KR101453258B1 - Health Functional Food Composition Comprising Red Ginseng Extract, Benincasa Hispida Extract, and Apios Americana Extract for Improving Skin Beauty - Google Patents
Health Functional Food Composition Comprising Red Ginseng Extract, Benincasa Hispida Extract, and Apios Americana Extract for Improving Skin Beauty Download PDFInfo
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- KR101453258B1 KR101453258B1 KR1020120157619A KR20120157619A KR101453258B1 KR 101453258 B1 KR101453258 B1 KR 101453258B1 KR 1020120157619 A KR1020120157619 A KR 1020120157619A KR 20120157619 A KR20120157619 A KR 20120157619A KR 101453258 B1 KR101453258 B1 KR 101453258B1
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- skin
- extract
- red ginseng
- apiose
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- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/25—Araliaceae (Ginseng family), e.g. ivy, aralia, schefflera or tetrapanax
- A61K36/258—Panax (ginseng)
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- A23V2200/318—Foods, ingredients or supplements having a functional effect on health having an effect on skin health and hair or coat
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- A23V2250/00—Food ingredients
- A23V2250/20—Natural extracts
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Abstract
본 발명은 피부미용증진 용도를 갖는 건강기능식품 조성물에 관한 것으로, 자세하게는 발효홍삼, 동과자 및 아피오스 추출물의 혼합물을 유효성분으로 포함하는 피부미용증진 건강기능식품 조성물 및 이를 이용하여 제조된 건강기능식품에 관한 것이다. 본 발명의 발효홍삼, 동과자 및 아피오스 추출물의 혼합물은 MMP-1 활성 억제를 통하여 피부 주름개선 효과를 가지며, 티로시나아제 활성 저해 활성 및 멜라닌 생성 억제를 통하여 피부 미백 효과를 가질 뿐만 아니라 항산화 활성을 통하여 피부 노화 방지 효과를 동시에 가진다. 따라서 이를 유효성분으로 포함하는 본 발명의 조성물은 피부 주름개선, 피부 미백 및 피부 노화 방지를 통해 우수한 피부미용증진 효과를 나타낼 수 있어 기능성 건강기능식품 또는 건강기능 보조식품의 소재로서 유용하게 사용될 수 있다. 특히 본 발명의 발효홍삼, 동과자 및 아피오스 추출물의 혼합물은 천연물로부터 유래한 추출물로서, 인체에 매우 안전하다는 장점이 있다.The present invention relates to a health functional food composition having skin beauty enhancing use, and more particularly, to a skin health enhancing functional food composition comprising a fermented red ginseng, a frozen plant and a mixture of apiose extract as an active ingredient and a health function It is about food. The mixture of fermented red ginseng, frog and apiose extract of the present invention has a skin wrinkle-reducing effect through inhibition of MMP-1 activity, has a skin whitening effect through inhibition of tyrosinase activity inhibition and melanin formation inhibition, Thereby simultaneously preventing skin aging. Therefore, the composition of the present invention containing the active ingredient as an active ingredient can exhibit an excellent skin cosmetic enhancement effect by improving skin wrinkles, skin whitening, and skin aging, and thus can be useful as a material for functional health food or health supplement . In particular, the mixture of fermented red ginseng, frog and apiose extract of the present invention is an extract derived from a natural product and has an advantage that it is very safe for human body.
Description
본 발명은 피부미용증진 용도를 갖는 건강기능식품 조성물에 관한 것으로, 자세하게는 발효홍삼, 동과자 및 아피오스 추출물을 유효성분으로 포함하는 피부미용증진 건강기능식품 조성물 및 이를 이용하여 제조된 건강기능식품에 관한 것이다.The present invention relates to a health functional food composition having skin beauty enhancement use, and more particularly, to a skin health improving functional food composition comprising fermented red ginseng, frogs and apiose extract as an active ingredient, and a health functional food .
오늘날 삶의 질이 향상되면서 과거와는 달리 외모에 대한 관심이 크게 증가하고 있다. 이는 기존의 피부에 바르는 화장품에 대한 만족을 넘어 새로운 형태의 화장품에 대한 요구로 이어져, 최근 식품을 통하여 피부노화를 방지하고, 피부 미백 및 피부 보습을 증진시키고자 하는 움직임이 크게 증가하는 추세이다.Today, as the quality of life improves, unlike the past, interest in appearance has increased greatly. This leads to a demand for a new type of cosmetics beyond the satisfaction of existing skin cosmetics. Recently, there has been a tendency to prevent skin aging through food, and to increase skin whitening and skin moisturizing.
피부는 신체를 둘러싼 조직으로서 신체를 보호하고, 체온을 조절하며, 배설과 피부호흡 등의 기능을 담당한다. 피부는 바깥층에서부터 표피(epidermis), 진피(dermis) 및 피하지방(subcutaneous fat)의 3층으로 나누어지고, 털, 땀샘, 손발톱, 피지샘 등의 표피부속기(epidermal appendage)로 구성되어 있다. 상피조직으로 구성되어 있는 표피는 다시 각질층, 과립층, 가시층 및 기저층으로 나누어지고, 주로 각질형성세포(keratinocyte)와 멜라닌형성세포(melanocyte)로 구성되어 있다. 표피층 아래에 있는 섬유성 결합조직인 진피는 상부에 유두 모양으로 돌출된 유두진피와 하부에 그물 모양으로 분포된 그물진피로 구성되어 있다. 진피는 아교섬유(collagenous fibers)와 일부 탄력섬유(elastic fibers)로 구성되어있어 질기고 탄력적이며, 모세혈관과 신경 등이 분포되어 있다. 피하지방층은 지방세포로 구성되어 있으며, 외부의 충격으로부터 내부 장기를 보호하고 열을 차단하여 체온을 유지하며 에너지를 저장하는 역할을 담당하는 것으로 알려져 있다.Skin is the surrounding tissue that protects the body, regulates body temperature, excretion and skin respiration. The skin is divided into three layers from the outer layer to the epidermis, dermis and subcutaneous fat and consists of epidermal appendages such as hair, sweat glands, nails, and sebaceous glands. The epidermis is divided into the horny layer, the granular layer, the visible layer and the basal layer. The epidermis consists mainly of keratinocyte and melanocyte. The dermis, a fibrous connective tissue below the epidermal layer, is composed of a papillary dermis protruding from the upper part and a net dermis distributed in the lower part of the net. The dermis consists of collagenous fibers and some elastic fibers, which are vaginal and elastic, and have capillaries and nerves. The subcutaneous fat layer is composed of adipocytes and is known to protect internal organs from external shocks, block heat, maintain body temperature, and store energy.
피부 노화는 크게 내인성 노화(intrinsic aging)와 광노화(photo aging)로 구분할 수 있는데, 내인성 노화는 시간이 흐름에 따라 자연적으로 나타나는 피할 수 없는 노화 현상이며, 광노화는 햇빛에 노출된 피부 부위에서 관찰되는 내인성 노화 현상과 자외선의 복합적인 요인에 의해 발생한다. 내인성 노화의 특징은 경미한 잔주름, 피부건조증, 탄력감소 현상 등이 있으며, 광노화의 특징은 굵고 깊은 주름이나 많은 잔주름, 피부상의 불규칙한 색소침착이나 여러 색소 질환의 증가, 피부가 거칠고 건조한데 따른 탄력감소 현상 등을 들 수 있다.Skin aging can be divided into intrinsic aging and photo aging. Endogenous aging is an inevitable aging phenomenon that naturally occurs over time, and photoaging is observed in areas exposed to sunlight It is caused by a combination of endogenous aging and ultraviolet rays. The characteristics of endogenous aging are slight wrinkles, dryness of skin, reduction of elasticity, etc. The characteristics of photoaging are coarse and deep wrinkles and many fine wrinkles, irregular pigmentation on skin, increase of pigment diseases, decrease of elasticity due to rough and dry skin And the like.
피부의 노화과정은 표피와 진피가 서로 다르게 진행되는데, 먼저 표피에서는 두께와 각질층 수분함량이 감소하면서 탄력성이 떨어지고 일광화상을 입은 것과 같은 세포 형태의 변화가 일어난다. 또한 색소 세포가 감소하여 특정 부위에 탈색반이 나타나며 피부면역을 담당하는 랑게르한스세포의 수가 감소하여 피부암과 같은 질환이 발생한다. 진피에서는 피부 탄력을 유지하는 교원섬유와 탄력섬유가 감소하여 피부 탄력이 떨어지고, 잔주름이 증가한다. 특히 진피 내에 존재하는 콜라겐은 피부노화와 관련된 주요 성분으로서, 지방을 제외한 피부 전체 건조중량의 77%, 진피섬유 성분의 90%를 차지하고 있다. 콜라겐은 피부의 강도, 탄력성 및 유연성을 유지하는 역할을 담당하기 때문에 콜라겐의 합성 증진 및 분해 억제는 피부미용 및 피부 노화억제와 관련하여 매우 중요하게 여겨지고 있다.The aging process of the skin progresses differently from the epidermis to the dermis. First, the epidermis decreases in the thickness and the water content of the stratum corneum, resulting in a decrease in elasticity and a change in cell shape such as a sunburn. In addition, the pigment cells are reduced, the decolorizing part appears at a specific site, and the number of Langerhans cells responsible for skin immunity is decreased, resulting in diseases such as skin cancer. In the dermis, collagen fibers and elastic fibers that maintain skin elasticity decrease, skin elasticity decreases, and fine lines increase. In particular, collagen in the dermis is a major component of skin aging, accounting for 77% of the dry weight of the entire skin, excluding fat, and 90% of the dermis fiber component. Since collagen plays a role in maintaining the strength, elasticity and flexibility of the skin, inhibition of synthesis and inhibition of collagen synthesis is considered to be very important in relation to skin cosmetic and skin aging inhibition.
