weight was placed in a flask, and extracted with 1000 g of an extraction solvent (distilled water) for 3 days by cooling. The frozen extract was filtered with a filter having a pore size of 0.2 탆 to prepare a cucumber extract.
<< 실시예Example 5> 생지황 추출물의 제조 5> Preparation of raw persimmon extract
생지황을 세절한 후, 중량 100g을 플라스크에 넣고 추출 용매(증류수) 1000g으로 3일간 냉침하여 추출하였다. 냉침된 추출물을 0.2㎛의 기공 크기를 가진 필터로 여과하여 생지황 추출물을 제조하였다.After the raw persimmon was cut three times, 100 g of the weight was placed in a flask, and the mixture was extracted by cooling with 1000 g of an extraction solvent (distilled water) for 3 days. The frozen extract was filtered with a filter having a pore size of 0.2 탆 to prepare a raw persimmon extract.
<< 실시예Example 6> 지부자 추출물의 제조 6> Manufacture of leaf extract
지부자를 잘 건조하여 세절한 후, 건조 중량 100g을 플라스크에 넣고 추출 용매(증류수) 1000g으로 3일간 냉침하여 추출하였다. 냉침된 추출물을 0.2㎛의 기공 크기를 가진 필터로 여과하여 지부자 추출물을 제조하였다.The papers were dried and cut three times. 100 g of dry weight was placed in a flask, and extracted with 1000 g of an extraction solvent (distilled water) for 3 days by cooling. The frozen extract was filtered with a filter having a pore size of 0.2 mu m to prepare a leaf extract.
<< 실시예Example 7> 7> 백선피White line blood 추출물의 제조 Preparation of extract
백선피를 잘 건조하여 세절한 후, 건조 중량 100g을 플라스크에 넣고 추출 용매(증류수) 1000g으로 3일간 냉침하여 추출하였다. 냉침된 추출물을 0.2㎛의 기공 크기를 가진 필터로 여과하여 백선피 추출물을 제조하였다.The white blood was well-dried, and after three-fold drying, 100 g of dry weight was added to the flask and extracted with 1000 g of an extraction solvent (distilled water) for 3 days by cooling. The frozen extract was filtered with a filter having a pore size of 0.2 mu m to prepare a white line extract.
<< 실시예Example 8> 8> 황정Hwangje 추출물의 제조 Preparation of extract
황정을 잘 건조하여 세절한 후, 건조 중량 100g을 플라스크에 넣고 추출 용매(증류수) 1000g으로 3일간 냉침하여 추출하였다. 냉침된 추출물을 0.2㎛의 기공 크기를 가진 필터로 여과하여 황정 추출물을 제조하였다.Hwangjeong was dried well, and after finishing 3 times, 100 g of dry weight was put into a flask and extracted with 1000 g of extraction solvent (distilled water) for 3 days by cooling. The frozen extract was filtered with a filter having a pore size of 0.2 mu m to prepare Huang Qing extract.
<< 실시예Example 9> 9> 노근Fagot 추출물의 제조 Preparation of extract
노근을 잘 건조하여 세절한 후, 건조 중량 100g을 플라스크에 넣고 추출 용매(증류수) 1000g으로 3일간 냉침하여 추출하였다. 냉침된 추출물을 0.2㎛의 기공 크기를 가진 필터로 여과하여 백강잠 추출물을 제조하였다.The brow were carefully dried, and then 100 g of dry weight was added to the flask and extracted with 1000 g of an extraction solvent (distilled water) for 3 days by cooling. The frozen extract was filtered with a filter having a pore size of 0.2 mu m to prepare Baitake extract.
