KR102326086B1 - Composition for Improving Skin Conditions Having Skin Whitening, Moisturizing, Anti-Wrinkle and Skin Barrier Property Comprising Complex Extracts of Broussonetia kazinoki as Active Ingredient - Google Patents
Composition for Improving Skin Conditions Having Skin Whitening, Moisturizing, Anti-Wrinkle and Skin Barrier Property Comprising Complex Extracts of Broussonetia kazinoki as Active Ingredient Download PDFInfo
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- KR102326086B1 KR102326086B1 KR1020210005856A KR20210005856A KR102326086B1 KR 102326086 B1 KR102326086 B1 KR 102326086B1 KR 1020210005856 A KR1020210005856 A KR 1020210005856A KR 20210005856 A KR20210005856 A KR 20210005856A KR 102326086 B1 KR102326086 B1 KR 102326086B1
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Abstract
Description
본 발명은 닥나무 복합 추출물을 유효성분으로 포함하는 피부 미백, 주름개선, 보습, 및 피부장벽 강화용 조성물에 관한 것으로, 더욱 구체적으로 닥나무(Broussonetia kazinoki), 비타민나무(Hippophae rhamnoides), 및 석류나무(Punica granatum)의 복합 추출물을 유효성분으로 포함하는 피부 미백, 주름개선, 보습, 및 피부장벽 강화용 조성물에 관한 것이다.The present invention relates to a composition for skin whitening, wrinkle improvement, moisturizing, and skin barrier strengthening comprising a mulberry complex extract as an active ingredient, and more particularly, mulberry ( Broussonetia kazinoki ), vitamin tree ( Hippophae rhamnoides ), and pomegranate ( Punica granatum ) It relates to a composition for skin whitening, wrinkle improvement, moisturizing, and skin barrier strengthening comprising a complex extract of an active ingredient.
생물체의 항상성(homeostasis)을 유지하기 위한 과정에서 중요한 역할을 담당하고 있는 물질 중 일부가 각종 생물체 유래의 생리 활성 물질이다. 지금까지 수많은 생리 활성 물질에 관해 많은 연구가 진행되고 있으며, 그 중, 미생물, 식물, 또는 동물체 등에서 분리된 생리 활성을 갖는 물질에 관한 연구는 생명과학 및 의학 분야에서 매우 중요한 영역이다.Some of the substances that play an important role in the process for maintaining homeostasis of living organisms are physiologically active substances derived from various living organisms. Until now, many studies have been conducted on numerous physiologically active substances, and among them, research on substances having physiological activity isolated from microorganisms, plants, or animals, etc. is a very important field in life science and medicine.
한편, 종래 개발된 비타민C, 레티노익산, 형질전환생장인자(transforming growth factor, TGF) 등은 피부 주름 개선, 미백, 항산화 등에 효과가 있는 것으로 알려져 있다. 또한, 대표적인 피부 미백 성분으로는 코지산(Kojic acid), 알부틴(Arbutin) 등과 같은 티로시나제 효소 활성을 억제하는 물질로 잘 알려져 있다. 그러나, 종래 개발된 비타민C, 레티노익산, 형질전환 생장인자(transforming growth factor, TGF), 동물태반 유래의 단백질(JP8-231370), 베툴린산(betulinic acid, JP8-208424), 클로렐라 추출물(JP9-40523, JP10-36283, 섬유아세포 증식 촉진작용) 등의 소재들은 피부에 적용할 때 발생하는 자극과 발적 등의 안전성 문제로 사용량에 제한이 있거나, 그 효과가 미미하여 실질적으로 피부 탄력 및 주름, 미백 등의 개선에 효과를 기대할 수 없다는 문제점이 있었다.On the other hand, conventionally developed vitamin C, retinoic acid, transforming growth factor (TGF), etc. are known to be effective in skin wrinkle improvement, whitening, antioxidant, and the like. In addition, as a representative skin whitening component, it is well known as a substance that inhibits tyrosinase enzyme activity, such as kojic acid and arbutin. However, conventionally developed vitamin C, retinoic acid, transforming growth factor (TGF), animal placenta-derived protein (JP8-231370), betulinic acid (JP8-208424), chlorella extract (JP9- 40523, JP10-36283, fibroblast proliferation promoting action), etc., have limited usage due to safety issues such as irritation and redness that occur when applied to the skin, or their effect is insignificant, resulting in substantial skin elasticity, wrinkles, whitening, etc. There was a problem that the effect could not be expected in the improvement of
또한, 최근에는 천연물 등으로부터 얻은 생리 활성 물질을 화장료에 도입하여 피부 자체의 고유 방어 기능을 더욱 강화함으로써 피부에 작용하는 기능성을 증가시키는 화장품의 개발이 활발하게 진행되는 추세이며, 생체에 안전하고, 유효성분이 안정하며, 무엇보다도 기존의 피부 미백, 피부 주름 개선, 보습, 및 피부 재생 효과가 있는 물질보다 효과가 더욱 우수한 천연소재의 개발이 절실히 요망되고 있다.In addition, in recent years, the development of cosmetics that increase the functionality acting on the skin by introducing physiologically active substances obtained from natural products into cosmetics to further strengthen the skin's own natural defense function is actively progressing, and it is safe to the living body, The active ingredient is stable, and above all, there is an urgent need to develop a natural material that is more effective than the existing substances that have skin whitening, skin wrinkle improvement, moisturizing, and skin regeneration effects.
한편, 화장품 소재는 안전성, 효능, 물성 등이 균형을 이루어야 한다. 또한 경제성 확보도 중요한 요인으로서 화장품용 소재는 다양한 개발 방법, 천연물 추출과 미생물 등에 의한 발효를 이용하여 피부 투과, 노화 억제, 생리활성, 안전성 및 물성을 최적화하여 개발하게 된다. 화장품과 의약품의 경계가 무너지면서 화장품의 안정성과 의약품의 효과성을 겸비한 복합 기능성 원료 및 바이오 소재를 통해 다양한 코스메슈티컬 제품들이 높은 성장세를 이어가고 있다.On the other hand, cosmetic materials must be balanced in safety, efficacy, and physical properties. In addition, securing economic feasibility is an important factor, and cosmetic materials are developed by optimizing skin permeation, aging inhibition, physiological activity, safety and physical properties using various development methods, extraction of natural products and fermentation by microorganisms. As the boundary between cosmetics and pharmaceuticals is collapsing, various cosmeceutical products are continuing to grow rapidly through complex functional raw materials and biomaterials that combine the stability of cosmetics and the effectiveness of pharmaceuticals.
이러한 배경 아래에서, 본 발명자들은 피부 탄력 또는 피부 주름 개선에 효과가 우수한 천연물 소재의 발굴을 위해 예의 연구노력한 결과, 천연소재들의 조합 중 닥나무, 비타민나무, 및 석류나무의 복합 추출물이 피부 미백, 주름개선, 보습, 및 피부장벽 강화 활성에 있어서 우수한 시너지 효과를 나타냄을 규명하여, 본 발명을 완성하게 되었다.Under this background, the present inventors have made intensive research efforts to discover natural materials that are effective in improving skin elasticity or skin wrinkles. As a result, complex extracts of mulberry tree, vitamin tree, and pomegranate tree among the combinations of natural materials are effective for skin whitening and wrinkles. The present invention was completed by finding that it exhibits an excellent synergistic effect in improvement, moisturizing, and skin barrier strengthening activity.
따라서, 본 발명의 주된 목적은 닥나무, 비타민나무, 및 석류나무의 복합 추출물을 유효성분으로 포함하는 피부 미백, 주름개선, 보습, 및 피부장벽 강화용 화장료 조성물을 제공하는 데 있다.Accordingly, the main object of the present invention is to provide a cosmetic composition for skin whitening, wrinkle improvement, moisturizing, and skin barrier strengthening comprising a complex extract of mulberry tree, vitamin tree, and pomegranate tree as an active ingredient.
본 발명의 다른 목적은 닥나무, 비타민나무, 및 석류나무의 복합 추출물을 유효성분으로 포함하는 피부 미백, 주름개선, 보습, 및 피부장벽 강화용 식품 조성물을 제공하는 데 있다.Another object of the present invention is to provide a food composition for skin whitening, wrinkle improvement, moisturizing, and skin barrier strengthening comprising a complex extract of mulberry tree, vitamin tree, and pomegranate tree as an active ingredient.
본 발명의 다른 목적 및 이점은 하기의 발명의 상세한 설명, 청구범위 및 도면에 의해 보다 명확하게 된다.Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.
