KR102561151B1 - Composition for improving skin condition comprising herb extracts mixture - Google Patents
Composition for improving skin condition comprising herb extracts mixture Download PDFInfo
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- KR102561151B1 KR102561151B1 KR1020160069528A KR20160069528A KR102561151B1 KR 102561151 B1 KR102561151 B1 KR 102561151B1 KR 1020160069528 A KR1020160069528 A KR 1020160069528A KR 20160069528 A KR20160069528 A KR 20160069528A KR 102561151 B1 KR102561151 B1 KR 102561151B1
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- extract
- skin
- effect
- present
- composition
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- Medicines Containing Plant Substances (AREA)
Abstract
본 발명은 피부 적용 시 안전하고, 피부 개선 효과가 우수한 복합 생약 추출물을 유효성분으로 포함하는 조성물에 관한 것이다. 보다 구체적으로, 본 발명에 따른 조성물은 황금 추출물, 박하 추출물, 치자 추출물, 연교 추출물 및 대황 추출물을 유효성분으로 포함하는 조성물로, 항당화 효과, 피부 미백 효과, 피부 탄력 및 주름 개선 효과, 항산화 효과 및 피부 트러블 개선 효과를 포함한 피부 개선에 탁월한 효과를 갖는다. The present invention relates to a composition containing as an active ingredient a composite herbal medicine extract that is safe when applied to the skin and has excellent skin improvement effects. More specifically, the composition according to the present invention is a composition containing golden extract, peppermint extract, gardenia extract, yeongyo extract, and rhubarb extract as active ingredients, and has excellent effects on skin improvement, including antiglycation effect, skin whitening effect, skin elasticity and wrinkle improvement effect, antioxidant effect, and skin trouble improvement effect.
Description
본 발명은 피부 적용 시 안전하고, 피부 개선 효과가 우수한 복합 생약 추출물을 유효성분으로 포함하는 조성물에 관한 것이다. The present invention relates to a composition containing as an active ingredient a composite herbal medicine extract that is safe when applied to the skin and has excellent skin improvement effects.
피부는 외부 환경으로부터 인체를 보호하고, 내부의 수분 및 유용 성분이 밖으로 유출되는 것을 막아주는 장벽 기능, 체온조절 기능, 배설 기능 등 다양한 생리적 기능을 담당하고 있는 중요한 기관이다. 그러나, 다음과 같은 이유 등에 의해 피부 세포의 활성이 저하되고 피부 상태가 악화되는 현상이 일어날 수 있다.The skin is an important organ that protects the human body from the external environment and is in charge of various physiological functions such as a barrier function that prevents internal moisture and useful components from leaking out, a body temperature control function, and an excretion function. However, the activity of skin cells is reduced and the skin condition is deteriorated due to the following reasons.
피부의 진피층에 존재하는 콜라겐과 엘라스틴은 피부의 기계적 견고성, 결합조직의 저항력, 조직의 결합력 유지, 및 세포접착의 지탱 등의 기능을 수행하여 피부의 구성성분에 많은 영향을 미친다. 이러한 콜라겐은 스트레스, 고령화 또는 자외선 조사에 의한 광 노화에 의하여 감소되며, 한편, 엘라스틴은 자외선에 노출된 후 활성이 증가된 분해효소 엘라스타제에 의해 3차원 구조가 뒤틀려진다고 알려져있다. 이에 따라 피부 조직이 느슨해지고 탄력을 잃게 된다. Collagen and elastin present in the dermal layer of the skin have a great effect on the components of the skin by performing functions such as mechanical robustness of the skin, resistance of connective tissue, maintenance of tissue cohesiveness, and support of cell adhesion. It is known that such collagen is reduced by stress, aging, or photoaging caused by ultraviolet irradiation, and, on the other hand, elastin is distorted in its three-dimensional structure by an increased activity of elastin after exposure to ultraviolet rays. As a result, the skin tissue becomes loose and loses its elasticity.
표피의 세포 기능이 저하됨으로써 신진대사 작용이 원활하지 못하게 되어, 각질 탈락이 잘 일어나지 않아 각질이 과하게 축적된 상태에서 자외선을 받으면 탄력이 저하되어 주름 발생을 초래하게 된다. 또는 표피에 피지막이 형성되지 않을 경우 수분과 피지 분비가 감소하고 피부가 건조해지면서 주름이 형성되기도 한다. As the cell function of the epidermis is lowered, the metabolism is not smooth, and the exfoliation of the dead skin cells does not occur easily. When UV rays are applied in a state in which the dead skin cells are excessively accumulated, the elasticity is lowered, resulting in the occurrence of wrinkles. Or, if the sebaceous film is not formed on the epidermis, moisture and sebum secretion decreases and the skin becomes dry, resulting in wrinkles.
피부가 자외선을 받으면 피부를 보호하기 위해 멜라닌을 합성한다. 합성된 멜라닌은 멜라노좀을 통해 피부의 각질세포로 옮겨진다. 이 각질 세포는 28일을 주기로 턴-오버(turn over) 현상이 일어난다. 따라서 생성된 멜라닌은 각질세포에 의해 28일을 주기로 소실되는 것이 일반적이나, 스트레스, 피부 노화 등에 의해 피부 세포 주기가 잘 조절되지 않게 되면 각질세포가 제시기에 탈락하지 않아 기미, 주근깨, 검버섯 등의 색소 침착이 일어나게 된다. When the skin is exposed to UV rays, melanin is synthesized to protect the skin. The synthesized melanin is transferred to the keratinocytes of the skin through melanosomes. These keratinocytes turn over every 28 days. Therefore, it is common for the generated melanin to be lost every 28 days by keratinocytes, but if the skin cell cycle is not well regulated due to stress, skin aging, etc., keratinocytes do not drop out in time, resulting in pigmentation such as spots, freckles, and age spots.
피부에서는 피지, 땀 성분 및 화장품 성분들이 피부 상재균에 의해 독성이 강한 물질로 분해되어 피부에 자극을 일으키고 염증을 유발시킬 수 있다. 또한 자외선으로 인한 자극으로 피부에 염증 매개 물질인 일산화질소(NO)의 생성을 증가시켜 피부 트러블이 유발될 수 있다. 일산화질소의 생성은 대부분 iNOS에 의한 것인데, iNOS는 LPS, 사이토카인 같은 자극에 의해 급격하게 유도되어 과량의 NO를 생성하는 것으로 알려져있다.In the skin, sebum, sweat components, and cosmetic ingredients are decomposed into highly toxic substances by skin flora, which can cause skin irritation and inflammation. In addition, irritation caused by ultraviolet rays may cause skin trouble by increasing the production of nitric oxide (NO), an inflammatory mediator in the skin. Nitric oxide is mostly produced by iNOS, and iNOS is known to be rapidly induced by stimuli such as LPS and cytokines to produce excess NO.
유해산소(toxic oxygen species)라고도 하는 활성산소종(reactive oxygen species, ROS)은 호흡 등과 같은 생리작용에 의해 세포에서 생성되는 독성물질로 끊임없이 생산되고 소멸하며, 정상적인 상태에서는 3-5% 정도로 존재한다. 이런 활성산소종은 수퍼옥시드 라디칼(Superoxide radical, O2-), 하이드록시라디칼 (hydroxyl radical, HO+)과 같은 자유라디칼(free radical: 화학적으로 최외각 전자궤도에 쌍을 이루고 있지 않은 원자나 분자로 매우 불안정하여 높은 반응성을 가지는 형태)로 존재하거나 혹은 과산화수소(hydorgen peroxide, H2O2)나 일중항산소(Singlet radical)와 같이 쌍을 이룬 전자를 가진 화합물의 형태로 존재한다. 활성산소종은 생리계 내에서 세균을 살균하는 생체 방어 작용을 하는 장점도 있지만, 일반적으로 생체 내에서 산화를 일으켜 질병의 원인이 되는 유해한 작용을 한다. 이러한 활성산소종은 생물분자를 공격하여 세포나 조직에 피해를 주며, 노화나 각종 성인병 질환에 관여하는 여러 종류의 질병을 야기한다는 보고가 있다. Reactive oxygen species (ROS), also called toxic oxygen species, are toxic substances generated in cells by physiological processes such as respiration and are constantly produced and destroyed, and exist at about 3-5% in normal conditions. These reactive oxygen species exist as free radicals such as superoxide radical (O2-) and hydroxyl radical (HO+), or in the form of compounds with paired electrons such as hydrogen peroxide (H 2 O 2 ) or singlet oxygen (singlet radical). exists as Reactive oxygen species have the advantage of having a biological defense function of sterilizing bacteria in the physiological system, but generally cause harmful effects that cause diseases by causing oxidation in vivo. It has been reported that these reactive oxygen species attack biomolecules, damage cells or tissues, and cause various diseases related to aging or various adult diseases.
상술한 여러 요인들에 의한 피부 노화를 억제하고, 미백, 주름, 탄력 또는 트러블을 개선하고, 항산화 효과가 있는 물질을 개발하고자 하는 연구가 계속되어 왔다. Research has been conducted to suppress skin aging caused by the above-mentioned factors, to improve whitening, wrinkles, elasticity or trouble, and to develop substances having antioxidant effects.
상술한 효과가 있는 물질은 자연계에 널리 분포되어 있는데, 주로 식물로부터 유래된 물질을 사용해 식품, 화장품, 의약품 등의 원료로서 사용해 왔다. 그러나, 이 같은 자연계에서 유래된 물질은 효과가 크지 않아 유의미한 효과를 얻기 위해서는 다량으로 사용해야 하고, 이로 인한 독성 및 가격 상승 문제가 대두되었다. Substances having the above effects are widely distributed in the natural world, and have been mainly used as raw materials for foods, cosmetics, and pharmaceuticals using substances derived from plants. However, such materials derived from nature do not have significant effects and must be used in large amounts to obtain significant effects, resulting in toxicity and price increase problems.
이러한 천연 물질의 문제점을 해결하기 위해, 화학적으로 합성된 물질의 개발이 이어졌다. 이들은 천연 물질에 비해 소량 사용으로도 월등히 우수한 효과를 발휘한다는 장점이 있으나, 반면 인체에 크고 작은 부작용을 유발시킬 수 있다는 치명적인 문제점이 있어 사용이 제한된다. In order to solve these problems of natural materials, chemically synthesized materials have been developed. These have the advantage of exerting a much superior effect even with a small amount compared to natural substances, but on the other hand, there is a fatal problem that can cause large and small side effects in the human body, so their use is limited.