<< 실시예Example 10> 목단피, 당귀, 황기, 천궁, 생지황, 지부자, 10> Mulberry, Angelica, Mulberry, 백선피White line blood , , 황정Hwangje , , 노근Fagot (9가지) 혼합 추출물의 제조(9 kinds) Preparation of mixed extract
상술한 실시예와 같이 각각 추출물 형태로 제조한 목단피, 당귀, 황기, 천궁, 생지황, 지부자, 백선피, 황정, 노근 추출물을 동량 주입하여 목단피, 당귀, 황기, 천궁, 생지황, 지부자, 백선피, 황정, 노근 혼합 추출물을 제조하였다.In the same manner as in the above-mentioned examples, the extracts of Mulberry, Angelica japonica, Angelica keiskei, Cryptomeria japonica, Cryptomeria japonica, Cryptomeria japonica, Hwangjeong, and Root blend extracts were prepared.
<< 실험예Experimental Example 1> 1> 항당화Antialcification 효과 effect
본 발명자들은 항당화(anti-glycation) 효능을 확인하기 위하여, L-arginine과 포도당을 이용하여 당화 저해 활성을 측정하였다.In order to confirm anti-glycation efficacy, the present inventors measured glycosylation inhibitory activity using L-arginine and glucose.
먼저, 1M 인산 완충용액(pH 7.4)을 이용하여 1M L-아르기닌(arginine), 1M 포도당을 녹여 준비하고 1M 인산 완충용액을 이용하여 시료를 50ppm이 되도록 희석해서 준비하였다. 1M L-아르기닌과 1M 인산 완충용액을 1 대 4의 비율로 섞은 다음 96-웰 플레이트에 80 μl씩 분주하였다. 여기에 각각 50ppm으로 희석한 시료와 양성대조군으로 사용될 0.01M 아미노구아니딘(aminoguanidin)을 100 μl씩 첨가하였다. 이 시료들을 잘 섞어준 다음, 마지막으로 포도당의 최종 농도가 0.1M이 되도록 1M 인산 완충용액으로 희석한 포도당을 넣은 후, 70℃에서 4시간 동안 반응 시켰다. 96-웰 플레이트를 분광 광도계를 이용하여 420 nm에서 흡광도를 측정하여 당화 정도를 측정하였다.First, 1M L-arginine and 1M glucose were dissolved by using 1M phosphate buffer (pH 7.4), and prepared by diluting the sample to 50ppm with 1M phosphate buffer solution. 1M L-arginine and 1M phosphate buffer solution were mixed at a ratio of 1: 4, and then 80 [mu] l each was added to a 96-well plate. To each sample, 100 μl of 0.01 M aminoguanidine was added to each sample to be diluted to 50 ppm and used as a positive control. These samples were mixed well and finally glucose diluted with 1M phosphate buffer solution was added to the final concentration of glucose to 0.1 M and reacted at 70 ° C for 4 hours. The degree of saccharification was measured by measuring the absorbance of the 96-well plate at 420 nm using a spectrophotometer.
하기 식의 Glycation 실험군은 1M L-아르기닌과 1M 포도당을 넣어 당화를 유발시킨 실험군이며, 시료 자체의 흡광도를 측정하기 위해서 포도당을 넣지 않고 1M L-아르기닌과 시료만을 넣어 420 nm에서 흡광도를 측정하였다. 당화 저해 활성은 다음과 같은 식으로 구할 수 있다. 실험은 각각 3회씩 수행하여 평균값으로 나타내었다.Glycation group of the following formula was an experimental group in which 1M L-arginine and 1M glucose were added to induce glycation. Absorbance was measured at 420 nm by adding 1M L-arginine and sample alone without glucose to measure the absorbance of the sample itself. The saccharide inhibitory activity can be obtained by the following formula. Experiments were performed three times each and expressed as a mean value.
[수학식 1][Equation 1]
(상기 수학식 1에서의 'Glycation 실험군'은 'Glycation 실험군의 흡광도'를 의미함)('Glycation test group' in the above equation (1) means 'Absorbance of Glycation test group')
상기 표 1의 결과에서 볼 수 있듯이, 혼합 추출물은 항당화 물질로 알려진 아미노구아니딘(Aminoguanidine)과 비교할 때, 항당화 효과가 더욱 뛰어남을 알 수 있다.As can be seen from the results shown in Table 1, the mixed extract showed better anticarcinogenic effect than aminoguanidine, which is known as an anticarcinogenic substance.