본 발명의 한 양태에 따르면, 본 발명은 닥나무, 비타민나무, 및 석류나무의 복합 추출물을 유효성분으로 포함하는 피부 미백, 주름개선, 보습, 및 피부장벽 강화용 화장료 조성물을 제공한다.According to one aspect of the present invention, the present invention provides a cosmetic composition for skin whitening, wrinkle improvement, moisturizing, and skin barrier strengthening comprising a complex extract of mulberry tree, vitamin tree, and pomegranate tree as an active ingredient.
본 발명자들은 피부 미백, 주름개선, 보습, 및 피부장벽 강화 효과가 우수한 천연 식물체 추출물의 조합에 관하여 연구를 수행하였으며, 본 발명의 닥나무, 비타민나무, 및 석류나무의 복합 추출물이 세포 독성이 없고, 멜라닌 색소의 생성을 억제하며, 히알루론산의 합성을 촉진하고, 필라그린 합성을 촉진하는데 시너지 효과가 있는 것을 확인할 수 있었다(도 1 내지 도 5 참조). The present inventors conducted a study on a combination of natural plant extracts having excellent skin whitening, wrinkle improvement, moisturizing, and skin barrier strengthening effects, and the complex extract of mulberry, vitamin tree, and pomegranate tree of the present invention has no cytotoxicity, It was confirmed that there is a synergistic effect in inhibiting the production of melanin pigment, promoting the synthesis of hyaluronic acid, and promoting the filaggrin synthesis (see FIGS. 1 to 5).
본 발명의 "닥나무(Broussonetia kazinoki)"는 쌍떡잎식물 쐐기풀목 뽕나무과의 낙엽활엽 관목을 의미하고, "비타민나무(Hippophae rhamnoides)"는 보리수나뭇과(Elaeagnaceae)의 긴 가시로 덮여 있는 낙엽 관목으로 갈매보리수나무라고도 하며, "석류나무(Punica granatum)"는 쌍떡잎식물 도금양목 석류나무과의 낙엽소교목을 의미한다. 본 발명의 상기 닥나무, 비타민나무, 및 석류나무의 추출물은, 천연, 잡종 또는 변종 식물로부터 추출될 수 있다. "Mugwort (Broussonetia kazinoki )" of the present invention means a deciduous broad-leaved shrub of the dicotyledonous nettle family, and "vitamin tree ( Hippophae rhamnoides )" is a deciduous shrub covered with long thorns of the Elaeagnaceae family, and the gull tree Also called a tree, "pomegranate tree ( Punica granatum )" means a deciduous small arboreous tree of the dicotyledonous plant Pomegranate family. The extracts of the mulberry tree, the vitamin tree, and the pomegranate tree of the present invention may be extracted from natural, hybrid or mutant plants.
본 발명에서 사용되는 용어, "추출물"은 닥나무, 비타민나무, 및 석류나무의 추출 처리로 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. 본 발명의 상기 추출물은 바람직하게 추출 후 건조 분말 형태로 제조되어 사용될 수 있다. 상기 닥나무, 비타민나무, 및 석류나무의 추출에 있어서, 상기 추출물을 추출하는 방법은 특별히 제한되지 아니하며, 당해 기술 분야에서 통상적으로 사용하는 방법에 따라 추출할 수 있다. 상기 추출방법의 비제한적인 예로는, 열수 추출법, 초음파 추출법, 여과법, 환류 추출법 등을 들 수 있으며, 이들은 단독으로 수행되거나 2종 이상의 추출방법을 병용하여 수행될 수 있다. As used herein, the term "extract" refers to an extract obtained by extraction treatment of mulberry, vitamin tree, and pomegranate tree, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, a prepared or purified product of the extract, Or a mixture thereof, and the like, including extracts of all formulations that can be formed using the extract itself and the extract. The extract of the present invention may be prepared and used in the form of a dry powder after extraction. In the extraction of the mulberry tree, the vitamin tree, and the pomegranate tree, the method of extracting the extract is not particularly limited, and may be extracted according to a method commonly used in the art. Non-limiting examples of the extraction method include hot water extraction, ultrasonic extraction, filtration, reflux extraction, and the like, and these may be performed alone or in combination of two or more extraction methods.
본 발명에서 상기 닥나무, 비타민나무, 및 석류나무를 추출하는데 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, 부틸렌글리콜, 프로필렌글리콜 등의 다가 알코올; 및 메틸아세테이트, 에틸아세테이트, 아세톤, 벤젠, 헥산, 디에틸에테르, 디클로로메탄 등의 탄화수소계 용매; 또는 이들의 혼합물을 사용할 수 있으며, 바람직하게, 물, 저급알코올, 1,3-부틸렌글리콜, 에틸아세테이트를 단독으로 사용하거나 2종 이상 혼합하여 사용할 수 있다. In the present invention, the type of extraction solvent used for extracting the mulberry tree, the vitamin tree, and the pomegranate tree is not particularly limited, and any solvent known in the art may be used. Non-limiting examples of the extraction solvent include water; C1-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; Alternatively, a mixture thereof may be used, and preferably, water, lower alcohol, 1,3-butylene glycol, and ethyl acetate may be used alone or in a mixture of two or more.
본 발명에서 열수 추출 또는 냉침 추출한 추출물은 부유하는 고체 입자를 제거하기 위해 여과, 예를 들어 나일론 등을 이용해 입자를 걸러내거나 냉동여과법 등을 이용해 여과한 후, 그대로 사용하거나 이를 동결건조, 열풍건조, 분무건조 등을 이용해 건조하여 사용할 수 있다.In the present invention, the extract extracted with hot water or chilled extract is filtered to remove floating solid particles, for example, using nylon or the like to filter the particles, or filtered using freeze filtration, etc. It can be used by drying it using spray drying or the like.
본 발명에서 사용되는 용어, "분획물"은 여러 다양한 구성성분들을 포함하는 혼합물로부터 특정 성분 또는 특정 성분 그룹을 분리하기 위하여 분획을 수행하여 얻어진 결과물을 의미한다. As used herein, the term "fraction" refers to a result obtained by performing fractionation in order to separate a specific component or a specific group of components from a mixture including various components.
상기 분획 방법의 비제한적인 예로는, 닥나무, 비타민나무, 및 석류나무를 추출하여 얻은 추출물에 소정의 용매를 처리하여 상기 추출물로부터 분획물을 얻는 방법을 들 수 있다. 본 발명에서 상기 분획물을 얻는 데에 사용되는 용매의 종류는 특별히 제한되지 아니하며, 당해 기술 분야에서 공지된 임의의 용매를 사용할 수 있다. 상기 분획 용매의 비제한적인 예로는 물, 알코올 등의 극성 용매; 헥산(Hexane), 에틸아세테이트(Ethyl acetate), 클로로포름(Chloroform), 디클로로메탄(Dichloromethane) 등의 비극성 용매 등을 들 수 있다. 이들은 단독으로 사용되거나 2종 이상 혼합하여 사용될 수 있다. 상기 분획 용매 중 알코올을 사용하는 경우에는 구체적으로는 C1 내지 C4의 알코올을 사용할 수 있다.Non-limiting examples of the fractionation method include a method of obtaining a fraction from the extract by treating the extract obtained by extracting the mulberry tree, the vitamin tree, and the pomegranate tree with a predetermined solvent. The kind of solvent used to obtain the fraction in the present invention is not particularly limited, and any solvent known in the art may be used. Non-limiting examples of the fractionation solvent include polar solvents such as water and alcohol; and non-polar solvents such as hexane, ethyl acetate, chloroform, and dichloromethane. These may be used alone or in combination of two or more. In the case of using alcohol in the fractionation solvent, specifically C1 to C4 alcohol may be used.
본 발명의 화장료 조성물은 상기 닥나무, 비타민나무, 및 석류나무 추출물을 조합하여 사용할 수 있는데, 상기 이들 성분들을 조합하는 경우, 조합비는 특별히 이에 제한되지 않으나, 상기 닥나무, 비타민나무, 및 석류나무 추출물의 혼합비율은 1 : 0.5 - 2 : 0.5 - 2의 중량비로 조합하여 사용할 수 있다.The cosmetic composition of the present invention may be used in combination with the mulberry tree, vitamin tree, and pomegranate extract, and when combining these ingredients, the combination ratio is not particularly limited thereto, but the extract of the mulberry tree, vitamin tree, and pomegranate tree The mixing ratio can be used in combination in a weight ratio of 1: 0.5 - 2: 0.5 - 2.
본 발명의 닥나무, 비타민나무, 및 석류나무의 복합 추출물은 멜라닌(Melanin) 색소의 생성을 억제하여 피부 미백 활성을 나타내는 것을 특징으로 한다. 본 발명의 바람직한 실시예에서는 3종 식물 추출물의 조합으로 멜라닌 함량이 현저하게 감소되고 피부 미백에 대한 시너지 효과가 있음을 확인할 수 있었다(도 2 참조). The complex extract of mulberry tree, vitamin tree, and pomegranate tree of the present invention is characterized in that it suppresses the production of melanin pigment to exhibit skin whitening activity. In a preferred embodiment of the present invention, it was confirmed that the melanin content was significantly reduced by the combination of the three plant extracts and there was a synergistic effect on skin whitening (see FIG. 2 ).
