KR20180060609A - Composition for improving skin - Google Patents
Composition for improving skin Download PDFInfo
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- KR20180060609A KR20180060609A KR1020160160307A KR20160160307A KR20180060609A KR 20180060609 A KR20180060609 A KR 20180060609A KR 1020160160307 A KR1020160160307 A KR 1020160160307A KR 20160160307 A KR20160160307 A KR 20160160307A KR 20180060609 A KR20180060609 A KR 20180060609A
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- extract
- organic solvent
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Abstract
Description
본 발명은 피부 미백, 탄력 증진, 주름 개선, 항염증, 항산화, 항당화 효과를 나타내는 피부 개선용 조성물에 관한 것이다. The present invention relates to a skin improving composition exhibiting skin whitening, elasticity enhancement, wrinkle improvement, antiinflammation, antioxidant, anticarcinogenic effect.
희고 고운 피부를 갖고자 하는 것은 일반적인 소망이다. 피부의 색깔 또는 밝기는 사람의 피부 내 멜라닌(melanin)의 농도와 분포에 따라 유전적으로 결정되나, 태양 자외선, 피로 또는 스트레스 등의 환경적 또는 생리적 조건에 의해서도 영향을 받는다. 멜라닌은 아미노산의 일종인 티로신(tyrosine)에 티로시나제(tyrosinase)라는 효소가 촉매로 작용하여 도파(DOPA) 및 도파퀴논(dopaquinone)으로 순차적으로 바뀐 후, 비효소적인 산화반응을 거쳐 만들어진다. 이와 같이 멜라닌이 만들어지는 경로는 알려져 있으나, 멜라닌 합성을 유도하는 메커니즘에서 티로시나제가 촉발되는 원인이 무엇인지에 대해서는 아직도 자세히 밝혀지지 않고 있다. It is a common desire to have white skin. The color or brightness of the skin is genetically determined by the concentration and distribution of melanin in the human skin but is also influenced by environmental or physiological conditions such as sunlight, fatigue or stress. Melanin is produced by a nonenzymatic oxidation reaction in which tyrosine, an amino acid, is catalyzed by an enzyme called tyrosinase, which is subsequently converted to dopa (DOPA) and dopaquinone. The pathway for the formation of melanin is known, but the mechanism by which the melanin synthesis induces tyrosinase is not yet elucidated.
한편, 일반적으로 알려진 미백 성분으로서, 코지산(Kojic acid) 또는 알부틴(Arbutin) 등과 같은 티로시나제 효소활성을 억제하는 물질, 하이드로퀴논(Hydroquinone), 비타민 C(L-Ascorbic acid) 또는 이들의 유도체와 각종 식물 추출물이 있다. 이들은 멜라닌 색소의 합성을 저해함으로써, 피부 톤을 밝게 하여 피부 미백을 실현할 수 있을 뿐만 아니라, 자외선, 호르몬 또는 유전에 기인한 기미나 주근깨 등의 피부 과색소 침착증의 개선이 가능하다. 그러나 피부 적용 시, 자극과 발적 등의 안전성의 문제로 사용량의 제한이 있거나, 효과가 미미하여 실질적인 효과를 기대할 수 없는 문제점이 있다.On the other hand, commonly known whitening ingredients include substances inhibiting the activity of tyrosinase enzymes such as kojic acid or arbutin, hydroquinone, vitamin C (L-ascorbic acid) There are plant extracts. By inhibiting the synthesis of melanin pigment, they can brighten the skin tone to realize skin whitening, and it is possible to improve skin hypercholesterolemia such as stain or freckles due to ultraviolet rays, hormones or heredity. However, when applied to skin, there is a problem that the use amount is limited due to safety problems such as irritation and redness, or the effect is insignificant, so that a practical effect can not be expected.
또한, 콜라겐은 피부의 섬유아세포에서 생성되는 주요 기질 단백질로서 세포외 간질에 존재하고, 중요한 기능으로는 피부의 기계적 견고성, 결합조직의 저항력과 조직의 결합력, 세포접착의 지탱, 세포분할과 분화(유기체의 성장 혹은 상처 치유시)의 유도 등이 알려져 있다. 이러한 콜라겐은 연령 및 자외선 조사에 의한 광 노화에 의해 감소하며, 이는 피부의 주름 형성과 밀접한 연관이 있다고 알려져 있다. 또한, 근래에 들어 피부 노화에 대한 광범위한 연구가 발전되면서 피부에서의 콜라겐의 중요한 기능이 밝혀지고 있다.In addition, collagen is a major substrate protein produced in fibroblasts of the skin and exists in extracellular epilepsy. Its important functions are mechanical rigidity of skin, resistance of connective tissues and binding force of tissues, support of cell adhesion, division of cells and differentiation Induction of growth of an organism or wound healing) are known. Such collagen is reduced by aging and photo aging caused by ultraviolet irradiation, which is known to be closely related to the wrinkling of the skin. Also, in recent years, extensive research on skin aging has developed, and important functions of collagen in skin have been revealed.
콜라겐 합성을 촉진하여 주름 개선 효과를 나타내는 유효성분들이 알려져 있다. 예를 들어, 레티노산(retinoic acid), TGF(transforming growth factor)[비특허문헌 1], 동물 태반 유래의 단백질[특허문헌 1], 베튤린산(betulinic acid)[특허문헌 2], 클로렐라 추출물[특허문헌 3, 4] 등이 콜라겐 합성 촉진 물질로서 알려져 있다. 그러나, 상기 유효성분들은 피부 적용 시 자극과 발적 등의 안전성의 문제로 사용량의 제한이 있거나, 효과가 미미하여 실질적으로 피부의 콜라겐 합성을 촉진하여 피부 기능을 개선하는 효과를 기대할 수 없는 문제점이 있다.Effective ingredients promoting collagen synthesis and exhibiting wrinkle-reducing effects are known. For example, retinoic acid, transforming growth factor (TGF) [non-patent document 1], animal placenta-derived protein [Patent document 1], betulinic acid [Patent document 2], chlorella extract [ Patent Documents 3 and 4] are known as collagen synthesis promoting substances. However, the above-mentioned effective ingredients are limited in the use amount due to safety problems such as irritation and redness when applied to the skin, or have insufficient effect, so that the effect of improving the skin function by promoting the collagen synthesis of the skin can not be expected.
또한, 염증은 상처나 질병에 반응하는 인체의 면역 반응으로, 자외선이나 활성산소, 자유라디칼 등의 산화적 스트레스 등이 염증성 인자를 활성화시켜 각종 질병 및 피부의 노화를 일으킨다. 혈관 활성 폴리펩타이드인 키닌(kinin), 플라스민(plasmin) 또는 보체 (complement) 등이 혈관 확장과 수축 및 주화성(chemotaxis) 작용을 하고, 그 외에 인터루킨-6(IL-6) 등과 같은 림포카인과 아라키돈산(arachidonic acid) 등이 염증 반응을 담당한다. 아라키돈산은 싸이클로옥시게나아제(cyclooxygenase) 혹은 리포옥시게나아제(lipooxygenase)의 2가지 경로를 거쳐 염증 매개체인 프로스타글란딘(prostaglandin) 또는 류코트리엔(lukotriene)들로 대사되어 다양한 염증 반응을 매개한다. In addition, inflammation is an immune response of a human body in response to a wound or disease, and oxidative stress such as ultraviolet rays, active oxygen, free radicals, etc. activate inflammatory factors and cause aging of various diseases and skin. The vasoactive polypeptide, kinin, plasmin, or complement, has vasodilatation, contraction, and chemotaxis, as well as lymphokines such as interleukin-6 (IL-6) Phosphorus and arachidonic acid are responsible for inflammation. Arachidonic acid is metabolized through inflammatory mediators such as prostaglandin or lukotrienes via two pathways: cyclooxygenase or lipooxygenase, mediating a variety of inflammatory responses.
염증을 소실시키기 위해 염증원의 제거, 생체 반응 및 증상을 감소시키는 작용을 하는 것을 항염제라 한다. 현재까지 항염의 목적으로 이용되고 있는 물질로는 비스테로이드계로 플루폐나믹산(flufenamic acid), 이부프로펜(ibuprofen), 벤지다민(benzydamine) 또는 인도메타신(indomethacin) 등이 있고 스테로이드계통으로 프레드니솔론(prednisolone) 또는 덱사메타손(dexamethasone) 등이 있다. 또한, 알란토인, 아즈엔 또는 하이드로코티손 등이 항염증에 효과가 있는 것으로 알려져 있으나, 이들 물질은 피부에 대한 안전성의 문제로 사용량의 제한이 있거나, 효과가 미미하여 실질적으로 염증 완화 효과를 기대할 수 없는 문제점이 있다.Removal of inflammation to eliminate inflammation, the action of reducing vital signs and symptoms is called anti-inflammatory. To date, substances used for antiinflammatory purposes include non-steroidal drugs such as flufenamic acid, ibuprofen, benzydamine or indomethacin, prednisolone in the steroid line, Or dexamethasone. In addition, it is known that allantoin, azene or hydrocortisone are effective for antiinflammation. However, these substances are limited in the use amount due to the problem of safety to the skin, or the effect is insignificant, .
한편, 생체 외부로부터 유입되거나, 생체 내에서 발생하는 활성 산소는 생체의 노화를 촉진시키거나, 암을 발생시키는 등 많은 문제의 원인이 된다. 따라서 활성 산소에 의한 산화를 억제하는 항산화 물질에 대한 개발 및 연구가 많이 이루어 지고 있다. 항산화 물질은 동, 식물계에 널리 분포되어 있으며 과일과 채소에 많은 페놀성 화합물, 플라보노이드, 토코페롤, 비타민 C, 셀레늄 등이 알려져 있다. 다만, 천연에 존재하는 항산화 물질은 피부 적용 시 실질적으로 충분한 효과를 기대할 수 없는 실정이다. 따라서, 항산화력이 뛰어나고 가격이 저렴한 합성 항산화제가 많이 사용되고 있으나, 인체 부작용 등 안전성에 대한 우려로 그 사용이 제한된다. On the other hand, active oxygen introduced from the outside of the living body or generated in the living body causes many problems such as promoting aging of the living body, cancer, and the like. Therefore, the development and research of antioxidants that inhibit oxidation by active oxygen have been performed. Antioxidants are widely distributed in copper and plants. Many phenolic compounds, flavonoids, tocopherols, vitamin C and selenium are known in fruits and vegetables. However, the antioxidant substances present in nature can not be expected to have practically sufficient effects in skin application. Therefore, although synthetic antioxidants having excellent antioxidant ability and low cost are widely used, their use is restricted due to safety concerns such as human side effects.
당화(Glycation)는 일반적으로 효소의 관여 없이 일어나는 단백질 또는 지방에 포도당 또는 과당과 같은 단순당이 공유결합을 형성하는 반응을 의미한다. 최종당화산물의 축적은 단백질을 단단하고 더욱 부서지기 쉬운 상태로 변화시키며, 당화된 콜라겐은 진피층의 세포 외 기질에서 콜라겐이 적절한 구조를 형성하지 못하도록 함으로써 피부의 탄력을 잃게 하고 주름 생성을 촉진한다[비특허문헌 2]. 또한 당화로 인해 생성된 최종당화산물은 갈색 빛을 띠는 물질로써, 피부 노화가 진행되면서 점차 얼굴빛이 노랗게 변하는 원인 물질로 생각되고 있으며, 특정 최종당화산물이 쌓일수록 피부에서 반사되는 반사 빛이 줄어든다고 알려져 있다[비특허문헌 3]. 당화를 억제하는 물질로 아미노구아니딘(Aminoguanidine), 피리독사민(Pyridoxamine), 아스피린(Aspirin) 등이 알려져 있으나, 이들 물질은 피부에 대한 안전성의 문제로 사용량의 제한이 있거나, 효과가 미미하여 실질적으로 효과를 기대할 수 없는 문제점이 있다.Glycation generally refers to a reaction in which a simple sugar such as glucose or fructose forms a covalent bond to a protein or fat that occurs without involvement of the enzyme. The accumulation of the final glycation product changes the protein into a harder and more fragile state, and glycated collagen prevents the collagen from forming a proper structure in the extracellular matrix of the dermal layer, thereby losing skin elasticity and promoting wrinkle formation [ Non-Patent Document 2]. In addition, the final saccharification product produced by the saccharification is a brownish substance, which is considered to be a cause of yellowing of the face gradually as the skin ages, and as the specific final saccharification product accumulates, the reflection light reflected from the skin is reduced [Non-Patent Document 3]. Aminoguanidine, pyridoxamine, and aspirin are known to inhibit glycation. However, these substances are limited in their use due to the safety of the skin, There is a problem that can not be expected.