희고 맑고 투명한 피부를 갖고자 하는 것 역시 현대인들의 강한 욕망 중에 하나라고 할 정도로 깨끗한 피부는 피부미용의 또 다른 주요 관심사이다. 사람의 피부색은 피부 내 표피에 존재하는 멜라닌(melanin) 농도와 분포에 따라 결정되는데, 이는 유전적인 요인 외에도 자외선이나 피로, 스트레스 등의 환경적 또는 생리적 조건에 의해서도 영향을 받는다. 멜라닌은 아미노산의 일종인 티로신(tyrosine)이 티로시나제(tyrosinase)라는 효소에 의해 도파(DOPA), 도파퀴논(Dopaquinine)으로 전환된 후, 비효소적인 산화반응을 거쳐 만들어진다. 멜라닌은 태양의 자외선으로부터 신체를 보호하는 역할을 하지만, 피부 내의 과도한 멜라닌 합성은 피부 톤을 어둡게 하고, 기미나 주근깨를 발생시킬 수도 있다. 따라서 피부 내 멜라닌 색소의 합성을 저해하는 것이 피부미백의 위한 주요 접근방법이라 할 수 있다.Clean skin is another major concern of skin care, as it is one of the strong desires of modern people to have white, clear and transparent skin. The skin color of a person is determined by the concentration and distribution of melanin present in the epidermis of the skin. It is affected not only by genetic factors but also by environmental or physiological conditions such as ultraviolet rays, fatigue, and stress. Melanin is produced by a nonenzymatic oxidation reaction after tyrosine, an amino acid, is converted to DOPA and Dopaquinine by an enzyme called tyrosinase. Melanin protects the body from the sun's ultraviolet rays, but excessive melanin synthesis in the skin can darken skin tone and cause spots or freckles. Therefore, inhibiting the synthesis of melanin pigment in the skin is a major approach for skin whitening.
한편, 불완전한 이온 상태로 운동성이 큰 활성산소는 불포화 지방산을 공격하여 과산화반응을 일으켜 체내 과산화지질을 축척함으로써, 생체기능을 저하시키고 노화를 유발하는 것으로 잘 알려져 있다. 따라서 항산화 효과가 있는 음식물의 섭취는 체내 활성산소를 제거하여 노화를 방지하고, 나아가 피부미용에도 큰 도움이 될 것으로 여겨지고 있다.On the other hand, it is well known that active oxygen having a high mobility in an incomplete ion state attacks an unsaturated fatty acid to cause a peroxidation reaction to accumulate lipid peroxidation in the body, resulting in deterioration of biological functions and aging. Therefore, intake of food having antioxidant effect is considered to prevent aging by removing active oxygen in the body, and also to be very helpful for skin beauty.
상기에서 살펴본 바와 같이, 피부미용을 증진시키기 위해서는 피부 주름개선 및 피부 미백 및 노화 방지 등 다각적인 방향의 접근이 필요하며, 이러한 기능을 동시에 가지는 소재의 발굴이야말로 피부미용 증진을 위한 기능성 식품에 유용하게 사용될 수 있을 것으로 판단된다.As described above, in order to improve skin beauty, it is necessary to approach various directions such as skin wrinkle improvement, skin whitening and anti-aging, and excavation of materials having such functions simultaneously is useful for functional foods for skin care enhancement It can be used.
이에 본 발명자들은 천연물질로부터 인체에 안전하면서도, 탁월한 피부미용증진 효과를 나타낼 수 있는 소재를 발굴하고자 약 400 여종의 천연물로부터 스크리닝을 실시하였다. 그 결과, 발효홍삼, 동과자 및 아피오스 추출물이 항산화, 주름 개선 및 미백에 효과적임을 확인하고 이를 일정비율로 혼합한 후, 항산화 효과, MMP-1 발현 억제를 통한 피부 주름개선 효과, 티로시나아제 활성 억제 및 멜라닌 합성 저해를 통한 미백 효과를 확인함으로써, 본 발명을 완성하였다. Accordingly, the present inventors have screened about 400 kinds of natural products from natural materials in order to find a material which is safe to humans and exhibits excellent skin-beauty-enhancing effect. As a result, it was confirmed that the fermented red ginseng, frog and apiose extract were effective for antioxidation, wrinkle improvement and whitening, and after mixing them at a certain ratio, the antioxidative effect, the effect of improving skin wrinkles by inhibiting MMP-1 expression, Inhibiting and inhibiting melanin synthesis, thereby completing the present invention.
따라서 본 발명의 목적은 피부 주름개선, 피부 미백 및 노화 방지에 효과적인 피부미용증진용 건강기능식품 조성물을 제공하는 것이다.Accordingly, an object of the present invention is to provide a health functional food composition for enhancing skin beauty which is effective for improving skin wrinkles, skin whitening, and prevention of aging.
또한 본 발명의 다른 목적은 피부 주름개선, 피부 미백 및 노화 방지에 효과적인 피부미용증진용 건강기능식품을 제공하는 것이다.Another object of the present invention is to provide a health functional food for enhancing skin beauty, which is effective for improving skin wrinkles, skin whitening and preventing aging.
상기와 같은 본 발명의 목적을 달성하기 위해서, 본 발명은 발효홍삼, 동과자 및 아피오스 추출물을 유효성분으로 포함하는 피부미용증진용 건강기능식품 조성물을 제공한다.In order to accomplish the above object, the present invention provides a health functional food composition for promoting skin beauty comprising fermented red ginseng, frogs and apiose extract as an active ingredient.
본 발명의 일실시예에 있어서, 상기 발효홍삼, 동과자 및 아피오스 추출물은 각각의 물, 유기용매 또는 이들의 혼합 용매 추출물일 수 있다.In one embodiment of the present invention, the fermented red ginseng, frog and apiose extract may be water, an organic solvent or a mixed solvent extract thereof.
본 발명의 일실시예에 있어서, 상기 유기용매는 메탄올(methanol), 에탄올(ethanol), 프로판올(propanol), 이소프로판올(isopropanol) 및 부탄올(butanol)로 이루어진 군으로부터 선택될 수 있다.In one embodiment of the present invention, the organic solvent may be selected from the group consisting of methanol, ethanol, propanol, isopropanol, and butanol.
본 발명의 일실시예에 있어서, 상기 발효홍삼, 동과자 및 아피오스 추출물은 발효홍삼, 동과자 및 아피오스 각각의 70% 에탄올 추출물일 수 있다.In one embodiment of the present invention, the fermented red ginseng, frog and apiose extract may be 70% ethanol extracts of fermented red ginseng, frozen ginseng, and apiose, respectively.
본 발명의 일실시예에 있어서, 상기 혼합물은 발효홍삼 추출물, 동과자 추출물 및 아피오스 추출물 각각이 1:1:1, 2:1:1, 1:2:1 또는 1:1:2의 중량비(w:w)로 혼합된 혼합물일 수 있다.In one embodiment of the present invention, the mixture is prepared by mixing a fermented red ginseng extract, a frog extract and an apiose extract in a weight ratio of 1: 1: 1, 2: 1: 1, 1: 2: 1 or 1: w: w).
본 발명의 일실시예에 있어서, 상기 혼합물은 조성물에 0.1 내지 10,000 μg/ml의 농도로 포함될 수 있다.In one embodiment of the present invention, the mixture may be included in the composition at a concentration of from 0.1 to 10,000 μg / ml.
본 발명의 일실시예에 있어서, 상기 피부미용증진용은 피부 주름개선용, 피부 미백용 또는 피부 노화 방지용일 수 있다.In one embodiment of the present invention, the skin cosmetic enhancer may be used for improving skin wrinkles, for whitening skin or for preventing skin aging.
본 발명의 일실시예에 있어서, 상기 피부 주름개선은 MMP-1 발현 억제를 통해 이루어질 수 있다.In one embodiment of the present invention, the skin wrinkle improvement can be achieved through inhibition of MMP-1 expression.
본 발명의 일실시예에 있어서, 상기 피부 미백은 티로시나아제 활성 저해 및 멜라닌 생성 억제를 통해 이루어질 수 있다.In one embodiment of the present invention, the skin whitening can be achieved through inhibition of tyrosinase activity and inhibition of melanin formation.
본 발명의 일실시예에 있어서, 상기 피부 노화 방지는 항산화 활성을 통해 이루어질 수 있다.In one embodiment of the present invention, the anti-aging of the skin can be achieved through antioxidant activity.
또한, 본 발명은 상기 조성물로 제조된 피부미용증진용 건강기능식품을 제공한다.In addition, the present invention provides a health functional food for enhancing skin beauty prepared from the composition.
본 발명의 일실시예에 있어서, 상기 식품은 음료류, 육류, 초코렛, 식품류, 과자류, 피자, 라면, 기타 면류, 껌류, 사탕류, 아이스크림류, 알코올 음료류, 비타민 복합제 및 건강보조식품류로 이루어진 군으로부터 선택될 수 있다.In one embodiment of the present invention, the food is selected from the group consisting of beverage, meat, chocolate, foods, confectionery, pizza, ram noodles, gums, candy, ice cream, alcoholic beverages, .
본 발명의 발효홍삼, 동과자 및 아피오스 추출물의 혼합물은 MMP-1 발현 억제를 통한 피부 주름개선 효과, 티로시나아제 활성 저해 및 멜라닌 생성 억제를 통한 피부 미백 개선 효과를 가질 뿐만 아니라, 항산화 활성을 통하여 피부 노화 방지 효과를 동시에 가진다. 따라서 이를 유효성분으로 포함하는 본 발명의 조성물은 피부 주름개선, 피부 미백 및 노화 방지를 통하여 우수한 피부미용증진 효과를 나타낼 수 있기 때문에, 기능성 건강기능식품 또는 건강기능 보조식품의 소재로서 유용하게 사용될 수 있다. 또한 본 발명의 발효홍삼, 동과자 및 아피오스 추출물의 혼합물은 천연추출물로서, 인체에 안전하다는 이점을 가진다.The mixture of fermented red ginseng, frost and apiose extract of the present invention not only has a skin whitening effect by inhibiting MMP-1 expression, an inhibition of tyrosinase activity and a skin whitening effect by inhibiting melanin formation, It has anti-aging effect at the same time. Therefore, the composition of the present invention containing it as an active ingredient can exhibit an excellent skin cosmetic enhancement effect by improving skin wrinkles, skin whitening and anti-aging, so that it can be usefully used as a material for functional health food or health supplement have. Further, the mixture of the fermented red ginseng, frog and apiose extract of the present invention is a natural extract and has an advantage of being safe for human body.