이에, 피부 개선에 우수한 효과를 발휘하면서도 천연물로부터 유래되어 안정성이 확보되는 물질의 개발이 절실한 실정이다. Accordingly, there is an urgent need to develop a substance derived from a natural product and ensuring stability while exhibiting an excellent skin improvement effect.
본 발명이 해결하고자 하는 과제는 피부 개선, 특히 미백, 주름, 탄력, 트러블 개선, 항산화에 우수한 효과를 발휘하면서도 천연물로부터 유래되어 안정성이 확보되는 복합 생약 추출물을 제공하는데 있다. The problem to be solved by the present invention is to provide a complex herbal extract derived from natural products and ensuring stability while exhibiting excellent effects on skin improvement, in particular, whitening, wrinkles, elasticity, trouble improvement, and antioxidation.
상술한 과제를 해결하기 위하여, 본 발명은 황금 추출물, 박하 추출물, 치자 추출물, 연교 추출물 및 대황 추출물을 유효성분으로 포함하는 피부 미백, 주름, 탄력, 트러블, 또는 항산화용 조성물 (예를 들어, 화장료 조성물)을 제공한다. In order to solve the above problems, the present invention provides a composition for skin whitening, wrinkles, elasticity, trouble, or antioxidant (e.g., a cosmetic composition) comprising golden extract, peppermint extract, gardenia extract, yeongyo extract and rhubarb extract as active ingredients.
[추출물] [extract]
황금(黃芩)은 라틴명 Scutellariae Radix인, 속썩은풀 Scutellaria baicalensis Georgi (꿀풀과 Labiatae)의 뿌리로서 그대로 또는 주피를 제거한 것이다. 뿌리는 원뿔 모양이고 비틀어져 굽어 있으며, 바깥 면은 황갈색~진한 노란색이고, 질은 단단하면서 취약하고 절단하기 쉽다. 자른 면은 노란색이고, 중심부는 적갈색이다. 냄새가 거의 없고 맛은 약간 쓰다. 항균작용, 항염증작용, 혈압강하, 지혈작용, 간치료작용, 해열, 진토, 이뇨작용이 보고되었다. 주요 화학성분으로는 포르시톨(forsythol), 스테롤(sterol) 화합물, 사포닌(saponin), 올레아놀산(oleanolic acid) 등이 있다.Golden (黃芩) is a rotten grass with the Latin name Scutellariae Radix. It is the root of Scutellaria baicalensis Georgi (Lamiaceae Labiatae) intact or with the pericarp removed. The root is conical, twisted and curved, the outer surface is yellowish brown to dark yellow, and the quality is hard and fragile and easy to cut. The cut surface is yellow, and the center is reddish brown. It has almost no odor and the taste is slightly bitter. Antibacterial, anti-inflammatory, blood pressure lowering, hemostasis, liver treatment, antipyretic, antiemetic, and diuretic effects have been reported. Major chemical components include forsythol, sterol compounds, saponin, and oleanolic acid.
박하(薄荷, Mentha Herb)는 박하 Mentha arvensis Linne var. piperascens Malinvaud ex Holmes (꿀풀과 Labiatae)의 지상부이다. 지상부는 줄기와 여기에 마주 난 잎으로 되어 있다. 줄기는 네모기둥 모양이, 길이 15∼ 40 cm, 지름 0.2 ∼∼ 0.4 cm이다. 바깥면은 자갈색 또는 연한 녹색이고 모서리 근처에는 짧은 털이 있으며 마디사이는 길이 2 ∼ 5 cm이다. 질은 약하다. 잘린 면은 흰색이고 수부는 가운데가 비어 있다. 잎은 마주나며 짧은 잎자루가 있고, 잎조각은 찌그러지고 함께 말려있다. 온전한 잎을 펼쳐보면 긴 타원형 또는 달걀모양이고, 길이 2 ∼ 7 cm, 폭 1 ∼ 3 cm이다. 잎의 윗면은 진한 녹색이고, 아랫면은 회녹색이며 드물게 짧은 털이 덮여있고, 오목한 점모양의 선린이 있다. 꽃은 윤산화서로 액생하며, 꽃받침은 종 모양이고 끝은 5개로 갈라지며, 꽃부리는 연한 보라색이다. 손으로 비비면 특유하게 맑은 향기가 나며, 맛은 맵고 시원하다. 습기가 있는 들에서 자란다. 종자는 달걀 모양의 연한 갈색으로 가볍고 작으며 20 → 30 → 15℃의 변온에서 발아가 잘 된다.Peppermint (薄荷, Mentha Herb) is Mint Mentha arvensis Linne var. It is the aerial part of piperascens Malinvaud ex Holmes (Lamiaceae Labiatae). The above-ground part consists of a stem and a leaf opposite to it. Stem is square columnar, 15-40 cm long, 0.2-0.4 cm in diameter. The outer surface is purplish brown or light green, and there are short hairs near the edges, and the length between nodes is 2 ~ 5 cm. Quality is weak. The cut side is white, and the middle of the neck is hollow. The leaves are opposite and have short petioles, and the leaf pieces are crushed and rolled together. When whole leaves are unfolded, they are long oval or egg-shaped, 2 to 7 cm long and 1 to 3 cm wide. The upper surface of the leaf is dark green, and the lower surface is grayish green, rarely covered with short hairs, and has concave dot-shaped glands. Flowers are axillary, calyx is bell-shaped, and the end is split into 5 pieces, corolla is light purple. When you rub it with your hands, it has a unique clear scent, and the taste is spicy and cool. It grows in damp fields. Seeds are egg-shaped, light brown, light and small, and germinate well at room temperature of 20 → 30 → 15℃.
치자 (梔子, Gardenia Fruit)는 치자나무 Gardenia jasminoides Ellis (꼭두서니과 Rubiaceae)의 잘 익은 열매로서 그대로 또는 끓는 물에 데치거나 찐 것이다. 열매는 타원형으로 길이 3. 5cm 내외이고 보통 세로로 6개의 모서리가 붙어 있다. 열매의 바깥면은 적갈색 또는 황갈색을 띠고 있으나 내면은 황갈색이다. 열매의 내부는 두개의 방으로 나누어져 있고 종자는 편평하며 5mm 정도인데 덩어리로 엉겨 있다. 9월에 황홍색으로 익는다. 사화(瀉火), 제번(除煩), 청열(淸熱), 이뇨(利尿), 양혈(凉血), 해독(解毒) 작용을 한다. 일본 오키나와, 대만, 중국에 분포한다. Gardenia (梔子, Gardenia Fruit) is a well-ripe fruit of Gardenia jasminoides Ellis (Rubiaceae), either as it is or by boiling or steaming. The fruit is oval, about 3.5 cm long, and usually has 6 edges attached vertically. The outer surface of the fruit is reddish brown or yellowish brown, but the inner surface is yellowish brown. The inside of the fruit is divided into two chambers, and the seeds are flat, about 5mm, and are clumped together. It ripens to yellowish red in September. It has the functions of extinguishing fire, suppressing heat, clearing heat, diuretic, purifying blood, and detoxifying. Distributed in Okinawa, Taiwan, and China.
연교(連翹, Forsythia Fruit)는 물푸레나무과의 의성개나리 Forsythia viridissima Lindley 또는 당개나리 Forsythia suspense Vahl (물푸레나무과 Oleaceae)의 열매이다. 열매가 막 익기 시작하여 녹색 빛이 남아있을 때 채취하여 쪄서 말린 것을 청교 (靑翹)라 하고, 완전히 익었을 때 채취하여 말린 것을 노교 (老翹)라 한다. 열매는 달걀모양에 가깝고 약간 넓으며 납작하고 길이 10 ∼ 17 mm, 지름 5 ∼ 12 mm이다. 끝은 매우 뾰족하고 새부리처럼 벌어졌다. 아랫쪽은 약간 둥글고 열매자루는 남아있거나 떨어져있다. 바깥면은 갈색 또는 녹색이며 약간 볼록하고 고르지 않은 주름살이 있다. 약간 특유한 향기가 있고 맛은 쓰다. Forsythia Fruit is the fruit of Forsythia viridissima Lindley or Forsythia suspense Vahl (Oleaceae). When the fruit is just beginning to ripen and the green light remains, it is called Cheonggyo (靑翹), and when it is fully ripe and dried, it is called Nogyo (Nogyo). The fruit is close to egg shape, slightly wide and flat, 10 ~ 17 mm long, 5 ~ 12 mm in diameter. The tip is very pointed and flared like a bird's beak. The lower part is slightly round, and the fruit stalk remains or is separated. The outer surface is brown or green, slightly convex, and has uneven wrinkles. It has a slightly peculiar aroma and a bitter taste.
대황(大黃, Rhubarb)은 장엽대황 (掌葉大黃) Rheum palmatum Linne, 탕구트대황 Rheum tanguticum Maximowicz ex Balf. 또는 약용대황 (藥用大黃) Rheum officinale Baillon (여뀌과 Polygonaceae)의 뿌리 및 뿌리줄기로서 주피를 제거한 것이다. 뿌리줄기는 달걀모양, 긴 달걀모양 또는 원기둥모양이며, 때로 가로 및 세로로 잘려서 다듬어져 있으며, 길이 5 ∼ 15 cm, 지름 4 ∼ 10 cm이다. 바깥쪽은 껍질이 거의 벗겨져 있다. 특유한 냄새가 있고 맛은 떫고 쓰다. 이를 씹으면 가는 모래를 씹는 느낌이 있고 침을 노랗게 물들인다. 약리작용으로 대장운동촉진, 해열, 체온강하, 담즙분비 촉진, 혈액응고 시간 단축, 항균, 이뇨, 간기능 보호 등이 알려져 있다.Rhubarb (大黃, Rhubarb) is long-leaved rhubarb (掌葉大黃) Rheum palmatum Linne, Tangut rhubarb Rheum tanguticum Maximowicz ex Balf. Or medicinal rhubarb (藥用大黃) Rheum officinale Baillon (Polygonaceae) root and rhizome from which the pericarp is removed. The rhizome is egg-shaped, long egg-shaped or cylindrical, sometimes trimmed by cutting horizontally and vertically, and is 5 to 15 cm long and 4 to 10 cm in diameter. The outer shell is almost peeled off. It has a peculiar smell and tastes bitter and bitter. Chewing it gives the feeling of chewing fine sand and stains the saliva yellow. As for pharmacological action, it is known to promote colon movement, reduce fever, lower body temperature, promote bile secretion, shorten blood coagulation time, antibacterial, diuretic, and protect liver function.