또한, 생체에 안전하고, 유효성분이 안정하며, 무엇보다도 기존의 피부 미백, 주름 개선, 탄력 증진, 항염증 및/또는 항산화 효과가 있는 물질보다 효과가 우수한 피부 미백, 주름 개선, 탄력 증진, 항염증 및/또는 항산화 활성을 지닌 성분의 개발이 절실히 요망되고 있다.In addition, the present invention provides a cosmetic composition which is safe and biologically safe, has a superior effect on skin whitening, wrinkle improvement, elasticity enhancement, antiinflammation, anti-inflammation, And / or a component having antioxidant activity is strongly desired.
한편, 녹각(Cervi Cornu)은 매화록(梅花鹿) Cervus nippon Temminck, 마록(馬鹿) Cervus elaphus Linne 또는 대록(大鹿) Cervus canadensis Erxleben (사슴과 Cervidae)의 골질화된 뿔이다. 혈액순환을 촉진시키고 어혈(瘀血)을 없애주며 신장기능과 간 기능을 도와준다. 또 칼슘을 다량 함유하고 있어 뼈를 튼튼하게 해준다. On the other hand, Cervi Cornu is the osolated horn of Cervus nippon Temminck, Cervus elaphus Linne or Cervus canadensis Erxleben (deer and Cervidae). It promotes blood circulation, eliminates eosinophilia, helps kidney function and liver function. It also contains a large amount of calcium, making it stronger.
또한, 토사자(Cuscuta japonica Chois)는 메꽃과에 속하는 한해살이 덩굴성 식물인 새삼의 씨앗으로, 새삼씨라고도 한다. 주로 간과 신장을 보호하며 눈을 밝게 해주고, 양기(陽氣)를 도우며 신장 기능을 튼튼하게 해주는 약재로 알려져 있다. 신장이 허약하여 생긴 남성의 성교불능증, 저절로 정액이 흐르는 경우, 몽정(夢精) 등에 효과가 있다. 뼈를 튼튼하게 해주고 허리 힘을 세게 해주며, 신장 기능이 허약하여 허리와 무릎이 시리고 아픈 것을 치료한다. 또한, 오줌소태와 소변을 잘 보지 못하는 질병과 설사를 낫게 하며 당뇨병 치료에도 효과가 있는 것으로도 알려져 있다.Also, Tusa (Cuscuta japonica Chois) is a seed of Seongam, a year-old vine plant belonging to the family Myrosea, and is also called Seisham seed. It is known as a medicine that protects the liver and kidneys, brightens the eyes, helps the kidneys, and strengthens the kidney function. It is effective for men with sexual intercourse due to fragility of the kidneys, when semen flows by itself, and by the mungjeong (夢 精). It strengthens the bones, strengthens the back strength, and weakens the kidneys and heals back and knees and sore. It is also known to be effective in the treatment of diabetes, as well as in the treatment of illness and diarrhea that can not see urine and urine.
또한, 숙지황(REHMANNIAE RADIX PREPARAT)은 지황 Rehmannia glutinosa Liboschitz ex Steudel (현삼과 Scrophulariaceae)의 뿌리를 포제 가공한 것이다. 숙지황은 사물탕(四物湯)의 주요 약재이며 각종 만성병 중 몸이 허약하여 나타나는 내열(內熱), 인후건조(咽喉乾燥), 갈증 등의 증상에 쓰인다In addition, REHMANNIAE RADIX PREPARAT is a frosted process for the roots of Rehmannia glutinosa Liboschitz ex Steudel (Hyosung and Scrophulariaceae). Sukjihwang is the main medicinal material of Sangmatsu (hot water) and it is used for various symptoms of chronic diseases such as internal heat, throat drying and thirst.
또한, 백자인은 백과 식물 Platycladus orientalis, Thuja orientalis(=Biota orientalis)의 종인(種仁)으로, 정신을 안정되게 하고 대소변을 잘 나오게 하는 약재이다.In addition, white porcelain Platycladus orientalis, Thuja orientalis (= Biota orientalis) is a kind of seed (仁 仁), the spirit is stable and the feces is a good way to get out.
이에, 본 발명자들은 녹각 추출물, 토사자 추출물, 숙지황 추출물 및 백자인 추출물을 혼합한 혼합 추출물이 멜라닌 총량을 현저히 감소시켜 미백 효과를 나타내고, 피부의 섬유아세포의 콜라겐 합성을 촉진하며, 엘라스테이즈 활성을 저해하여 주름을 개선하며 탄력을 증진시키고, 산화질소(NO) 생성을 억제하여 항염증 효과를 나타내며, 자유 라디칼을 소거하는 항산화 효과 및 당화를 억제하는 항당화 효과를 확인함으로써 본 발명을 완성하게 되었다.Thus, the inventors of the present invention found that a mixed extract comprising a mixture of a green tea extract, a Tozai extract, a Sukjunghwang extract and a white porcelain extract significantly reduced the total amount of melanin, exhibited a whitening effect, promoted collagen synthesis of fibroblasts of the skin, inhibited Elastase activity Thereby improving wrinkles, improving elasticity, suppressing the production of nitric oxide (NO), exhibiting an anti-inflammatory effect, antioxidative effect of eliminating free radicals and anti-glycation effect inhibiting glycation.
따라서, 본 발명의 목적은 녹각 추출물, 토사자 추출물, 숙지황 추출물 및 백자인 추출물을 혼합한 혼합 추출물을 유효성분으로 포함하는 피부 미백, 주름 개선, 탄력 증진, 항염증, 항산화 및/또는 항당화용 조성물을 제공하는데 있다.Accordingly, an object of the present invention is to provide a composition for skin whitening, wrinkle improvement, elasticity enhancement, antiinflammation, antioxidant and / or antialcification, which comprises as an active ingredient, a mixed extract comprising a green tea extract, a Sorghum extract, .
상기 과제를 해결하기 위한 수단으로서, 본 발명은 의약, 화장료 또는 식품 제조를 위한, 녹각 추출물, 토사자 추출물, 숙지황 추출물 및 백자인 추출물을 혼합한 혼합 추출물을 유효성분으로 포함하는 피부 미백, 주름 개선, 탄력 증진, 항염증, 항산화 및/또는 항당화용 조성물을 제공한다.As a means for solving the above problems, the present invention relates to a skin whitening, wrinkle-improving, elasticity-improving agent for skin, hair, skin, An anti-inflammatory, antioxidant, and / or antialgalytic composition.
본 발명에 따른 녹각 추출물, 토사자 추출물, 숙지황 추출물 및 백자인 추출물을 혼합한 혼합 추출물은 멜라닌 총량을 현저히 감소시켜 미백 효과를 나타내고, 피부의 섬유아세포의 콜라겐 합성을 촉진하며, 엘라스테이즈 활성을 저해하여 주름을 개선하며 탄력을 증진시키고, 산화질소(NO) 생성을 억제하여 항염증 효과를 나타내며, 자유 라디칼을 소거하여 항산화 효과 및 당화를 억제하는 항당화 효과를 나타냄으로써, 의약, 화장료 또는 식품 제조에 사용할 수 있다. According to the present invention, the mixed extract of a mixture of the extracts of green tea extract, sorghum extract, sorghum extract and porcine garnet extract significantly reduces the total amount of melanin to exhibit whitening effect, promotes collagen synthesis of fibroblasts of the skin, inhibits elastase activity It exhibits antioxidative and anti-glycosylation effects by improving wrinkles and improving elasticity, inhibiting NO production, inhibiting free radicals, and inhibiting antioxidation and glycation. Can be used.
이하, 본 발명의 구성을 구체적으로 설명한다.Hereinafter, the configuration of the present invention will be described in detail.
피부 미백, 주름 개선, 탄력 증진, 항염증, 항산화 및/또는 항당화 성분이 실제 피부에 적용 시 우수한 효과를 발휘하기 위해서는 저농도에서 고활성의 피부 미백, 주름 개선, 탄력 증진, 항염증, 항산화 및/또는 항당화 활성을 나타내고, 피부를 투과하여 흡수되는 능력이 우수하고, 피부 미백, 주름 개선, 탄력 증진, 항염증, 항산화 및/또는 항당화 효과를 나타내기에 충분한 시간 동안 머무를 수 있도록 휘발성이 낮고, 조성물이나 피부 상에서 활성 성분이 안정하게 유지되고, 의약, 화장료 또는 식품 등으로의 제조가 용이하며, 또한 피부에 안전한 것이 바람직하다. 그러나, 공지의 성분 중 상기 특성을 모두 만족시키는 성분은 흔치 않다. 예를 들어, 몇몇 피부 미백, 주름 개선, 탄력 증진, 항염증, 항산화 및/또는 항당화 성분들은 시험관 내 실험 시 저농도에서도 피부 미백, 주름 개선, 탄력 증진, 항염증, 항산화 및/또는 항당화 활성은 우수하나, 피부를 투과하여 흡수되는 능력이 떨어져 실제 피부에 적용하기엔 어렵다. 또 다른 활성 성분들은 친수성이 낮아 의약이나 화장품, 식품으로 제형화가 어렵다. 또한, 몇몇 피부 미백, 주름 개선, 탄력 증진, 항염증, 항산화 및/또는 항당화 성분들은 열, 광, 또는 산소에 노출되었을 때 상기 활성 성분이 분해되거나 다른 화합물로 변형되어 피부에 적용하기 전에 이미 효과가 사라지는 경우도 있다. Skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammation, antioxidant and / or anti-glycation components are applied to skin in order to exert excellent effects, skin whitening, wrinkle improvement, elasticity enhancement, anti- / Or exhibits an antihyperglycosylating activity and is excellent in the ability to permeate and absorb the skin and has a low volatility so as to be able to stay for a sufficient time to exhibit skin whitening, wrinkle improvement, elasticity enhancement, antiinflammation, antioxidant and / , It is preferable that the active ingredient is kept stably on the composition or on the skin, and it is easy to manufacture into medicines, cosmetics or foods, and is safe for the skin. However, the components satisfying all of the above-mentioned characteristics among the known components are not common. For example, some skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammatory, antioxidant and / or antialgalytic ingredients may be used for skin whitening, wrinkle improvement, elasticity enhancement, anti- inflammation, antioxidant and / But it is difficult to apply to real skin because of its ability to penetrate through the skin and be absorbed. Other active ingredients have low hydrophilicity, making them difficult to formulate into medicines, cosmetics, and foods. In addition, some skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammatory, antioxidant and / or antialcification components can be decomposed or converted into other compounds when exposed to heat, light or oxygen, Sometimes the effect disappears.
