KR102211942B1 - Cosmetic composition for whitening skin comprising extract of Sorbus aucuparia berry or fermented product of Sorbus aucuparia berry - Google Patents
Cosmetic composition for whitening skin comprising extract of Sorbus aucuparia berry or fermented product of Sorbus aucuparia berry Download PDFInfo
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Abstract
본 발명은 정공실 추출물을 유산균을 이용하여 발효한 정공실 발효물을 유효성분으로 함유하는 피부 미백용 화장료 조성물에 관한 것이다.
본 발명에 따르면, 정공실 추출물 또는 발효한 정공실 발효물을 함유하는 피부 미백용 화장료 조성물은 마가목 줄기 및 수피 추출물보다 더 효과적으로 티로시나아제(tyrosinase) 및 MITF((microphthalmia-associated transcription factor)의 유전자 발현을 억제하고, LXR(Liver X Receptor) 활성을 촉진시킴으로써 멜라닌 생합성을 억제하여, 결론적으로 피부 미백 효과를 나타낼 수 있다.The present invention relates to a cosmetic composition for skin whitening containing the fermented Jeonggongsil fermented product by using lactic acid bacteria as an active ingredient.
According to the present invention, the cosmetic composition for skin whitening containing the extract or fermented hole extract is more effective than the extract of rowan stem and bark, the gene of tyrosinase and MITF (microphthalmia-associated transcription factor) By suppressing the expression and promoting LXR (Liver X Receptor) activity, melanin biosynthesis is suppressed, and consequently, it can exhibit skin whitening effect.
Description
본 발명은 정공실 추출물 또는 정공실 발효물을 함유하는 피부 미백용 화장료 조성물에 관한 것으로, 더욱 구체적으로 정공실 추출물 또는 정공실 추출물을 유산균을 이용하여 발효한 정공실 발효물을 유효성분으로 함유하는 피부 미백용 화장료 조성물에 관한 것이다.The present invention relates to a cosmetic composition for skin whitening containing a Jeonggongsil extract or a Jeonggongsil fermented product, and more specifically, containing a Jeonggongsil fermented product obtained by fermenting the Jeonggongsil extract or the Jeonggongsil extract using lactic acid bacteria as an active ingredient. It relates to a cosmetic composition for skin whitening.
사람의 피부색을 결정하는 요인들에는 멜라닌, 각질층의 두께, 말초혈관과 혈색소, 카로틴 등이 있지만 이들 중 피부 색깔을 결정하는 가장 중요한 요인은 멜라닌이다. 멜라닌 색소를 생성하는 멜라닌생성세포의 수는 민족이나 피부색에 관계없이 일정하지만, 멜라닌화의 정도, 멜라닌 소체의 크기와 수 그리고 분포상태, 각질형성 세포 내에서의 멜라닌 소체의 분해능 등이 달라서 피부색의 차이가 나타난다. (이승헌 등, 피부 장벽학, 여문각, 2010) The factors that determine a person's skin color include melanin, the thickness of the stratum corneum, peripheral blood vessels, hemoglobin, and carotene, but the most important factor in determining the color of the skin is melanin. The number of melanogenic cells that produce melanin pigments is constant regardless of ethnicity or skin color, but the degree of melanization, the size and number and distribution of melanosomes, and the resolution of melanosomes in keratinocytes are different. A difference appears. (Lee Seung-heon et al., Skin Barrier Studies, Yeo Mun-gak, 2010)
멜라닌 생성시에 멜라닌 세포를 감싸는 다양한 환경에 반응하여 그 생합성량이 증가 또는 감소한다. 이러한 조절 구조의 연구는 자외선에 의한 피부의 멜라닌 생성 증가 기전 중심으로 진행되어 왔다. 자외선에 의해 각질형성세포에서 basic fibroblast growth factor (bFGF),alpha-melanocyte stimulating hormone (α-MSH),endothelin-1 (ET-1), prostaglandin E 2 (PGE 2 ) 등의 cytokine이 생성/분비 된다. 이러한 인자들에 반응하는 수용체는 멜라닌 세포의 세포막상에 존재하고, 수용체로부터 전달된 신호는 MITF (Microphthalmia-associated transcription factor)에 영향을 준다. MITF는 LXR (Liver X Receptor)의 영향도 받는데, 발현자체의 조절뿐만 아니라 세포외 신호 조절 인산화 효소(extracellular signal-regulated kinase, ERK) 경로의 활성화에도 영향을 받는다. 핵수용체의 일종인 Liver X receptor (LXR)는 리간드에 의해 활성화되는 핵 수용체로서, 지질 대사 및 콜레스테롤 항상성 유지에 중심 역할을 한다고 알려져 있다. 최근 연구에 의하면 LXR의 활성화가 멜라닌생성세포에서 멜라닌 생합성과정을 억제한다는 보고가 있었다(Lee CS et al., Liver X Receptor Activation Inhibits Melanogenesis through the Acceleration of ERK-Mediated MITF. Degradation J Invest Dermatol., 2013). 