<< 실험예Experimental Example 2> 멜라닌 생성 저해 효과 2> Melanin formation inhibitory effect
멜라닌 생성 저해를 통한 미백 효과를 확인하기 위하여, Lotan R. 외(Cancer Res. 40:3345-3350, 1980)에 기재된 방법에 따라 쥐의 멜라노마 세포(B-16 mouse melanoma cell)의 배양액에, 추출물을 첨가하여 멜라닌 총량을 측정하였다. 실험 시, 먼저 쥐의 멜라노마 세포에 대하여 독성을 평가하여 독성이 없는 농도에서 미백평가를 수행하였다. 음성 대조군으로는 DMSO를, 양성 대조군으로는 알부틴(albutin)을 사용하였다.In order to confirm the whitening effect through inhibition of melanin formation, a culture solution of B-16 mouse melanoma cells in rat was cultured according to the method described in Lotan R. et al. (Cancer Res. 40: 3345-3350, 1980) The total amount of melanin was measured by adding an extract. In the experiment, first the toxicity of melanoma cells in rats was evaluated and the whitening evaluation was carried out at a concentration that is not toxic. DMSO was used as a negative control group, and albutin was used as a positive control group.
구체적으로, 시료를 최종 농도가 100 ppm이 되도록 배지에 첨가하고, 알부틴은 100 ppm이 되도록 배지에 첨가한 후 멜라노마 세포를 3일간 배양하였다. 이후, 세포들을 트립신(trypsin) 처리하여 배양용기로부터 떼어내 원심분리한 후, 멜라닌을 추출하였다. 떼어낸 세포는 수산화나트륨 용액(1N 농도) 1 ml를 가하여 10분간 끓여 멜라닌을 녹이고 분광 광도계를 이용하여, 400 nm에서 흡광도를 측정하여 생성된 멜라닌의 양을 측정하였다.Specifically, the sample was added to the medium to a final concentration of 100 ppm, arbutin was added to the medium to be 100 ppm, and melanoma cells were cultured for 3 days. Cells were then trypsinized, detached from the culture, centrifuged, and then melanin was extracted. The removed cells were incubated with 1 ml of sodium hydroxide solution (1N concentration), boiled for 10 minutes to dissolve melanin, and the absorbance was measured at 400 nm using a spectrophotometer to measure the amount of melanin produced.
상기 멜라닌 양은 단위 세포수당(1×106 cell)의 흡광도로 나타내는 방법으로 측정하였으며, 대조군에 대한 상대적인 멜라닌 총량을 저해율(%)로 계산하고 결과를 하기 표 2에 나타내었으며, 실험은 각각 3회씩 수행하여 평균값으로 나타내었다.The amount of melanin was measured by an absorbance of 1 × 10 6 cells per unit cell, and the total amount of melanin relative to the control group was calculated as the inhibition rate (%). The results are shown in Table 2 below. As shown in FIG.
(abs)Melanin production
(abs)
(100ppm)Example 10
(100 ppm)
상기 표 2의 결과에서 볼 수 있듯이, 혼합 추출물은 뛰어난 멜라닌 총량 감소 효과를 나타내어 미백 용도로 유용함을 알 수 있었다.As can be seen from the results of Table 2 above, the mixed extract showed excellent melanin total amount reduction effect and was found to be useful for whitening purposes.
<< 실험예Experimental Example 3> 항염 효과 3> Anti-inflammatory effect
항염증 효과 및 피부트러블 개선 효과를 확인하기 위하여, RAW264.7 세포주 (ATCC number: CRL-2278)를 이용한 GRIESS 법으로 nitric oxide(NO) 생성 억제력 실험을 실시하였다.In order to confirm the anti-inflammatory effect and the improvement of skin trouble, nitric oxide (NO) production inhibition experiment was performed by GRIESS method using RAW264.7 cell line (ATCC number: CRL-2278).