하기 실시예에서 확인할 수 있는 바와 같이, 녹각 추출물, 토사자 추출물, 숙지황 추출물 및 백자인 추출물은 저농도에서 월등히 우수한 멜라닌 총량 감소 효과, 항당화 효과, 콜라겐 합성 촉진 효과, 항염증, 항산화 및/또는 항당화를 위한 의약, 화장료 또는 식품 조성물의 유효성분으로 사용할 수 있다.As can be seen from the following examples, the extracts of green tea extract, sorghum extract, sorghum extract and porcine garnet extract have remarkably excellent total melanin reduction effect, anti-glycation effect, collagen synthesis promotion effect, anti-inflammation, antioxidant and / A cosmetic composition, or a food composition.
따라서, 본 발명은 녹각 추출물, 토사자 추출물, 숙지황 추출물 및 백자인 추출물을 혼합한 혼합 추출물을 유효성분으로 포함하는 피부 미백, 주름 개선, 탄력 증진, 항염증, 항산화 및/또는 항당화용 조성물을 제공한다.Accordingly, the present invention provides a composition for skin whitening, wrinkle improvement, elasticity enhancement, antiinflammation, antioxidant and / or antialcification, which comprises as an active ingredient, a mixed extract of a green algae extract, a sesame extract, a Suji purple extract and a white porcelain extract .
본 발명에서 녹각은 매화록 Cervus nippon Temminck, 마록 Cervus elaphus Linne 또는 대록 Cervus canadensis Erxleben (사슴과 Cervidae)의 골질화된 뿔로, 학명은 Cervi Cornu이다. 또한, 토사자는 갯실새삼 Cuscuta chinensis Lamark (메꽃과 Convolvulaceae)의 씨로, 학명은 Cuscuta japonica Chois이다. 또한, 숙지황은 지황(地黃)의 뿌리를 쪄서 말린 한약재로서, 학명은 Rehmannia glutinosa (Gaertner) Libosch이다. 백자인은 약은 측백나무 Thuja orientalis Linne (측백나무과 Cupressaceae)의 씨로서 씨껍질을 제거한 것이다.In the present invention, overgrown antler is solvated lock Cervus nippon Temminck, Marcus Cervus elaphus Linne or Larry Cervus canadensis Erxleben (deer and Cervidae), the scientific name is Cervi Cornu. Also, it is a seed of Cuscuta chinensis Lamark (Convolvulaceae), and its scientific name is Cuscuta japonica Chois. In addition, Sukjipu is a Chinese medicinal herb that is steamed and dried at the roots of Geumgang (地 黄), and its scientific name is Rehmannia glutinosa (Gaertner) Libosch. White porcine is the seed of the seed of Thuja orientalis Linne (Cupressaceae).
상기 녹각 추출물, 토사자 추출물, 숙지황 추출물 또는 백자인 추출물은 당업계에 공지된 방법에 의해 추출될 수 있으며, 그 방법은 특별히 한정되지 않는다. 또는, 시판되고 있는 추출물을 이용할 수 있다.The extracts of green tea extract, Sorghum extract, Sambrook extract, or White porcelain extract may be extracted by a method known in the art, and the method is not particularly limited. Alternatively, commercially available extracts can be used.
바람직하기로는 상기 녹각 추출물, 토사자 추출물, 숙지황 추출물 또는 백자인 추출물은 녹각, 토사자, 숙지황 또는 백자인을 물 및/또는 유기용매로 추출하여 수득한 추출물을 사용할 수 있으며, 상기 유기용매는 극성 유기용매, 비극성 유기용매 또는 이들의 혼합용매일 수 있다. 상기 극성 유기용매는 탄소수 1 내지 5의 저급 알코올, 에틸아세테이트 또는 아세톤일 수 있으며, 비극성 유기용매는 에테르, 클로로포름, 벤젠, 헥산 또는 디클로로메탄일 수 있다. 예를 들어, 상기 탄소수 1 내지 5의 저급 알코올은 메탄올, 에탄올, 프로판올, 부탄올 또는 이소프로판올일 수 있다. Preferably, the extract of green tea extract, green tea extract, green tea extract or white porcelain extract may be obtained by extracting green tea, green tea, sorghum or white porcelain with water and / or organic solvent, and the organic solvent may be polar organic solvent, nonpolar Organic solvents or mixtures thereof. The polar organic solvent may be a lower alcohol having 1 to 5 carbon atoms, ethyl acetate or acetone, and the nonpolar organic solvent may be ether, chloroform, benzene, hexane or dichloromethane. For example, the lower alcohol having 1 to 5 carbon atoms may be methanol, ethanol, propanol, butanol or isopropanol.
일 구체예에서, 상기 녹각 추출물, 토사자 추출물, 숙지황 추출물 또는 백자인 추출물은 전술한 추출용매를 이용하여 추출된 1차 추출물을 극성이 다른 추출용매를 이용하여 분획한 분획물을 포함할 수 있다. 예를 들어, 녹각 추출물, 토사자 추출물, 숙지황 추출물 또는 백자인 추출물은 탄소수 1 내지 5의 알코올로 추출한 후, 에테르, 벤젠, 헥산 등의 극성이 다른 용매로 다시 분획한 분획물 일 수 있다. 상기 분획 시 용매는 2종 이상 사용할 수 있으며, 용매의 극성에 따라 순차적으로 사용하거나 혼합하여 사용하여, 각 용매 추출물을 제조할 수 있다. In one embodiment, the extracts of green tea extract, Sorghum extract, Sambrook extract, or White porcelain extract may include fractions obtained by fractionating the primary extract extracted using the above-described extraction solvent with an extraction solvent having a different polarity. For example, the extracts of green tea extract, extracts of Sorghum extract, extract of Sukhiteol, or extract of Phellodendrir may be fractions obtained by extracting with alcohol having 1 to 5 carbon atoms and then fractionating again with solvents having different polarity such as ether, benzene, hexane and the like. Two or more kinds of solvents may be used for the fractionation, and the solvent extracts may be sequentially used or mixed according to the polarity of the solvent.
본 발명에서, 상기 제조된 추출물 또는 상기 분획과정을 수행하여 수득한 분획물에 대해 여과하거나 농축 또는 건조과정을 수행하여 용매를 제거할 수 있으며, 상기 여과, 농축 및 건조를 모두 수행할 수 있다. 구체적으로 상기 여과는 여과지를 이용하거나 감압여과기를 이용할 수 있고, 농축은 감압 농축기 등을 이용하여 감압 농축할 수 있으며, 건조는 동결건조법을 수행할 수 있다. In the present invention, the extracted extract or the fraction obtained by performing the fractionation process may be filtered, concentrated or dried to remove the solvent, and the filtration, concentration, and drying may all be performed. Specifically, the filtration can be performed using a filter paper or a vacuum filter, and the concentration can be reduced by using a reduced pressure concentrator or the like, and the freeze-drying method can be carried out for drying.
또한, 상기 제조된 추출물 또는 상기 분획과정을 수행하여 수득한 분획물에 대해 실리카겔 컬럼 크로마토그래피(silica gel column chromatography), 박층 크로마토그래피(thin layer chromatography) 또는 고성능 액체 크로마토그래피(high performance liquid chromatography) 등의 다양한 크로마토그래피를 이용하여 정제함으로써 추가로 정제된 분획을 얻을 수 있다. The fractions obtained by performing the above-described extract or the fractionation process can be further purified by silica gel column chromatography, thin layer chromatography or high performance liquid chromatography Further purification can be achieved by purification using various chromatographic techniques.
본 발명의 녹각 추출물, 토사자 추출물, 숙지황 추출물 및 백자인 추출물의 혼합 추출물은 녹각 추출물, 토사자 추출물, 숙지황 추출물과 백자인 추출물이 1: 1~5 : 1~5 : 1~5 중량비로 혼합된 것이 바람직하다. 특히, 녹각 추출물, 토사자 추출물, 숙지황 추출물과 백자인 추출물이 1: 1: 1: 1의 중량비로 혼합되는 것이 보다 바람직하다.In the present invention, it is preferable that the mixture of the extracts of green tea extract, sorghum extract, sorghum extract and porcine garnet extract is mixed at a weight ratio of 1: 1 ~ 5: 1 ~ 5: 1 ~ 5 . Particularly, it is more preferable that the extracts of green tea extract, sorghum extract, safflower extract and white porcelain extract are mixed at a weight ratio of 1: 1: 1: 1.
본 발명의 조성물은 의약 제형 제조를 위해 사용할 수 있다.The compositions of the present invention may be used for the manufacture of pharmaceutical formulations.
상기 의약 제형은 오일 또는 수성 매질중의 용액, 현탁액 또는 유화액의 형태이거나, 엑스제, 분말제, 과립제, 정제 또는 캅셀제의 형태일 수 있다.The pharmaceutical formulations may be in the form of solutions, suspensions or emulsions in oils or aqueous media, or in the form of excipients, powders, granules, tablets or capsules.
또한, 상기 조성물은 추가로 동일 또는 유사한 기능을 나타내는 유효성분을 1종 이상 함유할 수 있다. 예컨대, 공지의 피부 미백, 주름 개선, 탄력 증진, 항염증, 항산화 및/또는 항당화 성분을 포함할 수 있을 것이다. 추가적인 피부 미백, 주름 개선, 탄력 증진, 항염증, 항산화 및/또는 항당화 성분을 포함하게 되면 본 발명의 조성물의 피부 재생, 주름 개선, 탄력 증진, 항염증, 항산화 및/또는 항당화 개선 효과는 더욱 증진될 수 있을 것이다. 상기 성분 추가 시에는 복합 사용에 따른 피부 안전성, 제형화의 용이성, 유효성분들의 안정성을 고려할 수 있다. 본 발명의 한 구체예에서, 상기 조성물은 당업계에 공지된 피부 재생 성분으로서, 레티노산, TGF, 동물 태반 유래의 단백질, 베튤린산 및 클로렐라 추출물, 당업계에 공지된 항산화 성분으로서, 토코페롤, 셀레늄, 비타민 C 및 페놀성 화합물, 당업계에 공지된 미백 성분으로서, 코지산(Kojic acid), 알부틴(Arbutin) 등과 같은 티로시나제 효소활성을 억제하는 물질, 하이드로퀴논(Hydroquinone), 비타민-C(L-Ascorbic acid) 및 이들의 유도체와 각종 식물 추출물로 구성되는 군으로부터 선택되는 1종 또는 2종 이상의 성분을 추가로 포함할 수 있다. 추가의 성분은 전체 조성물 중량에 대하여 0.0001 중량% 내지 10 중량%로 포함될 수 있을 것이며, 상기 함량 범위는 피부 안전성, 상기 유효성분의 제형화 시의 용이성 등의 요건에 따라 조절될 수 있을 것이다.In addition, the composition may further contain one or more active ingredients showing the same or similar functions. For example, it may contain known skin whitening, wrinkle enhancement, elasticity enhancement, antiinflammation, antioxidant and / or antialcification components. The skin regeneration, wrinkle improvement, elasticity enhancement, anti-inflammation, antioxidant, and / or antialgia improvement effect of the composition of the present invention when the composition contains the additional skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammation, antioxidant and / It can be further improved. When the above ingredients are added, skin safety, easiness of formulation, and stability of effective ingredients can be considered according to the combined use. In one embodiment of the present invention, the composition is a skin regeneration component known in the art comprising retinoic acid, TGF, protein from animal placenta, betulinic acid and chlorella extract, antioxidant components known in the art, such as tocopherol, selenium , Vitamin C and phenolic compounds, whitening ingredients known in the art, substances inhibiting the activity of tyrosinase enzymes such as kojic acid and arbutin, hydroquinone, vitamin C (L- Ascorbic acid, derivatives thereof, and various plant extracts. The additional ingredient may be included in an amount of 0.0001 to 10% by weight based on the total weight of the composition, and the content range may be adjusted according to requirements such as skin safety and easiness in formulating the active ingredient.