본 발명의 닥나무, 비타민나무, 및 석류나무의 복합 추출물은 MMP-1 단백질의 발현을 억제하여 피부 주름 개선 활성을 나타내는 것을 특징으로 한다. 본 발명의 바람직한 실시예에서는 상기 닥나무, 비타민나무, 및 석류나무의 복합 추출물의 처리로 MMP-1 단백질 발현이 각각 21%, 53% 및 62% 감소되는 것을 확인한 바 있으며, 3종 식물 추출물의 조합으로 피부 주름 개선에 대한 시너지 효과가 있음을 확인할 수 있었다(도 3 참조).The complex extract of mulberry tree, vitamin tree, and pomegranate tree of the present invention is characterized in that it exhibits skin wrinkle improvement activity by suppressing the expression of MMP-1 protein. In a preferred embodiment of the present invention, it was confirmed that the MMP-1 protein expression was reduced by 21%, 53%, and 62%, respectively, by the treatment of the complex extract of the mulberry tree, the vitamin tree, and the pomegranate tree, and the combination of the three plant extracts It was confirmed that there is a synergistic effect on skin wrinkle improvement (see FIG. 3).
본 발명의 닥나무, 비타민나무, 및 석류나무의 복합 추출물은 히알루론산 합성을 촉진하여 피부 보습 활성을 나타내는 것을 특징으로 한다. 상기 용어 "피부 보습"은 피부에 수분감을 증가시켜주고, 촉촉한 상태를 유지하는 것을 의미한다. 본 발명의 바람직한 실시예에서는 상기 3종 식물 복합 추출물의 처리로 히알루론산의 생성이 현저하게 증가되는 시너지 효과가 있음을 확인할 수 있었으며, 이러한 효과는 각각의 단일 추출물 대비 현격하게 증가된 효과임을 확인할 수 있었다(도 4 참조). The complex extract of mulberry tree, vitamin tree, and pomegranate tree of the present invention is characterized in that it exhibits skin moisturizing activity by promoting hyaluronic acid synthesis. The term "moisturizing the skin" means to increase the feeling of moisture in the skin and to maintain a moist state. In a preferred embodiment of the present invention, it was confirmed that there is a synergistic effect in which the production of hyaluronic acid is significantly increased by the treatment of the three plant complex extracts, and it can be confirmed that this effect is a significantly increased effect compared to each single extract. There was (see Fig. 4).
또한, 본 발명의 닥나무, 비타민나무, 및 석류나무의 복합 추출물은 필라그린 유전자 발현을 촉진하여 피부 장벽 강화 활성을 나타내는 것을 특징으로 한다. 필라그린은 표피 과립층에서 케라토히알린 과립(keratohyalin granule)을 형성하는 단백질인 프로필라그린(profilaggrin)의 형태로 존재하다가, 각질형성세포의 최종분화과정에서 필라그린으로 분해된 후에 각질세포막을 형성할 때 케라틴 필라멘트를 응집하여 각질세포의 단단하고 편평한 구조를 형성함으로써 피부 장벽에서 벽돌(brick)의 역할을 한다. 본 발명의 바람직한 실시예에서는 상기 닥나무, 비타민나무, 및 석류나무의 복합 추출물을 처리한 처리군은 필라그린(filaggrin) 유전자의 mRNA 발현량이 각각 118%(1μg/mL), 135%(10μg/mL) 및 172%(100μg/mL) 증가한 것으로 관찰되었으며, 각각의 단일 추출물 대비 현격하게 증가된 필라그린(filaggrin) 유전자 발현을 확인할 수 있었다(도 5 참조). In addition, the complex extract of mulberry tree, vitamin tree, and pomegranate tree of the present invention is characterized in that it exhibits skin barrier strengthening activity by promoting filaggrin gene expression. Filaggrin exists in the form of profilagrin, a protein that forms keratohyalin granules in the granular layer of the epidermis. It acts as a brick in the skin barrier by aggregating keratin filaments to form a hard and flat structure of keratinocytes. In a preferred embodiment of the present invention, in the treatment group treated with the complex extract of mulberry tree, vitamin tree, and pomegranate tree, mRNA expression levels of filaggrin gene were 118% (1 μg/mL) and 135% (10 μg/mL), respectively. ) and 172% (100 μg/mL) was observed to increase, and it was possible to confirm significantly increased filaggrin gene expression compared to each single extract (see FIG. 5 ).
본 발명의 화장료 조성물은 천연 식물 소재에서 유래하여 인체에 해가 없으며, 그 첨가로 인해 화장품의 물질의 품질을 손상하지 않으므로 화장료 조성물에 포함되어 사용되는데 적합하다. 특히, 피부 미백, 주름개선, 보습, 및 피부장벽 강화와 관련된 기능이 있으므로, 기능성 화장료 조성물에 포함되어 사용되는 것이 바람직하다.The cosmetic composition of the present invention is derived from natural plant materials and is harmless to the human body, and does not impair the quality of the cosmetic material due to its addition, so it is suitable for being included in the cosmetic composition. In particular, since it has functions related to skin whitening, wrinkle improvement, moisturizing, and skin barrier strengthening, it is preferably included in the functional cosmetic composition.
본 발명의 화장료 조성물은 스킨로션, 스킨소프너, 스킨토너, 아스트린젠트, 로션, 밀크로션, 모이스쳐 로션, 영양로션, 마사지크림, 영양크림, 모이스처크림, 핸드크림, 파운데이션, 에센스, 영양에센스, 마스크팩, 비누, 클렌징폼, 클렌징로션, 클렌징크림, 바디로션 및 바디클린저에서 선택된 어느 하나의 제형으로 제조할 수 있으나, 이에 제한되는 것은 아니다.The cosmetic composition of the present invention includes skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisture cream, hand cream, foundation, essence, nourishing essence, mask pack, It may be prepared in any one formulation selected from soap, cleansing foam, cleansing lotion, cleansing cream, body lotion and body cleanser, but is not limited thereto.
본 발명의 상기 화장료 조성물은 일반 피부 화장료에 배합되는 화장품학적으로 허용 가능한 담체를 1종 이상 추가로 포함할 수 있으며, 통상의 성분으로 예를 들면 유분, 물, 계면 활성제, 보습제, 저급 알코올, 증점제, 킬레이트제, 색소, 방부제, 향료 등을 적절히 배합할 수 있으나, 이에 제한되는 것은 아니다.The cosmetic composition of the present invention may further include one or more cosmetically acceptable carriers to be formulated in general skin cosmetics, and common ingredients include, for example, oil, water, surfactant, humectant, lower alcohol, and thickener. , a chelating agent, a colorant, a preservative, a fragrance, and the like may be appropriately blended, but the present invention is not limited thereto.
본 발명의 화장료 조성물에 포함되는 화장품학적으로 허용 가능한 담체는 화장료 조성물의 제형에 따라 다양하다.The cosmetically acceptable carrier included in the cosmetic composition of the present invention varies depending on the formulation of the cosmetic composition.
본 발명의 제형이 연고, 페이스트, 크림 또는 젤인 경우에는, 담체 성분으로서 동물성 유, 식물성 유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크, 산화아연 등이 이용될 수 있으나, 이에 제한되는 것은 아니다. 이들은 단독으로 사용되거나 2종 이상 혼합되어 사용될 수 있다.When the formulation of the present invention is an ointment, paste, cream, or gel, as a carrier component, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, etc. may be used, but is not limited thereto. These may be used alone or in combination of two or more.
본 발명의 제형이 파우더 또는 스프레이인 경우에는, 담체 성분으로서 락토스, 탈크, 실리카, 알루미늄 히드록사이드, 칼슘 실케이트, 폴리아미드 파우더 등이 이용될 수 있고, 특히 스프레이인 경우에는 추가적으로 클로로플루오로하드로카본, 프로판/부탄 또는 디메틸 에테르와 같은 추진제를 포함할 수 있으나, 이에 제한되는 것은 아니다. 이들은 단독으로 사용되거나 2종 이상 혼합되어 사용될 수 있다.When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, etc. may be used as a carrier component, and in particular, in the case of a spray, additional chlorofluorohard propellants such as, but not limited to, locarbon, propane/butane or dimethyl ether. These may be used alone or in combination of two or more.