본 발명자들은 상기 추출물의 모두가 피부에 유익한 생물학적 활성을 가지며, 상기 추출물 중 2이상의 복합 추출물, 바람직하게 황금 추출물, 박하 추출물, 치자 추출물, 연교 추출물 및 대황 추출물의 복합 추출물은 시너지적 생물학적 활성을 가짐을 발견하였다. The present inventors have found that all of the above extracts have biological activities beneficial to the skin, and that a combination of two or more of the above extracts, preferably a combination of golden extract, peppermint extract, gardenia extract, perilla extract and rhubarb extract, has synergistic biological activity.
본 발명에 있어서, 용어 "추출물"은 상기 추출처리에 의하여 얻어지는 추출액, 상기 추출액의 희석액이나 농축액, 상기 추출액을 건조하여 얻어지는 건조물, 상기 추출액의 조정제물이나 정제물, 또는 이들의 혼합물 등, 추출액 자체 및 추출액을 이용하여 형성 가능한 모든 제형의 추출물을 포함한다.In the present invention, the term "extract" includes an extract obtained by the extraction treatment, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, a crude or purified product of the extract, or a mixture thereof, and the extract itself and extracts of all formulations that can be formed using the extract.
상기 추출물에는 각각의 식물을 설명하며 특정된 부위 외에도 그의 잎, 줄기, 나무껍질, 뿌리, 꽃 또는 꽃눈, 과실, 종자, 수액 및 전체 식물이 포함될 수 있다. The extract describes each plant and may include its leaves, stems, barks, roots, flowers or flower buds, fruits, seeds, sap and whole plants in addition to the specified parts.
상기 추출물을 제조하기 위하여, 통상의 기술자는 당업계에 공지된 임의의 적합한 방법을 사용할 수 있다. 예를 들어, 용매 추출법에 의할 수 있다. 식물 전체 또는 임의의 부분을 분쇄한 다음(예를 들어, 블렌더), 추출 용매를 처리하여, 용매 추출물을 수득할 수 있다. 분쇄 전에 건조 과정(예를 들어, 40 내지 70℃에서 15 내지 50시간 동안의 건조)을 거친 다음 분쇄할 수도 있다. 또한, 용매 추출물은 감압 증류 및 동결건조 또는 분무 건조 등과 같은 추가적인 과정에 의해 분말 상태로 제조될 수 있다. To prepare the extract, one skilled in the art can use any suitable method known in the art. For example, it may be by a solvent extraction method. A solvent extract can be obtained by crushing the whole plant or any part thereof (eg, in a blender) and then treating the extraction solvent. It may be pulverized after undergoing a drying process (for example, drying at 40 to 70° C. for 15 to 50 hours) before pulverization. In addition, the solvent extract can be prepared in a powder state by additional processes such as distillation under reduced pressure and lyophilization or spray drying.
상기 사용되는 추출 용매의 종류는 특별히 제한되지 아니하며, 당해 기술 분야에서 공지된 임의의 용매를 사용할 수 있다. 상기 추출 용매의 비제한적인 예로는 물; 메탄올, 에탄올, 프로필알코올, 부틸알코올 등의 C1 내지 C4의 저급 알코올; 글리세린, 부틸렌글라이콜, 프로필렌글라이콜 등의 다가 알코올; 및 메틸아세테이트, 에틸아세테이트, 아세톤, 벤젠, 헥산, 디에틸에테르, 디클로로메탄 등의 탄화수소계 용매; 또는 이들의 혼합물을 사용할 수 있으며, 바람직하게, 물, 저급알코올을 단독으로 사용하거나 2종 이상 혼합하여 사용할 수 있다. 상기 용매를 사용하여 1회 이상 추출하여 용매 추출물을 제조할 수 있으며, 상기 용매 추출물을 감압 증류한 후 동결건조 또는 분무 건조하여 얻은 건조 추출물을 제조할 수 있다. The type of extraction solvent used is not particularly limited, and any solvent known in the art may be used. Non-limiting examples of the extraction solvent include water; C1 to C4 lower alcohols such as methanol, ethanol, propyl alcohol and butyl alcohol; polyhydric alcohols such as glycerin, butylene glycol, and propylene glycol; and hydrocarbon solvents such as methyl acetate, ethyl acetate, acetone, benzene, hexane, diethyl ether, and dichloromethane; Or a mixture thereof may be used, and preferably, water and lower alcohol may be used alone or in combination of two or more. A solvent extract may be prepared by extracting at least once using the solvent, and a dried extract obtained by distilling the solvent extract under reduced pressure and then freeze-drying or spray-drying may be prepared.
상기 추출 용매의 양은 이용되는 추출 용매의 종류에 따라 다양할 수 있으나, 예를 들어 대상 식물의 건조 중량에 대하여 1 내지 20배, 또는 5 내지 20배로 사용할 수 있다. The amount of the extraction solvent may vary depending on the type of extraction solvent used, but may be used, for example, 1 to 20 times, or 5 to 20 times the dry weight of the target plant.
이외에도, 당업계 공지된 다양한 추출 공정, 예를 들어, 온침(maceration), 인퓨전(infusion), 퍼콜레이션(percolation), 소화, 전즙(decoction), 고온 연속 추출(hot continuous extraction), 수성-알코올성 추출, 역류 추출, 마이크로파 보조 추출, 초음파 추출, 초임계 유체 추출, 조직편 추출(phytonic extract) (예를 들어, 하이드로-플루오로-카본 용매) 등) 등을 선택하여 사용할 수 있으며, 이들은 단독으로 수행되거나 2 종 이상의 방법을 병용하여 수행될 수 있다.In addition, various extraction processes known in the art, for example, maceration, infusion, percolation, digestion, decoction, hot continuous extraction, aqueous-alcoholic extraction, countercurrent extraction, microwave assisted extraction, ultrasonic extraction, supercritical fluid extraction, phytonic extract (eg, hydro-fluoro-carbon solvent), etc.) may be selected and used, and these may be performed alone or Two or more methods may be used in combination.
[조성물][composition]
본 발명에 따른 조성물은 황금 추출물, 박하 추출물, 치자 추출물, 연교 추출물 및 대황 추출물을 유효성분으로 포함한다. The composition according to the present invention contains golden extract, peppermint extract, gardenia extract, yeongyo extract and rhubarb extract as active ingredients.
본 발명에 있어서 "유효성분으로 포함된다"는 의미는 본 발명에 따른 조성물로부터 피부 개선 효과를 나타낼 수 있는 정도로, 추출물이 첨가되는 것을 의미하고, 피부 전달 및 안정화 등을 위하여 다양한 성분을 부성분으로 첨가하여 다양한 형태로 제형화(formulation) 가능함을 포함하는 의미이다.In the present invention, the meaning of "included as an active ingredient" means that an extract is added to the extent that can exhibit a skin improvement effect from the composition according to the present invention, and various ingredients are added as subcomponents for skin delivery and stabilization. It means that it can be formulated in various forms.
본 발명에 따른 조성물에 각각의 식물 추출물은 다음과 같은 비율로 포함될 수 있다. 예를 들어, 황금 추출물, 박하 추출물, 치자 추출물, 연교 추출물 및 대황 추출물의 중량비는 1: 0.01 ~ 10: 0.01 ~ 10: 0.01 ~ 10: 0.01 ~ 10 또는 1: 0.1 ~ 10: 0.1 ~ 10: 0.1 ~ 10: 0.1 ~ 10, 1: 1 ~ 5: 1 ~ 5: 1 ~ 5: 1 ~ 5, 또는 1: 1 ~ 3: 1 ~ 3: 1 ~ 3: 1 ~ 3으로 포함될 수 있다. Each plant extract in the composition according to the present invention may be included in the following ratio. For example, the weight ratio of gold extract, peppermint extract, gardenia extract, perilla extract and rhubarb extract is 1: 0.01 to 10: 0.01 to 10: 0.01 to 10: 0.01 to 10 or 1: 0.1 to 10: 0.1 to 10: 0.1 to 10: 0.1 to 10, 1: 1 to 5: 1 to 5: 1 to 5: 1 to 5, or 1: 1 to 3: 1 to 3: 1 to 3: 1 to 3.