또한, 본 발명의 조성물은 약학적으로 허용 가능한 담체를 더 포함할 수 있다.In addition, the composition of the present invention may further comprise a pharmaceutically acceptable carrier.
약학적으로 허용 가능한 담체는 완충액, 주사용 멸균수, 일반 식염수 또는 인산염 완충 식염수, 수크로스, 히스티딘, 염 및 폴리솔베이트 등과 같은 여러 성분을 함유할 수 있다.The pharmaceutically acceptable carrier may contain various components such as buffer, injectable sterile water, normal saline or phosphate buffered saline, sucrose, histidine, salts and polysorbates.
본 발명의 조성물은 경구 또는 비경구로 투여할 수 있으며, 일반 약학 제제의 형태, 예를 들어, 임상 투여 시 경구 및 비경구의 여러 가지 제형으로 투여될 수 있는데, 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제될 수 있다.The composition of the present invention can be administered orally or parenterally, and can be administered in the form of a general pharmaceutical preparation, for example, various forms of oral and parenteral administration at the time of clinical administration. In the case of formulation, a filler, , A binder, a wetting agent, a disintegrant, a surfactant, and the like.
경구 투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 본 발명의 의약 조성물에 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘 카보네이트(Calcium carbonate), 수크로스(Sucrose) 또는 락토오스(Lactose), 젤라틴 등을 섞어 조제될 수 있다.Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like. These solid preparations can be prepared by mixing the pharmaceutical composition of the present invention with at least one excipient such as starch, calcium carbonate, Sucrose, lactose, gelatin and the like can be prepared.
단순한 부형제 이외에 마그네슘 스티레이트 탈크 같은 윤활제들도 사용된다. 경구를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순희석제인 물, 리퀴드 파라핀 이외에 여러가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다.In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Examples of liquid formulations for oral administration include suspensions, solutions, emulsions, syrups and the like. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used.
비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조제제, 좌제가 포함된다. 비수성용제, 현탁용제로는 프로필렌 글리콜(Propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세로제라틴 등이 사용될 수 있다.Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used as the non-aqueous solvent and suspension agent. As a base for suppositories, witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like can be used.
본 발명에 있어서, '미백 효과'라 함은 멜라닌 색소의 합성을 저해함으로써 피부 톤을 밝게 할 뿐만 아니라, 자외선, 호르몬 또는 유전에 기인한 기미나 주근깨 등의 피부 과색소 침착을 개선하는 것을 말한다.In the present invention, the 'whitening effect' refers to not only brightening the skin tone by inhibiting the synthesis of the melanin pigment but also improving skin hypercholesterolemia due to ultraviolet rays, hormones or heredity, such as spots or freckles.
본 발명에 있어서, '주름 개선 효과'라 함은 피부에 주름이 생성되는 것을 억제 또는 저해하거나, 이미 생성된 주름을 완화시키는 것을 말한다. In the present invention, the term " wrinkle-reducing effect " refers to inhibiting or inhibiting the generation of wrinkles on the skin, or alleviating already-generated wrinkles.
본 발명에 있어서, '탄력 증진 효과'라 함은 피부에 대한 탄력성이 증가되는 것으로, 피부 탄력의 손실을 억제 또는 저해하거나, 이미 감소된 탄력을 완화시키는 것을 말한다. In the present invention, the 'elasticity-enhancing effect' refers to an increase in elasticity to the skin, which suppresses or inhibits the loss of elasticity of the skin, or alleviates the already-reduced elasticity.
본 발명에 있어서, '항염증 효과'라 함은 염증을 억제하는 것을 말한다. 상기 염증은 어떤 자극에 대한 생체조직의 방어반응의 하나로, 조직 변질, 순환 장애와 삼출 및 조직 증식의 세 가지를 병발하는 복잡한 병변을 말한다. 보다 구체적으로 염증은 선천성 면역의 일부이며 다른 동물에서처럼 인간의 선천성 면역은 병원체에 특이적으로 존재하는 세포 표면의 패턴을 인식한다. 식세포는 그런 표면을 가진 세포를 비자기로 인식하고 병원체를 공격한다. 만일 병원균이 신체의 물리적 장벽을 깨고 들어온다면 염증반응이 일어난다. 염증반응은 상처 부위에 침입한 미생물들에 대한 적대 환경을 만드는 비특이적인 방어작용이다. 염증반응에서, 상처가 나거나 외부 감염체가 체내로 들어왔을 때, 초기단계 면역반응을 맡고 있는 백혈구들이 몰려들어 사이토카인을 발현한다. 따라서 세포 내 사이토카인의 발현양이 염증반응 활성화의 지표가 된다. 염증과 관련된 피부질환의 예로는 아토피 피부염, 건선, 방사선, 화학물질 또는 화상 등에 의한 홍반성 질환; 산 화상, 수포성 피부병, 태선 모양 종류 질환 또는 알레르기 등에 의한 가려움증; 지루성 습진, 장미 여드름, 심상성 천포창, 다형 삼출성 홍반, 결절 홍반, 귀두염, 음문염 또는 원형 탈모증과 같은 염증성 모발 손실; 피부 T-세포 림프종 등이 있으나 이에 제한되는 것은 아니다.In the present invention, the term 'anti-inflammatory effect' refers to inhibition of inflammation. The inflammation is one of biological tissue defense responses to a certain stimulus and refers to complicated lesions involving tissue degeneration, circulatory disorders, exudates and tissue proliferation. More specifically, inflammation is part of congenital immunity and, like in other animals, human congenital immunity recognizes a pattern of cell surfaces that are specifically present in a pathogen. Phagocytes recognize cells with such surfaces as non-magnetic and attack pathogens. If pathogens break through the physical barriers of the body, an inflammatory reaction occurs. Inflammation is a nonspecific defense that creates hostile environments for microorganisms entering the wound. In the inflammatory response, white blood cells that are responsible for the early stage of immune response, when wounded or infected, enter the body to express cytokines. Therefore, the expression level of intracellular cytokine is an index of inflammatory response activation. Examples of skin diseases associated with inflammation include, but are not limited to, atopic dermatitis, psoriasis caused by psoriasis, radiation, chemicals or burns; Itching caused by acid burns, watery skin diseases, psoriasis type diseases or allergies; Inflammatory hair loss such as seborrhoeic eczema, rose acne, pemphigus pemphigus, polymorphous exudative erythema, erythema nodosum, acuminate, pharyngitis or alopecia areata; Skin T-cell lymphoma, and the like.
본 발명에 있어서, '항산화 효과'라 함은 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.In the present invention, the term 'antioxidative effect' refers to the action of free radicals or reactive oxygen species (ROS), which are highly reactive according to oxidative stress caused by intracellular metabolism or ultraviolet rays, Refers to inhibition of oxidation, and includes removal of free radical or reactive oxygen species, thereby reducing damage to the cells.
본 발명에 있어서, '항당화 효과'라 함은 세포에 존재하는 포도당과 단백질이 반응하여 신체 내 단백질 기능이 저하되는 당화(Glycation) 현상을 예방하거나 억제하는 것을 뜻한다.In the present invention, 'antagonism effect' means preventing or suppressing glycation phenomenon in which glucose and protein present in cells react with each other to lower protein function in the body.
본 발명에 있어서, '유효량'이라 함은 손상된 미백 효과를 나타내거나, 주름을 개선하거나, 탄력을 증진시키거나, 염증을 억제하거나, 세포의 산화를 억제 또는 완화하거나, 당화의 생성을 억제할 수 있는 추출물의 양을 의미한다. 본 발명의 조성물이 유효량의 상기 녹각 추출물, 토사자 추출물, 숙지황 추출물 및 백자인 추출물을 혼합한 혼합 추출물을 포함할 때 바람직한 피부 미백 효과, 주름 개선 효과, 탄력 증진 효과, 항염증 효과, 항산화 효과, 항당화 효과를 제공할 수 있다. 본 발명의 조성물에 포함되는 상기 녹각 추출물, 토사자 추출물, 숙지황 추출물 및 백자인 추출물을 혼합한 혼합 추출물의 유효량은 조성물이 제품화되는 형태, 상기 화합물이 피부에 적용되는 방법 및 피부에 머무르는 시간 등에 따라 달라질 것이다. 예컨대, 상기 조성물이 약학 제형으로 제품화되는 경우에는 일상적으로 피부에 적용하게 되는 화장품으로 제품화되는 경우에 비해 높은 농도로 상기 유효성분을 포함할 수 있을 것이다. 따라서, 일일 투여량은 상기 녹각 추출물, 토사자 추출물, 숙지황 추출물 및 백자인 추출물을 혼합한 혼합 추출물의 양을 기준으로 0.1 내지 100 ㎎/㎏이고, 바람직하게는 30 내지 80 ㎎/㎏이고, 더욱 바람직하게는 50 내지 60 mg/kg이며, 하루 1 ∼ 6 회 투여될 수 있다. In the present invention, the term "effective amount" refers to an effective amount of a compound capable of exhibiting a damaged whitening effect, improving wrinkles, improving elasticity, suppressing inflammation, inhibiting or alleviating oxidation of cells, Means the amount of extract present. When the composition of the present invention contains an effective amount of a mixed extract obtained by mixing the extracts of the green tea extract, the extract of Sorghum extract, the extract of Sukjiro, and the extract of White porcelain, the skin whitening effect, wrinkle improving effect, elasticity improving effect, anti- Effect can be provided. The effective amount of the mixed extract of the composition of the present invention, which is obtained by mixing the extracts of green tea extract, Sorghum extract, Sambrook extract, and White porcelain extract, will vary depending on the form of the composition, how the compound is applied to the skin, . For example, when the composition is produced into a pharmaceutical formulation, the active ingredient may be contained at a higher concentration than in the case where it is commercialized as cosmetics that are routinely applied to the skin. Therefore, the daily dosage is 0.1 to 100 mg / kg, preferably 30 to 80 mg / kg, based on the amount of the mixed extract obtained by mixing the above-mentioned Green peach extract, Sorghum extract, Is 50 to 60 mg / kg, and can be administered 1 to 6 times a day.
본 발명의 조성물은 단독으로, 또는 수술, 방사선 치료, 호르몬 치료, 화학 치료 및 생물학적 반응조절제를 사용하는 방법들과 병용하여 사용할 수 있다.The composition of the present invention may be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers.
본 발명의 의약 제형은 피부 외용제 제형을 포함할 수 있다. The pharmaceutical formulations of the present invention may include external preparation for skin.
상기 녹각 추출물, 토사자 추출물, 숙지황 추출물 및 백자인 추출물을 혼합한 혼합 추출물을 피부외용제로 사용하는 경우, 추가로 지방 물질, 유기 용매, 용해제, 농축제 및 겔화제, 연화제, 항산화제, 현탁화제, 안정화제, 발포제(foaming agent), 방향제, 계면활성제, 물, 이온형 또는 비이온형 유화제, 충전제, 금속이온봉쇄제 및 킬레이트화제, 보존제, 비타민, 차단제, 습윤화제, 필수 오일, 염료, 안료, 친수성 또는 친유성 활성제, 지질 소낭 또는 피부용 외용제에 통상적으로 사용되는 임의의 다른 성분과 같은 피부 과학 분야에서 통상적으로 사용되는 보조제를 함유할 수 있다. 또한 상기 성분들은 피부 과학 분야에서 일반적으로 사용되는 양으로 도입될 수 있다. In the case of using the mixed extract obtained by mixing the above extract of green tea extract, the extract of Sorghum, the extract of Suhwangjang, and the extract of Ishigaki extract, as the external preparation for skin, it is further possible to use a lipid, an organic solvent, a solubilizing agent, a thickening agent and a gelling agent, a softening agent, an antioxidant, Surfactants, water, ionic or non-ionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic Or lipophilic active agents, lipid vesicles, or any other ingredient conventionally used in external preparations for skin. The components can also be introduced in amounts commonly used in the field of dermatology.