도 1a는 본 발명의 혼합물의 사람 섬유아세포에 대한 세포독성을 나타낸 그래프이다.
도 1b는 본 발명의 혼합물의 마우스 흑색종 세포주에 대한 세포독성을 나타낸 그래프이다.
도 2는 본 발명의 혼합물의 MMP-1 생성 저해 활성을 나타낸 그래프이다.FIG. 1A is a graph showing cytotoxicity of a mixture of the present invention to human fibroblasts. FIG.
FIG. 1B is a graph showing cytotoxicity of the mixture of the present invention on mouse melanoma cell line. FIG.
Fig. 2 is a graph showing MMP-1 production inhibitory activity of the mixture of the present invention.
본 발명은 발효홍삼, 동과자 및 아피오스 추출물의 혼합물을 유효성분으로 포함하는 피부미용증진용 건강기능식품 조성물을 제공함에 그 특징이 있다.The present invention is characterized by providing a health functional food composition for enhancing skin beauty comprising a mixture of fermented red ginseng, frogs and apiose extract as an active ingredient.
본 발명의 발효홍삼에 사용된‘홍삼(Panax ginseng C.A. Meyer)'은 인삼을 증기 또는 기타 방법으로 쪄서 익혀 말린 것으로써, 예로부터 한반도를 중심으로 민간에 널리 이용되어 왔다. 홍삼은 원재료인 인삼과 마찬가지로 면역증강, 심혈관계 조절 및 항암 효능이 있는 것으로 보고되어 있으며, 홍삼의 주요 생리활성물질로는 steroidal saponin인 여러 종류의 진세노사이드가 존재하는 것으로 잘 알려져 있다.The "red ginseng (Panax ginseng used in the fermentation of the present invention ginseng CA Meyer 'has been widely used in the private sector, especially in the Korean peninsula, since it has been steamed or steamed by ginseng or other methods. Red ginseng has been reported to have immune enhancement, cardiovascular control and anticancer effects like ginseng, which is a raw material. It is well known that ginsenoside, which is steroidal saponin, is present as a major physiologically active substance of red ginseng.
본 발명의 ‘동과자(Benincasa hispida Cogniaux)’는 박과에 속하는 한해살이 덩굴식물로써, 동아호박의 성숙한 종자를 일컫는다. 동과자는 우레아제(urease)를 다량 함유하고 있으며, 이뇨완화, 배농제 및 피부미용에 효과가 있다고 잘 알려져 있다.Of the present invention, donggwaja (Benincasa hispida Cogniaux) is an annual plant belonging to the genus Bacillus, and refers to the mature seed of the amber pumpkin. It is well known that frogs contain a large amount of urease and are effective for diuretic relief, drainage and skin care.
본 발명의 ‘아피오스(Apios americana Medikus)’는 콩과의 덩굴성 식물로서, ‘인디언 감자’라 불린다. 아피오스는 고혈압 및 고콜레스테롤에 효과적이고, 피로를 회복시키고 스태미나를 증강시키며, 아토피와 같은 피부염 치료에도 효능이 있는 것으로 잘 알려져 있다. 또한 아피오스는 감자와 비교하여 철분은 4배, 섬유직은 5배, 단백질은 6배, 칼슘은 30배에 이를 정도로 영양학적으로도 매우 우수하다.The term " Apios " americana Medikus) is a vine plant in soybean, called 'Indian potato'. Apiosis is known to be effective for hypertension and high cholesterol, restoring fatigue, enhancing stamina, and also for treating dermatitis such as atopy. In addition, Apiose has excellent nutritional value compared to potatoes such as 4 times for iron, 5 times for fiber, 6 times for protein, and 30 times for calcium.
본 발명의 ‘발효홍삼, 동과자 및 아피오스 추출물의 혼합물’은 발효홍삼, 동과자 및 아피오스 각각의 추출물이 혼합된 혼합물을 의미한다.The 'mixture of fermented red ginseng, frost and apiose extract' of the present invention means a mixture of fermented red ginseng, frozen ginseng, and apiose, respectively.
본 발명에 따른 발효홍삼, 동과자 및 아피오스 각각의 추출물은 당업계에 공지된 추출 및 분리하는 방법을 사용하여 천연으로부터 추출 및 분리하여 수득한 것을 사용할 수 있으며, 본 발명에서 정의된‘추출물’은 적절한 용매를 이용하여 발효홍삼, 동과자 및 아피오스 각각으로부터 추출한 것이며, 예를 들어, 발효홍삼, 동과자 및 아피오스 각각의 조추출물, 극성용매 가용 추출물 또는 비극성용매 가용 추출물을 모두 포함한다.The extracts of fermented red ginseng, frozen ginseng, and apiose according to the present invention can be obtained by extracting and isolating from natural substances by using extraction and separation methods known in the art, and the 'extract' For example, crude extracts of fermented red ginseng, frog and apiose, polar solvent-soluble extracts, or nonpolar solvent-soluble extracts, all of which are extracted from fermented red ginseng, frostbite and apiose using an appropriate solvent.
상기 발효홍삼, 동과자 및 아피오스 각각으로부터 추출물을 추출하기 위한 적절한 용매로는 약제학적으로 허용되는 유기용매라면 어느 것을 사용해도 무방하며, 물 또는 유기용매를 사용할 수 있으며, 이에 제한되지는 않으나, 예를 들어, 정제수, 메탄올(methanol), 에탄올(ethanol), 프로판올(propanol), 이소프로판올(isopropanol), 부탄올(butanol) 등을 포함하는 탄소수 1 내지 4의 알코올, 아세톤(acetone), 에테르(ether), 벤젠(benzene), 클로로포름(chloroform), 에틸아세테이트(ethyl acetate), 메틸렌클로라이드(methylene chloride), 헥산(hexane) 및 시클로헥산(cyclohexane) 등의 각종 용매를 단독으로 혹은 혼합하여 사용할 수 있다.As an appropriate solvent for extracting the extract from each of the fermented red ginseng, frozen ginseng, and apiose, any pharmaceutically acceptable organic solvent may be used, and water or an organic solvent may be used. Examples thereof include, but are not limited to, For example, alcohols having 1 to 4 carbon atoms, acetone, ether, etc., including purified water, methanol, ethanol, propanol, isopropanol, butanol, Various solvents such as benzene, chloroform, ethyl acetate, methylene chloride, hexane and cyclohexane may be used alone or in combination.
본 발명의 일실시예에서, 본 발명의 발효홍삼, 동과자 및 아피오스 각각의 추출물은 발효홍삼의 70% 에탄올 추출물, 동과자의 70% 에탄올 추출물 및 아피오스의 70% 에탄올 추출물일 수 있다.In one embodiment of the present invention, the extracts of fermented red ginseng, frozen fruits and apiose of the present invention may be 70% ethanol extract of fermented red ginseng, 70% ethanol extract of frogs and 70% ethanol extract of apiose.
추출 방법으로는 열수추출법, 냉침추출법, 환류냉각추출법, 용매추출법, 수증기증류법, 초음파추출법, 용출법, 압착법 등의 방법 중 어느 하나를 선택하여 사용할 수 있다. 또한, 목적하는 추출물은 추가로 통상의 분획 공정을 수행할 수도 있으며, 통상의 정제 방법을 이용하여 정제될 수도 있다. 본 발명의 발효홍삼, 동과자 및 아피오스 추출물의 제조방법에는 제한이 없으며, 공지되어 있는 어떠한 방법도 이용될 수 있다.As the extraction method, any one of the methods such as hot water extraction method, cold extraction method, reflux cooling extraction method, solvent extraction method, steam distillation method, ultrasonic extraction method, elution method and compression method can be selected and used. In addition, the desired extract may be further subjected to a conventional fractionation process or may be purified using a conventional purification method. There is no limitation to the method for producing the fermented red ginseng, frosty and apiose extract of the present invention, and any known method may be used.
예를 들면, 본 발명의 조성물에 포함되는 발효홍삼, 동과자 및 아피오스 각각의 추출물은 상기한 열수 추출 또는 용매 추출법으로 추출된 1차 추출물을, 감압 증류 및 동결 건조 또는 분무 건조 등과 같은 추가적인 과정에 의해 분말 상태로 제조할 수 있다. 또한 상기 1차 추출물을 실리카겔 컬럼 크로마토그래피(silica gel column chromatography), 박층 크로마토그래피(thin layer chromatography), 고성능 액체 크로마토그래피(high performance liquid chromatography) 등과 같은 다양한 크로마토그래피를 이용하여 추가로 정제된 분획을 얻을 수도 있다.For example, each of the extracts of fermented red ginseng, frozen ginseng and apiose contained in the composition of the present invention is subjected to an additional step such as vacuum distillation, freeze drying, spray drying, or the like, And can be produced in a powder state. Further, the primary extract can be further purified by using various chromatographies such as silica gel column chromatography, thin layer chromatography, high performance liquid chromatography and the like, You can get it.
따라서 본 발명에 있어서 발효홍삼, 동과자 및 아피오스 각각의 추출물은 추출, 분획 또는 정제의 각 단계에서 얻어지는 모든 추출액, 분획 및 정제물, 그들의 희석액, 농축액 또는 건조물을 모두 포함하는 개념이다.Therefore, in the present invention, each of the extracts of fermented red ginseng, frozen ginseng, and apiose is a concept including all the extract, fraction and purified product obtained in each step of extraction, fractionation or purification, and diluted solution, concentrate or dried product thereof.
본 발명의 일실시예에 따른 발효홍삼, 동과자 및 아피오스 각각의 추출물을 제조하는 방법을 조금 더 구체적으로 살펴보면 다음과 같다.The method for preparing the extracts of fermented red ginseng, frozen persimmon, and apiose according to an embodiment of the present invention will be described in more detail as follows.