본 발명에 따른 조성물에는 본 발명의 1 이상의 추출물이 조성물의 최종 형태 중에 적어도 약 0.0001%, 0.0002%, 0.0003%, 0.0004%, 0.0005%, 0.0006%, 0.0007%, 0.0008%, 0.0009%, 0.0010%, 0.0011%, 0.0012%, 0.0013%, 0.0014%, 0.0015%, 0.0016%, 0.0017%, 0.0018%, 0.0019%, 0.0020%, 0.0021%, 0.0022%, 0.0023%, 0.0024%, 0.0025%, 0.0026%, 0.0027%, 0.0028%, 0.0029%, 0.0030%, 0.0031%, 0.0032%, 0.0033%, 0.0034%, 0.0035%, 0.0036%, 0.0037%, 0.0038%, 0.0039%, 0.0040%, 0.0041%, 0.0042%, 0.0043%, 0.0044%, 0.0045%, 0.0046%, 0.0047%, 0.0048%, 0.0049%, 0.0050%, 0.0051%, 0.0052%, 0.0053%, 0.0054%, 0.0055%, 0.0056%, 0.0057%, 0.0058%, 0.0059%, 0.0060%, 0.0061%, 0.0062%, 0.0063%, 0.0064%, 0.0065%, 0.0066%, 0.0067%, 0.0068%, 0.0069%, 0.0070%, 0.0071%, 0.0072%, 0.0073%, 0.0074%, 0.0075%, 0.0076%, 0.0077%, 0.0078%, 0.0079%, 0.0080%, 0.0081%, 0.0082%, 0.0083%, 0.0084%, 0.0085%, 0.0086%, 0.0087%, 0.0088%, 0.0089%, 0.0090%, 0.0091%, 0.0092%, 0.0093%, 0.0094%, 0.0095%, 0.0096%, 0.0097%, 0.0098%, 0.0099%, 0.0100%, 0.0200%, 0.0250%, 0.0275%, 0.0300%, 0.0325%, 0.0350%, 0.0375%, 0.0400%, 0.0425%, 0.0450%, 0.0475%, 0.0500%, 0.0525%, 0.0550%, 0.0575%, 0.0600%, 0.0625%, 0.0650%, 0.0675%, 0.0700%, 0.0725%, 0.0750%, 0.0775%, 0.0800%, 0.0825%, 0.0850%, 0.0875%, 0.0900%, 0.0925%, 0.0950%, 0.0975%, 0.1000%, 0.1250%, 0.1500%, 0.1750%, 0.2000%, 0.2250%, 0.2500%, 0.2750%, 0.3000%, 0.3250%, 0.3500%, 0.3750%, 0.4000%, 0.4250%, 0.4500%, 0.4750%, 0.5000%, 0.5250%, 0.550%, 0.5750%, 0.6000%, 0.6250%, 0.6500%, 0.6750%, 0.7000%, 0.7250%, 0.7500%, 0.7750%, 0.8000%, 0.8250%, 0.8500%, 0.8750%, 0.9000%, 0.9250%, 0.9500%, 0.9750%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8.0%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9.0%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% 또는 99% 이상의 범위로 본 발명의 1 이상의 추출물을 포함할 수 있다. A composition according to the present invention comprises at least about 0.0001%, 0.0002%, 0.0003%, 0.0004%, 0.0005%, 0.0006%, 0.0007%, 0.0008%, 0.0009%, 0.0010%, 0.0011%, 0 .0012%, 0.0013%, 0.0014%, 0.0015%, 0.0016%, 0.0017%, 0.0018%, 0.0019%, 0.0020%, 0.0021%, 0.0022%, 0.0023%, 0.0024%, 0.0025% , 0.0026%, 0.0027%, 0.0028%, 0.0029%, 0.0030%, 0.0031%, 0.0032%, 0.0033%, 0.0034%, 0.0035%, 0.0036%, 0.0037%, 0.0038%, 0.00 39%, 0.0040%, 0.0041%, 0.0042%, 0.0043%, 0.0044%, 0.0045%, 0.0046%, 0.0047%, 0.0048%, 0.0049%, 0.0050%, 0.0051%, 0.0052%, 0 .0053%, 0.0054%, 0.0055%, 0.0056%, 0.0057%, 0.0058%, 0.0059%, 0.0060%, 0.0061%, 0.0062%, 0.0063%, 0.0064%, 0.0065%, 0.0066% , 0.0067%, 0.0068%, 0.0069%, 0.0070%, 0.0071%, 0.0072%, 0.0073%, 0.0074%, 0.0075%, 0.0076%, 0.0077%, 0.0078%, 0.0079%, 0.00 80%, 0.0081%, 0.0082%, 0.0083%, 0.0084%, 0.0085%, 0.0086%, 0.0087%, 0.0088%, 0.0089%, 0.0090%, 0.0091%, 0.0092%, 0.0093%, 0 .0094%, 0.0095%, 0.0096%, 0.0097%, 0.0098%, 0.0099%, 0.0100%, 0.0200%, 0.0250%, 0.0275%, 0.0300%, 0.0325%, 0.0350%, 0.0375% , 0.0400%, 0.0425%, 0.0450%, 0.0475%, 0.0500%, 0.0525%, 0.0550%, 0.0575%, 0.0600%, 0.0625%, 0.0650%, 0.0675%, 0.0700%, 0.07 25%, 0.0750%, 0.0775%, 0.0800%, 0.0825%, 0.0850%, 0.0875%, 0.0900%, 0.0925%, 0.0950%, 0.0975%, 0.1000%, 0.1250%, 0.1500%, 0 .1750%, 0.2000%, 0.2250%, 0.2500%, 0.2750%, 0.3000%, 0.3250%, 0.3500%, 0.3750%, 0.4000%, 0.4250%, 0.4500%, 0.4750%, 0.5000% , 0.5250%, 0.550%, 0.5750%, 0.6000%, 0.6250%, 0.6500%, 0.6750%, 0.7000%, 0.7250%, 0.7500%, 0.7750%, 0.8000%, 0.8250%, 0.850 0%, 0.8750%, 0.9000%, 0.9250%, 0.9500%, 0.9750%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2. 2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6. 3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8.0%, 8.1%, 8.2%, 8.3%, 8.4% 8.5% 8.6% 8.7% 8.8% 8.9% 9.0% 9.1% 9.2% 9.3% 9.4% 9.5% 9.6% 9.7% 9.8% 9.9% 10% 11% 12% 13% 14% 1 5%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 99% or more of one or more extracts of the present invention.
또한, 본 발명에 따른 조성물에는 본 발명의 각각의 추출물이 조성물의 최종 형태 중에 적어도 약 0.0001% ~ 90%, 0.001% ~ 90%, 0.01% ~ 90%, 0.1% ~ 90%, 0.1% ~ 90%, 0.1% ~ 80%, 0.1% ~ 70%, 0.1% ~ 60%, 0.1% ~ 50%, 0.1% ~ 40%, 0.1% ~ 30%, 0.1% ~ 20% 또는 0.1% ~ 10%의 범위로 포함될 수 있다. In addition, in the composition according to the present invention, each extract of the present invention in the final form of the composition is at least about 0.0001% to 90%, 0.001% to 90%, 0.01% to 90%, 0.1% to 90%, 0.1% to 90%, 0.1% to 80%, 0.1% to 70%, 0.1% to 60%, 0.1% to 50%, 0.1% to 40%, 0.1% to 30%, 0.1% to 20%, or 0.1% to 10%.
상기 %는 조성물의 총 중량 대비 중량 또는 총 부피 대비 부피를 기준으로 계산할 수 있으며, 농도는 조성물의 목적하는 효과 또는 조성물이 혼입되는 제품에 따라 조정될 수 있다. The % may be calculated based on the total weight of the composition or the total volume of the composition, and the concentration may be adjusted according to the desired effect of the composition or the product into which the composition is incorporated.
본 발명의 조성물은 모든 유형의 비히클 내로 제형화될 수 있다. 적합한 비히클의 예로는 에멀젼 (예를 들어, 수중유, 유중수, 수중실리콘, 실리콘중수, 수중유중수, 수중유, 유중수중유, 실리콘중수중유 등), 크림, 로션, 용액 (예컨대, 수성 및 하이드로-알코올 용액), 무수 베이스 (예를 들어, 립스틱 및 파우더), 젤, 연고, 페이스트, 밀크, 액체, 에어로졸, 고체 형태 또는 아이 젤리를 포함하나, 이에 제한되지 않는다. Compositions of the present invention may be formulated into all types of vehicles. Examples of suitable vehicles include, but are not limited to, emulsions (e.g., oil in water, water in oil, silicone in water, water in silicone, water in oil in water, oil in water, oil in water in oil, oil in water in silicone, etc.), creams, lotions, solutions (e.g., aqueous and hydro-alcoholic solutions), anhydrous bases (e.g., lipsticks and powders), gels, ointments, pastes, milks, liquids, aerosols, solid forms, or eye jellies.
또한, 본 발명의 조성물은 피부와 같은 표적 영역으로의 운반을 위해 캡슐화될 수도 있다. 캡슐화 기술은 예를 들어, 피부에 성분을 전달하는 운반 비히클로서 사용될 수 있는 리포좀, 소낭, 및/또는 나노입자 (예를 들어, 성분이 트랩핑(trap되고, 캡슐화되고/되거나 흡수되는 폴리머 물질을 포함하는 생분해성 및 비생분해성 콜로이드성 입자, 예를 들어 나노스피어(nanosphere) 및 나노캡슐을 포함)의 사용을 포함할 수 있으나, 이에 제한되지 않는다.Compositions of the present invention may also be encapsulated for delivery to a target area such as the skin. Encapsulation techniques may include, but are not limited to, the use of liposomes, vesicles, and/or nanoparticles (including, for example, biodegradable and non-biodegradable colloidal particles comprising polymeric materials into which an ingredient is trapped, encapsulated, and/or absorbed, such as nanospheres and nanocapsules), which can be used as a delivery vehicle to deliver ingredients to the skin, for example.
본 발명에 따른 조성물은 다양하게 제품화될 수 있다. 예를 들어, 화장료 조성물, 식품 조성물 등으로 제품화될 수 있다. The composition according to the present invention can be variously commercialized. For example, it can be commercialized as a cosmetic composition, a food composition, and the like.
상기 화장료 조성물은 예를 들어, 일반적인 유화 제형 및 가용화 제형의 형태로 제조된 것일 수 있다. 예를 들어, 유연 화장수 또는 영양 화장수 등과 같은 화장수, 훼이셜 로션, 바디로션 등과 같은 유액, 영양 크림, 수분 크림, 아이크림, 마사지 크림 등과 같은 크림, 에센스, 세럼, 화장연고, 스프레이, 오일젤, 젤, 팩, 선 스크린, 메이크업베이스, 액체 타입, 고체 타입 또는 스프레이 타입 등의 파운데이션, 파우더, 클렌징 크림, 클렌징 로션, 클렌징 오일과 같은 메이크업 제거제, 클렌징 폼, 비누, 바디 워쉬 등과 같은 세정제 등의 제형을 가질 수 있으나, 이에 제한되지 않고 당업계 공지된 다양한 형태로 제형화될 수 있다. The cosmetic composition may be prepared in the form of, for example, a general emulsified formulation and a solubilized formulation. For example, lotion such as flexible lotion or nutrient lotion, emulsion such as facial lotion, body lotion, cream such as nourishing cream, moisture cream, eye cream, massage cream, essence, serum, cosmetic ointment, spray, oil gel, gel, pack, sunscreen, makeup base, liquid type, solid type, or spray type foundation, powder, cleansing cream, cleansing lotion, makeup remover such as cleansing oil, cleanser such as cleansing foam, soap, body wash, etc. It is not limited thereto and may be formulated in various forms known in the art.