상기 녹각 추출물, 토사자 추출물, 숙지황 추출물 및 백자인 추출물을 혼합한 혼합 추출물이 피부 외용제 제형으로 제공될 경우, 이에 제한되는 것은 아니나, 연고, 패취, 겔, 크림 또는 분무제와 같은 제형을 가질 수 있다.When the mixed extract obtained by mixing the green tea extract, the Sorghum extract, the Sukhwajyang extract, and the Phellodendron chinense extract is provided as an external preparation for skin, it may have a form such as ointment, patch, gel, cream or spray.
또한, 본 발명의 피부 미백, 주름 개선, 탄력 증진, 항염증, 항산화 및/또는 항당화용 조성물은 화장료 제형 제조를 위해 사용될 수 있다.Further, the composition for skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammation, antioxidant and / or antialcification of the present invention can be used for the production of cosmetic formulations.
상기 화장료 제형은 일반적인 유화 제형 및 가용화 제형의 형태일 수 있다. 예컨대, 유연 화장수 또는 영양 화장수 등과 같은 화장수, 훼이셜 로션, 바디로션 등과 같은 유액, 영양 크림, 수분 크림, 아이 크림 등과 같은 크림, 에센스, 화장연고, 스프레이, 젤, 팩, 선 스크린, 메이크업 베이스, 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 있다. The cosmetic formulation may be in the form of a conventional emulsion formulation and a solubilized formulation. For example, creams, essences, cosmetic creams, sprays, gels, packs, sunscreens, make-up bases, liquids such as lotions such as lotion, facial lotion, body lotion, A powder, a cleansing lotion, a makeup removing agent such as a cleansing oil, a cleaning agent such as a cleansing foam, a soap, a body wash and the like.
또한, 상기 화장품은 녹각 추출물, 토사자 추출물, 숙지황 추출물 및 백자인 추출물을 혼합한 혼합 추출물에 추가로 지방 물질, 유기 용매, 용해제, 농축제 및 겔화제, 연화제, 항산화제, 현탁화제, 안정화제, 발포제(foaming agent), 방향제, 계면활성제, 물, 이온형 또는 비이온형 유화제, 충전제, 금속이온봉쇄제 및 킬레이트화제, 보존제, 비타민, 차단제, 습윤화제, 필수 오일, 염료, 안료, 친수성 또는 친유성 활성제, 지질 소낭 또는 화장품에 통상적으로 사용되는 임의의 다른 성분과 같은 화장품학 분야에서 통상적으로 사용되는 보조제를 함유할 수 있다.In addition, the above-mentioned cosmetic product may further contain a fatty substance, an organic solvent, a solubilizing agent, a thickening agent and a gelling agent, a softening agent, an antioxidant, a suspending agent, a stabilizer, a foaming agent a foaming agent, a fragrance, a surfactant, water, an ionic or nonionic emulsifier, a filler, a sequestering and chelating agent, a preservative, a vitamin, a barrier agent, a wetting agent, Active agents, lipid vesicles, or any other ingredient commonly used in cosmetics, as well as adjuvants commonly used in the cosmetics field.
상기 화장료 제형은 유효성분이 단기간 내에 피부에 머무르게 되는 메이크업 제거제, 세정제 등과 같은 워쉬-오프(wash-off) 타입의 화장품의 경우에는 비교적 높은 농도의 상기 유효성분을 포함할 수 있을 것이다. 반면, 유효성분이 장기간 동안 피부에 머무르게 되는 화장수, 유액, 크림, 에센스 등의 리브-온(leave-on) 타입의 화장품의 경우에는 워쉬-오프 타입의 화장품에 비해 낮은 농도의 상기 유효성분을 포함해도 무방할 것이다. 이에 제한되는 것은 아니나, 본 발명의 한 구체예에서, 상기 조성물은 상기 유효성분을 전체 조성물 중량에 대하여 0.0001 중량% 내지 10 중량%(바람직하게는 0.0001 중량% 내지 1 중량%)로 포함할 수 있다. 본 발명의 조성물이 상기 녹각 추출물, 토사자 추출물, 숙지황 추출물 및 백자인 추출물을 혼합한 혼합 추출물을 0.0001 중량% 미만으로 포함할 경우에는 충분한 피부 미백, 주름 개선, 탄력 증진, 항염증, 항산화 및/또는 항당화 효과를 기대할 수 없고, 10 중량%를 초과하여 포함할 경우에는 알러지 등 원치 않는 반응이 발생하거나 피부 안전성에 문제가 있을 수 있으므로 이를 방지하기 위한 것이다.The cosmetic formulation may contain a relatively high concentration of the active ingredient in the case of a wash-off type cosmetic such as a make-up remover, a cleanser, etc. in which the active ingredient remains on the skin in a short period of time. On the other hand, in the case of leave-on type cosmetics such as lotion, cream, essence and the like in which the active ingredient remains on the skin for a long period of time, It will be acceptable. In one embodiment of the present invention, although not limited thereto, the composition may contain the active ingredient in an amount of 0.0001 wt% to 10 wt% (preferably 0.0001 wt% to 1 wt%) based on the total weight of the composition . When the composition of the present invention contains less than 0.0001% by weight of the mixed extract of the green tea extract, the Sorghum extract, the Sambrook extract, and the white porcelain extract, sufficient skin whitening, wrinkle improvement, elasticity enhancement, antiinflammation, antioxidant and / A saccharifying effect can not be expected. If it is contained in an amount exceeding 10% by weight, undesired reactions such as allergies may occur or there may be a problem in skin safety.
또한, 본 발명의 피부 미백, 주름 개선, 탄력 증진, 항염증, 항산화 및/또는 항당화용 조성물은 식품 제형 제조를 위해 사용될 수 있다.In addition, the composition for skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammation, antioxidant and / or antialcification of the present invention can be used for manufacturing food formulations.
상기 식품 제형은 상기 녹각 추출물, 토사자 추출물, 숙지황 추출물 및 백자인 추출물을 혼합한 혼합 추출물을 음료, 차류, 향신료, 껌, 과자류 등의 식품 소재에 첨가하거나, 캡슐화, 분말화, 현탁액 등으로 제조한 식품을 의미한다. The food formulation may be prepared by adding a mixed extract obtained by mixing the green algae extract, alum extract, Succinic acid extract and porcine green extract to a food material such as beverage, tea, spice, gum and confectionery, or a food product prepared by encapsulation, .
상기 식품 제형은 일상적으로 섭취하는 것이 가능하기 때문에 높은 피부 미백, 주름 개선, 탄력 증진, 항염증, 항산화 및/또는 항당화 효과를 기대할 수 있어 매우 유용하다.Since the food preparation can be routinely ingested, it is very useful because high skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammation, antioxidant and / or anticarcinogenic effect can be expected.
상기 유효성분을 식품첨가물로 사용하는 경우, 상기 녹각 추출물, 토사자 추출물, 숙지황 추출물 및 백자인 추출물을 혼합한 혼합 추출물을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 유효성분의 혼합양은 그의 사용 목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있다. 일반적으로, 식품 또는 음료의 제조시에 본 발명의 조성물은 원료에 대하여 15 중량부 이하, 바람직하게는 10 중량부 이하의 양으로 첨가된다. 그러나, 건강 및 위생을 목적으로 하거나 또는 건강 조절을 목적으로 하는 장기간의 섭취의 경우에는 상기 양은 상기 범위 이하일 수 있으며, 안전성 면에서 아무런 문제가 없기 때문에 유효성분은 상기 범위 이상의 양으로도 사용될 수 있다.When the above-mentioned effective ingredient is used as a food additive, a mixed extract obtained by mixing the above-mentioned Green peach extract, Sorghum extract, Sambrook extract and White Pepper extract may be directly added or used together with other food or food ingredients, Can be used. The amount of the active ingredient to be mixed can be suitably determined according to its intended use (prevention, health or therapeutic treatment). Generally, the composition of the present invention is added in an amount of not more than 15 parts by weight, preferably not more than 10 parts by weight, based on the raw material, when the food or beverage is produced. However, in the case of long-term consumption intended for health and hygiene purposes or for health control purposes, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range .
상기 식품의 종류에는 특별한 제한은 없다. 상기 물질을 첨가할 수 있는 식품의 예로는 육류, 소세지, 빵, 초콜릿, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알코올 음료 및 비타민 복합제 등이 있으며, 통상적인 의미에서의 건강식품을 모두 포함한다.There is no particular limitation on the kind of the food. Examples of the food to which the above substance can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include health foods in a conventional sense.
식품 제형이 음료인 경우, 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상술한 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드, 말토스, 수크로스와 같은 디사카라이드, 및 덱스트린, 사이클로덱스트린과 같은 폴리사카라이드, 자일리톨, 소르비톨, 에리트리톨 등의 당알코올이다. 감미제로서는 타우마틴, 스테비아 추출물과 같은 천연 감미제나, 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 조성물 100 mL 당 일반적으로 약 0.01 ∼ 0.04 g, 바람직하게는 약 0.02 ∼ 0.03 g 이다.When the food is a beverage, various flavors or natural carbohydrates may be added as an additional ingredient such as a normal drink. The above-mentioned natural carbohydrates are sugar alcohols such as monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 mL of the composition of the present invention.
상기 외에 식품 제형은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산 음료에 사용되는 탄산화제 등을 함유할 수 있다. 그 밖에 식품 제형은 천연 과일주스, 과일주스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 혼합하여 사용할 수 있다. 이러한 첨가제의 비율은 크게 중요하진 않지만 본 발명의 조성물 100 중량부당 0.01 ~ 0.1 중량부의 범위에서 선택되는 것이 일반적이다.In addition to the above, the food formulations may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, And the like. Other food formulations may contain flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not critical, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.
이하, 본 발명을 실시예에 의해 상세히 설명한다. 단, 하기 실시예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예에 한정되는 것은 아니다. Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.
제조예 1: 녹각 추출물의 제조Preparation Example 1: Preparation of a green algae extract
녹각을 잘 건조하여 세절한 후, 건조중량 100g을 플라스크에 넣고 추출용매(증류수) 1000g으로 3일간 냉침하여 추출하였다. 냉침된 추출물을 0.2㎛의 기공 크기를 가진 필터로 여과하여 녹각 추출물을 제조하였다.The dough was dried thoroughly, and then 100 g of dry weight was placed in a flask and extracted with 1000 g of an extraction solvent (distilled water) for 3 days by cooling. The frozen extract was filtered with a filter having a pore size of 0.2 mu m to prepare a green leaf extract.
제조예 2: 토사자 추출물의 제조Preparation Example 2: Preparation of soil extract
토사자를 잘 건조하여 세절한 후, 건조중량 100g을 플라스크에 넣고 추출용매(증류수) 1000g으로 3일간 냉침하여 추출하였다. 냉침된 추출물을 0.2㎛의 기공 크기를 가진 필터로 여과하여 토사자 추출물을 제조하였다.The soil was dried well, and the dried weight of 100 g was placed in a flask and extracted with 1000 g of an extraction solvent (distilled water) for 3 days by cooling. The frozen extract was filtered with a filter having a pore size of 0.2 mu m to prepare a soil extract.