먼저, 발효홍삼, 동과자 및 아피오스 100 g씩을 각각 70% 에탄올 1 L에 첨가한 후 가열 추출한 다음 여과하여 각각의 여과액을 수득하고, 이렇게 수득한 각각의 여과액을 감압농축 후 열풍 또는 동결 건조하여 분말화하는 과정을 통해 제조될 수 있다.First, 100 g of each of fermented red ginseng, frogs, and apiose was added to 1 L of 70% ethanol, followed by heating and extracting the mixture. Each of the filtrates thus obtained was concentrated under reduced pressure, Followed by pulverization.
본 발명의 피부미용증진용 건강기능식품 조성물은 상기 발효홍삼, 동과자 및 아피오스 각각의 추출물이 혼합한 형태의 혼합물을 유효성분으로 포함한다.The health functional food composition for enhancing skin beauty of the present invention contains as an active ingredient, a mixture of the fermented red ginseng, the frog and the apiose, respectively.
본 발명의 일실시예에 있어서, 상기 혼합물은 발효홍삼 추출물, 동과자 추출물 및 아피오스 추출물이 1:1:1, 2:1:1, 1:2:1 또는 1:1:2의 중량비(w:w)를 가질 수 있다.In one embodiment of the present invention, the mixture comprises a mixture of fermented red ginseng extract, frog extract and apiose extract in a weight ratio of w: 1: 1: 1, 2: 1: 1, 1: 2: : w).
본 발명자들은 발효홍삼, 동과자 및 아피오스 추출물의 혼합물이 MMP-1 활성 억제를 통한 피부 주름개선 효과와 티로시나아제 활성 저해 및 멜라닌 생성 억제를 통한 피부 미백 개선 효과를 가질 뿐만 아니라, 항산화 활성을 통하여 피부 노화 방지 효과를 동시에 가진다는 사실을 확인하였으며, 이를 통해 발효홍삼, 동과자 및 아피오스 추출물의 혼합물이 우수한 피부미용증진 효과를 나타낼 수 있어 기능성 건강기능식품 또는 건강기능 보조식품의 소재로서 유용하게 사용될 수 있다는 사실을 최초로 규명하였다.The present inventors have found that a mixture of fermented red ginseng, frogs, and apiose extract has an effect of improving skin wrinkle through inhibition of MMP-1 activity, inhibiting tyrosinase activity and improving skin whitening through inhibition of melanin production, It has been confirmed that the mixture of fermented red ginseng, frozen ginseng and apiose extract has a good skin cosmetic enhancing effect and is useful as a material for functional health food or health supplement food It was the first time that I could do that.
본 발명의 하기 실험예 1에서는, 본 발명의 혼합물의 자유 라디칼 소거 활성을 측정한 결과, 본 발명의 혼합물의 처리 농도에 의존적으로 DPPH 자유 라디칼 소거 활성이 증대되는 것을 확인할 수 있었다(표 1 참조).In the following Experimental Example 1 of the present invention, the free radical scavenging activity of the mixture of the present invention was measured and it was confirmed that the DPPH free radical scavenging activity was increased depending on the treatment concentration of the mixture of the present invention (see Table 1) .
또한, 본 발명의 하기 실험예 2에서는 본 발명의 혼합물의 사람 섬유아세포 및 마우스 흑색종 세포주에 대한 세포독성을 확인한 결과, 2가지 세포 모두에서 세포독성을 유발하지 않는 것을 확인할 수 있었다(도 1a 및 1b 참조)In the following Experimental Example 2 of the present invention, the cytotoxicity of the mixture of the present invention to human fibroblast and mouse melanoma cell line was confirmed, and it was confirmed that cytotoxicity was not induced in both of the two cells (FIGS. 1A and 1B) 1b)
또한 본 발명의 하기 실험예 3에서는 본 발명의 혼합물의 MMP-1 발현 저해율을 측정한 결과, 본 발명의 혼합물 모두에서 처리농도에 의존적으로 사람 섬유아세포 MMP-1 발현이 억제되는 것을 확인할 수 있었다(도 2 참조).In Experiment 3 of the present invention, the inhibitory rate of MMP-1 expression of the mixture of the present invention was measured. As a result, it was confirmed that the expression of human fibroblast MMP-1 was inhibited in all the mixtures of the present invention depending on the treatment concentration 2).
또한 본 발명의 하기 실험예 4 및 5에서는 본 발명의 혼합물의 피부 미백 활성을 측정한 결과, 본 발명의 혼합물 모두에서 처리농도에 의존적으로 티로시나아제 활성이 저해되었으며(표 2 참조), 멜라닌 생성 또한 억제되는 것을 확인할 수 있었다(표 3 참조).In the following Experimental Examples 4 and 5 of the present invention, the skin whitening activity of the mixture of the present invention was measured. As a result, tyrosinase activity was inhibited depending on the treatment concentration in all of the inventive mixtures (see Table 2) (See Table 3).
이와 같은 결과를 통해, 본 발명자들은 발효홍삼, 동과자 및 아피오스 추출물의 혼합물이 피부 주름개선, 피부 미백과 더불어 피부 노화를 방지함에 있어서 매우 효과적이며, 따라서 피부미용증진에 우수하게 작용할 수 있을 것 이라는 사실을 실험적으로 입증하였다.The present inventors have found that a mixture of fermented red ginseng, frogs and apiose extract is very effective in preventing skin wrinkles and skin whitening as well as preventing skin aging, and thus can exert an excellent effect on skin beauty enhancement I have experimentally proved the facts.
본 발명의 조성물은 건강기능식품 조성물일 수 있는데, 이러한 건강기능식품 조성물은 유효성분인 상기 발효홍삼, 동과자 및 아피오스 추출물의 혼합물을 함유하는 것 외에 통상의 식품 조성물과 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다.The composition of the present invention may be a health functional food composition. In addition to containing the above fermented red ginseng, frog and apiose extract as active ingredients, the health functional food composition may contain various flavors or natural Carbohydrates and the like as additional components.
상술한 천연 탄수화물의 예는 모노사카라이드, 예를 들어, 포도당, 과당 등; 디사카라이드, 예를 들어 말토스, 슈크로스 등; 및 폴리사카라이드, 예를 들어 덱스트린, 시클로덱스트린 등과 같은 통상적인 당, 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 상술한 향미제는 천연 향미제 (타우마틴), 스테비아 추출물(예를 들어 레바우디오시드 A, 글리시르히진 등) 및 합성 향미제 (사카린, 아스파르탐 등)를 유리하게 사용할 수 있다.Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. The above-described flavors can be advantageously used as natural flavorings (tau martin), stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.).
상기 건강기능식품 조성물은 상기 기재한 유효성분 이외에 추가로 약제학적으로 허용 가능한 담체를 1종 이상 포함하여 건강기능식품 조성물로 바람직하게 제제화할 수 있다.The health functional food composition may further comprise one or more pharmaceutically acceptable carriers in addition to the above-described active ingredients, and may be suitably formulated into a health functional food composition.
상기 건강기능식품 조성물의 제제 형태는 정제, 캅셀, 분말, 과립, 액상, 환 등이 될 수 있다. 예를 들어, 정제 또는 캡슐제의 형태로의 제제화를 위해, 유효성분은 에탄올, 글리세롤, 물 등과 같은 경구, 무독성의 약제학적으로 허용 가능한 불활성 담체와 결합될 수 있다. 또한, 원하거나 필요한 경우, 적합한 결합제, 윤활제, 붕해제 및 발색제 또한 혼합물로 포함될 수 있다. 적합한 결합제는 이에 제한되는 것은 아니나, 녹말, 젤라틴, 글루코스 또는 베타-락토오스와 같은 천연 당, 옥수수 감미제, 아카시아, 트래커캔스 또는 소듐올레이트와 같은 천연 및 합성 검, 소듐 스테아레이트, 마그네슘 스테아레이트, 소듐 벤조에이트, 소듐 아세테이트, 소듐 클로라이드 등을 포함한다. 붕해제는 이에 제한되는 것은 아니나, 녹말, 메틸 셀룰로스, 아가, 벤토니트, 잔탄 검 등을 포함한다. 액상 용액으로 제제화되는 조성물에 있어서 허용 가능한 약제학적 담체로는, 멸균 및 생체에 적합한 것으로서, 식염수, 멸균수, 링거액, 완충 식염수, 알부민 주사용액, 덱스트로즈 용액, 말토 덱스트린 용액, 글리세롤, 에탄올 및 이들 성분 중 1 성분 이상을 혼합하여 사용할 수 있으며, 필요에 따라 항산화제, 완충액, 정균제 등 다른 통상의 첨가제를 첨가할 수 있다.The form of the health functional food composition may be a tablet, a capsule, a powder, a granule, a liquid, a ring, or the like. For example, for formulation into tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Also, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included as a mixture. Suitable binders include, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, natural and synthetic gums such as corn sweeteners, acacia, tracker candles or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum and the like. Acceptable pharmaceutical carriers for compositions that are formulated into a liquid solution include sterile solutions suitable for the living body such as saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, One or more of these components may be mixed and used. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added.
본 발명의 일실시예에 있어서, 본 발명의 상기 발효홍삼, 동고자 및 아피오스 추출물의 혼합물은 조성물에 0.1 내지 10,000 ㎍/ml의 농도로 포함될 수 있으며, 조성물 총 중량에 대하여 0.1 ~ 95 중량%로 포함될 수 있다.In one embodiment of the present invention, the mixture of fermented red ginseng, corn steep liquor and apiose extract of the present invention may be contained in the composition at a concentration of 0.1 to 10,000 μg / ml, and may be 0.1 to 95% ≪ / RTI >
본 발명의 건강기능식품 조성물은 상기와 같이 제제화되어 기능성 식품으로 이용하거나, 각종 식품에 첨가할 수 있다. 본 발명의 조성물을 첨가할 수 있는 식품으로는 예를 들어, 음료류, 육류, 초코렛, 식품류, 과자류, 피자, 라면, 기타 면류, 껌류, 사탕류, 아이스크림류, 알코올 음료류, 비타민 복합제 및 건강보조식품류 등이 있다.The health functional food composition of the present invention can be formulated as described above and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, meat, chocolates, foods, confectionery, pizza, ram noodles, other noodles, gums, candy, ice cream, alcoholic beverages, vitamin complexes, .