상기 화장료 조성물에는 본 발명의 목적을 해하지 않는 이상, 본 발명에 따른 추출물 외에 당업계에서 화장료 조성물의 성분으로 공지된 통상의 성분을 임의로 선택하여 추가로 포함할 수 있다. 예를 들어, 방향제 (합성 및 천연), 염료 및 색 성분, 흡수제, 에멀젼화제, 안정화제, 윤활제, 용매, 보습제 (예: 연화제(emollient), 습윤제(humectant), 피막형성제, 폐색제(occlusive agent), 및 피부의 천연 보습 메커니즘에 영향을 주는 작용제(agent)를 포함함), 발수제(water-repellant), UV 흡수제 (물리적 및 화학적 흡수제, 예를 들어 파라아미노벤조산("PABA") 및 상응하는 PABA 유도체, 이산화티탄, 산화아연 등), 에센셜 오일(essential oil), 비타민 (예: A, B, C, D, E 및 K), 미량 금속 (예: 아연, 칼슘 및 셀레늄), 항-자극제(anti-irritant) (예: 스테로이드 및 비스테로이드성 항염증제), 항-미생물제(anti-microbial agent), 항산화제 (예: BHT 및 토코페롤), 킬레이트제 (예: 다이소듐 EDTA 및 테트라소듐 EDTA), 보존제 (예: 메틸파라벤 및 프로필파라벤), pH 조정제 (예: 수산화나트륨 및 시트르산), 흡수제(예를 들어, 알루미늄 전분 옥테닐석시네이트, 카올린, 옥수수 전분, 귀리 전분, 사이클로덱스트린, 탈크(talc), 및 제올라이트), 피부 블리칭(bleaching) 및 라이트닝(lightening) 작용제 (예를 들어, 하이드로퀴논 및 니아신아미드 락테이트), 습윤제 (예: 글리세린, 프로필렌 글리콜, 부틸렌 글리콜, 펜틸렌 글리콜, 소르비톨, 우레아 및 만니톨), 각질제거제(exfoliant) (예: 알파-하이드록시산 및 베타-하이드록시산, 예를 들어 락트산, 글리콜산 및 살리실산; 및 이들의 염), 발수제(waterproofing agent) (예: 마그네슘/알루미늄 하이드록시드 스테아레이트), 피부 컨디셔닝제(skin conditioning agent) (예: 알로에 추출물, 알란토인(allantoin), 비사볼롤(bisabolol), 세라마이드(ceramide), 다이메티콘, 히알루론산, 및 다이포타슘 글리시리제이트), 및 증점제(thickening agent) (예:조성물의 점도를 증가시킬 수 있는 물질, 예를 들어 카복실산 폴리머, 가교된 폴리아크릴레이트 폴리머, 폴리아크릴아미드 폴리머, 다당류 및 검(gum)) 및 실리콘 함유 화합물(예: 실리콘 오일 및 폴리오르가노실록산) 등이 포함될 수 있다. The cosmetic composition may further include, in addition to the extract according to the present invention, a conventional component known as a component of a cosmetic composition in the art, optionally selected, as long as the object of the present invention is not impaired. For example, fragrances (synthetic and natural), dyes and color ingredients, absorbents, emulsifiers, stabilizers, lubricants, solvents, humectants (including emollients, humectants, film formers, occlusive agents, and agents that affect the skin's natural moisturizing mechanism), water-repellants, UV absorbers (physical and chemical absorbers such as para Aminobenzoic acid ("PABA") and its corresponding PABA derivatives, titanium dioxide, zinc oxide, etc.), essential oils, vitamins (e.g. A, B, C, D, E and K), trace metals (e.g. zinc, calcium and selenium), anti-irritants (e.g. steroid and non-steroidal anti-inflammatory drugs), anti-microbial agents, antioxidants (e.g. BHT and tocopherol), chelating agents (e.g., disodium EDTA and tetrasodium EDTA), preservatives (e.g., methylparaben and propylparaben), pH adjusting agents (e.g., sodium hydroxide and citric acid), absorbents (e.g., aluminum starch octenylsuccinate, kaolin, corn starch, oat starch, cyclodextrins, talc, and zeolites), skin bleaching and lightening agents (e.g., hydroquinone and niacinamide lactate), humectants (eg glycerin, propylene glycol, butylene glycol, pentylene glycol, sorbitol, urea and mannitol), exfoliants (eg alpha-hydroxy acids and beta-hydroxy acids such as lactic acid, glycolic acid and salicylic acid; and salts thereof), waterproofing agents (eg magnesium/aluminum hydroxy de stearate), skin conditioning agents (e.g., aloe extract, allantoin, bisabolol, ceramide, dimethicone, hyaluronic acid, and dipotassium glycyrrhizate), and thickening agents (e.g., substances capable of increasing the viscosity of the composition, such as carboxylic acid polymers, crosslinked polyacrylate polymers, poly acrylamide polymers, polysaccharides and gums) and silicone-containing compounds such as silicone oils and polyorganosiloxanes.
상기 식품 조성물은 예를 들어, 음료, 강화수(fortified water), 에너지 드링크(energy drink), 영양 드링크(nutritional drink), 고형 식품, 비타민, 보조제(Supplement) 등) 등으로 제형화 될 수 있으나, 이에 제한되지 않는다. 상기 보조제에는 비타민, 무기질, 허브 또는 기타 식물, 아미노산, 효소 및 대사산물을 포함할 수 있다. 이러한 보조제는 경구 이용(oral consumption)에 적합하며, 경구로 투여될 수 있다.The food composition may be formulated into, for example, beverages, fortified water, energy drinks, nutritional drinks, solid foods, vitamins, supplements, etc.), but is not limited thereto. Such supplements may include vitamins, minerals, herbs or other botanicals, amino acids, enzymes and metabolites. These adjuvants are suitable for oral consumption and can be administered orally.
본 발명에 따른 조성물을 포함하는 키트로 제공될 수 있다. 본 발명에 따른 조성물은 용기에 포함되며, 용기는 병, 금속 튜브, 라미네이트 튜브, 플라스틱 튜브, 디스펜서, 압력 용기, 장벽 용기, 패키지, 컴파트먼트, 립스틱 용기, 컴팩트 용기, 화장용 조성물을 담을 수 있는 화장용 팬, 또는 다른 타입의 용기 예를 들어, 분산매체(dispersions) 또는 조성물 또는 유지되는 바람직한 병, 디스펜서, 또는 패키지 내로 주입 또는 블로우-몰드된 플라스틱 용기 등이 포함될 수 있으며, 이에 제한되지 않는다. 상기 키트에는 키트 또는 조성물을 사용하기 위한 지시서가 포함될 수 있으며, 지시서는 별개 용지에 기재될 수 있고, 또는 용기 표면, 용기 포장지의 표면 상에 기재될 수 있다. 지시사항은 글자, 어구, 약어, 그림 또는 기호 등이 포함되며, 이에 제한되지 않는다. 지시서에는 예를 들어, 키트 또는 조성물의 사용 방법, 적용 방법 및 유지 방법 등에 관한 지시사항이 포함될 수 있다. 용기에는 사전에 정해진 양에 따라 분배하여 담을 수 있다. It may be provided as a kit comprising a composition according to the present invention. The composition according to the present invention is contained in a container, which may include, but is not limited to, a bottle, a metal tube, a laminate tube, a plastic tube, a dispenser, a pressure vessel, a barrier container, a package, a compartment, a lipstick container, a compact container, a cosmetic pan capable of containing the cosmetic composition, or other types of containers such as, but not limited to, a dispersion or composition or preferred bottle, dispenser, or plastic container that is injected or blow-molded into a package. The kit may include instructions for using the kit or composition, and the instructions may be written on a separate sheet or may be written on the surface of the container or the surface of the container wrapper. Instructions include, but are not limited to, letters, phrases, abbreviations, pictures, or symbols. Instructions may include, for example, instructions on how to use, apply, and maintain the kit or composition. The containers may be distributed according to a pre-determined amount.
본 발명에 따른 조성물은 국소용 피부 조성물로 제공될 수 있다. Compositions according to the present invention may be provided as topical skin compositions.
또한, 본 발명에 따른 조성물을 피부에 국소 적용하는 방법을 제공할 수 있다. In addition, a method of topically applying the composition according to the present invention to the skin can be provided.
본 발명에 있어서, 용어 "국소 적용"은 조성물을 각질 조직의 표면상에 적용하거나 도포하는 것을 의미하고, "국소용 피부 조성물"은 각질 조직상에 국소 적용 또는 도포하기에 적합한 조성물을 포함한다. 이러한 조성물은 피부에 적용되는 경우 과도한 독성, 부적합성(incompatibility), 불안정성, 알러지 반응 등을 갖지 않는다는 점에서, 전형적으로 피부과학적으로 허용된다. 본 발명의 국소 피부 케어 조성물은, 피부에 적용된 후 현저한 적하(dripping) 또는 풀링(pooling)을 회피하기 위해, 선택된 점도를 지닐 수 있다.In the present invention, the term "topical application" means applying or applying a composition onto the surface of keratinous tissue, and "topical skin composition" includes a composition suitable for topical application or application onto keratinous tissue. Such compositions are typically dermatologically acceptable in that they do not exhibit excessive toxicity, incompatibility, instability, allergic reactions, etc. when applied to the skin. Topical skin care compositions of the present invention may have a selected viscosity to avoid significant dripping or pooling after application to the skin.
[피부 개선 효과][Skin improvement effect]
본 발명에 따른 조성물은 생물학적 활성을 가지며, 피부 개선에 우수한 효과를 발휘한다. The composition according to the present invention has biological activity and exhibits excellent effects on skin improvement.
보다 구체적으로, 본 발명에 따른 조성물은 피부 미백 효과가 있다. More specifically, the composition according to the present invention has a skin whitening effect.
상기 피부 미백은 멜라닌 형성 과정을 억제 시켜 멜라닌 생성을 억제하는 것을 말한다. The skin whitening refers to inhibiting melanin production by inhibiting the melanin formation process.
본 발명자들은 본 발명에 따른 복합 추출물을 멜라노마 세포에 처리하여 생성된 멜라닌의 양을 측정한 결과, 각각의 추출물을 처리하는 경우에 비해 상승된 멜라닌 생성 억제 효과를 보임을 확인할 수 있었고, 피부 미백 효과가 공지되어 있는 알부틴에 상응하는 효과를 발휘함을 알 수 있었다. 본 발명에 따른 조성물은 멜라닌 생성 저해 효과가 뛰어나, 우수한 미백 효과를 나타냄을 확인하였다. The present inventors measured the amount of melanin produced by treating the complex extract according to the present invention to melanoma cells, and as a result, it was confirmed that the melanin production inhibitory effect was increased compared to the case of treating each extract, and the skin whitening effect was known. It was found to exert an effect corresponding to arbutin. It was confirmed that the composition according to the present invention has an excellent melanin production inhibitory effect and exhibits an excellent whitening effect.
또한, 본 발명에 따른 조성물은 주름 및 탄력 개선 효과가 있다. In addition, the composition according to the present invention has an effect of improving wrinkles and elasticity.
상기 피부 주름이란 피부가 쇠하여 생긴 잔줄을 의미하며, 유전자에 의한 원인, 피부 진피에 존재하는 콜라겐과 엘라스틴의 감소, 외부환경 등에 의해 유발될 수 있다. The skin wrinkles refer to fine lines caused by skin deterioration, and may be caused by genes, reduction of collagen and elastin present in the skin dermis, external environment, and the like.