제조예Manufacturing example 3: 숙지황 추출물의 제조 3: Preparation of Sukjwanghwang Extract
숙지황(숙건지황)을 잘 건조하여 세절한 후, 건조중량 100g을 플라스크에 넣고 추출용매(증류수) 1000g으로 3일간 냉침하여 추출하였다. 냉침된 추출물을 0.2㎛의 기공 크기를 가진 필터로 여과하여 숙지황 추출물을 제조하였다Sukjungghwang (Sukjunggwanghang) was well dried and finely divided, and 100 g of dry weight was put into a flask and extracted with 1000 g of an extraction solvent (distilled water) for 3 days by cooling. The frozen extract was filtered with a filter having a pore size of 0.2 [micro] m to prepare a Sukhwangju extract
제조예Manufacturing example 4: 백자인 추출물의 제조 4: Preparation of white porcelain extract
백자인을 잘 건조하여 세절한 후, 건조중량 100g을 플라스크에 넣고 추출용매(증류수) 1000g으로 3일간 냉침하여 추출하였다. 냉침된 추출물을 0.2㎛의 기공 크기를 가진 필터로 여과하여 백자인 추출물을 제조하였다The white porcelain was dried well and cut into three pieces. 100 g of dry weight was put into a flask and extracted with 1000 g of an extraction solvent (distilled water) for 3 days by cooling. The frozen extract was filtered with a filter having a pore size of 0.2 mu m to prepare a white porcelain extract
제조예Manufacturing example 5: 녹각, 토사자, 숙지황 및 5: Tapestry, tobacco, 백자인의White porcelain 혼합 추출물 제조 Mixed Extract Manufacture
제조예 1의 녹각 추출물 10g, 제조예 2의 토사자 추출물 10g, 제조예 3의 숙지황 추출물 10g과 제조예 4의 백자인 추출물 10g을 혼합하여 녹각, 토사자, 숙지황 및 백자인 혼합 추출물을 제조하였다.10 g of the green oak extract of Preparation Example 1, 10 g of the Sorghum extract of Preparation Example 2, 10 g of the Sukhwajo Extract of Preparation Example 3, and 10 g of the Pahojang Extract of Preparation Example 4 were mixed to prepare a mixed extract of green tea, sorghum,
실시예Example 1: 멜라닌 생성 저해 효과 1: Melanin formation inhibitory effect
멜라닌 생성 저해를 통한 미백 효과를 확인하기 위하여, Lotan R. 외(Cancer Res. 40:3345-3350, 1980)에 기재된 방법에 따라 쥐의 멜라노마 세포(B-16 mouse melanoma cell)의 배양액에, 추출물을 첨가하여 멜라닌 총량을 측정하였다. 실험 시, 먼저 쥐의 멜라노마 세포에 대하여 독성을 평가하여 독성이 없는 농도에서 미백 평가를 수행하였다. 음성 대조군으로는 DMSO를, 양성 대조군으로는 알부틴(albutin)을 사용하였다. In order to confirm the whitening effect through inhibition of melanin formation, a culture solution of B-16 mouse melanoma cells in rat was cultured according to the method described in Lotan R. et al. (Cancer Res. 40: 3345-3350, 1980) The total amount of melanin was measured by adding an extract. In the experiment, first the toxicity of melanoma cells in rats was evaluated and the whitening evaluation was carried out at a concentration that is not toxic. DMSO was used as a negative control group, and albutin was used as a positive control group.
구체적으로, 시료를 최종 농도가 100 ppm이 되도록 배지에 첨가하고, 알부틴은 100 ppm이 되도록 배지에 첨가한 후 멜라노마 세포를 3일간 배양하였다. 이후, 세포들을 트립신(trypsin) 처리하여 배양용기로부터 떼어내 원심분리한 후, 멜라닌을 추출하였다. 떼어낸 세포는 수산화나트륨 용액(1N 농도) 1 ml를 가하여 10분간 끓여 멜라닌을 녹이고 분광 광도계를 이용하여, 400 nm에서 흡광도를 측정하여 생성된 멜라닌의 양을 측정하였다. Specifically, the sample was added to the medium to a final concentration of 100 ppm, arbutin was added to the medium to be 100 ppm, and melanoma cells were cultured for 3 days. Cells were then trypsinized, detached from the culture, centrifuged, and then melanin was extracted. The removed cells were incubated with 1 ml of sodium hydroxide solution (1N concentration), boiled for 10 minutes to dissolve melanin, and the absorbance was measured at 400 nm using a spectrophotometer to measure the amount of melanin produced.
상기 멜라닌 양은 단위 세포수당(1×106 cell)의 흡광도로 나타내는 방법으로 측정하였으며, 대조군에 대한 상대적인 멜라닌 총량을 저해율(%)로 계산하고 결과를 하기 표 1에 나타내었으며, 실험은 각각 3회씩 수행하여 평균값으로 나타내었다. The amount of melanin was measured by an absorbance of 1 × 10 6 cells per unit cell, and the total amount of melanin relative to the control group was calculated as the inhibition rate (%). The results are shown in Table 1 below. As shown in FIG.
상기 표 1의 결과에서 볼 수 있듯이, 제조예 5의 혼합 추출물은 저농도에서도 뛰어난 멜라닌 총량 감소 효과를 나타내어 미백 용도로 사용할 수 있음을 알 수 있었다. As can be seen from the results of Table 1, the mixed extract of Preparation Example 5 exhibited an excellent effect of reducing the total amount of melanin even at a low concentration, and thus it could be used as a whitening agent.
실시예Example 2: 콜라겐 합성 촉진 효과 2: promoting collagen synthesis
시료를 인간 유래의 섬유아세포의 배양액에 첨가하여 세포수준에서 제1형 콜라겐 합성 촉진 효과를 확인하였다. 합성된 콜라겐의 측정은 PICP EIA kit (Procollagen Type I C-Peptide Enzyme ImmunoAssay KIT)를 이용하여 정량하였다.The sample was added to a culture solution of human fibroblasts to confirm the promoting effect of type I collagen synthesis at the cellular level. The synthesized collagen was quantitated using a PICP EIA kit (Procollagen Type I C-Peptide Enzyme Immunoassay Kit).
시료를 최종 농도 10 ppm이 되도록 하여 섬유아세포의 배양배지(DMEM 배지)에 첨가하여 48시간 배양한 후 배양액을 취하여 PICP EIA 키트로 각 농도에서 제1형 콜라겐 합성 정도를 분광광도계를 이용하여 450 nm에서 측정하였다. Samples were added to a culture medium of fibroblasts (DMEM medium) to a final concentration of 10 ppm, and cultured for 48 hours. The culture broth was taken and the degree of type I collagen synthesis at each concentration was measured with a PICP EIA kit using a spectrophotometer at 450 nm Respectively.
효과 비교를 위하여 시료를 처리하지 않은 섬유아세포의 배양 배지(음성대조군)와 비타민 C(양성대조군)를 최종 농도 52.85 ㎍/ml가 되도록 첨가한 시료에 대하여 동일한 방법으로 콜라겐 합성 정도를 측정하였다. For comparison, the degree of collagen synthesis was measured in the same manner for the samples in which the culture medium of the untreated fibroblasts (negative control) and the vitamin C (positive control) were added to a final concentration of 52.85 / / ml.
콜라겐 생성 증가율은 음성 대조군에 대한 상대적인 콜라겐 생성량의 비율로 계산하고 결과를 하기 표 2에 나타내었으며, 실험은 4회씩 수행하여 평균값으로 나타내었다.The increase rate of collagen production was calculated by the ratio of relative collagen production to the negative control, and the results are shown in Table 2 below.
상기 표 2에 나타난 바와 같이, 제조예 5의 혼합 추출물을 처리한 경우에 콜라겐 합성을 증가시켰고, 일반적으로 콜라겐 합성을 유도하는 것으로 잘 알려진 비타민C를 적용한 경우와 유사한 콜라겐 합성 효과를 나타내었다. As shown in the above Table 2, when the mixed extract of Preparation Example 5 was treated, collagen synthesis was increased and collagen synthesis effect similar to that of vitamin C, which is generally known to induce collagen synthesis, was exhibited.
실시예Example 3: 3: 엘라스테이즈Ella Stays 활성 저해 효과 Active inhibitory effect
엘라스틴(Elastin)을 분해하는 효소인 엘라스테이즈(Elastase)의 활성 저해 효과를 다음과 같이 확인하였다.The activity inhibitory effect of Elastase, an enzyme that degrades elastin, was confirmed as follows.
엘라스테이즈(Elastase)는 사람의 백혈구 세포로부터 유래한 엘라스테이즈를 사용하였고, 엘라스테이즈의 기질로 합성 기질인 MeOSuc-Ala-Ala-Pro-Val-pNA를 사용하였다. 완충 용액은 100 mM의 Tris(pH 7.5) 용액을 사용하였다. 엘라스테이즈는 완충용액을 이용하여 최종적으로 0.2 mU을 사용하였다. 또한, 엘라스테이즈의 합성 기질은 DMSO를 이용하여 100 mM 용액을 만든 후 최종 농도가 0.5 mM이 되도록 완충용액을 이용하여 희석하였다. 이때, 양성 대조군은 엘라스테이즈 저해 물질로 알려진 쿼세틴(Quercetin)을 10 ppm 농도로 넣은 것으로 설정하였다. 엘라스테이즈 저해 후보는 최종 농도가 10 ppm이 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 다음 수학식 1과 같이 계산하였다. Elastase used Elastase from human leukocyte cells and MeOSuc-Ala-Ala-Pro-Val-pNA as synthetic substrate for Elastase. The buffer solution used was 100 mM Tris (pH 7.5) solution. Finally, 0.2 mU was used for the ELASTASES using a buffer solution. In addition, the synthetic substrate of Elastinase was diluted with a buffer solution to a final concentration of 0.5 mM after making a 100 mM solution using DMSO. At this time, the positive control group was set to contain 10 ppm of quercetin, which is known as Elastase inhibitor. Ella stase inhibition candidates were added to give a final concentration of 10 ppm. The reaction was carried out in a 96-well plate and allowed to react at room temperature for 20 minutes. Absorbance was measured at 405 nm using a spectrophotometer at intervals of 1 minute, and the slope of the absorbance versus time was determined as the activity of the enzyme. The inhibition rate of the ELA stasis was calculated by the following equation (1).
[수학식 1] [Equation 1]
상기 표 3에서 볼 수 있듯이, 제조예 5의 혼합 추출물을 처리한 경우, 양성대조군 보다 효과는 적으나, 엘라스테이즈 활성 저해가 뛰어남을 알 수 있다. 즉, 혼합 추출물은 우수한 엘라스테이즈 활성 저해 효과를 발휘하였다. 따라서, 혼합 추출물은 탄력 증진, 주름 개선을 위한 용도로 사용할 수 있음을 알 수 있었다. As can be seen from the above Table 3, when the mixed extract of Preparation Example 5 was treated, the effect was less than that of the positive control, but the inhibition of the Elastase activity was excellent. That is, the mixed extract exerted a superior inhibitory effect on the activity of Elastase. Therefore, it was found that the mixed extract can be used for the purpose of improving the elasticity and improving the wrinkles.
실시예Example 4: 항염 효과 4: anti-inflammatory effect
항염증 효과 및 피부 트러블 개선 효과를 확인하기 위하여, RAW264.7 세포주 (ATCC number: CRL-2278)를 이용한 GRIESS 법으로 nitric oxide(NO) 생성 억제력 실험을 실시하였다. In order to confirm the anti-inflammatory effect and the improvement of skin trouble, nitric oxide (NO) production inhibition experiment was performed by GRIESS method using RAW264.7 cell line (ATCC number: CRL-2278).