또한 상기 건강기능식품 조성물은 유효성분인 상기 발효홍삼, 동과자 및 아피오스 추출물의 혼합물 외에 여러 가지 영양제, 비타민, 광물 (전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 중진제 (치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 그밖에 본 발명의 건강기능식품 조성물은 천연 과일 쥬스 및 과일 쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다.In addition, the health functional food composition may further comprise a flavoring agent such as various nutrients, vitamins, minerals (electrolytes), a synthetic flavor and a natural flavor, a coloring agent and an aging agent Cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols and carbonating agents used in carbonated beverages. In addition, the health functional food composition of the present invention may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks.
본 발명은 또한 상기 발효홍삼, 동과자 및 아피오스 추출물의 혼합물을 유효성분으로 포함하는 피부미용증진용 건강기능식품을 제공한다.The present invention also provides a health functional food for enhancing skin beauty comprising a mixture of fermented red ginseng, frozen persimmon and apiose extract as an active ingredient.
상기 피부미용증진은 피부 주름개선, 피부 미백 또는 피부 노화 방지를 통한 피부미용증진일 수 있다.The skin cosmetic enhancement may be skin cosmetic enhancement through skin wrinkle improvement, skin whitening or skin aging prevention.
본 발명의 건강기능식품은 피부미용증진을 목적으로, 정제, 캅셀, 분말, 과립, 액상, 환 등의 형태로 제조 및 가공할 수 있다.The health functional food of the present invention can be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, and circles for the purpose of enhancing skin beauty.
본 발명에서 “건강기능식품”이라 함은 건강기능식품에 관한 법률에 따른 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 제조 및 가공한 식품을 말하며, 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건 용도에 유용한 효과를 얻을 목적으로 섭취하는 것을 의미한다.In the present invention, the term " health functional food " refers to a food prepared or processed using raw materials or ingredients having useful functions in accordance with the Act on Health Functional Foods, and the nutrients are controlled Physiological action, etc., for the purpose of obtaining a beneficial effect.
본 발명의 건강기능식품은 통상의 식품 첨가물을 포함할 수 있으며, 식품 첨가물로서의 적합 여부는 다른 규정이 없는 한, 식품의약품안전청에 승인된 식품 첨가물 공전의 총칙 및 일반시험법 등에 따라 해당 품목에 관한 규격 및 기준에 의하여 판정한다.The health functional foods of the present invention may contain conventional food additives and, unless otherwise specified, whether or not they are suitable as food additives are classified according to the General Rules for Food Additives approved by the Food and Drug Administration, Standards and standards.
상기 “식품 첨가물 공전”에 수재된 품목으로는 예를 들어, 케톤류, 글리신, 구연산칼슘, 니코틴산, 계피산 등의 화학적 합성물; 감색소, 감초추출물, 결정셀룰로오스, 고량색소, 구아검 등의 천연첨가물; L-글루타민산나트륨제제, 면류첨가알칼리제, 보존료제제, 타르색소제제 등의 혼합제제류 등을 들 수 있다.Examples of the items listed in the above-mentioned "food additives" include chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; Natural additives such as persimmon extract, licorice extract, crystalline cellulose, high color pigment and guar gum; L-glutamic acid sodium preparations, noodle-added alkalis, preservative preparations, tar coloring preparations and the like.
예를 들어, 정제 형태의 건강기능식품은 본 발명의 유효성분인 상기 발효홍삼, 동과자 및 아피오스 추출물의 혼합물을 부형제, 결합제, 붕해제 및 다른 첨가제와 혼합한 혼합물을 통상의 방법으로 과립화한 다음, 활택제 등을 넣어 압축성형하거나, 상기 혼합물을 직접 압축 성형할 수 있다. 또한 상기 정제 형태의 건강기능식품은 필요에 따라 교미제 등을 함유할 수도 있다.For example, a health functional food in the form of tablets may be prepared by granulating a mixture of the fermented red ginseng, frog and apiose extract, which is an effective ingredient of the present invention, with an excipient, a binder, a disintegrant and other additives in a usual manner Next, a lubricant or the like may be put in a compression mold, or the mixture may be directly compression molded. In addition, the health functional food of the tablet form may contain a mating agent or the like if necessary.
캅셀 형태의 건강기능식품 중 경질 캅셀제는 통상의 경질 캅셀에 본 발명의 유효성분인 상기 발효홍삼, 동과자 및 아피오스 추출물의 혼합물을 부형제 등의 첨가제와 혼합한 혼합물을 충진하여 제조할 수 있으며, 연질 캅셀제는 상기 유효성분을 부형제 등의 첨가제와 혼합한 혼합물을 젤라틴과 같은 캅셀기제에 충진하여 제조할 수 있다. 상기 연질 캅셀제는 필요에 따라 글리세린 또는 소르비톨 등의 가소제, 착색제, 보존제 등을 함유할 수 있다.The hard capsule of the capsule-type health functional food can be prepared by filling a mixture of the fermented red ginseng, the frog and the apiose extract, which are the active ingredients of the present invention, with an additive such as an excipient into a normal hard capsule, The capsule can be prepared by filling a mixture of the active ingredient with an excipient such as an excipient into a capsule base such as gelatin. The soft capsule may contain a plasticizer such as glycerin or sorbitol, a coloring agent, a preservative and the like, if necessary.
환 형태의 건강기능식품은 본 발명의 유효성분인 상기 발효홍삼, 동과자 및 아피오스 추출물의 혼합물과 부형제, 결합제, 붕해제 등을 혼합한 혼합물을 기존에 공지된 방법으로 성형하여 조제할 수 있으며, 필요에 따라 백당이나 다른 제피제로 제피할 수 있으며, 또는 전분, 탈크와 같은 물질로 표면을 코팅할 수도 있다.The ring-shaped health functional food can be prepared by molding a mixture of the fermented red ginseng, frozen duck, and apiose extract, which is an effective ingredient of the present invention, together with excipients, binders, disintegrants, etc. by a known method, If necessary, it may be coated with white sugar or other skin care agent, or the surface may be coated with a substance such as starch or talc.
과립 형태의 건강기능식품은 본 발명의 유효성분인 상기 발효홍삼, 동과자 및 아피오스 추출물의 혼합물과 부형제, 결합제, 붕해제 등을 혼합한 혼합물을 기존에 공지된 방법으로 입상으로 제조할 수 있으며, 필요에 따라 착향제, 교미제 등을 함유할 수 있다.The granular health functional food may be prepared by granulating a mixture of the above fermented red ginseng, frozen gruel and apiose extract, excipient, binder, disintegrant, etc., which are effective ingredients of the present invention, by a conventionally known method, If necessary, a flavoring agent, a mating agent, and the like may be contained.
상기 건강기능식품은 음료류, 육류, 초코렛, 식품류, 과자류, 피자, 라면, 기타 면류, 껌류, 사탕류, 아이스크림류, 알코올 음료류, 비타민 복합제 및 건강보조식품류 등일 수 있다.The health functional food may be a beverage, a meat, a chocolate, a food, a confectionery, a pizza, a ramen, a noodle, a gum, a candy, an ice cream, an alcoholic beverage, a vitamin complex and a health supplement food.
본 발명은 또한 발효홍삼, 동과자 및 아피오스 추출물의 혼합물을 유효성분으로 포함하는 피부미용증진용 화장료 조성물을 제공한다.The present invention also provides a cosmetic composition for enhancing skin beauty comprising a mixture of fermented red ginseng, frogs and apiose extract as an active ingredient.
본 발명의 화장료 조성물은 상술한 발효홍삼, 동과자 및 아피오스 추출물의 혼합물뿐만 아니라, 화장료 조성물에 통상적으로 이용되는 성분들을 포함할 수 있으며, 예컨대 항산화제, 안정화제, 용해화제, 비타민, 안료 및 향료와 같은 통상적인 보조제, 그리고 담체를 포함할 수 있다.The cosmetic composition of the present invention may contain not only the above-mentioned mixture of fermented red ginseng, frogs and apiose extract, but also components commonly used in cosmetic compositions such as antioxidants, stabilizers, dissolving agents, vitamins, , ≪ / RTI > and carriers.
또한, 본 발명의 조성물은 상술한 발효홍삼, 동과자 및 아피오스 추출물의 혼합물 이외에, 상기 혼합물과 반응하여 피부보호 효과를 손상시키지 않는 한도에서 종래부터 사용되어오던 유기 자외선 차단제를 혼합하여 사용할 수도 있다.In addition to the above-mentioned mixture of fermented red ginseng, frog and apiose extract, the composition of the present invention may be mixed with an organic ultraviolet screening agent which has been used in the past so long as it does not impair the skin protecting effect by reacting with the above-mentioned mixture.
상기 유기 자외선 차단제로는 글리세릴파바, 드로메트리졸트리실록산, 드로메트리졸, 디갈로일트리올리에이트, 디소듐페닐디벤즈이미다졸테트라설포네이트, 디에칠헥실부타미도트리아존, 디에칠아미노하이드록시벤조일헥실벤조에이트, 디이에이-메톡시신나메이트, 로우손과 디하이드록시아세톤의 혼합물, 메칠렌비스-벤조트리아졸릴테트라메칠부틸페놀, 4-메칠벤질리덴캠퍼, 멘틸안트라닐레이트, 벤조페논-3(옥시벤존),벤조페논-4, 벤조페논-8(디옥시페벤존), 부틸메톡시디벤조일메탄, 비스에칠헥실옥시페놀메톡시페닐트리아진, 시녹세이트, 에칠디하이드록시프로필파바, 옥토크릴렌, 에칠헥실디메칠파바, 에칠헥실메톡시신나메이트, 에칠헥실살리실 레이트, 에칠헥실트리아존, 이소아밀-p-메톡시신나메이트, 폴리실리콘-15(디메치코디에칠벤잘말로네이트), 테레프탈릴리덴디캠퍼설포닉애씨드 및 그 염류, 티이에이-살리실레이트 및 아미노벤조익애씨드(파바)로 이루어진 군으로부터 선택된 1종 이상을 사용할 수 있다.Examples of the organic ultraviolet screening agent include glyceryl paraben, drometrizol trisiloxane, drometrizol, dicaloyl triolate, disodium phenyldibenzimidazole tetrasulfonate, diethylhexylbutamidotriazone, Hydroxybenzoylhexyl benzoate, di-methoxy cinnamate, a mixture of Rawson and dihydroxyacetone, methylenebis-benzotriazolyltetramethylbutylphenol, 4-methylbenzylidene camphor, menthyl anthranylate, benzophenone Benzophenone-4, benzophenone-8 (dioxyphenyl) benzene, butylmethoxydibenzoylmethane, bishexyloxyphenol methoxyphenyltriazine, synoxate, , Octocrylene, ethylhexylmethylmethylphenyl, ethylhexylmethoxycinnamate, ethylhexylsalicylate, ethylhexyltriazone, isoamyl-p-methoxy cinnamate, polysilicon-15 (dimethicone ethyl benzene A carbonate), terephthalic ylidene dikaem peoseol sulphonic Acid and their salts, tea yieyi - may be used salicylate and at least one member selected from the group consisting of amino-benzo ripening Acid (paba).