본 발명의 제형이 용액 또는 유탁액인 경우에는, 담체 성분으로서 용매, 용해화제 또는 유탁화제 등이 이용될 수 있으며, 예컨대 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌 글리콜, 1,3-부틸글리콜 오일 등이 이용될 수 있고, 특히, 목화씨 오일, 땅콩 오일, 옥수수 배종 오일, 올리브 오일, 피마자 오일 및 참깨 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜 또는 소르비탄의 지방산 에스테르가 이용될 수 있으나, 이에 제한되는 것은 아니다. 이들은 단독으로 사용되거나 2종 이상 혼합되어 사용될 수 있다.When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer, or emulsifier may be used as a carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, Propylene glycol, 1,3-butylglycol oil and the like can be used, and in particular, cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol aliphatic ester, fatty acid ester of polyethylene glycol or sorbitan may be used, but is not limited thereto. These may be used alone or in combination of two or more.
본 발명의 제형이 현탁액인 경우에는, 담체 성분으로서 물, 에탄올 또는 프로필렌 글리콜과 같은 액상의 희석제, 에톡실화 이소스테아릴 알코올, 폴리옥시에틸렌 소르비톨 에스테르 및 폴리옥시에틸렌 소르비탄 에스테르와 같은 현탁제, 미소결정성 셀룰로오스, 알루미늄 메타하이드록시드, 벤토나이트, 아가 또는 트라칸트 등이 이용될 수 있으나, 이에 제한되는 것은 아니다. 이들은 단독으로 사용되거나 2종 이상 혼합되어 사용될 수 있다.When the formulation of the present invention is a suspension, as a carrier component a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters; Crystalline cellulose, aluminum metahydroxide, bentonite, agar or tracanth may be used, but is not limited thereto. These may be used alone or in combination of two or more.
본 발명의 제형이 비누인 경우에는, 담체 성분으로서 지방산의 알칼리 금속 염, 지방산 헤미에스테르 염, 지방산 단백질 히드롤리제이트, 이세티오네이트, 라놀린 유도체, 지방족 알코올, 식물성 유, 글리세롤, 당 등이 이용될 수 있으나, 이에 제한되는 것은 아니다. 이들은 단독으로 사용되거나 2종 이상 혼합되어 사용될 수 있다.When the formulation of the present invention is a soap, alkali metal salts of fatty acids, fatty acid hemiester salts, fatty acid protein hydrolysates, isethionate, lanolin derivatives, aliphatic alcohols, vegetable oils, glycerol, sugar, etc. are used as carrier components. may be, but is not limited thereto. These may be used alone or in combination of two or more.
본 발명의 화장료 조성물에서, 상기 닥나무, 비타민나무, 및 석류나무 추출물은 구체적으로는 건조 중량으로, 화장료 조성물 총 중량의 0.001 중량% 내지 50 중량%로 포함될 수 있으며, 보다 구체적으로는 0.005 중량% 내지 10 중량%로 포함될 수 있다. 상기 범위 내에서 피부 미백, 주름개선, 보습, 및 피부장벽 강화에 대한 우수한 효능을 나타내는 이점이 있으며, 조성물의 제형이 안정화되는 이점이 있다.In the cosmetic composition of the present invention, the extracts of mulberry tree, vitamin tree, and pomegranate tree may be specifically included by dry weight, in an amount of 0.001% to 50% by weight of the total weight of the cosmetic composition, and more specifically, 0.005% by weight to It may be included in 10% by weight. Within the above range, there is an advantage of showing excellent efficacy for skin whitening, wrinkle improvement, moisturizing, and skin barrier strengthening, and there is an advantage in that the formulation of the composition is stabilized.
본 발명의 다른 양태에 따르면, 본 발명은 닥나무, 비타민나무, 및 석류나무의 복합 추출물을 유효성분으로 포함하는 피부 미백, 주름개선, 보습, 및 피부장벽 강화용 식품 조성물을 제공한다. According to another aspect of the present invention, the present invention provides a food composition for skin whitening, wrinkle improvement, moisturizing, and skin barrier strengthening comprising a complex extract of mulberry tree, vitamin tree, and pomegranate tree as an active ingredient.
본 발명의 상기 닥나무, 비타민나무, 및 석류나무의 복합 추출물은 세포 독성이 없고, 멜라닌 색소의 생성을 억제하며, 피부 주름을 개선하고, 히알루론산의 합성을 촉진하며, 필라그린 합성을 촉진하는데 시너지 효과가 있으므로, 피부 미백, 주름개선, 보습, 및 피부 장벽 강화와 관련된 피부 상태의 개선을 위한 식품 조성물로 유용하게 사용될 수 있다. The complex extract of the mulberry tree, vitamin tree, and pomegranate tree of the present invention has no cytotoxicity, inhibits the production of melanin pigment, improves skin wrinkles, promotes the synthesis of hyaluronic acid, and promotes filaggrin synthesis. Since it is effective, it can be usefully used as a food composition for skin whitening, wrinkle improvement, moisturizing, and improvement of skin conditions related to skin barrier strengthening.
본 발명의 닥나무, 비타민나무, 및 석류나무의 복합 추출물과 피부 미백, 주름개선, 보습, 및 피부장벽 강화에 대한 효과는 상기에서 설명한 바와 같다. 상기 식품 조성물은 건강기능식품의 형태로 사용될 수 있으나, 이에 제한되는 것은 아니다. The complex extract of mulberry tree, vitamin tree, and pomegranate tree of the present invention and the effect on skin whitening, wrinkle improvement, moisturizing, and skin barrier strengthening are as described above. The food composition may be used in the form of health functional food, but is not limited thereto.
본 발명의 식품 조성물은 닥나무, 비타민나무, 및 석류나무의 복합 추출물의 분획물 또는 이의 가공물의 형태로 포함될 수 있다. 또한, 상기 조성물은 유효성분 이외에 식품학적으로 허용 가능한 식품보조첨가제를 포함할 수 있다.The food composition of the present invention may be included in the form of a fraction of a complex extract of mulberry tree, vitamin tree, and pomegranate tree, or a processed product thereof. In addition, the composition may include a food pharmaceutically acceptable food supplement additive in addition to the active ingredient.
본 발명에 있어서, "식품보조첨가제"란 식품에 보조적으로 첨가될 수 있는 구성요소를 의미하며, 각 제형의 건강기능식품을 제조하는데 첨가되는 것으로서 당업자가 적절히 선택하여 사용할 수 있다. 식품보조첨가제의 예로는 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 충진제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산음료에 사용되는 탄산화제 등이 포함되지만, 상기 예들에 의해 본 발명의 식품보조첨가제의 종류가 제한되는 것은 아니다.In the present invention, "food supplementary additive" means a component that can be added to food as an auxiliary, added to the manufacture of health functional food of each formulation can be appropriately selected and used by those skilled in the art. Examples of food supplement additives include various nutrients, vitamins, minerals (electrolytes), synthetic flavoring agents and flavoring agents such as natural flavoring agents, coloring agents and fillers, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners , pH adjuster, stabilizer, preservative, glycerin, alcohol, carbonation agent used in carbonated beverages, etc., but the above examples are not limited to the type of food supplement additive of the present invention.
본 발명의 식품 조성물에는 건강기능식품이 포함될 수 있다. 본 발명에 있어서, "건강기능식품"이란 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 정제, 캅셀, 분말, 과립, 액상 및 환 등의 형태로 제조 및 가공한 식품을 말한다. 여기서 “기능성”이라 함은 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건 용도에 유용한 효과를 얻는 것을 의미하며, 구체적으로 피부 미백, 주름개선, 보습, 및 피부장벽 강화의 효과를 의미한다. 본 발명의 건강기능식품은 당업계에서 통상적으로 사용되는 방법으로 제조 가능하며, 상기 제조시에는 당업계에서 통상적으로 첨가하는 원료 및 성분을 첨가하여 제조할 수 있다. The food composition of the present invention may include a health functional food. In the present invention, "health functional food" refers to a food manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc. using raw materials or ingredients useful for the human body. Here, “functionality” refers to obtaining useful effects for health purposes such as regulating nutrients or physiological actions with respect to the structure and function of the human body, specifically, the effects of skin whitening, wrinkle improvement, moisturizing, and skin barrier strengthening. means The health functional food of the present invention can be prepared by a method commonly used in the art, and at the time of manufacture, it can be prepared by adding raw materials and components commonly added in the art.
또한, 상기 건강기능식품의 제형 또한 건강기능식품으로 인정되는 제형이면 제한 없이 제조될 수 있다. 본 발명의 식품 조성물은 다양한 형태의 제형으로 제조될 수 있으며, 일반 약품과는 달리 식용가능한 식물 추출물을 원료로 하여 약품의 장기 복용 시 발생할 수 있는 부작용 등이 없는 장점이 있고, 휴대성이 뛰어나며, 멜라닌 색소의 생성을 억제하고, 피부 주름을 개선하며, 히알루론산의 합성을 촉진하고, 필라그린 합성을 촉진하는 효과를 증진시키기 위한 보조제로 섭취할 수 있다.In addition, the dosage form of the health functional food may also be manufactured without limitation as long as it is a dosage form recognized as a health functional food. The food composition of the present invention can be prepared in various types of dosage forms, and unlike general drugs, it uses an edible plant extract as a raw material and has the advantage of not having side effects that may occur during long-term administration of the drug, and has excellent portability, It can be taken as an adjuvant to suppress the production of melanin pigment, improve skin wrinkles, promote the synthesis of hyaluronic acid, and enhance the effect of promoting the synthesis of filaggrin.