구체적으로, 생쥐의 대식세포인 RAW264.7 세포를 수차례 계대 배양하고, 웰 하나에 3×105 개씩 들어가도록 24-웰 프레이트에 넣은 후, 24시간 동안 배양하였다. 이어서, 최종 농도 10 ppm의 농도로 시료를 함유한 세포 배지로 교체하였다. 이때, NO-생성 억제 물질인 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, 540 nm에서의 흡광도를 측정함으로써 NO 억제 효과를 판단하고, NO 생성 저해율(%)은 하기 수학식 2를 이용하여 계산하여 하기 표 4에 나타내었으며, 실험은 각각 3회씩 수행하여 평균값으로 나타내었다. Specifically, RAW264.7 cells, macrophages of mice, were subcultured several times, placed in 24-well plates so as to enter 3x10 5 wells into each well, and cultured for 24 hours. Subsequently, the cell culture medium containing the sample was replaced with a final concentration of 10 ppm. At this time, L-NMMA (L-NG-Monomethylarginine), which is an inhibitor of NO production, was treated together as a positive control and cultured for 30 minutes. Lipopolysaccharide (LPS) 100 μl of the supernatant was transferred to a 96-well plate, and 100 μl of GRIESS solution was added thereto at room temperature for 10 minutes. The absorbance at 540 nm was measured to determine the NO inhibitory effect. The results are shown in Table 4 below, and the experiments were carried out three times each, and the results were shown as average values.
[수학식 2] &Quot; (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 4에서 볼 수 있듯이, 제조예 5의 혼합 추출물은 대표적인 항염 의약물질인 L-NMMA와 비교하였을 때, 그 활성은 낮으나 천연물질로서 우수한 활성을 나타내어, 피부 트러블 개선에 있어, 항염 효과가 보조적인 역할로 작용하여 상승 효과를 기대할 수 있다.As shown in Table 4, the mixed extract of Preparation Example 5 exhibited excellent activity as a natural substance when compared with L-NMMA, which is a representative anti-inflammatory drug, and its activity is low, It can act as a role and a synergistic effect can be expected.
실시예Example 5: 항산화 효과- 자유라디칼 소거율 5: Antioxidant effect - Free radical scavenging rate
시료의 항산화 작용을 확인하기 위해 자유라디칼 소거 활성을 측정하였다. 자유라디칼 소거 활성은 DPPH를 이용하여 측정하였다 (Blois, Nature 181, 1190, 1958). Free radical scavenging activity was measured to confirm the antioxidative activity of the sample. Free radical scavenging activity was measured using DPPH (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 사용하였다.DPPH is a relatively stable free radical, which shows maximum absorption at 517 nm in the presence of radicals and loses its absorbance when radicals are eliminated. DPPH was purchased from Sigma.
먼저, 1.5 mM (0.06 mg/ml)의 표준 DPPH 에탄올 용액을 만들었다. 그리고, 3차 증류수를 이용하여 1000 μg/ml (0.1%) 비타민 C 용액을 만든 다음 1/2씩 연속 희석하여 1000, 500, 250, 125, 62.5, 31.25, 15.63, 7.81, 3.91, 1.95, 0.98, 0.49 μg/ml 농도의 비타민 C 시료를 만들었다. 또한, 비타민 C와 동일한 농도로 제조예 1~5의 희석액을 준비하였다. 그 다음, 상기 시료와 표준 DPPH 용액을 같은 비율로 첨가하여 잘 교반한 후, 37 ℃에서 30분간 반응시키고 마이크로플레이트 리더(BioTek EL-340)를 이용하여 517㎚에서의 흡광도를 측정하여 IC50을 구하여 하기 표 5에 나타내었다. First, a standard DPPH ethanol solution of 1.5 mM (0.06 mg / ml) was prepared. Then, 1000 μg / ml (0.1%) vitamin C solution was prepared using the third distilled water and then serially diluted by 1/2, and then diluted to 1000, 500, 250, 125, 62.5, 31.25, 15.63, 7.81, 3.91, 1.95, , And a vitamin C sample at a concentration of 0.49 μg / ml. In addition, a diluted solution of Preparation Examples 1 to 5 was prepared at the same concentration as that of vitamin C. Then, after stirring well was added to the samples and standards DPPH solution at the same rate, from 37 ℃ 30 minutes of reaction and a microplate reader (BioTek EL-340) by measuring the absorbance at 517㎚ using the IC 50 The results are shown in Table 5 below.
IC50(half maximal inhibitory concentration)은 무첨가 대조군(이 경우 DPPH)의 자유라디칼을 50% 제거하는데 필요한 물질의 농도로서, 자유라디칼 소거능을 표현하는 일반적인 방법이다.The IC 50 (half maximal inhibitory concentration) is the concentration of the substance required to remove 50% of the free radicals of the no-added control (DPPH in this case) and is a common method of expressing the free radical scavenging ability.
상기 표 5와 같이, 제조예 5의 혼합 추출물은 제조예 1~4와 비교하였을 때 자유라디칼 소거능이 뛰어났으며, 양성대조군인 비타민 C와 유사한 항산화 효과를 지님을 확인하였다.As shown in Table 5, the mixed extract of Preparation Example 5 had excellent free radical scavenging ability as compared with Preparation Examples 1 to 4, and it was confirmed that the extract had an antioxidative effect similar to that of the positive control vitamin C.
실시예Example 6: 6: 항당화Antialcification 효과 effect
항당화(anti-glycation) 효능을 확인하기 위하여, L-아르기닌(arginine)과 포도당을 이용하여 당화 저해 활성을 측정하였다. In order to confirm anti-glycation efficacy, glycation inhibition activity was measured using L-arginine and glucose.
먼저, 1M 인산 완충용액(pH 7.4)을 이용하여 1M L-아르기닌, 1M 포도당을 녹여 준비하고 1M 인산 완충용액을 이용하여 시료를 50 ppm이 되도록 희석해서 준비하였다. 1M L-아르기닌과 1M 인산 완충용액을 1 대 4의 비율로 섞은 다음 96-웰 플레이트에 80 μl씩 분주하였다. 여기에 각각 50ppm으로 희석한 시료와 양성대조군으로 사용될 0.01M 아미노구아니딘(aminoguanidin)을 100 μl씩 첨가하였다. 이 시료들을 잘 섞어준 다음, 마지막으로 포도당의 최종 농도가 0.1M이 되도록 1M 인산 완충용액으로 희석한 포도당을 넣은 후, 70 ℃에서 4시간 동안 반응시켰다. 96-웰 플레이트를 분광 광도계를 이용하여 420 nm에서 흡광도를 측정하여 당화 정도를 측정하였다. First, 1 M L-arginine and 1 M glucose were prepared by dissolving 1 M L-arginine and 1 M glucose in 1M phosphate buffer (pH 7.4) and diluted to 50 ppm with 1M phosphate buffer solution. 1M L-arginine and 1M phosphate buffer solution were mixed at a ratio of 1: 4, and then 80 [mu] l each was added to a 96-well plate. To each sample, 100 μl of 0.01 M aminoguanidine was added to each sample to be diluted to 50 ppm and used as a positive control. These samples were mixed well and finally glucose diluted with 1M phosphate buffer solution was added to the final concentration of glucose to 0.1 M and reacted at 70 ° C for 4 hours. The degree of saccharification was measured by measuring the absorbance of the 96-well plate at 420 nm using a spectrophotometer.
하기 수학식 1의 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. In the Glycation test group of the following equation 1, 1M L-arginine and 1M glucose were added to induce glycation. To measure the absorbance of the sample itself, absorbance was measured at 420 nm by adding 1M L-arginine and sample without glucose Respectively.
당화 저해 활성은 다음과 같은 수학식 3으로 구할 수 있다. 실험은 각각 3회씩 수행하여 평균값으로 나타내었다. The saccharide inhibitory activity can be obtained by the following formula (3). Experiments were performed three times each and expressed as a mean value.
[수학식 3] &Quot; (3) "
상기 표 6의 결과에서 볼 수 있듯이, 본 발명에 따른 제조예 5의 혼합 추출물은 항당화 물질로 알려진 Aminoguanidine과 비교할 때, 항당화 효과가 더욱 뛰어남을 알 수 있다. 즉, 혼합추출물은 우수한 항당화 효과를 나타내어 미백 또는 주름 개선에 있어, 항당화 효과가 보조적인 역할로 작용하여 상승 효과를 기대할 수 있다. As can be seen from the results of Table 6, the mixed extract of Preparation Example 5 according to the present invention is superior to the aminoguanidine known as an anticarious substance, and thus has an excellent anticarcinogenic effect. That is, the mixed extract has excellent antihyperglycosylation effect, and antagonism effect plays an auxiliary role in whitening or wrinkle improvement, so that a synergistic effect can be expected.
제제예Formulation example 1: 의약 제제의 제조 1: Manufacture of pharmaceutical preparations
1. 정제의 제조1. Preparation of tablets
제조예 5의 혼합 추출물 0.2㎎0.2 mg of the mixed extract of Preparation Example 5
옥수수전분 100㎎100 mg of corn starch
유 당 100㎎100 mg of milk
스테아린산 마그네슘 2㎎ 2 mg of magnesium stearate
상기의 성분을 혼합한 후, 통상의 정제의 제조방법에 따라서 타정하여 정제를 제조하였다.After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.
제제예Formulation example 2: 화장품의 제조 2: Manufacture of cosmetics
1. 영양크림의 제조1. Manufacture of nutritional cream
하기 조성과 같이, 제조예 5의 혼합 추출물을 유효성분으로 포함하는 영양크림을 통상의 방법에 따라 제조하였다.A nutritional cream containing the mixed extract of Preparation Example 5 as an active ingredient was prepared according to a conventional method.
제조예 5의 혼합 추출물 0.2중량%0.2% by weight of the mixed extract of Preparation Example 5
베타-1,3-글루칸 5.0 중량%Beta-1,3-glucan 5.0 wt%
밀납 10.0 중량%Wax 10.0 wt%
폴리솔베이트60 1.5 중량%Polysorbate 60 1.5 wt%
피이지 60 경화피마자유 2.0 중량%≪ tb > < tb > < tb >
솔비탄세스퀴올레이트 0.5 중량%0.5% by weight of sorbitan sesquioleate
유동파라핀 10.0 중량%Liquid paraffin 10.0 wt%
스쿠알란 5.0 중량%Squalane 5.0 wt%
카프릴릭/카프릭트리글리세라이드 5.0 중량%Caprylic / capric triglyceride 5.0 wt%
글리세린 5.0 중량%Glycerin 5.0 wt%
부틸렌글리콜 3.0 중량%3.0% by weight of butylene glycol
프로필렌글리콜 3.0 중량%3.0% by weight of propylene glycol
트리에탄올아민 0.2 중량%0.2% by weight triethanolamine
방부제 0.05 중량%Preservative 0.05 wt%
색소 0.05 중량%0.05% by weight of pigment
향료 0.05 중량%0.05% by weight fragrance
정제수 to 100 중량%Purified water to 100%
제제예Formulation example 3: 피부 외용제의 제조 3: Preparation of external preparation for skin
1. 연고의 제조1. Manufacture of ointment
하기 조성과 같이, 제조예 5의 혼합 추출물을 유효성분으로 포함하는 연고를 통상의 방법에 따라 제조하였다.An ointment containing the mixed extract of Preparation Example 5 as an active ingredient was prepared according to a conventional method as shown in the following composition.
제조예 5의 혼합 추출물 0.5 중량%0.5% by weight of the mixed extract of Preparation Example 5
베타-1,3-글루칸 10.0 중량%Beta-1,3-glucan 10.0 wt%
밀납 10.0 중량%Wax 10.0 wt%
폴리솔베이트60 5.0 중량%Polysorbate 60 5.0 wt%
피이지 60 경화피마자유 2.0 중량%≪ tb > < tb > < tb >
솔비탄세스퀴올레이트 0.5 중량%0.5% by weight of sorbitan sesquioleate
바셀린 5.0 중량%Vaseline 5.0 wt%
유동파라핀 10.0 중량%Liquid paraffin 10.0 wt%
스쿠알란 5.0 중량%Squalane 5.0 wt%
쉐어버터 3.0 중량%SHARE BUTTER 3.0 wt%
카프릴릭/카프릭트리글리세라이드 5.0 중량%Caprylic / capric triglyceride 5.0 wt%
글리세린 10.0 중량%Glycerin 10.0 wt%
프로필렌글리콜 10.2 중량%Propylene glycol 10.2 wt%
트리에탄올아민 0.2 중량%0.2% by weight triethanolamine
방부제 0.05 중량%Preservative 0.05 wt%
색소 0.05 중량%0.05% by weight of pigment
향료 0.05 중량%0.05% by weight fragrance
정제수 to 100 중량%Purified water to 100%
Claims (20)
A composition for skin whitening comprising, as an active ingredient, a mixed extract obtained by mixing a green tea extract, a peel extract, a safflower extract and a white porcelain extract.