구체적으로, 생쥐의 대식세포인 RAW264.7 세포를 수차례 계대 배양하고, 웰 하나에 3×105 개씩 들어가도록 24-웰 프레이트에 넣은 후, 24 시간 동안 배양하였다. 이어서, 최종농도 10ppm의 농도로 시료를 함유한 세포 배지로 교체하였다. 이때, NO-생성 억제물질인 L-NMMA(L-NG-Monomethylarginine)을 양성 대조군으로 함께 처리하여 30분 동안 배양하였고, 자극원으로 LPS(Lipopolysaccharide)를 1 ㎍씩 처리하여 24시간 동안 배양하였다. 상층액을 100㎕씩 취해 96-웰 프레이트에 옮기고, GRIESS 용액을 100㎕씩 가해 상온에서 10분간 반응시키고, 540nm에서의 흡광도를 측정함으로써 NO 억제 효과를 판단하고, NO 생성 저해율(%)은 하기 수학식 2를 이용하여 계산하여 하기 표 3에 나타내었으며, 실험은 각각 3회씩 수행하여 평균값으로 나타내었다.Specifically, RAW264.7 cells, macrophages of mice, were subcultured several times, placed in 24-well plates so as to enter 3x10 5 wells into each well, and cultured for 24 hours. Subsequently, the cell culture medium containing the sample was replaced with a final concentration of 10 ppm. At this time, L-NMMA (L-NG-Monomethylarginine), which is an inhibitor of NO production, was treated together as a positive control and cultured for 30 minutes. Lipopolysaccharide (LPS) 100 μl of the supernatant was transferred to a 96-well plate, 100 μl of GRIESS solution was added thereto, and the reaction was allowed to proceed at room temperature for 10 minutes. The absorbance at 540 nm was measured to determine the NO inhibitory effect. Calculated using Equation (2) and shown in Table 3 below. Experiments were performed three times each and expressed as average values.
[수학식 2]&Quot; (2) "
NO 생성 저해율(%)={(음성대조군의 흡광도 - 각 추출물의 흡광도)/음성 대조군의 흡광도} x 100NO production inhibition rate (%) = {(absorbance of negative control - absorbance of each extract) / absorbance of negative control} x 100
(10ppm)Example 10
(10 ppm)
상기 표 3의 결과에서 알 수 있듯이, 혼합 추출물은 대표적인 항염 의약물질인 L-NMMA와 비교하였을 때, 활성이 더 높아 천연물질로써 우수한 활성을 나타냄을 알 수 있었다.As can be seen from the results in Table 3, the mixed extracts showed higher activity as a natural substance than the representative anti-inflammatory drug L-NMMA.
<< 실험예Experimental Example 4> 콜라겐 합성 촉진 효과 4> Promoting collagen synthesis
시료를 인간 유래 섬유아세포의 배양액에 첨가하여 세포수준에서 제1형 콜라겐 합성 촉진 효과를 확인하였다. 합성된 콜라겐의 측정은 PICP EIA kit(Procollagen Type I C-Peptide Enzyme Immuno Assay KIT)를 이용하여 정량하였다. 콜라겐 합성량을 측정하기 위해 시료를 최종 농도가 10 ppm이 되도록 섬유아세포의 배양배지(DMEM 배지)에 첨가하여 48 시간 배양한 후 배양액을 취하여 PICP EIA 키트로 각 농도에서 제1형 콜라겐 합성 정도를 분광광도계를 이용하여 450 nm에서 측정하였다.The sample was added to the culture medium of human - derived fibroblasts to confirm the promoting effect of type I collagen synthesis at the cellular level. The synthesized collagen was quantitated using a PICP EIA kit (Procollagen Type I C-Peptide Enzyme Immuno Assay Kit). In order to measure the amount of collagen synthesis, the sample was added to a fibroblast culture medium (DMEM medium) at a final concentration of 10 ppm and cultured for 48 hours. The culture broth was taken and the degree of type 1 collagen synthesis was measured at each concentration using a PICP EIA kit And measured at 450 nm using a spectrophotometer.