본 발명의 건강기능식품이 취할 수 있는 형태에는 제한이 없으며, 통상적인 의미의 식품을 모두 포함할 수 있고, 기능성 식품 등 당업계에 알려진 용어와 혼용하여 사용할 수 있다. 아울러 본 발명의 건강기능식품은 당업자의 선택에 따라 식품에 포함될 수 있는 적절한 기타 보조 성분과 공지의 첨가제를 혼합하여 제조할 수 있다. 첨가할 수 있는 식품의 예로는 육류, 소세지, 빵, 초콜릿, 캔디류, 스낵류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알코올 음료 및 비타민 복합제 등이 있으며, 본 발명에 따른 닥나무, 비타민나무, 및 석류나무의 복합 추출물을 주성분으로 하여 제조한 즙, 차, 젤리 및 주스 등에 첨가하여 제조할 수 있다. 또한, 동물을 위한 사료로 이용되는 식품도 포함된다.There is no limitation on the form that the health functional food of the present invention can take, and it may include any food in a conventional sense, and may be used in combination with terms known in the art, such as functional food. In addition, the health functional food of the present invention can be prepared by mixing known additives with other suitable auxiliary ingredients that may be included in the food according to the selection of those skilled in the art. Examples of foods that can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages and There are vitamin complexes, and the like, and it can be prepared by adding the complex extract of mulberry, vitamin tree, and pomegranate according to the present invention as a main component to juice, tea, jelly, juice, and the like. Also included are foods used as feed for animals.
본 발명의 특징 및 이점을 요약하면 다음과 같다:The features and advantages of the present invention are summarized as follows:
(1) 본 발명은 닥나무, 비타민나무, 및 석류나무의 복합 추출물을 유효성분으로 포함하는 피부 미백, 주름개선, 보습, 및 피부장벽 강화용 조성물을 제공한다.(1) The present invention provides a composition for skin whitening, wrinkle improvement, moisturizing, and skin barrier strengthening comprising a complex extract of mulberry tree, vitamin tree, and pomegranate tree as an active ingredient.
(2) 본 발명의 닥나무, 비타민나무, 및 석류나무로 구성된 3종 복합 추출물은 세포 독성이 없고, 멜라닌 색소의 생성을 억제하며, 피부 주름을 개선하고, 히알루론산의 합성을 촉진하며, 필라그린 합성을 촉진하는데 시너지 효과가 있으므로, 화장품, 식품, 및 의약품 분야에서 피부 미백, 주름개선, 보습, 및 피부장벽 강화의 목적으로 유용하게 이용 가능하다.(2) Three kinds of complex extracts composed of mulberry tree, vitamin tree, and pomegranate tree of the present invention have no cytotoxicity, inhibit the production of melanin pigment, improve skin wrinkles, promote the synthesis of hyaluronic acid, filaggrin Since it has a synergistic effect in promoting synthesis, it can be usefully used for the purpose of skin whitening, wrinkle improvement, moisturizing, and skin barrier strengthening in cosmetics, food, and pharmaceutical fields.
(3) 또한, 본 발명의 조성물은 인체에 매우 안전할 뿐만 아니라, 안정성도 매우 탁월하다.(3) In addition, the composition of the present invention is not only very safe for the human body, but also has excellent stability.
도 1은 닥나무, 비타민나무, 및 석류나무의 단일 또는 복합 추출물을 인간각질형성세포(HaCaT)에 농도별로 처리하고, 세포 생존률을 측정하여 나타낸 그래프이다.
도 2는 닥나무, 비타민나무, 및 석류나무의 단일 또는 복합 추출물을 멜란-A 세포에 농도별로 처리하고, 멜라닌 함량을 측정하여 나타낸 그래프이다.
도 3은 닥나무, 비타민나무, 및 석류나무의 단일 또는 복합 추출물을 인간섬유아세포(NHDFs, normal human dermal fibroblast)에 농도별로 처리하고, MMP-1 단백질의 발현량 변화를 ELISA 방법으로 측정하여 나타낸 그래프이다.
도 4는 닥나무, 비타민나무, 및 석류나무의 단일 또는 복합 추출물을 HaCaT 세포에 농도별로 처리하고, 히알루론산 생성량 변화를 측정하여 나타낸 그래프이다.
도 5는 닥나무, 비타민나무, 및 석류나무의 단일 또는 복합 추출물을 HaCaT 세포에 농도별로 처리하고, 필라그린 유전자 발현 변화를 측정하여 나타낸 그래프이다.1 is a graph showing the human keratinocytes (HaCaT) treated with single or complex extracts of mulberry tree, vitamin tree, and pomegranate tree by concentration, and the cell viability was measured.
FIG. 2 is a graph showing single or complex extracts of mulberry tree, vitamin tree, and pomegranate tree by concentration in melan-A cells and measuring the melanin content.
3 is a graph showing changes in the expression level of MMP-1 protein measured by ELISA method by treating single or complex extracts of mulberry tree, vitamin tree, and pomegranate tree by concentration in human fibroblasts (NHDFs, normal human dermal fibroblasts); am.
4 is a graph showing changes in the amount of hyaluronic acid produced by treating single or complex extracts of mulberry tree, vitamin tree, and pomegranate tree by concentration in HaCaT cells.
5 is a graph showing changes in filaggrin gene expression by treating single or complex extracts of mulberry tree, vitamin tree, and pomegranate tree by concentration in HaCaT cells.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하기로 한다. 이들 실시예는 단지 본 발명을 예시하기 위한 것이므로, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지는 않는다.Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and therefore, the scope of the present invention should not be construed as being limited by these examples.
실시예 1. 닥나무, 비타민나무, 및 석류나무의 추출물 제조Example 1. Preparation of extracts of mulberry tree, vitamin tree, and pomegranate tree
1-1. 닥나무 추출물의 제조1-1. Preparation of mulberry extract
건조된 닥나무 전초 100g을 분쇄한 후 3L의 정제수를 용매로 사용하여 12시간 가온 환류추출한 후, 냉침 시키고, 1.2㎛ 투과 사이즈를 갖는 여과지로 여과하여 여과액을 수득하였다. 수득된 여과액을 감압농축 및 감압 건조로 농축하여 닥나무 건조 분말을 제조하였다. After pulverizing 100 g of dried mulberry seedlings, using 3 L of purified water as a solvent, heating and refluxing for 12 hours, followed by cooling, and filtering with filter paper having a 1.2 μm permeation size to obtain a filtrate. The obtained filtrate was concentrated under reduced pressure and dried under reduced pressure to prepare dry mulberry powder.
1-2. 비타민나무 추출물의 제조1-2. Preparation of vitamin tree extract
건조된 비타민나무 전초 100g을 분쇄한 후 3L의 정제수를 용매로 사용하여 12시간 가온 환류추출한 후, 냉침 시키고, 1.2㎛ 투과 사이즈를 갖는 여과지로 여과하여 여과액을 수득하였다. 수득된 여과액을 감압농축 및 감압 건조로 농축하여 비타민나무 건조 분말을 제조하였다. After crushing 100 g of dried vitamin tree outpost, using 3 L of purified water as a solvent, heating and refluxing for 12 hours, followed by cooling, and filtering with filter paper having a 1.2 μm permeation size to obtain a filtrate. The obtained filtrate was concentrated under reduced pressure and dried under reduced pressure to prepare a dry vitamin tree powder.
1-3. 석류나무 추출물의 제조1-3. Preparation of pomegranate extract
건조된 석류나무 전초 100g을 분쇄한 후 3L의 정제수를 용매로 사용하여 12시간 가온 환류추출한 후, 냉침 시키고, 1.2㎛ 투과 사이즈를 갖는 여과지로 여과하여 여과액을 수득하였다. 수득된 여과액을 감압농축 및 감압 건조로 농축하여 석류나무 건조 분말을 제조하였다. After pulverizing 100 g of dried pomegranate outpost, using 3 L of purified water as a solvent, heating and refluxing extraction for 12 hours, cooling and filtering with filter paper having a 1.2 μm permeation size to obtain a filtrate. The obtained filtrate was concentrated under reduced pressure and dried under reduced pressure to prepare pomegranate dry powder.