녹각 추출물은 녹각을 물 및 유기용매로 이루어진 군에서 선택된 하나 이상으로 추출하여 얻은 추출물이고,
토사자 추출물은 녹각을 물 및 유기용매로 이루어진 군에서 선택된 하나 이상으로 추출하여 얻은 추출물이고,
숙지황 추출물은 숙지황을 물 및 유기용매로 이루어진 군에서 선택된 하나 이상으로 추출하여 얻은 추출물이고,
백자인 추출물은 백자인을 물 및 유기용매로 이루어진 군에서 선택된 하나 이상으로 추출하여 얻은 추출물인 피부 미백용 조성물.
The method according to claim 1,
The extract of green tea extract is an extract obtained by extracting the green tea with at least one selected from the group consisting of water and organic solvent,
The extract of Tozan extract is an extract obtained by extracting a green bean with at least one selected from the group consisting of water and an organic solvent,
The Sukjwiphang extract is an extract obtained by extracting Sukjibulgi with at least one selected from the group consisting of water and an organic solvent,
The white cane extract is an extract obtained by extracting white porphyry with at least one selected from the group consisting of water and an organic solvent.
유기 용매는 탄소수 1 내지 5의 저급 알코올, 에틸아세테이트, 또는 아세톤을 포함하는 극성용매; 에테르, 클로로포름, 벤젠, 헥산 또는 디클로로헥산을 포함하는 비극성용매; 또는 이들의 혼합용매인 피부 미백용 조성물.
3. The method of claim 2,
The organic solvent may be a polar solvent containing a lower alcohol having 1 to 5 carbon atoms, ethyl acetate, or acetone; A nonpolar solvent comprising ether, chloroform, benzene, hexane or dichlorohexane; Or a mixed solvent thereof.
조성물은 피부 미백용 의약, 화장료 또는 식품 제조를 위한 것인, 피부 미백용 조성물.
The method according to claim 1,
Wherein the composition is for skin whitening medicines, cosmetics or foods.
A composition for enhancing skin elasticity or improving wrinkles, comprising as an active ingredient, a mixed extract of a green tea extract, a watery extract, and a white porcelain extract.
녹각 추출물은 녹각을 물 및 유기용매로 이루어진 군에서 선택된 하나 이상으로 추출하여 얻은 추출물이고
토사자 추출물은 녹각을 물 및 유기용매로 이루어진 군에서 선택된 하나 이상으로 추출하여 얻은 추출물이고,
숙지황 추출물은 숙지황을 물 및 유기용매로 이루어진 군에서 선택된 하나 이상으로 추출하여 얻은 추출물이고,
백자인 추출물은 백자인을 물 및 유기용매로 이루어진 군에서 선택된 하나 이상으로 추출하여 얻은 추출물인 피부 탄력 증진 또는 주름 개선용 조성물.
6. The method of claim 5,
The extract of the green peach is an extract obtained by extracting the green peel with at least one selected from the group consisting of water and an organic solvent
The extract of Tozan extract is an extract obtained by extracting a green bean with at least one selected from the group consisting of water and an organic solvent,
The Sukjwiphang extract is an extract obtained by extracting Sukjibulgi with at least one selected from the group consisting of water and an organic solvent,
The composition for enhancing skin elasticity or improving wrinkles is an extract obtained by extracting white porcelain with at least one selected from the group consisting of water and an organic solvent.
유기 용매는 탄소수 1 내지 5의 저급 알코올, 에틸아세테이트, 또는 아세톤을 포함하는 극성용매; 에테르, 클로로포름, 벤젠, 헥산 또는 디클로로헥산을 포함하는 비극성용매; 또는 이들의 혼합용매인 피부 탄력 증진 또는 주름 개선용 조성물.
The method according to claim 6,
The organic solvent may be a polar solvent containing a lower alcohol having 1 to 5 carbon atoms, ethyl acetate, or acetone; A nonpolar solvent comprising ether, chloroform, benzene, hexane or dichlorohexane; Or a mixed solvent thereof, for improving skin elasticity or improving wrinkles.
조성물은 피부 탄력 증진 또는 주름 개선용 의약, 화장료 또는 식품 제조를 위한 것인, 피부 탄력 증진 또는 주름 개선용 조성물.
6. The method of claim 5,
Wherein the composition is for the manufacture of a medicament for the enhancement of skin elasticity or wrinkle, a cosmetic preparation or a food product.
Wherein the composition comprises as an active ingredient, a mixed extract of a green tea extract, a green tea extract, a safflower extract, and a white porcelain extract.
녹각 추출물은 녹각을 물 및 유기용매로 이루어진 군에서 선택된 하나 이상으로 추출하여 얻은 추출물이고,
토사자 추출물은 녹각을 물 및 유기용매로 이루어진 군에서 선택된 하나 이상으로 추출하여 얻은 추출물이고,
숙지황 추출물은 숙지황을 물 및 유기용매로 이루어진 군에서 선택된 하나 이상으로 추출하여 얻은 추출물이고,
백자인 추출물은 백자인을 물 및 유기용매로 이루어진 군에서 선택된 하나 이상으로 추출하여 얻은 추출물인 항염증용 조성물.
10. The method of claim 9,
The extract of green tea extract is an extract obtained by extracting the green tea with at least one selected from the group consisting of water and organic solvent,
The extract of Tozan extract is an extract obtained by extracting a green bean with at least one selected from the group consisting of water and an organic solvent,
The Sukjwiphang extract is an extract obtained by extracting Sukjibulgi with at least one selected from the group consisting of water and an organic solvent,
Wherein the white porcelain extract is an extract obtained by extracting white porcelain with at least one selected from the group consisting of water and an organic solvent.
유기 용매는 탄소수 1 내지 5의 저급 알코올, 에틸아세테이트, 또는 아세톤을 포함하는 극성용매; 에테르, 클로로포름, 벤젠, 헥산 또는 디클로로헥산을 포함하는 비극성용매; 또는 이들의 혼합용매인 항염증용 조성물.
11. The method of claim 10,
The organic solvent may be a polar solvent containing a lower alcohol having 1 to 5 carbon atoms, ethyl acetate, or acetone; A nonpolar solvent comprising ether, chloroform, benzene, hexane or dichlorohexane; Or a mixed solvent thereof.
조성물은 항염용 의약, 화장료 또는 식품 제조를 위한 것인, 항염증용 조성물.
10. The method of claim 9,
Wherein the composition is for anti-inflammatory medicines, cosmetics or foods.
Which comprises a mixture of a green tea extract, a green tea extract, a safflower extract and a white porcelain extract as an active ingredient.
녹각 추출물은 녹각을 물 및 유기용매로 이루어진 군에서 선택된 하나 이상으로 추출하여 얻은 추출물이고,
토사자 추출물은 녹각을 물 및 유기용매로 이루어진 군에서 선택된 하나 이상으로 추출하여 얻은 추출물이고,
숙지황 추출물은 숙지황을 물 및 유기용매로 이루어진 군에서 선택된 하나 이상으로 추출하여 얻은 추출물이고,
백자인 추출물은 백자인을 물 및 유기용매로 이루어진 군에서 선택된 하나 이상으로 추출하여 얻은 추출물인 항산화용 조성물.
14. The method of claim 13,
The extract of green tea extract is an extract obtained by extracting the green tea with at least one selected from the group consisting of water and organic solvent,
The extract of Tozan extract is an extract obtained by extracting a green bean with at least one selected from the group consisting of water and an organic solvent,
The Sukjwiphang extract is an extract obtained by extracting Sukjibulgi with at least one selected from the group consisting of water and an organic solvent,
Wherein the extract is obtained by extracting white porphyry with at least one selected from the group consisting of water and an organic solvent.
유기 용매는 탄소수 1 내지 5의 저급 알코올, 에틸아세테이트, 또는 아세톤을 포함하는 극성용매; 에테르, 클로로포름, 벤젠, 헥산 또는 디클로로헥산을 포함하는 비극성용매; 또는 이들의 혼합용매인 항산화용 조성물.
15. The method of claim 14,
The organic solvent may be a polar solvent containing a lower alcohol having 1 to 5 carbon atoms, ethyl acetate, or acetone; A nonpolar solvent comprising ether, chloroform, benzene, hexane or dichlorohexane; Or a mixed solvent thereof.
조성물은 항산화용 의약, 화장료 또는 식품 제조를 위한 것인, 항산화용 조성물.
14. The method of claim 13,
Wherein the composition is for the production of antioxidant medicines, cosmetics or foods.
Which comprises as an active ingredient, a mixed extract of a green tea extract, a green tea extract, a safflower extract and a white porcelain extract.
녹각 추출물은 녹각을 물 및 유기용매로 이루어진 군에서 선택된 하나 이상으로 추출하여 얻은 추출물이고,
토사자 추출물은 녹각을 물 및 유기용매로 이루어진 군에서 선택된 하나 이상으로 추출하여 얻은 추출물이고,
숙지황 추출물은 숙지황을 물 및 유기용매로 이루어진 군에서 선택된 하나 이상으로 추출하여 얻은 추출물이고,
백자인 추출물은 백자인을 물 및 유기용매로 이루어진 군에서 선택된 하나 이상으로 추출하여 얻은 추출물인 항당화용 조성물.
18. The method of claim 17,
The extract of green tea extract is an extract obtained by extracting the green tea with at least one selected from the group consisting of water and organic solvent,
The extract of Tozan extract is an extract obtained by extracting a green bean with at least one selected from the group consisting of water and an organic solvent,
The Sukjwiphang extract is an extract obtained by extracting Sukjibulgi with at least one selected from the group consisting of water and an organic solvent,
Wherein the white sugar extract is an extract obtained by extracting white porcelain with at least one selected from the group consisting of water and an organic solvent.
유기 용매는 탄소수 1 내지 5의 저급 알코올, 에틸아세테이트, 또는 아세톤을 포함하는 극성용매; 에테르, 클로로포름, 벤젠, 헥산 또는 디클로로헥산을 포함하는 비극성용매; 또는 이들의 혼합용매인 항당화용 조성물.
19. The method of claim 18,
The organic solvent may be a polar solvent containing a lower alcohol having 1 to 5 carbon atoms, ethyl acetate, or acetone; A nonpolar solvent comprising ether, chloroform, benzene, hexane or dichlorohexane; Or a mixed solvent thereof.
조성물은 항당화용 의약, 화장료 또는 식품 제조를 위한 것인, 항당화용 조성물.18. The method of claim 17,
Wherein the composition is for the manufacture of a medicament for the anti-glycation, a cosmetic preparation or a food.
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CN109673900A (en) * | 2019-02-25 | 2019-04-26 | 黑龙江阳光工业大麻研究院 | Anti- saccharification sports drink of one kind and preparation method thereof |
CN112168475A (en) * | 2020-09-29 | 2021-01-05 | 汇云聚美(广州)科技有限公司 | Preparation method of steam eye patch for eye relief |
CN112168475B (en) * | 2020-09-29 | 2022-03-15 | 汇云聚美(广州)科技有限公司 | Preparation method of steam eye patch for eye relief |
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