또한, LXR 활성화제가 멜라닌 생성을 저해하여 뛰어난 미백효과를 나타낸다는 것이 보고되어 있다(10-2012-0078675 : 간 X 수용체 활성제를 포함하는 피부 미백제).During the production of melanin, the amount of biosynthesis increases or decreases in response to various environments surrounding melanocytes. Research on this regulatory structure has been focused on the mechanism of increasing skin melanin production by UV rays. Cytokines such as basic fibroblast growth factor (bFGF), alpha-melanocyte stimulating hormone (α-MSH), endothelin-1 (ET-1), and prostaglandin E 2 (PGE 2) are produced/secreted in keratinocytes by UV light. . Receptors that respond to these factors exist on the cell membrane of melanocytes, and signals transmitted from the receptors affect the microphthalmia-associated transcription factor (MITF). MITF is also affected by LXR (Liver X Receptor), which not only regulates its expression itself, but also activates the extracellular signal-regulated kinase (ERK) pathway. Liver X receptor (LXR), a kind of nuclear receptor, is a nuclear receptor that is activated by a ligand, and is known to play a central role in lipid metabolism and maintenance of cholesterol homeostasis. According to a recent study, it has been reported that activation of LXR inhibits the process of melanogenesis in melanogenic cells (Lee CS et al., Liver X Receptor Activation Inhibits Melanogenesis through the Acceleration of ERK-Mediated MITF. Degradation J Invest Dermatol., 2013 ). In addition, it has been reported that an LXR activator inhibits melanin production and exhibits an excellent whitening effect (10-2012-0078675: skin whitening agent containing a liver X receptor activator).
전사인자로 작용하는 MITF는 티로시나아제(tyrosinase) 뿐만 아니라 티로시나아제 관련 단백질 1 (tyrosinase related protein 1, TRP-1), 티로시나아제 관련 단백질 2 (tyrosinase related protein 2, TRP-2), DCT (dopachrome tautomerase)와 같은 멜라닌 합성에 관여하는 일련의 효소들의 전사를 유도한다(Saha B et al., Activation of the Mitf promoter by lipid-stimulated activation of p38-stress signalling to CREB. Pigment Cell Res., 2006). MITFs that act as transcription factors are not only tyrosinase, but also tyrosinase related protein 1 (TRP-1), tyrosinase related protein 2 (TRP-2), DCT (Saha B et al., Activation of the Mitf promoter by lipid-stimulated activation of p38-stress signaling to CREB. Pigment Cell Res., 2006), such as (dopachrome tautomerase). ).
생성된 멜라닌 합성효소들은 멜라닌생성세포에서 타이로신(L-tyrosine)을 가수분해시켜 도파(L-dopa), 도파퀴논(L-dopaquinone)으로의 반응을 유발한다. 티로시나아제(tyrosinase)는 이 2개 반응의 촉매가 되는 효소로 미백제로 알려진 하이드로퀴논, 알부틴 같은 물질이 반응을 방해하여 멜라닌 생성을 억제하는 것으로 미백 효과를 가져온다. 생성된 도파퀴논(L-dopaquinone)은 DCT (dopachrome tautomerase), TRP-1 (tyrosinase related protein 1), TRP-2 (tyrosinase related protein 2)의 일련의 효소 작용을 거쳐 흑색의 유멜라닌(Eumelanin)으로 합성된다. The produced melanin synthases hydrolyze L-tyrosine in melanogenic cells, causing reactions to L-dopa and L-dopaquinone. Tyrosinase (tyrosinase) is an enzyme that catalyzes these two reactions. Substances such as hydroquinone and arbutin, known as whitening agents, interfere with the reaction to inhibit melanin production, thereby bringing whitening effect. The produced dopaquinone is converted to black eumelanin through a series of enzyme actions of DCT (dopachrome tautomerase), TRP-1 (tyrosinase related protein 1), and TRP-2 (tyrosinase related protein 2). Is synthesized.
한편, 정공실(Sorbus Aucuparia Berry)은 장미과에 속하는 낙엽활엽소교목인 마가목의 열매로써, 가을에 붉게 익은 것을 채취하여 햇볕에 말려 사용한다. 시큼한 맛의 정공실은 기침과 가래에 특효약이다. 종래의 연구에서 마가목의 줄기 및 수피의 미백 효능에 대하여 밝혔으나(10-2005-0110149 : 마가목 추출물을 함유하는 화장료 조성물), 마가목의 열매인 정공실에 대한 미백 효과는 어디에도 기재되어 있지 않다. Meanwhile, Sorbus Aucuparia Berry ) is the fruit of the rowan tree, a deciduous broad-leaved small arboreous tree belonging to the Rosaceae family. It is used after harvesting red ripe ones in autumn and drying them in the sun. The sour taste of Jeonggongsil is a special medicine for coughing and phlegm. In a previous study, the whitening effect of the stem and bark of rowan has been revealed (10-2005-0110149: cosmetic composition containing rowan extract), but the whitening effect on Jeonggongsil, the fruit of rowan tree, is not described anywhere.