1-4. 닥나무, 비타민나무, 및 석류나무의 복합 추출물 제조1-4. Manufacture of complex extracts of mulberry tree, vitamin tree, and pomegranate tree
건조된 닥나무, 비타민나무, 및 석류나무의 전초를 동량으로 혼합한 혼합소재 100g을 분쇄한 후 3L의 정제수를 용매로 사용하여 12시간 가온 환류추출한 후, 냉침 시키고, 1.2㎛ 투과 사이즈를 갖는 여과지로 여과하여 여과액을 수득하였다. 수득된 여과액을 감압농축 및 감압 건조로 농축하여 닥나무, 비타민나무, 및 석류나무의 3종 식물 복합소재의 건조 분말을 제조하였다. 100 g of mixed material mixed in equal amounts of dried mulberry tree, vitamin tree, and pomegranate plant was pulverized, heated and refluxed for 12 hours using 3 L of purified water as a solvent, then cooled and cooled and filtered with a filter paper having a penetration size of 1.2 μm Filtration gave a filtrate. The obtained filtrate was concentrated under reduced pressure and dried under reduced pressure to prepare dry powder of a composite material of three types of plants: mulberry tree, vitamin tree, and pomegranate tree.
실시예 2. 식물 추출물 첨가에 따른 세포 생존 시험Example 2. Cell viability test according to the addition of plant extracts
닥나무, 비타민나무, 및 석류나무의 단일 또는 복합 추출물들이 인간각질형성세포(HaCaT, Human immortalized keratinocyte)의 생장에 미치는 효과를 아래와 같은 방법으로 확인하였다. The effect of single or complex extracts of mulberry tree, vitamin tree, and pomegranate tree on the growth of human immortalized keratinocytes (HaCaT) was confirmed by the following method.
우선, 10% FBS(fetal bovine serum, Cambrex)가 포함된 전용 배지인 DMEM(Dulbeccos modified Eagles medium) 배지에 인간각질형성세포를 24 well 세포 배양 접시에 1×105 농도로 접종한 후 24시간 동안, 37℃, 5% CO2 습윤 조건 하에서 배양하였다. 이후, 배지를 제거하고 무혈청 DMEM 배지로 희석한 실시예 1-1 내지 1-4에서 제조한 닥나무, 비타민나무, 및 석류나무의 단일 또는 복합 추출물들을 농도별로 처리하여 24시간 동안 배양하였다. 24시간 후, 1mg/mL의 MTT를 처리하고 2시간 후, 세포에 MTT 처리로 인해 생성된 포마잔(formazan)을 DMSO로 녹여 570nm에서 흡광도를 측정하였다. First, human keratinocytes were inoculated in a 24-well cell culture dish at a concentration of 1×10 5 in DMEM (Dulbeccos modified Eagles medium) medium, which is a dedicated medium containing 10% FBS (fetal bovine serum, Cambrex), for 24 hours. , 37° C., 5% CO 2 Incubated under humid conditions. Thereafter, the medium was removed and the single or complex extracts of mulberry trees, vitamin trees, and pomegranates prepared in Examples 1-1 to 1-4 diluted with serum-free DMEM medium were treated at different concentrations and cultured for 24 hours. 24 hours later, 1 mg/mL of MTT was treated, and 2 hours later, formazan generated by MTT treatment in cells was dissolved in DMSO and absorbance was measured at 570 nm.
그 결과, 인간각질형성세포에 닥나무, 비타민나무, 및 석류나무의 단일 또는 복합 추출물들을 처리한 경우, 세포 생존율에 크게 영향을 미치지 않는 것으로 확인되었으며, 오히려 닥나무, 비타민나무, 및 석류나무의 복합 추출물을 처리한 실험군에서는 인간각질형성세포의 성장이 농도 의존적으로 증가하는 것으로 나타났다. As a result, when human keratinocytes were treated with single or complex extracts of mulberry tree, vitamin tree, and pomegranate tree, it was confirmed that the cell viability was not significantly affected, rather, complex extracts of mulberry tree, vitamin tree, and pomegranate tree In the experimental group treated with , it was found that the growth of human keratinocytes increased in a concentration-dependent manner.
상기 결과를 통해 닥나무, 비타민나무, 및 석류나무의 단일 또는 복합 추출물들이 인체에 독성을 나타내지 않음을 확인할 수 있었다(도 1 참조).From the above results, it was confirmed that single or complex extracts of mulberry tree, vitamin tree, and pomegranate tree did not show toxicity to the human body (see FIG. 1 ).
실시예 3. 닥나무, 비타민나무, 및 석류나무 복합 추출물의 미백 활성Example 3. Whitening activity of mulberry tree, vitamin tree, and pomegranate tree complex extract
본 발명의 닥나무, 비타민나무, 및 석류나무의 복합 추출물이 갖는 미백효과를 검증하기 위해, 선행문헌(Hosoi et al., 1985)에 보고된 방법을 수정하여 멜라닌 함량을 측정하였다. In order to verify the whitening effect of the complex extract of mulberry tree, vitamin tree, and pomegranate tree of the present invention, the method reported in the prior literature (Hosoi et al., 1985) was modified to measure the melanin content.
먼저, 멜란-A(Melan-a) 세포에 상기 실시예 1에서 제조한 닥나무, 비타민나무, 및 석류나무의 단일 또는 복합 추출물을 1, 10, 및 100ug/ml의 농도로 3일 동안 처리한 후, 배지를 제거하고, PBS(phosphate buffer saline)로 두 번씩 세척하였다. 이후, 멜라닌을 용해하기 위해, 1N의 NaOH 400㎕를 첨가하고, 마이크로플레이트 리더를 사용하여 405㎚의 흡수 파장에서 멜라닌 함량을 측정하였다. First, after treating melan-A (Melan-a) cells with single or complex extracts of mulberry tree, vitamin tree, and pomegranate tree prepared in Example 1 at a concentration of 1, 10, and 100ug/ml for 3 days , the medium was removed, and washed twice with PBS (phosphate buffer saline). Then, in order to dissolve the melanin, 400 μl of 1N NaOH was added, and the melanin content was measured at an absorption wavelength of 405 nm using a microplate reader.
실험 결과, 도 2를 참조하면, 본 발명의 3종 복합 추출물이 처리된 시험군에서 농도 의존적으로 멜라닌 색소의 생성이 억제되는 것을 확인할 수 있었다. As a result of the experiment, referring to FIG. 2 , it was confirmed that the production of melanin pigment was inhibited in a concentration-dependent manner in the test group treated with the three complex extracts of the present invention.
상기 복합 추출물 처리군의 멜라닌 색소 생성 효과는 단일 추출물 처리군에 비해 현저하게 생성이 억제되는 경향을 나타내었는데, 닥나무 단일 추출물 처리군과 비교할 때 1, 10, 및 100ug/ml의 농도에서 각각 8%, 17%, 및 16% 멜라닌 색소의 생성이 감소하였고, 비타민나무 단일 추출물 처리군과 비교할 때 1, 10, 및 100ug/ml의 농도에서 각각 7%, 24%, 및 27% 멜라닌 색소의 생성이 감소하였으며, 석류나무 단일 추출물 처리군과 비교할 때 1, 10, 및 100ug/ml의 농도에서 각각 9%, 27%, 및 29% 멜라닌 색소의 생성이 감소하였다. The melanin production effect of the complex extract treatment group showed a tendency to be significantly inhibited compared to the single extract treatment group, respectively, 8% at concentrations of 1, 10, and 100ug/ml compared to the single extract treatment group of mulberry tree. , 17%, and 16% of the production of melanin was reduced, and compared with the vitamin tree single extract treatment group, the production of 7%, 24%, and 27% of melanin at concentrations of 1, 10, and 100ug/ml, respectively, was was decreased, and the production of melanin pigment was reduced by 9%, 27%, and 29% at concentrations of 1, 10, and 100ug/ml, respectively, when compared to the pomegranate single extract treatment group.
상기와 같은 결과를 통해, 본 발명의 닥나무, 비타민나무, 및 석류나무의 복합 추출물은 피부 세포에서 멜라닌 생성을 효과적으로 억제하여 피부 미백과 관련된 용도로 매우 효과적으로 사용될 수 있음을 확인할 수 있었고, 각각의 단일 추출물 대비 피부 미백에 시너지 효과가 있음을 확인할 수 있었다. Through the above results, it was confirmed that the complex extract of mulberry tree, vitamin tree, and pomegranate tree of the present invention can be used very effectively for purposes related to skin whitening by effectively inhibiting melanin production in skin cells, and each single It was confirmed that there was a synergistic effect on skin whitening compared to the extract.