본 발명의 화장료 조성물을 첨가할 수 있는 제품으로는, 예를 들어, 수렴화장수, 유연화장수, 영양화장수, 각종크림, 에센스, 팩, 파운데이션 등과 같은 화장품류와 클렌징, 세안제, 비누, 트리트먼트, 미용액 등이 있다.Examples of products to which the cosmetic composition of the present invention can be added include cosmetics such as astringent lotion, softening longevity lotion, nutrition lotion, various creams, essences, packs, foundation and the like, cleansing, cleanser, soap, .
본 발명의 화장료 조성물의 구체적인 제형으로서는 스킨로션, 스킨 소프너, 스킨토너, 아스트린젠트, 로션, 밀크로션, 모이스처 로션, 영양로션, 맛사지크림, 영양크림, 모이스처 크림, 핸드크림, 에센스, 영양에센스, 팩, 비누, 샴푸, 클렌징폼, 클렌징로션, 클렌징크림, 바디로션, 바디클렌저, 유액, 립스틱, 메이컵 베이스, 파운데이션, 프레스파우더, 루스파우더, 아이섀도 등의 제형을 포함한다.As a specific formulation of the cosmetic composition of the present invention, there may be mentioned a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutrition lotion, a massage cream, It includes formulations such as soap, shampoo, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, latex, lipstick, makeup base, foundation, press powder, loose powder, eye shadow.
본 발명의 일 구체예에 있어서, 본 발명의 발효홍삼, 동과자 및 아피오스 추출물의 혼합물의 함량은 조성물 총 중량에 대하여 0.0001-20중량%이며, 바람직하게는 0.5-20%이며, 보다 바람직하게는 1.0-20중량%이다.
In one embodiment of the present invention, the content of the mixture of the fermented red ginseng, frozen fruits and apiose extract of the present invention is 0.0001-20% by weight, preferably 0.5-20%, based on the total weight of the composition, 1.0 to 20% by weight.
이하, 실시예를 통하여 본 발명을 보다 상세히 설명하고자 한다. 이들 실시예는 본 발명을 보다 구체적으로 설명하기 위한 것으로, 본 발명의 범위가 이들 실시예에 한정되는 것은 아니다.
Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are for further illustrating the present invention, and the scope of the present invention is not limited to these examples.
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실시예Example
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비교예Comparative Example 1: One: 발효홍삼Fermented red ginseng 추출물의 제조 Preparation of extract
발효홍삼(충북 음성군 소재 세종고려인삼, 건조물) 100g에 10배 가량의 물 또는 주정을 넣고 90-100℃에서 4시간 동안 환류추출하였다. 이 추출액을 여과지 (Whatmann사 제조)로 여과한 후 60℃ 회전 증발기를 사용하여 감압 농축하였다. 이후 농축액을 동결 건조하여 건조 중량으로 발효홍삼 추출물 16.4 g을 얻은 다음, 이를 하기의 실험용 시료로 사용하였다.
About 10 times of water or alcohol was added to 100 g of fermented red ginseng (Sejong Korean ginseng, dried ginseng, Hyanggun, Chungbuk province) and refluxed at 90-100 ℃ for 4 hours. The extract was filtered with a filter paper (Whatmann), and then concentrated under reduced pressure using a rotary evaporator at 60 ° C. Thereafter, the concentrate was lyophilized to obtain 16.4 g of fermented red ginseng extract as a dry weight, and this was used as the experimental sample described below.
비교예Comparative Example 2: 2: 동과자Frog 추출물의 제조 Preparation of extract
동과자(인천시 부평구 소재 아센코리아, 건조물) 100 g에 10배 가량의 물 또는 주정을 넣고 90-100℃에서 4시간 동안 환류추출하였다. 이 추출액을 여과지 (Whatmann사 제조)로 여과한 후 60℃ 회전 증발기를 사용하여 감압 농축하였다. 이후 농축액을 동결 건조하여 건조 중량으로 동과자 추출물 18.1 g을 얻은 다음, 이를 하기의 실험용 시료로 사용하였다.
About 10 times as much water or alcohol was added to 100 g of frozen fish (Asen Korea, Bupyung-gu, Incheon city) and the mixture was refluxed at 90-100 ℃ for 4 hours. The extract was filtered with a filter paper (Whatmann), and then concentrated under reduced pressure using a rotary evaporator at 60 ° C. Thereafter, the concentrate was lyophilized to obtain 18.1 g of a frog extract as a dry weight, and this was used as the following experimental sample.
비교예Comparative Example 3: 3: 아피오스Apios 추출물의 제조 Preparation of extract
아피오스(인천시 부평구 소재 아센코리아, 건조물) 100 g에 10배 가량의 물 또는 주정을 넣고 90-100℃에서 4시간 동안 환류추출하였다. 이 추출액을 여과지 (Whatmann사 제조)로 여과한 후 60℃ 회전 증발기를 사용하여 감압 농축하였다. 이후 농축액을 동결 건조하여 건조 중량으로 아피오스 추출물 21.7 g을 얻은 다음, 이를 하기의 실험용 시료로 사용하였다.
About 10 times water or alcohol was added to 100 g of Apiose (Asen Korea, Bupyung-gu, Incheon city) and the mixture was refluxed at 90-100 ℃ for 4 hours. The extract was filtered with a filter paper (Whatmann), and then concentrated under reduced pressure using a rotary evaporator at 60 ° C. Thereafter, the concentrate was lyophilized to obtain 21.7 g of Apiose extract as a dry weight, and this was used as the experimental sample described below.
실시예Example 1 One
비교예 1, 2 및 3의 발효홍삼 추출물, 동과자 추출물 및 아피오스 추출물을 1:1:1의 중량비로 혼합한 후, 이를 하기의 실험용 시료로 사용하였다.
Fermented red ginseng extracts, frog extracts and apiose extracts of Comparative Examples 1, 2 and 3 were mixed at a weight ratio of 1: 1: 1 and used as the experimental sample described below.
실시예Example 2 2
비교예 1, 2 및 3의 발효홍삼 추출물, 동과자 추출물 및 아피오스 추출물을 2:1:1의 중량비로 혼합한 후, 이를 하기의 실험용 시료로 사용하였다.
Fermented red ginseng extracts, frog extracts, and apiose extracts of Comparative Examples 1, 2 and 3 were mixed at a weight ratio of 2: 1: 1 and used as the following experimental sample.
실시예Example 3 3
비교예 1, 2 및 3의 발효홍삼 추출물, 동과자 추출물 및 아피오스 추출물을 1:2:1의 중량비로 혼합한 후, 이를 하기의 실험용 시료로 사용하였다.
Fermented red ginseng extracts, frog extracts, and apiose extracts of Comparative Examples 1, 2 and 3 were mixed at a weight ratio of 1: 2: 1 and used as the following experimental sample.
실시예Example 4 4
비교예 1, 2 및 3의 발효홍삼 추출물, 동과자 추출물 및 아피오스 추출물을 1:1:2의 중량비로 혼합한 후, 이를 하기의 실험용 시료로 사용하였다.
Fermented red ginseng extracts, frog extracts, and apiose extracts of Comparative Examples 1, 2 and 3 were mixed at a weight ratio of 1: 1: 2 and used as the following experimental sample.
실험예Experimental Example 1: One: DPPHDPPH 라디칼Radical 소거능Scatters 측정을 통한 피부 노화억제 효과 Effect of inhibiting skin aging by measuring
본 발명의 혼합물의 항산화 효과를 알아보기 위하여 Kilani 등의 방법(Kilani et al., Environ. Toxicol. Phar. 20: 478-484(2005) 참조)에 준하여 1,1-diphenyl-2-picrylhydrazyl(DPPH; Sigma Co., USA) 라디칼 소거능을 측정하였다. DPPH는 항산화 활성이 있는 물질과 반응하면 라디칼이 소거되어 짙은 자색으로 탈색되기 때문에, 흡광도 측정을 통하여 항산화 물질의 수소 공여능을 측정하는데 널리 이용되고 있다. 본 실험에서 양성대조군으로는 항산화 효능이 잘 알려진 비타민 C(아스코르브산)를 사용하였다.To investigate the antioxidative effect of the mixture of the present invention, a method of Kilani et al. (Kilani et al al ., Environ. Toxicol. Phar. 1,1-diphenyl-2-picrylhydrazyl (DPPH; Sigma Co., USA) was assayed for its radical scavenging activity. DPPH is widely used to measure the hydrogen donating ability of antioxidants through the measurement of absorbance because it reacts with a substance having an antioxidant activity to decolorize with a dark purple color due to radical scavenging. Vitamin C (ascorbic acid), a well-known antioxidant, was used as a positive control in this experiment.
96 웰 플레이트에 메탄올로 희석된 시험물질을 농도별(100, 500 및 1,000 ㎍/ml)로 100 ㎕씩 첨가한 후, DPPH 용액(0.2mM DPPH in Methanol)을 100 ㎕ 첨가하여 25분간 반응시켜 ELISA reader(BioTEK, USA)를 이용하여 517 nm에서 흡광도를 측정하였다. 각 시험물질의 라디칼 소거능은 아래의 식에 의해 전자공여능으로 계산하여 백분율로 나타내었다.