상기 주름 개선은 주름이 생성되는 것을 억제 또는 저해하거나, 이미 생성된 주름을 완화시키는 것을 말한다.The wrinkle improvement refers to suppressing or inhibiting the formation of wrinkles or alleviating wrinkles already formed.
상기 피부 탄력이란 진피층에 존재하는 엘라스틴(elastin)으로 구성된 탄력섬유 및 콜라겐(collagen)이라고 하는 교원섬유에 의해 나타나는 것으로, 상기 탄력 개선이란 탄력 감소를 억제 또는 저해하거나, 탄력을 유지 또는 향상시키는 것을 말한다.The skin elasticity is expressed by elastic fibers composed of elastin and collagen fibers present in the dermal layer, and the improvement of elasticity refers to suppressing or inhibiting a decrease in elasticity, maintaining or improving elasticity.
본 발명자들은 본 발명에 따른 복합 추출물을 처리한 결과, 각각의 추출물을 처리하는 경우에 비해 상승된 콜라겐 합성 촉진 효과 및 엘라스타제 활성 저해 효과를 보임을 확인할 수 있었고, 피부 콜라겐 합성 촉진 효과가 공지되어 있는 비타민 C에 상응하는 효과를 보이며, 피부 엘라스타제 활성 저해 효과가 공지되어 있는 퀘세틴에 상응하는 효과를 발휘함을 알 수 있었다. 본 발명에 따른 조성물은 콜라겐 합성 촉진 효과 및 엘라스타제 활성 저해 효과가 뛰어나, 우수한 피부 주름 및 탄력 개선 효과를 나타냄을 확인하였다. As a result of treating the complex extract according to the present invention, the present inventors confirmed that the collagen synthesis promoting effect and the elastase activity inhibitory effect were increased compared to the case of treating each extract, and the skin collagen synthesis promoting effect was known. It showed an effect corresponding to vitamin C, and it was found that the effect corresponding to quercetin, which is known to inhibit skin elastase activity, was exhibited. It was confirmed that the composition according to the present invention has an excellent collagen synthesis promoting effect and an elastase activity inhibiting effect, and exhibits excellent skin wrinkle and elasticity improvement effects.
또한, 본 발명에 따른 조성물은 트러블 개선 효과가 있다. In addition, the composition according to the present invention has a trouble improving effect.
상기 트러블은 피부자극 및 염증과 같은 증상을 말하며, 염증 반응에는 외부 자극에 의한 동통, 발열, 발적, 종창, 기능상실 등을 특징으로 하며, 조직학적으로는 소동맥, 모세혈관 및 소정맥의 투과성과 혈류증가를 동반한 확장, 혈장단백을 함유한 혈장의 삼출, 백혈구의 염증부위로의 이동 등을 포함한 복잡한 증상 등이 관찰되며, 피부자극 반응에는 홍반, 가려움, 발열 등을 특징으로 한다. The trouble refers to symptoms such as skin irritation and inflammation, and the inflammatory reaction is characterized by pain, fever, redness, swelling, loss of function, etc. caused by external stimuli. Histologically, complex symptoms including dilatation accompanied by increased permeability and blood flow of arterioles, capillaries, and venules, exudation of plasma containing plasma proteins, migration of leukocytes to inflamed areas, etc. are observed, and skin irritation reactions are characterized by erythema, itching, fever, etc.
상기 트러블 개선은 피부 자극 반응 및 염증 반응을 억제 또는 저해하거나, 이미 진행 중인 피부자극 반응 또는 염증 반응을 완화시키는 것을 말한다.The improvement of the trouble refers to inhibiting or inhibiting the skin irritation reaction and inflammatory reaction, or alleviating the skin irritation reaction or inflammatory reaction that is already in progress.
본 발명자들은 본 발명에 따른 복합 추출물을 처리하여 NO 생성 저해 효과를 측정한 결과, 각각의 추출물을 처리하는 경우에 비해 상승된 NO 생성 저해 효과를 보임을 확인할 수 있었고, NO 생성 저해 효과가 공지되어 있는 L-NMMA에 상응하는 효과를 발휘함을 알 수 있었다. 본 발명에 따른 조성물은 NO 생성 저해를 통해 우수한 피부 트러블 개선 효과를 나타냄을 확인하였다. The present inventors measured the NO production inhibitory effect by treating the composite extract according to the present invention, and as a result, it was confirmed that the NO production inhibitory effect was increased compared to the case of treating each extract, and the NO production inhibitory effect was known. It was found to exert an effect corresponding to L-NMMA. It was confirmed that the composition according to the present invention exhibits an excellent skin trouble improvement effect through inhibition of NO production.
또한, 본 발명에 따른 조성물은 항산화 효과가 있다. In addition, the composition according to the present invention has an antioxidant effect.
상기 항산화는 세포내 대사 또는 자외선의 영향으로 인한 산화적 스트레스에 따라 반응성이 높은 자유 라디칼(free radical) 또는 활성산소종(reactive oxygen species;ROS)에 의한 세포의 산화를 억제하는 것을 말하며, 자유 라디칼 또는 활성산소종을 제거하여 이로 인한 세포의 손상이 감소되는 것을 포함한다. The antioxidant refers to inhibiting oxidation of cells by free radicals or reactive oxygen species (ROS) that are highly reactive according to oxidative stress caused by intracellular metabolism or ultraviolet rays, and includes reducing damage to cells caused by removing free radicals or reactive oxygen species.
본 발명자들은 본 발명에 따른 복합 추출물을 처리하여 자유라디칼 소거능을 측정한 결과, 각각의 추출물을 처리하는 경우에 비해 상승된 자유라디칼 소거능을 보임을 확인할 수 있었고, 자유라디칼 소거능이 공지되어 있는 비타민 C에 상응하는 효과를 발휘함을 알 수 있었다. 이로써, 본 발명에 따른 조성물은 우수한 항산화 효과를 나타냄을 확인하였다. The present inventors measured the free radical scavenging activity by treating the complex extract according to the present invention, and as a result, it was confirmed that the free radical scavenging activity was increased compared to the case of treating each extract, and the free radical scavenging activity was known. It was found to exert an effect corresponding to vitamin C. Thus, it was confirmed that the composition according to the present invention exhibits an excellent antioxidant effect.
또한, 본 발명에 따른 조성물은 항당화 효과가 있다. In addition, the composition according to the present invention has an anti-glycation effect.
상기 당화는 과도한 당의 존재로 인해 생기는 당과 단백질이 결합해 단백질이 파괴되는 현상을 말한다. 당화가 진행되면 콜라겐과 엘라스틴에 영향을 미쳐 피부 탄력 저하 외 각종 피부손상을 일으키게 된다. The glycation refers to a phenomenon in which proteins are destroyed by binding sugars and proteins caused by the presence of excessive sugars. When glycation progresses, it affects collagen and elastin, causing various skin damages including loss of skin elasticity.
본 발명자들은 본 발명에 따른 복합 추출물을 처리하여 항당화 효과를 측정한 결과, 각각의 추출물을 처리하는 경우에 비해 상승된 항당화 효과를 보임을 확인할 수 있었고, 항당화 효과가 공지되어 있는 아미노구아니딘에 상응하는 효과를 발휘함을 알 수 있었다. 이로써, 본 발명에 따른 조성물은 우수한 항당화 효과를 나타냄을 확인하였다. As a result of measuring the antiglycation effect by treating the composite extract according to the present invention, the present inventors confirmed that the antiglycation effect was increased compared to the case of treating each extract, and it was found that the antiglycation effect exhibited an effect corresponding to aminoguanidine known. As a result, it was confirmed that the composition according to the present invention exhibits an excellent antiglycation effect.
본 발명에 따른 복합 추출물은 피부에 안전하며, 항당화 효과, 피부 미백 효과, 피부 탄력 및 주름 개선 효과, 항산화 효과 및 피부 트러블 개선 효과를 포함한 피부 개선에 탁월한 효과를 갖는다. 또한, 섬유아세포증식 효과를 갖는다. The complex extract according to the present invention is safe for the skin and has excellent effects on skin improvement including antiglycation effect, skin whitening effect, skin elasticity and wrinkle improvement effect, antioxidant effect and skin trouble improvement effect. In addition, it has a fibroblast proliferative effect.
이하, 본 발명을 구체적으로 설명하기 위해 실시예 및 실험예를 들어 상세하게 설명하기로 한다. 그러나 본 발명에 따른 실시예 및 실험예는 여러 가지 다른 형태로 변형될 수 있으며, 본 발명의 범위가 아래에서 상술하는 실시예 및 실험예에 한정되는 것으로 해석 되어서는 안 된다. 본 발명의 실시예 및 실험예는 당업계에서 평균적인 지식을 가진 자에게 본 발명을 보다 완전하게 설명하기 위해서 제공되는 것이다.Hereinafter, examples and experimental examples will be described in detail to explain the present invention in detail. However, the examples and experimental examples according to the present invention may be modified in many different forms, and the scope of the present invention should not be construed as being limited to the examples and experimental examples described below. Examples and experimental examples of the present invention are provided to more completely explain the present invention to those skilled in the art.
<실시예 1> 박하추출물의 제조<Example 1> Preparation of peppermint extract
박하를 잘 건조하여 세절한 후, 건조중량 100g을 플라스크에 넣고 추출용매(증류수) 1000g으로 3일간 냉침하여 추출하였다. 냉침된 추출물을 0.2㎛의 기공 크기를 가진 필터로 여과하여 박하추출물을 제조하였다.After mint was well dried and chopped, 100 g of dry weight was placed in a flask and extracted by cooling with 1000 g of an extraction solvent (distilled water) for 3 days. The chilled extract was filtered through a filter having a pore size of 0.2 μm to prepare a peppermint extract.
<실시예 2> 치자추출물의 제조<Example 2> Preparation of gardenia extract
치자를 잘 건조하여 세절한 후, 건조중량 100g을 플라스크에 넣고 추출용매(증류수) 1000g으로 3일간 냉침하여 추출하였다. 냉침된 추출물을 0.2㎛의 기공 크기를 가진 필터로 여과하여 치자추출물을 제조하였다.After drying and cutting the gardenia well, 100 g of dry weight was put in a flask and extracted by cooling with 1000 g of an extraction solvent (distilled water) for 3 days. The chilled extract was filtered through a filter having a pore size of 0.2 μm to prepare a gardenia extract.