실시예 3. 피부 주름 생성 억제 활성Example 3. Skin wrinkle-inhibiting activity
본 발명의 닥나무, 비타민나무, 및 석류나무로 구성된 3종 복합 추출물이 갖는 피부 주름 개선 효과를 검증하기 위해 인간섬유아세포(NHDFs, normal human dermal fibroblast)에서 MMP-1 단백질 생성 억제 효과를 측정하였다.In order to verify the skin wrinkle improvement effect of the three types of complex extracts composed of mulberry tree, vitamin tree, and pomegranate tree of the present invention, the inhibitory effect of MMP-1 protein production in normal human dermal fibroblasts (NHDFs) was measured.
실험을 위해 40mm 세포 배양 접시에 2mL의 DMEM 배양액을 넣고, 인간섬유아세포를 약 1.2x105 농도로 접종한 후, 37℃, 5% CO2 환경에서 24시간 동안 배양하였다. 그 후, 실시예 1에서 제조된 닥나무, 비타민나무, 및 석류나무의 단일 또는 복합 추출물(각각 1, 10, 및 100mg/mL)을 포함한 배지로 교체하여 3일간 배양하였다. 이후 배양액을 회수(harvest)하여 4℃, 7,500rpm 환경에서 5분간 원심분리(centrifuge)하고 ELISA 방법을 이용하여 MMP-1(Human Total MMP-1 kit, R&D Systems, Inc., Minneapolis, MN, USA)의 단백질 발현량 변화를 확인하였다. 모든 실험은 3회 반복하여 측정되었다.For the experiment, 2mL of DMEM culture medium was placed in a 40mm cell culture dish, and human fibroblasts were inoculated at a concentration of about 1.2x10 5 , and then cultured at 37° C., 5% CO 2 environment for 24 hours. Thereafter, the medium was replaced with a medium containing single or complex extracts (1, 10, and 100 mg/mL, respectively) of mulberry tree, vitamin tree, and pomegranate tree prepared in Example 1, and cultured for 3 days. Afterwards, the culture medium was harvested, centrifuged at 4° C. and 7,500 rpm for 5 minutes, and MMP-1 (Human Total MMP-1 kit, R&D Systems, Inc., Minneapolis, MN, USA) using an ELISA method. ) of the protein expression level was confirmed. All experiments were repeated three times.
실험 결과, 단일 추출물을 처리한 시험군들의 MMP-1 단백질 생성 억제 효과는 크지 않은 반면, 본 발명의 닥나무, 비타민나무, 및 석류나무로 구성된 3종 복합 추출물을 처리한 시험군에서는 MMP-1 단백질 생성량이 농도 의존적으로 크게 감소하는 양상을 확인할 수 있었다. As a result of the experiment, the MMP-1 protein production inhibitory effect of the test groups treated with a single extract was not significant, whereas the MMP-1 protein It was confirmed that the amount of production significantly decreased in a concentration-dependent manner.
구체적으로, 도 3을 참조하면, 본 발명의 닥나무, 비타민나무, 및 석류나무로 구성된 3종 복합 추출물을 처리한 시험군은 1, 10, 및 100mg/mL 농도에서 MMP-1 단백질 발현이 각각 21%, 53% 및 62% 감소하였다. Specifically, referring to FIG. 3, the test group treated with the three complex extracts consisting of mulberry tree, vitamin tree, and pomegranate tree of the present invention exhibited MMP-1 protein expression of 21 at concentrations of 1, 10, and 100 mg/mL, respectively. %, 53% and 62%.
3종 복합 추출물군은 MMP-1 단백질 발현이 닥나무 단일 추출물에 비해 각각 20%(1mg/mL), 42%(10mg/mL), 및 50%(100mg/mL) 감소하였고, 비타민나무 단일 추출물에 비해 각각 29%(1mg/mL), 45%(10mg/mL), 및 54%(100mg/mL) 감소하였으며, 석류나무 단일 추출물에 비해 각각 14%(1mg/mL), 45%(10mg/mL), 및 49%(100mg/mL) 감소하여 3종 식물 추출물의 조합으로 MMP-1 단백질 발현이 현저하게 감소하는 피부 주름 개선에 대한 시너지 효과가 있음을 확인할 수 있었다.In the three complex extract groups, MMP-1 protein expression was reduced by 20% (1mg/mL), 42% (10mg/mL), and 50% (100mg/mL), respectively, compared to the single extract of mulberry tree, and compared to 29% (1mg/mL), 45% (10mg/mL), and 54% (100mg/mL), respectively, compared to that of the pomegranate single extract, respectively, 14% (1mg/mL) and 45% (10mg/mL) ), and 49% (100mg/mL), it was confirmed that the combination of three plant extracts had a synergistic effect on skin wrinkle improvement, in which MMP-1 protein expression was significantly reduced.
상기와 같은 결과를 통해, 본 발명의 닥나무, 비타민나무, 및 석류나무로 구성된 3종 복합 추출물은 피부 세포에서 MMP-1 단백질 발현을 효과적으로 억제하여 피부 주름 개선과 관련된 용도로 효과적으로 사용될 수 있음을 확인할 수 있었다. Through the above results, it was confirmed that the three kinds of complex extracts composed of mulberry tree, vitamin tree, and pomegranate tree of the present invention can be effectively used for purposes related to skin wrinkle improvement by effectively inhibiting MMP-1 protein expression in skin cells. could
실시예 4. 피부 보습 활성Example 4. Skin moisturizing activity
본 발명의 닥나무, 비타민나무, 및 석류나무로 구성된 3종 복합 추출물의 피부 보습 효과를 검증하기 위해 이들이 히알루론산(Hyaluronan) 합성 촉진에 미치는 영향을 실험하였다.In order to verify the skin moisturizing effect of three types of complex extracts composed of mulberry tree, vitamin tree, and pomegranate tree of the present invention, their effect on promoting hyaluronan synthesis was tested.
히알루론산(HA, Hyaluronan, Hyaluronic acid)은 주로 표피 각질형성세포와 진피 섬유아세포에 의해 합성되며, 수분과 결합하여 세포가 정상적으로 활동할 수 있는 환경을 조성하는 중요한 역할을 하는 물질이다. 피부에서 히알루론산의 감소는 피부 탄력 저하 및 수분 함유량 감소의 직접적인 원인 중 하나이다. 따라서 본 실험에서는 인간 섬유아세포 내에서 히알루론산의 생성량을 확인하였다. Hyaluronic acid (HA, Hyaluronan, Hyaluronic acid) is mainly synthesized by epidermal keratinocytes and dermal fibroblasts, and is a substance that plays an important role in creating an environment in which cells can function normally by combining with water. A decrease in hyaluronic acid in the skin is one of the direct causes of a decrease in skin elasticity and a decrease in moisture content. Therefore, in this experiment, the amount of hyaluronic acid produced in human fibroblasts was confirmed.
인간각질형성세포(HaCaT)를 24 well 세포배양 접시에 1㎖의 배양액을 넣고 약 1×105의 농도로 세포를 접종한 후, 37℃, 5% CO2 환경에서 24시간 동안 배양하였다. 그 후, 상기 실시예 1에 따라 준비된 닥나무, 비타민나무, 및 석류나무의 단일 추출물과 이들의 3종 복합 추출물을 각각 1, 10, 및 100㎍/㎖를 포함한 배지로 교체하여 24시간 배양하였다. 이후 배양액을 회수(harvest)하여 4℃, 7,500rpm 환경에서 5분간 원심분리(centrifuge)한 뒤, ELISA 방법을 이용하여 히알루론산(Hyaluronan kit, R&D Systems, Inc., Minneapolis, MN, USA)의 발현량 변화를 확인하였다. Human keratinocytes (HaCaT) were placed in 1 ml of a culture solution in a 24-well cell culture dish and inoculated with cells at a concentration of about 1×10 5 , and then cultured at 37° C., 5% CO 2 environment for 24 hours. Thereafter, the single extracts of mulberry tree, vitamin tree, and pomegranate tree prepared according to Example 1 and the three complex extracts thereof were replaced with a medium containing 1, 10, and 100 μg/ml, respectively, and cultured for 24 hours. Thereafter, the culture medium was harvested and centrifuged for 5 minutes at 4° C., 7,500 rpm, and then hyaluronic acid (Hyaluronan kit, R&D Systems, Inc., Minneapolis, MN, USA) was expressed using an ELISA method. The amount change was confirmed.