100 μl of the test substance diluted with methanol (100, 500 and 1,000 μg / ml) was added to each well of a 96-well plate, 100 μl of DPPH solution (0.2 mM DPPH in Methanol) was added, Absorbance was measured at 517 nm using a reader (BioTEK, USA). The radical scavenging ability of each test substance was calculated as the percentage of electron donating ability by the following formula.
[수학식 1][Equation 1]
전자공여능(%) = (1-시험물질 첨가군의 흡광도/무첨가군의 흡광도) × 100
Electron donating ability (%) = (1 - absorbance of test substance addition group / absorbance of no addition group) x 100
상기 표 1의 결과에서 나타낸 바와 같이, 발효홍삼(비교예 1), 동과자(비교예 2), 아피오스(비교예 3) 또는 다양한 혼합물(실시예 1, 2, 3 및 4) 처리군은 양성대조군인 비타민 C에 비하여 라디칼 소거능이 떨어지는 것으로 확인되었다. 하지만, 발효홍삼을 제외한 나머지 시험물질 처리군에서는 대체적으로 라디칼 소거능이 우수한 것으로 확인되었으며, 특히 아피오스 처리군 및 아피오스가 상대적으로 많이 함유된 혼합물(실시예 4) 처리군은 라디칼 소거능이 매우 우수하다는 것을 확인할 수 있었다.
As shown in the results of the above Table 1, the group treated with fermented red ginseng (Comparative Example 1), frog (Comparative Example 2), Apiose (Comparative Example 3) or various mixtures (Examples 1, 2, 3 and 4) It was confirmed that the radical scavenging ability was lower than that of vitamin C, which is a control group. However, it was confirmed that the group treated with the test substances except fermented red ginseng generally had excellent radical scavenging ability. In particular, the group treated with the apiose-treated group and the apiose-treated group (Example 4) showed excellent radical scavenging ability .
실험예Experimental Example 2: 세포 생존율 측정 2: Measurement of cell survival rate
하기 피부 주름개선 및 피부 미백 효능을 평가하기에 앞서, 본 발명의 조성물에 대한 세포독성 효과를 확인하기 위하여 사람 섬유아세포(CCD-985sk cell; 한국세포주은행, 서울, 대한민국) 및 마우스 흑색종 세포주(B16F10 cell; 한국세포주은행)를 이용하여 MTT assay를 실시하였다.Prior to evaluating the skin wrinkle improvement and skin whitening efficacy, human fibroblasts (CCD-985sk cell; Korea Cell Line Bank, Seoul, Korea) and mouse melanoma cell line MTT assay was performed using B16F10 cell; Korean Cell Line Bank).
사람 섬유아세포 및 마우스 흑색종 세포주를 1.5×104 cells/well이 되도록 96 well plate에 접종한 후, 10%(v/v) 우태아혈청(FBS; Gibco사, USA)이 함유된 DMEM(Gibco사, USA) 배지를 이용하여 37 ℃, 5% CO2 인큐베이터에서 18시간 배양하였다. 이 후, 상기 각 시험물질을 우태아혈청이 함유되지 않은 DMEM 배지로 희석하여 다양한 농도별(0, 25, 50, 100 및 200 ㎍/ml)로 24시간 처리한 다음, 5 mg/ml의 MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; Sigma사, USA) 용액을 최종농도 1 mg/ml이 되도록 첨가하고 4시간 동안 추가로 배양하였다. 마지막으로 상등액을 버리고 dimethylsulfoxide(DMSO; Sigma사, USA)를 200 ㎕씩 각 well에 넣고 30분 동안 쉐이킹 인큐베이터(TW3, Fine PCR, 대한민국)에서 crystal violet을 녹인 다음, microplate reader를 이용하여 570 nm에서 흡광도를 측정하였다. 사람 섬유아세포 및 마우스 흑색종 세포의 생존율은 시험물질을 처리하지 않은 대조군의 생존율을 100%로 하여 이에 대한 %로 표시하였다.Human fibroblasts and mouse melanoma cell lines were inoculated into 96-well plates at a density of 1.5 × 10 4 cells / well. DMEM containing 10% (v / v) fetal bovine serum (FBS; Gibco, USA) USA) medium at 37 ° C in a 5% CO 2 incubator for 18 hours. Each of the test substances was diluted with DMEM medium containing no fetal bovine serum and treated for 24 hours at various concentrations (0, 25, 50, 100 and 200 / / ml). Then, 5 mg / ml of MTT (Sigma, USA) was added to a final concentration of 1 mg / ml and further cultured for 4 hours. Finally, the supernatant was discarded and 200 μl of dimethylsulfoxide (DMSO; Sigma, USA) was added to each well. The crystal violet was dissolved in a shaking incubator (TW3, Fine PCR, Korea) for 30 min. Absorbance was measured. The survival rate of human fibroblasts and mouse melanoma cells was expressed as a percentage of the survival rate of the control group not treated with the test substance as 100%.
도 1a와 1b에서 나타낸 바와 같이, 각 시험물질의 세포독성을 측정한 결과 사람 섬유아세포 및 마우스 흑색종 세포주 모두 최고농도인 200 ㎍/ml에서도 통계적으로 유의적인 세포독성은 관찰되지 않았다. 이러한 결과를 바탕으로 본 발명의 혼합물이 독성에 대하여 어느 정도 안전성을 갖고 있음을 확인할 수 있었다.
As shown in FIGS. 1A and 1B, the cytotoxicity of each test substance was measured, and no statistically significant cytotoxicity was observed even at the highest concentration of 200 μg / ml in both human fibroblast and mouse melanoma cell lines. Based on these results, it was confirmed that the mixture of the present invention has a certain degree of safety against toxicity.
실험예Experimental Example 3: 3: MMPMMP -1 발현 억제 분석을 통한 피부 주름개선 효과-1 wrinkle improvement effect by inhibition of expression
MMP(matrix metalloproteinase)는 세포외기질(extracellular matrix)과 기저막 분해에 관여하는 효소군으로써, 구조와 기능적 특성에 따라 간질성 콜라게나제(interstitial collagenase), 스트로멜리신(stromelysin), 젤라티나제 (gelatinase), 막-타입 MMP(membrane-type MMP, MT-MMP) 등 크게 4개의 subfamily로 나눌 수 있다. 피부에서는 주로 MMP-1, MMP-2 및 MMP-9이 존재하는 것으로 알려져 있으며, 이 중에서도 MMP-1이 콜라겐 분해와 관련하여 가장 중요하게 작용하는 것으로 알려져 있다. 따라서 본 연구자들은 MMP-1 단백질 발현 억제 분석을 통하여 상기 시험물질들의 주름개선 효과를 확인하고자 하였다.MMP (matrix metalloproteinase) is a group of enzymes involved in the degradation of extracellular matrix and basement membrane. It has been shown to be an interstitial collagenase, stromelysin, gelatinase gelatinase), membrane-type MMP (MT-MMP) and membrane-type MMP (membrane-type MMP) can be divided into four subfamily. It is known that MMP-1, MMP-2 and MMP-9 are mainly present in the skin, and MMP-1 is known to be most important for collagen degradation. Therefore, the present inventors tried to confirm the wrinkle-reducing effect of the test materials through inhibition of MMP-1 protein expression.
사람 섬유아세포(CCD-986sk cell)를 24 well plate에 1×105 cell/well로 접종하고, 10%(v/v) 우태아혈청이 함유된 DMEM 배지를 이용하여 37 ℃, 5% CO2 인큐베이터에서 24시간 배양하였다. PBS로 1회 세척하고 우태아혈청이 없는 DMEM 배지로 교체한 후, 자외선조사기(ELC-500, Lightning Enterprisis, USA)를 이용하여 자외선 A를 세포에 영향을 주지 않는 범위로 조사하였다. 각 시험물질을 농도별(0, 25, 50 및 100 ㎍/ml)로 처리한 다음, 2일간 추가로 배양하였다. 배양액을 수집하여 단백질 양을 정량한 다음, ELISA kit(Amersham, USA)를 이용하여 MMP-1 양을 측정하였다. 시험물질 처리에 따른 MMP-1 발현 억제 정도는 시험물질을 처리하지 않고 자외선만 조사한 대조군을 100 %로 하여 이에 대한 %로 표시하였으며, MMP-1 합성을 억제하는 것으로 잘 알려진 TFG-β(transforming growth factor-β)를 양성대조군으로 사용하였다.The human fibroblasts (CCD-986sk cell) in 24 well plate 1 × 10 5 cell / well inoculated and, 10% (v / v) fetal bovine serum by using a DMEM medium 37 ℃, 5% CO 2 And cultured in an incubator for 24 hours. The cells were washed once with PBS and replaced with DMEM medium lacking fetal bovine serum. Ultraviolet A was irradiated to the extent of not affecting the cells using an ultraviolet irradiator (ELC-500, Lightning Enterprisis, USA). Each test substance was treated with concentration (0, 25, 50 and 100 / / ml) and then incubated for 2 days. The amount of MMP-1 was measured using an ELISA kit (Amersham, USA). The degree of inhibition of MMP-1 expression by treatment with test substances was expressed as% of the control group in which only UV light was not irradiated and 100% of the control group was treated, and TFG-β (transforming growth factor-β) was used as a positive control.
도 2에 나타낸 바와 같이, 각 시험물질의 처리 농도에 의존적으로 MMP-1 발현이 저해되었다. 따라서 본 발명의 혼합물이 내인성 또는 광노화에 의해 발생되는 피부조직 분해효소인 MMP-1의 생합성을 저해함으로써, 피부노화를 억제하고 피부 주름개선에 효과적임을 알 수 있었다.
As shown in Fig. 2, expression of MMP-1 was inhibited depending on the treatment concentration of each test substance. Therefore, it was found that the mixture of the present invention inhibits biosynthesis of MMP-1, a skin tissue degrading enzyme generated by endogenous or photoaging, thereby suppressing skin aging and improving skin wrinkles.