효과의 비교를 위하여 시료를 처리하지 않은 섬유아세포의 배양배지(음성대조군)와 비타민 C(양성대조군)를 최종농도 52.85 ㎍/ml가 되도록 첨가한 시료에 대하여 동일한 방법으로 콜라겐 합성 정도를 측정하였다. 콜라겐 생성 증가율은 음성 대조군에 대한 상대적인 콜라겐 생성량의 비율로 계산하고 결과를 하기 표 4에 나타내었으며, 실험은 4회씩 수행하여 평균값으로 나타내었다.For the comparison of the effects, the degree of collagen synthesis was measured in the same manner for the samples in which the culture medium of the untreated fibroblasts (negative control) and vitamin C (positive control) were added to a final concentration of 52.85 / / ml. The increase rate of collagen production was calculated by the ratio of relative collagen production to the negative control, and the results are shown in Table 4 below.
상기 표 4의 결과에서 볼 수 있듯이, 혼합 추출물을 처리한 경우에 콜라겐 합성을 증가시켰고, 일반적으로 콜라겐 합성을 유도하는 것으로 잘 알려진 비타민C를 적용한 경우와 유사한 정도의 콜라겐 합성 효과를 나타내었다.As can be seen from the results in Table 4, the collagen synthesis was increased when the mixed extract was treated, and the collagen synthesis effect was similar to that of vitamin C, which is generally known to induce collagen synthesis.
<< 실험예Experimental Example 5> 5> 엘라스테이즈Ella Stays 활성 저해 효과 Active inhibitory effect
엘라스틴(Elastin)을 분해하는 효소인 엘라스테이즈(Elastase)의 활성 저해 효과를 다음과 같이 확인하였다.The activity inhibitory effect of Elastase, an enzyme that degrades elastin, was confirmed as follows.
엘라스테이즈(Elastase)는 사람의 백혈구 세포로부터 유래한 엘라스테이즈를 사용하였고, 엘라스테이즈의 기질로 합성 기질인 MeOSuc-Ala-Ala-Pro-Val-pNA를 사용하였다. 완충 용액은 100mM의 Tris(pH 7.5) 용액을 사용하였다. 엘라스테이즈는 완충용액을 이용하여 최종적으로 0.2 mU을 사용하였다. 또한, 엘라스테이즈의 합성 기질은 DMSO를 이용하여 100mM 용액을 만든 후 최종 농도가 0.5mM이 되도록 완충용액을 이용하여 희석하였다. 이 때, 양성 대조군은 엘라스테이즈 저해 물질로 알려진 쿼세틴(Quercetin)을 10ppm 농도로 넣은 것으로 설정하였다. 엘라스테이즈 저해 후보는 최종농도가 10ppm이 되도록 첨가하였다. 반응은 96-웰 플레이트에서 진행하였으며, 상온에서 20분간 반응시켰다. 분광 광도계를 이용하여 1분 간격으로 405 nm에서 흡광도를 측정하여, 시간 대비 흡광도의 기울기를 구하여 효소의 활성도로 정하였다. 엘라스테이즈 저해율은 다음과 같이 계산하였다.Elastase used Elastase from human leukocyte cells and MeOSuc-Ala-Ala-Pro-Val-pNA as synthetic substrate for Elastase. The buffer solution used was 100 mM Tris (pH 7.5) solution. Finally, 0.2 mU was used for the ELASTASES using a buffer solution. In addition, the synthetic substrate of Elastinase was diluted with a buffer solution to make a final concentration of 0.5 mM after making a 100 mM solution using DMSO. At this time, the positive control group was set to contain 10 ppm of quercetin, which is known as an inhibitor of Elastinase. Ellazease inhibition candidates were added to give a final concentration of 10 ppm. The reaction was carried out in a 96-well plate and allowed to react at room temperature for 20 minutes. Absorbance was measured at 405 nm using a spectrophotometer at intervals of 1 minute, and the slope of the absorbance versus time was determined as the activity of the enzyme. Ella stasis inhibition rates were calculated as follows.