실험 결과, 도 4를 참조하면, 닥나무, 비타민나무, 및 석류나무의 단일 추출물 처리군에 비해 이들의 3종 복합추출물 처리군은 1㎍/㎖ 용량에서 각각 119%, 128%, 및 131% 개선된 히알루론산 합성 증가 효과를 나타내었고, 10㎍/㎖ 용량에서 각각 117%, 133%, 및 138% 개선된 히알루론산 합성 증가 효과를 나타내었으며, 100㎍/㎖ 용량에서 각각 109%, 127%, 및 129% 개선된 히알루론산 합성 증가 효과를 나타내었다(도 3 참조).As a result of the experiment, referring to FIG. 4 , compared with the single extract treatment group of mulberry, vitamin tree, and pomegranate tree, these three complex extract treatment groups improved by 119%, 128%, and 131%, respectively, at the 1㎍/㎖ dose. showed the effect of increasing hyaluronic acid synthesis, respectively, 117%, 133%, and 138% improved hyaluronic acid synthesis at the dose of 10 μg/ml, respectively, at the dose of 100 μg/ml, 109%, 127%, and 129% improved hyaluronic acid synthesis increased effect (see FIG. 3 ).
상기와 같은 결과를 통해, 본 발명의 닥나무, 비타민나무, 및 석류나무 3종 혼합에 따라 히알루론산 합성 증가에 대한 시너지 효과가 발생하는 것을 확인할 수 있었다.Through the above results, it was confirmed that the synergistic effect on the increase in hyaluronic acid synthesis occurred according to the three types of mixture of mulberry tree, vitamin tree, and pomegranate tree of the present invention.
실시예 5. 피부 장벽 강화(Filaggrin 유전자 발현 분석)Example 5. Skin barrier strengthening (Filaggrin gene expression analysis)
닥나무, 비타민나무, 및 석류나무로 구성된 3종 복합 추출물이 필라그린(Filaggrin, FLG, 각질층 형성 인자) 유전자 발현에 미치는 영향을 실험하였다.The effect of three types of complex extracts composed of mulberry, vitamin tree, and pomegranate tree on the gene expression of filaggrin (FLG, stratum corneum) was tested.
실험을 위해 100mm 세포 배양 접시에 10% FBS를 첨가한 DMEM 배지를 첨가하고, 인간각질형성세포(HaCaT)를 1×106 세포 농도로 접종하여 37℃, 5% CO2 배양기에서 24시간 동안 배양하였다. 24시간 배양 후, 닥나무, 비타민나무, 및 석류나무의 단일 또는 복합 추출물이 각각 1, 100, 및 100㎍/㎖ 농도가 되도록 DMEM 배지에 희석하여 제조한 희석용액을 첨가하여 24시간 동안 동일 조건에서 추가로 배양하였다. 이후 Trizol reagent(invitrogen, USA)를 이용하여 세포를 회수하여 RNA를 분리하였다. RT-PCR(Step One Plus, Applied Biosystems, 미국) 방법을 이용하여 필라그린 유전자의 mRNA 발현량을 확인하였다. 사용한 프라이머는 다음과 같다:For the experiment, DMEM medium supplemented with 10% FBS was added to a 100 mm cell culture dish, and human keratinocytes (HaCaT) were inoculated at a concentration of 1×10 6 cells and cultured at 37° C., 5% CO 2 in an incubator for 24 hours. did. After culturing for 24 hours, a diluted solution prepared by diluting the single or complex extracts of mulberry tree, vitamin tree, and pomegranate tree in DMEM medium to a concentration of 1, 100, and 100 μg/ml, respectively, was added, and the solution was added under the same conditions for 24 hours. Further incubation. Thereafter, cells were recovered using Trizol reagent (invitrogen, USA) and RNA was isolated. RT-PCR (Step One Plus, Applied Biosystems, USA) was used to determine the mRNA expression level of the filaggrin gene. The primers used were as follows:
- Filaggrin : 정방향 5′-AAG CTT CAT GGT GAT GCG AC-3′, 역방향 5′-TCA AGC AGA AGA GGA AGG CA-3′- Filaggrin: Forward 5′-AAG CTT CAT GGT GAT GCG AC-3′, Reverse 5′-TCA AGC AGA AGA GGA AGG CA-3′
- GAPDH: 정방향 5'-ACC ACA GTC CAT GCC ATC AC-3', 역방향 5'-CCA CCA CCC TGT TGC TGT AG-3'.- GAPDH: forward 5'-ACC ACA GTC CAT GCC ATC AC-3', reverse 5'-CCA CCA CCC TGT TGC TGT AG-3'.
상기와 같은 프라이머를 이용하여 RT-PCR 수행하고, 그 결과를 도 4에 정리하였다. RT-PCR was performed using the primers as described above, and the results are summarized in FIG. 4 .
실험 결과, 시료 무처리군과 비교하여 본 발명의 닥나무, 비타민나무, 및 석류나무로 구성된 3종 복합 추출물을 처리한 시험군에서는 필라그린(filaggrin) 유전자의 mRNA 발현량이 각각 118%(1μg/mL), 135%(10μg/mL) 및 172%(100μg/mL) 증가한 것으로 관찰되었다(도 5 참조). As a result of the experiment, the mRNA expression level of the filaggrin gene was 118% (1 μg/mL) in the test group treated with the three complex extracts consisting of mulberry, vitamin tree, and pomegranate tree of the present invention compared to the sample untreated group, respectively. ), 135% (10 μg/mL) and 172% (100 μg/mL) increases were observed (see FIG. 5 ).
이러한 필라그린 유전자의 mRNA 발현 증가는 각각의 단일 추출물 처리군에 비해 현저하게 증가하였는데, 닥나무 단일 추출물 처리군에 비해 117%(1μg/mL), 117%(10μg/mL) 및 126%(100μg/mL) 증가하였고, 비타민나무 단일 추출물 처리군에 비해 115%(1μg/mL), 122%(10μg/mL) 및 134%(100μg/mL) 증가하였으며, 석류나무 단일 추출물 처리군에 비해 110%(1μg/mL), 112%(10μg/mL) 및 127%(100μg/mL) 증가한 것으로 나타났다(도 4 참조).The increase in mRNA expression of the filaggrin gene was significantly increased compared to each single extract treatment group, 117% (1 μg/mL), 117% (10 μg/mL) and 126% (100 μg/mL) compared to the single extract treatment group of mulberry tree. mL) increased, and 115% (1μg/mL), 122% (10μg/mL) and 134% (100μg/mL) increased compared to the group treated with the single extract of pomegranate tree, and 110% ( 1 μg/mL), 112% (10 μg/mL) and 127% (100 μg/mL) increase (see FIG. 4 ).
이상으로 본 발명의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서 이러한 구체적인 기술은 단지 바람직한 구현예일 뿐이며, 이에 본 발명의 범위가 제한되는 것이 아닌 점은 명백하다. 따라서, 본 발명의 실질적인 범위는 첨부된 청구항과 그의 등가물에 의하여 정의된다고 할 것이다.As described above in detail a specific part of the present invention, for those of ordinary skill in the art, this specific description is only a preferred embodiment, and it is clear that the scope of the present invention is not limited thereto. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.
Claims (7)
상기 닥나무, 비타민나무, 및 석류나무 추출물은 1:0.5-2:0.5-2의 중량비로 배합되고,
상기 조성물은 필라그린 유전자 발현을 촉진하여 피부 장벽 강화 활성을 나타내는 것을 특징으로 하는,
피부 미백, 주름개선, 보습, 및 피부장벽 강화용 화장료 조성물.A cosmetic composition comprising a complex extract of mulberry tree, vitamin tree, and pomegranate tree as an active ingredient,
The mulberry tree, vitamin tree, and pomegranate extract are blended in a weight ratio of 1:0.5-2:0.5-2,
The composition promotes filaggrin gene expression, characterized in that it exhibits skin barrier strengthening activity,
A cosmetic composition for skin whitening, wrinkle improvement, moisturizing, and skin barrier strengthening.
상기 조성물은 멜라닌 색소의 생성을 억제하여 피부 미백 활성을 나타내는 것을 특징으로 하는 피부 미백, 주름개선, 보습, 및 피부장벽 강화용 화장료 조성물.According to claim 1,
The composition is a cosmetic composition for skin whitening, wrinkle improvement, moisturizing, and skin barrier strengthening, characterized in that it exhibits skin whitening activity by inhibiting the production of melanin pigment.
상기 조성물은 MMP-1 단백질의 발현을 억제하여 피부 주름 개선 활성을 나타내는 것을 특징으로 하는 피부 미백, 주름개선, 보습, 및 피부장벽 강화용 화장료 조성물.According to claim 1,
The composition suppresses the expression of MMP-1 protein, and a cosmetic composition for skin whitening, wrinkle improvement, moisturizing, and skin barrier strengthening, characterized in that it exhibits skin wrinkle-improving activity.
상기 조성물은 히알루론산 합성을 촉진하여 피부 보습 활성을 나타내는 것을 특징으로 하는 피부 미백, 주름개선, 보습, 및 피부장벽 강화용 화장료 조성물.According to claim 1,
The composition promotes the synthesis of hyaluronic acid, and a cosmetic composition for skin whitening, wrinkle improvement, moisturizing, and skin barrier strengthening, characterized in that it exhibits skin moisturizing activity.
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