실험예Experimental Example 4: 4: 티로시나제Tyrosinase 저해 분석을 통한 피부 미백 효과 Skin whitening effect through inhibition analysis
티로신에서 멜라닌을 형성하는 연속적인 효소적 산화반응에서 티로시나제는 처음 tyrosine이 dopa로 산화되는 과정과 다음 단계인 dopa가 dopaquinone으로 산화되는 2가지 단계의 반응을 촉진하는 효소로 알려져 있다. 따라서 본 발명의 혼합물이 피부 미백에 관여하는 티로시나제를 저해함으로써, 미백에 효과적으로 작용하는지 확인하기 위하여 mushroom tyrosinase assay를 이용하여 in vitro 상에서 티로시나제 저해 활성을 측정하였다.In the continuous enzymatic oxidation reaction of tyrosine to form melanin, tyrosinase is known to be the enzyme that firstly oxidizes tyrosine to dopa and the next step, dopa, is a two step reaction that is oxidized to dopaquinone. Thus, in order to confirm that the mixture of the present invention effectively inhibits tyrosinase involved in skin whitening, it was tested for tyrosinase inhibitory activity in vitro using mushroom tyrosinase assay.
시험관에 pH 6.8의 67 mM sodium phosphate buffer 500 ㎕에 10 mM L-DOPA를 녹인 기질액 200 ㎕ 및 시료용액 100 ㎕를 넣은 혼합액에 110 Unit/ml mushroom tyrosinase 200 ㎕를 첨가하여 25 ℃에서 2분간 반응시켜 반응액 중에 생성된 DOPA chrome을 microplate reader를 이용하여 475 nm에서 흡광도를 측정하였다. 각 시험물질의 티로시나제 저해 활성은 아래의 식에 의해 티로시나제 저해율로 나타내었다.
200 μl of a substrate solution containing 10 mM L-DOPA dissolved in 500 μl of 67 mM sodium phosphate buffer (pH 6.8) and 100 μl of a sample solution were added to 200 μl of 110 Unit / ml mushroom tyrosinase, followed by reaction at 25 ° C for 2 minutes The absorbance of DOPA chrome produced in the reaction solution was measured at 475 nm using a microplate reader. The tyrosinase inhibitory activity of each test substance was expressed by the inhibition rate of tyrosinase by the following equation.
[수학식 2]&Quot; (2) "
티로시나제 저해율(%) = 100-(시험물질의 반응흡광도/대조군의 반응흡광도 × 100)
Tyrosinase inhibition rate (%) = 100- (reaction absorbance of test substance / reaction absorbance of control group x 100)
상기 표 2의 결과에서 나타낸 바와 같이, 시험물질의 처리 농도에 의존적으로 티로시나제 활성이 저해되었으며, 특히 비교예 2, 비교예 3, 실시예 3 및 실시예 4의 티로시나제 저해율이 상대적으로 높은 것으로 확인되었다. 이는 본 발명의 조성물 중 동과자 추출물 및 아피오스 추출물이 미백에 보다 효과적이며, 이들을 혼합하여 사용할 경우 이들의 시너지 효과로 인하여 미백 효과가 더욱 우수해지는 것을 알 수 있었다.
As shown in the results of Table 2, tyrosinase activity was inhibited depending on the treatment concentration of the test substance, and it was confirmed that the inhibition rate of tyrosinase in Comparative Example 2, Comparative Example 3, Example 3 and Example 4 was relatively high . It was found that the extract of Copper and Apis extract in the composition of the present invention is more effective for whitening, and when they are used in combination, the whitening effect is more excellent due to the synergy effect thereof.
실험예Experimental Example 5: 멜라닌 생성 억제 분석을 통한 피부 미백 효과 5: Skin whitening effect through inhibition of melanin formation
마우스 흑색종 세포주를 24 well plate에 1×105 cell/well로 접종하고, 10%(v/v) 우태아혈청이 함유된 DMEM 배지를 이용하여 37 ℃, 5% CO2 인큐베이터에서 80% 이상 되도록 배양하였다. PBS로 1회 세척하고 우태아혈청이 없는 DMEM 배지를 이용하여 각 시험물질을 농도별(0, 25, 50 및 100 ㎍/ml)로 처리한 다음, 2일간 추가로 배양하였다. 배양액을 제거하고 PBS로 세척한 후, 1 N 수산화나트륨으로 세포를 녹여 microplate reader를 이용하여 400 nm에서 흡광도를 측정하였다.
The mouse melanoma cell line was inoculated in a 24-well plate at 1 × 10 5 cells / well and cultured in DMEM medium containing 10% (v / v) fetal bovine serum at a concentration of 80% or more in a 5% CO 2 incubator at 37 ° C. Lt; / RTI > After washing once with PBS, each test substance was treated with concentrations (0, 25, 50 and 100 ㎍ / ml) using DMEM medium without fetal bovine serum, and then cultured for 2 days. After removing the culture medium and washing with PBS, the cells were dissolved with 1 N sodium hydroxide and absorbance was measured at 400 nm using a microplate reader.
[수학식 3]&Quot; (3) "
멜라닌 생성억제율(%) = 100-(시험물질의 흡광도/대조군의 흡광도 × 100)
Melanin production inhibition rate (%) = 100- (absorbance of test substance / absorbance of control x 100)
상기 표 3의 결과에서 나타낸 바와 같이, 모든 시험물질의 처리 농도에 의존적으로 멜라닌 생성이 억제되었으며, 앞선 티로시나제 저해 실험결과와 마찬가지로 동과자 추출물 및 아피오스 추출물 처리군에서 멜라닌 생성이 크게 억제되는 것을 확인할 수 있었다. 또한 이러한 멜라닌 생성 억제효과는 동과자 추출물 또는 아피오스 추출물을 단독으로 처리하였을 때보다 이들을 혼합하여 처리하였을 때 더욱 우수해지는 것을 알 수 있었다.
As shown in the results of Table 3, melanin production was inhibited depending on the treatment concentration of all the test substances, and it was confirmed that the melanin production was greatly suppressed in the group treated with copper extract and apiose extract as in the result of the previous tyrosinase inhibition experiment there was. It was also found that the inhibitory effect on melanogenesis is better when the extracts of Copper or Apiose are mixed with each other than when they are treated alone.
이제까지 본 발명에 대하여 그 바람직한 실시예들을 중심으로 살펴보았다. 본 발명이 속하는 기술 분야에서 통상의 지식을 가진 자는 본 발명이 본 발명의 본질적인 특성에서 벗어나지 않는 범위에서 변형된 형태로 구현될 수 있음을 이해할 수 있을 것이다. 그러므로 개시된 실시예들은 한정적인 관점이 아니라 설명적인 관점에서 고려되어야 한다. 본 발명의 범위는 전술한 설명이 아니라 특허청구범위에 나타나 있으며, 그와 동등한 범위 내에 있는 모든 차이점은 본 발명에 포함된 것으로 해석되어야 할 것이다.The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.
Claims (12)
상기 발효홍삼, 동과자 및 아피오스 추출물은 발효홍삼, 동과자 및 아피오스 각각을 물, 유기용매 또는 이들의 혼합 용매를 사용하여 추출한 추출물인 것을 특징으로 하는 피부미용증진용 건강기능식품 조성물.The method according to claim 1,
Wherein the fermented red ginseng, frog and apiose extract are extracts obtained by extracting each of fermented red ginseng, frozen ginseng and apiose with water, an organic solvent or a mixed solvent thereof.
상기 유기용매는 메탄올(methanol), 에탄올(ethanol), 프로판올(propanol), 이소프로판올(isopropanol) 및 부탄올(butanol)로 이루어진 군으로부터 선택된 것을 특징으로 하는 피부미용증진용 건강기능식품 조성물.3. The method of claim 2,
Wherein the organic solvent is selected from the group consisting of methanol, ethanol, propanol, isopropanol, and butanol.
상기 발효홍삼, 동과자 및 아피오스 추출물은 발효홍삼, 동과자 및 아피오스 각각의 70% 에탄올 추출물인 것을 특징으로 하는 피부미용증진용 건강기능식품 조성물.The method according to claim 1,
Wherein the fermented red ginseng, frog and apiose extract are 70% ethanol extracts of fermented red ginseng, frog ginseng, and apiose, respectively.
상기 혼합물은 조성물에 0.1 내지 10000μg/ml의 농도로 포함되는 것을 특징으로 하는 피부미용증진용 건강기능식품 조성물.The method according to claim 1,
Wherein the composition is contained in the composition at a concentration of 0.1 to 10000 g / ml.
상기 피부미용증진용은 피부 주름개선용, 피부 미백용 또는 피부 노화 방지용인 것을 특징으로 하는 건강기능식품 조성물.The method according to claim 1,
Wherein said skin beauty enhancing agent is for skin wrinkle improvement, skin whitening or skin aging prevention.
상기 피부 주름개선은 MMP-1 발현 저해를 통해 이루어지는 것을 특징으로 하는 피부미용증진용 건강기능식품 조성물.8. The method of claim 7,
Wherein said improvement of skin wrinkles is achieved through inhibition of MMP-1 expression.
상기 피부 미백은 티로시나아제 활성 저해 및 멜라닌 생성 억제를 통해 이루어지는 것을 특징으로 하는 피부미용증진용 건강기능식품 조성물.8. The method of claim 7,
Wherein said skin whitening is achieved through inhibition of tyrosinase activity and inhibition of melanin formation.
상기 피부 노화 방지는 항산화 활성을 통해 이루어지는 것을 특징으로 하는 피부미용증진용 건강기능식품 조성물.8. The method of claim 7,
Wherein said skin aging prevention is achieved through antioxidant activity.
상기 식품은 음료류, 육류, 초코렛, 식품류, 과자류, 피자, 라면, 기타 면류, 껌류, 사탕류, 아이스크림류, 알코올 음료류, 비타민 복합제 및 건강보조식품류로 이루어진 군으로부터 선택되는 것을 특징으로 하는 건강기능식품.12. The method of claim 11,
Wherein said food is selected from the group consisting of beverage, meat, chocolates, foods, confectionery, pizza, ramen, other noodles, gums, candy, ice cream, alcoholic beverages, vitamin complexes and health supplement foods.
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