[수학식 3]&Quot; (3) "
엘라스테이즈 저해율은 상기 수학식 3을 이용해 계산하여 하기 표 5에 나타내었으며, 실험은 각각 3회씩 수행하여 평균값으로 나타내었다.The inhibition rate of the ELA stase was calculated using the above equation (3) and shown in the following Table 5, and the experiment was performed three times each and expressed as an average value.
(10ppm)Example 10
(10 ppm)
상기 표 5의 결과에서 볼 수 있듯이, 혼합 추출물을 처리한 경우, 양성대조군인 쿼세틴과 비교하여서도 우수한 엘라스테이즈 활성 저해 효과를 발휘함을 알 수 있었다. 따라서, 혼합 추출물은 피부재생, 주름개선을 위한 용도로 사용할 수 있음을 알 수 있었다.As can be seen from the results of Table 5, when the mixed extract was treated, it was found that Elastase activity inhibitory effect excelling even in comparison with the positive control group, quercetin, was exerted. Therefore, it was found that the mixed extract can be used for skin regeneration and wrinkle improvement.
<< 실험예Experimental Example 6> 항산화 효과 6> Antioxidant effect
혼합 추출물에 대한 자유라디칼 소거능을 1,1-디페닐-2-피크릴히드라질(DPPH) 방법으로 측정하였다(Blois, Nature 181, 1190, 1958). DPPH는 비교적 안정한 자유라디칼로서 라디칼 상태로 존재 시 517㎚에서 최대 흡광을 보이며 라디칼이 소거되면 흡광성을 잃는다. DPPH는 시그마(Sigma)사의 것을 사용하였으며, 0.15 mM의 농도로 메틸 알코올에 녹여 사용하였다.The free radical scavenging ability of the mixed extract was measured by the 1,1-diphenyl-2-picryl hydrazyl (DPPH) method (Blois, Nature 181, 1190, 1958). DPPH is a relatively stable free radical, which shows maximum absorption at 517 nm in the presence of radicals and loses its absorbance when radicals are eliminated. DPPH was purchased from Sigma, and dissolved in methyl alcohol at a concentration of 0.15 mM.
먼저, 혼합 추출물 또는 양성 대조군인 비타민 C를 96-웰 플레이트의 각각의 웰에 100μl씩 넣었다. 여기에 DPPH 용액을 100μl씩 첨가한 다음 상온에서 30분간 방치하고 마이크로플레이트 리더(BioTek EL-340)를 이용하여 517㎚에서의 흡광도를 측정하였다.First, 100 μl of mixed extract or vitamin C, a positive control, was added to each well of a 96-well plate. 100 μl of DPPH solution was added thereto, and the mixture was allowed to stand at room temperature for 30 minutes, and the absorbance at 517 nm was measured using a microplate reader (BioTek EL-340).
시료를 처리한 것의 흡광도가 대조군의 흡광도의 절반이 될 때의 추출물의 농도를 IC50으로 표시한 결과를 하기 표 6에 나타내었다. 본 실험은 3회 반복하였다.IC 50 represents the concentration of the extract when the absorbance of the sample is half the absorbance of the control group. The results are shown in Table 6 below. This experiment was repeated 3 times.
상기 표 6과 같이, 혼합 추출물은 비타민 C와 비교할 때도 자유라디칼 소거능이 매우 강력하였으며, 이로써 항산화 효과가 우수함을 확인하였다.As shown in Table 6, the mixed extracts showed a strong free radical scavenging ability as compared with vitamin C, confirming that the antioxidative effect was excellent.
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