<실시예 3> 연교추출물의 제조<Example 3> Preparation of Yeongyo extract
연교를 잘 건조하여 세절한 후, 건조중량 100g을 플라스크에 넣고 추출용매(증류수) 1000g으로 3일간 냉침하여 추출하였다. 냉침된 추출물을 0.2㎛의 기공 크기를 가진 필터로 여과하여 연교추출물을 제조하였다.After well drying and slicing, 100 g of dry weight was placed in a flask and extracted by cooling with 1000 g of an extraction solvent (distilled water) for 3 days. The chilled extract was filtered through a filter having a pore size of 0.2 μm to prepare a Yeongyo extract.
<실시예 4> 대황추출물의 제조<Example 4> Preparation of rhubarb extract
대황을 잘 건조하여 세절한 후, 건조중량 100g을 플라스크에 넣고 추출용매(증류수) 1000g으로 3일간 냉침하여 추출하였다. 냉침된 추출물을 0.2㎛의 기공 크기를 가진 필터로 여과하여 대황추출물을 제조하였다.After drying and cutting the rhubarb well, 100 g of dry weight was put in a flask and extracted by cooling with 1000 g of an extraction solvent (distilled water) for 3 days. The chilled extract was filtered through a filter having a pore size of 0.2 μm to prepare a rhubarb extract.
<실시예 5> 황금추출물의 제조<Example 5> Preparation of golden extract
황금을 잘 건조하여 세절한 후, 건조중량 100g을 플라스크에 넣고 추출용매(증류수) 1000g으로 3일간 냉침하여 추출하였다. 냉침된 추출물을 0.2㎛의 기공 크기를 가진 필터로 여과하여 황금추출물을 제조하였다.After drying and cutting the gold well, 100 g of dry weight was put in a flask and extracted by cooling with 1000 g of an extraction solvent (distilled water) for 3 days. The chilled extract was filtered through a filter having a pore size of 0.2 μm to prepare a golden extract.
<실시예 6> 황금, 박하, 치자, 연교, 대황 혼합추출물의 제조<Example 6> Preparation of mixed extracts of gold, mint, gardenia, yeongyo, and rhubarb
상술한 실시예와 같이 각각 추출물 형태로 제조한 황금, 박하, 치자, 연교, 대황 을 동량을 황금, 박하, 치자, 연교, 대황 혼합추출물을 제조하였다.As in the above-described example, a mixed extract of gold, mint, gardenia, yeongyo, and rhubarb was prepared by using the same amount of gold, mint, gardenia, yeongyo, and rhubarb, each prepared in the form of an extract.
<실험예 1> 항당화 효과 <Experimental Example 1> Anti-glycation effect
항당화(anti-glycation) 효능을 확인하기 위하여, L-arginine과 포도당을 이용하여 당화 저해 활성을 측정하였다. In order to confirm anti-glycation efficacy, glycation inhibitory activity was measured using L-arginine and glucose.
먼저, 1M 인산 완충용액(pH 7.4)을 이용하여 1M L-아르기닌(arginine), 1M 포도당을 녹여 준비하고 1M 인산 완충용액을 이용하여 시료를 50ppm이 되도록 희석해서 준비하였다. 1M L-아르기닌과 1M 인산 완충용액을 1대 4의 비율로 섞은 다음 96-웰 플레이트에 80 μl씩 분주하였다. 여기에 각각 50ppm으로 희석한 시료와 양성대조군으로 사용될 0.01M 아미노구아니딘(aminoguanidin)을 100 μl씩 첨가하였다. 이 시료들을 잘 섞어준 다음, 마지막으로 포도당의 최종 농도가 0.1M이 되도록 1M 인산 완충용액으로 희석한 포도당을 넣은 후, 70℃에서 4시간 동안 반응 시켰다. 96-웰 플레이트를 분광 광도계를 이용하여 420 nm에서 흡광도를 측정하여 당화 정도를 측정하였다. First, 1M L-arginine and 1M glucose were dissolved in 1M phosphate buffer (pH 7.4) and prepared by diluting the sample to 50 ppm using 1M phosphate buffer. 1M L-arginine and 1M phosphate buffer were mixed at a ratio of 1:4, and then 80 μl was dispensed into a 96-well plate. Here, 100 μl of 0.01 M aminoguanidine to be used as a positive control and a sample diluted to 50 ppm, respectively, were added thereto. After mixing these samples well, finally, after adding glucose diluted with 1M phosphate buffer so that the final concentration of glucose is 0.1M, the mixture was reacted at 70°C for 4 hours. The degree of glycosylation was measured by measuring the absorbance at 420 nm of the 96-well plate using a spectrophotometer.
하기 식의 Glycation 실험군은 1M L-아르기닌과 1M 포도당을 넣어 당화를 유발시킨 실험군이며, 시료 자체의 흡광도를 측정하기 위해서 포도당을 넣지 않고 1M L-아르기닌과 시료만을 넣어 420 nm에서 흡광도를 측정하였다. 당화 저해 활성은 다음과 같은 식으로 구할 수 있다. The Glycation experimental group of the following formula is an experimental group in which 1M L-arginine and 1M glucose are added to induce glycation. Glycosylation inhibitory activity can be obtained by the following formula.
[수학식 1] [Equation 1]
(상기 수학식 1에서의 'Glycation 실험군'은 'Glycation 실험군의 흡광도'를 의미함.)(The 'Glycation experimental group' in Equation 1 means the 'absorbance of the Glycation experimental group'.)
상기 표 1의 결과에서 나타난 바와 같이, 혼합추출물은 항당화 물질로 알려진 아미노구아니딘(aminoguanidin)과 비교할 때, 항당화 효과가 더욱 뛰어남을 알 수 있다. As shown in the results of Table 1, it can be seen that the mixed extract has a more excellent anti-glycation effect compared to aminoguanidin, which is known as an anti-glycation substance.
<실험예 2> 멜라닌 생성 저해 효과 <Experimental Example 2> Melanin production inhibitory effect
멜라닌 생성 저해를 통한 미백 효과를 확인하기 위하여, Lotan R. 외(Cancer Res. 40:3345-3350, 1980)에 기재된 방법에 따라 쥐의 멜라노마 세포(B-16 mouse melanoma cell)의 배양액에, 추출물을 첨가하여 멜라닌 총량을 측정하였다. 실험 시, 먼저 쥐의 멜라노마 세포에 대하여 독성을 평가하여 독성이 없는 농도에서 미백평가를 수행하였다. 음성 대조군으로는 DMSO를, 양성 대조군으로는 알부틴(albutin)을 사용하였다.In order to confirm the whitening effect through inhibition of melanin production, according to the method described by Lotan R. et al. (Cancer Res. 40: 3345-3350, 1980), the extract was added to a culture medium of B-16 mouse melanoma cells to measure the total amount of melanin. In the experiment, whitening was evaluated at a non-toxic concentration by first evaluating toxicity to melanoma cells of mice. DMSO was used as a negative control and albutin was used as a positive control.
구체적으로, 시료를 최종 농도가 100 ppm이 되도록 배지에 첨가하고, 알부틴은 100 ppm이 되도록 배지에 첨가한 후 멜라노마 세포를 3일간 배양하였다. 이후, 세포들을 트립신(trypsin) 처리하여 배양용기로부터 떼어내 원심분리한 후, 멜라닌을 추출하였다. 떼어낸 세포는 수산화나트륨 용액(1N 농도) 1 ml를 가하여 10분간 끓여 멜라닌을 녹이고 분광 광도계를 이용하여, 400 nm에서 흡광도를 측정하여 생성된 멜라닌의 양을 측정하였다. Specifically, the sample was added to the medium so that the final concentration was 100 ppm, and arbutin was added to the medium so that the final concentration was 100 ppm, and then the melanoma cells were cultured for 3 days. Thereafter, the cells were treated with trypsin, detached from the culture vessel, centrifuged, and then melanin was extracted. The detached cells were boiled for 10 minutes by adding 1 ml of sodium hydroxide solution (1N concentration) to dissolve melanin, and the absorbance was measured at 400 nm using a spectrophotometer to measure the amount of melanin produced.
상기 멜라닌 양은 단위 세포수당(1×106 cell)의 흡광도로 나타내는 방법으로 측정하였으며, 대조군에 대한 상대적인 멜라닌 총량을 저해율(%)로 계산하고 결과를 하기 표 2에 나타내었으며, 실험은 각각 3회씩 수행하여 평균값으로 나타내었다.The amount of melanin was measured by the method of expressing the absorbance per unit cell number (1 × 10 6 cell), the total amount of melanin relative to the control group was calculated as an inhibition rate (%), and the results are shown in Table 2 below. Each experiment was performed three times and represented as an average value.
상기 표 2의 결과에서 볼 수 있듯이, 혼합추출물은 뛰어난 멜라닌 총량 감소 효과를 나타내어 미백 용도로 유용함을 알 수 있었다.As can be seen from the results of Table 2, the mixed extract was found to be useful for whitening because it exhibited an excellent effect of reducing the total amount of melanin.
<실험예 3> 항염 효과<Experimental Example 3> Anti-inflammatory effect
항염증 효과 및 피부트러블 개선 효과를 확인하기 위하여, RAW264.7 세포주 (ATCC number: CRL-2278)를 이용한 GRIESS 법으로 nitric oxide(NO) 생성 억제력 실험을 실시하였다.In order to confirm the anti-inflammatory effect and skin trouble improvement effect, nitric oxide (NO) production inhibition test was performed by the GRIESS method using the RAW264.7 cell line (ATCC number: CRL-2278).
구체적으로, 생쥐의 대식세포인 RAW264.7 세포를 수차례 계대 배양하고, 웰 하나에 3×105 개씩 들어가도록 24-웰 프레이트에 넣은 후, 24 시간 동안 배양하였다. 이어서, 최종농도 10ppm의 농도로 시료를 함유한 세포 배지로 교체하였다. 이때, NO-생성 억제물질인 L-NMMA(L-NG-Monomethylarginine)을 양성 대조군으로 함께 처리하여 30분 동안 배양하였고, 자극원으로 LPS(Lipopolysaccharide)를 1 ㎍씩 처리하여 24시간 동안 배양하였다. 상층액을 100㎕씩 취해 96-웰 프레이트에 옮기고, GRIESS 용액을 100㎕씩 가해 상온에서 10분간 반응시키고, 540nm에서의 흡광도를 측정함으로써 NO 억제 효과를 판단하고, NO 생성 저해율(%)은 하기 수학식 2를 이용하여 계산하여 하기 표 3에 나타내었으며, 실험은 각각 3회씩 수행하여 평균값으로 나타내었다.Specifically, RAW264.7 cells, which are mouse macrophages, were subcultured several times, placed in a 24-well plate so that 3×10 5 per well, and then cultured for 24 hours. It was then replaced with a cell medium containing the sample at a final concentration of 10 ppm. At this time, L-NMMA (L-NG-Monomethylarginine), an NO-production inhibitor, was treated as a positive control and cultured for 30 minutes, and 1 μg of LPS (Lipopolysaccharide) was treated as a stimulus and cultured for 24 hours. 100 μl of the supernatant was transferred to a 96-well plate, 100 μl of the GRIESS solution was added, reacted at room temperature for 10 minutes, and the absorbance at 540 nm was measured to determine the NO inhibition effect. The NO production inhibition rate (%) was calculated using Equation 2 below and shown in Table 3 below.
[수학식 2][Equation 2]
NO 생성 저해율(%)={(음성대조군의 흡광도 - 각 추출물의 흡광도)/음성 대조군의 흡광도} x 100 NO production inhibition rate (%) = {(absorbance of negative control - absorbance of each extract) / absorbance of negative control} x 100
상기 표 3의 결과에서 알 수 있듯이, 혼합추출물은 대표적인 항염 의약물질인 L-NMMA와 비교하였을 때, 상대적 활성은 다소 낮으나 천연물질로써 우수한 활성을 나타냄을 알 수 있었다. As can be seen from the results of Table 3, it was found that the mixed extract exhibited excellent activity as a natural substance, although the relative activity was somewhat lower when compared to L-NMMA, a representative anti-inflammatory drug.
<실험예 4> 콜라겐 합성 촉진 효과 <Experimental Example 4> Collagen synthesis promotion effect
시료를 인간 유래 섬유아세포의 배양액에 첨가하여 세포수준에서 제1형 콜라겐 합성 촉진 효과를 확인하였다. 합성된 콜라겐의 측정은 PICP EIA kit(Procollagen Type I C-Peptide Enzyme Immuno Assay KIT)를 이용하여 정량하였다. 콜라겐 합성량을 측정하기 위해 시료를 최종 농도가 10 ppm이 되도록 섬유아세포의 배양배지(DMEM 배지)에 첨가하여 48 시간 배양한 후 배양액을 취하여 PICP EIA 키트로 각 농도에서 제1형 콜라겐 합성 정도를 분광광도계를 이용하여 450 nm에서 측정하였다.The sample was added to the culture medium of human-derived fibroblasts to confirm the effect of promoting type 1 collagen synthesis at the cellular level. The measurement of synthesized collagen was quantified using a PICP EIA kit (Procollagen Type I C-Peptide Enzyme Immuno Assay KIT). In order to measure the amount of collagen synthesis, the sample was added to the culture medium (DMEM medium) of fibroblasts to a final concentration of 10 ppm, cultured for 48 hours, and then the culture medium was taken and the degree of type 1 collagen synthesis at each concentration with the PICP EIA kit was measured at 450 nm using a spectrophotometer.
효과의 비교를 위하여 시료를 처리하지 않은 섬유아세포의 배양배지(음성대조군)와 비타민 C(양성대조군)를 최종농도 52.85 ㎍/ml가 되도록 첨가한 시료에 대하여 동일한 방법으로 콜라겐 합성 정도를 측정하였다. 콜라겐 생성 증가율은 음성 대조군에 대한 상대적인 콜라겐 생성량의 비율로 계산하고 결과를 하기 표 4에 나타내었으며, 실험은 4회씩 수행하여 평균값으로 나타내었다.For comparison of the effect, the degree of collagen synthesis was measured in the same manner for the samples to which the culture medium of untreated fibroblasts (negative control) and vitamin C (positive control) were added to a final concentration of 52.85 μg/ml. The rate of increase in collagen production was calculated as the ratio of the amount of collagen production relative to the negative control group, and the results are shown in Table 4 below. The experiment was performed 4 times and presented as the average value.
상기 표 4의 결과에서 볼 수 있듯이, 혼합추출물을 처리한 경우에 콜라겐 합성을 증가시켰고, 일반적으로 콜라겐 합성을 유도하는 것으로 잘 알려진 비타민C를 적용한 경우와 대응한 정도의 콜라겐 합성 효과를 나타내었다.As can be seen from the results of Table 4, collagen synthesis was increased when the mixed extract was treated, and the effect of collagen synthesis was comparable to that of vitamin C, which is generally known to induce collagen synthesis.
<실험예 5> 엘라스타제 활성 저해 효과 <Experimental Example 5> Elastase activity inhibitory effect
엘라스틴(Elastin)을 분해하는 효소인 엘라스테이즈(Elastase)의 활성 저해 효과를 다음과 같이 확인하였다.The effect of inhibiting the activity of elastase, an enzyme that degrades elastin, was confirmed as follows.
엘라스테이즈(Elastase)는 사람의 백혈구 세포로부터 유래한 엘라스테이즈를 사용하였고, 엘라스테이즈의 기질로 합성 기질인 MeOSuc-Ala-Ala-Pro-Val-pNA를 사용하였다. 완충용액은 100mM의 Tris(pH 7.5) 용액을 사용하였다. 엘라스테이즈는 완충용액을 이용하여 최종적으로 0.2 mU을 사용하였다. 또한 엘라스테이즈의 합성 기질은 DMSO를 이용하여 100mM 용액을 만든 후 최종 농도가 0.5mM이 되도록 완충용액을 이용하여 희석하였다. 이 때, 양성 대조군은 엘라스테이즈 저해 물질로 알려진 쿼세틴(Quercetin)을 10ppm 농도로 넣은 것으로 설정하였다. 엘라스테이즈 저해 후보는 최종농도가 10ppm이 되도록 첨가하였다. 반응은 96-웰 플레이트에서 진행하였으며, 상온에서 20분간 반응시켰다. 분광 광도계를 이용하여 1분 간격으로 405 nm에서 흡광도를 측정하여, 시간 대비 흡광도의 기울기를 구하여 효소의 활성도로 정하였다. 엘라스테이즈 저해율은 다음과 같이 계산하였다.Elastase derived from human leukocytes was used, and a synthetic substrate MeOSuc-Ala-Ala-Pro-Val-pNA was used as a substrate for the elastase. A 100 mM Tris (pH 7.5) solution was used as the buffer solution. Elastase was finally used at 0.2 mU using a buffer solution. In addition, the synthetic substrate of Elastase was diluted with a buffer solution to a final concentration of 0.5 mM after making a 100 mM solution using DMSO. At this time, as a positive control group, Quercetin, known as an elastase inhibitor, was set at a concentration of 10 ppm. The elastase inhibitory candidate was added to a final concentration of 10 ppm. The reaction was carried out in a 96-well plate and reacted at room temperature for 20 minutes. The absorbance was measured at 405 nm at 1-minute intervals using a spectrophotometer, and the slope of the absorbance versus time was determined to determine the activity of the enzyme. The elastase inhibition rate was calculated as follows.
[수학식 3] [Equation 3]
엘라스타제 저해율은 상기 수학식 3을 이용해 계산하여 하기 표 5에 나타내었으며, 실험은 각각 3회씩 수행하여 평균값으로 나타내었다.The elastase inhibition rate was calculated using Equation 3 and shown in Table 5 below.
상기 표 5의 결과에서 볼 수 있듯이, 혼합추출물을 처리한 경우, 양성대조군 보다 상대적으로 효과가 다소 약하나, 우수한 엘라스타제 활성 저해 효과를 발휘함을 알 수 있었다. 따라서, 혼합추출물은 피부재생, 주름개선을 위한 용도로 사용할 수 있음을 알 수 있었다.As can be seen from the results of Table 5, when the mixed extract was treated, the effect was relatively weaker than that of the positive control group, but it was found to exhibit an excellent elastase activity inhibitory effect. Therefore, it was found that the mixed extract can be used for skin regeneration and wrinkle improvement.
<실험예 6> 항산화 효과 <Experimental Example 6> Antioxidant effect
혼합추출물에 대한 자유라디칼 소거능을 1,1-디페닐-2-피크릴히드라질(DPPH) 방법으로 측정하였다(Blois, Nature 181, 1190, 1958). DPPH는 비교적 안정한 자유라디칼로서 라디칼 상태로 존재 시 517㎚에서 최대 흡광을 보이며 라디칼이 소거되면 흡광성을 잃는다. DPPH는 시그마(Sigma)사의 것을 사용하였으며, 0.15 mM의 농도로 메틸 알코올에 녹여 사용하였다.The free radical scavenging ability of the mixed extract was measured by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) method (Blois, Nature 181, 1190, 1958). DPPH is a relatively stable free radical and exhibits maximum absorption at 517 nm when present in a radical state, and loses absorbance when the radical is scavenged. DPPH was used from Sigma, and was dissolved in methyl alcohol at a concentration of 0.15 mM.
먼저, 혼합추출물 또는 양성 대조군인 비타민 C를 96-웰 플레이트의 각각의 웰에 100μl씩 넣었다. 여기에 DPPH 용액을 100μl씩 첨가한 다음 상온에서 30분간 방치하고 마이크로플레이트 리더(BioTek EL-340)를 이용하여 517㎚에서의 흡광도를 측정하였다.First, 100 μl of the mixed extract or vitamin C as a positive control was added to each well of a 96-well plate. 100 μl of DPPH solution was added thereto, left at room temperature for 30 minutes, and absorbance at 517 nm was measured using a microplate reader (BioTek EL-340).
시료를 처리한 것의 흡광도가 대조군의 흡광도의 절반이 될 때의 추출물의 농도를 IC50으로 표시한 결과를 하기 표 6에 나타내었다. 본 실험은 3회 반복하였다. Table 6 shows the results of expressing the concentration of the extract as IC 50 when the absorbance of the treated sample becomes half of that of the control group. This experiment was repeated 3 times.
상기 표 6과 같이, 혼합추출물은 비타민 C와 비교할 때도 자유라디칼 소거능이 매우 강력하였으며, 이로써 항산화 효과가 우수함을 확인하였다. As shown in Table 6, the mixed extract had very strong free radical scavenging activity compared to vitamin C, and thus it was confirmed that the antioxidant effect was excellent.
Claims (4)
The cosmetic composition according to claim 1, wherein the cosmetic composition is anti-glycation, inhibits melanin production, promotes collagen synthesis, inhibits elastase activity, or scavenges free radicals.
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