이에, 본 발명자들은 상기 종래기술들의 문제점들을 극복하기 위하여 예의 연구노력한 결과, 마가목의 열매인 정공실만을 이용하여 발효공법을 거쳐 기존의 마가목 추출물(수피 및 줄기)보다 뛰어난 정공실 발효물을 확보할 수 있었으며, 이를 이용한 피부 미백용 화장료 조성물의 경우, 티로시나아제(tyrosinase) 발현 억제 및 멜라닌 생합성을 억제함으로써 피부 미백효과를 나타낼 수 있음을 확인하고, 본 발명을 완성하게 되었다.Accordingly, the present inventors have made intensive research efforts to overcome the problems of the prior art, and as a result of the fermentation method using only the hole of rowan tree, it is possible to secure a fermented product superior to the existing rowan extract (bark and stem). In the case of a cosmetic composition for skin whitening using the same, it was confirmed that the skin whitening effect could be exhibited by suppressing the expression of tyrosinase and melanin biosynthesis, and this invention was completed.
따라서, 본 발명의 주된 목적은 티로시나아제(tyrosinase) 발현 억제 및 멜라닌 생합성을 억제함으로써 피부 미백효과를 갖는 정공실 추출물 또는 정공실 발효물을 함유하는 피부 미백용 화장료 조성물을 제공하는 데 있다.Therefore, the main object of the present invention is to provide a cosmetic composition for skin whitening containing a hole-whitening effect or a hole-white fermented product having a skin whitening effect by inhibiting tyrosinase expression and melanin biosynthesis.
본 발명의 한 양태에 따르면, 본 발명은 정공실 추출물 또는 정공실 발효물을 유효성분으로 함유하는 피부 미백용 화장료 조성물을 제공한다.According to one aspect of the present invention, the present invention provides a cosmetic composition for skin whitening comprising an extract or a fermented hole in the hole as an active ingredient.
정공실(Sorbus Aucuparia Berry)은 장미과에 속하는 낙엽활엽소교목인 마가목의 열매로써, 종래에 화장료로 이용되던 마가목의 줄기와 수피의 추출물, 즉 마가목 추출물과는 구별된다. 본 발명자들은 마가목의 줄기와 수피에 비해 열매인 정공실의 멜라닌 생성억제능이 높음을 확인하였고, 더욱이 이를 발효과정을 거친 정공실 발효물의 경우, 티로시나아제 및 멜라닌 생합성 억제가 마가목 수피 및 줄기 추출물, 및 정공실 추출물보다 높아 더 우수한 피부 미백 효과를 나타낼 수 있음을 확인하고, 본 발명을 완성하게 되었다. Precision Office ( Sorbus Aucuparia Berry ) is the fruit of the rowan tree, a deciduous broad-leaved small arboreous tree belonging to the Rosaceae family, and is distinguished from the extract of the stem and bark of rowan , which was used as a cosmetic in the past, that is, rowan extract. The present inventors confirmed that the melanin production inhibitory ability of the fruit jeonggongsil is higher than that of the stem and bark of rowan, and furthermore, in the case of the fermented jeonggongsil fermented product, inhibiting tyrosinase and melanin biosynthesis, And it was confirmed that it can exhibit a more excellent skin whitening effect, higher than the extract of Jeonggongsil, and completed the present invention.
본 발명에 있어서, 상기 정공실 추출물은 종래에 식물, 생약, 열매 추출을 위해 사용되어진 어떠한 용매로도 추출될 수 있으며, 바람직하게는 물, 탄소수 1 내지 4의 저급알코올, 글리세린, 에틸아세테이트, 부틸렌글리콜, 프로필렌글리콜, 디클로로메탄 또는 핵산으로 추출하는 것을 특징으로 한다.In the present invention, the hole chamber extract can be extracted with any solvent conventionally used for extracting plants, herbal medicines, and fruits, preferably water, lower alcohol having 1 to 4 carbon atoms, glycerin, ethyl acetate, butyl It is characterized by extraction with ene glycol, propylene glycol, dichloromethane or nucleic acid.
상기 추출용매를 이용한 추출방법으로는 통상적인 식물의 추출방법 예를 들면, 에탄올 추출, 환류냉각 추출, 초음파 추출, 초임계 추출 등의 방법을 사용할 수 있으며, 본 발명의 실시예에서는 열수 추출물을 설명하고 있으나, 이에 한정되지는 않을 것이다. As the extraction method using the extraction solvent, a conventional plant extraction method, for example, ethanol extraction, reflux extraction, ultrasonic extraction, supercritical extraction, etc. can be used.In the embodiment of the present invention, a hot water extract is described. However, it will not be limited thereto.
본 발명에 있어서, 상기 정공실 발효물은 정공실 추출물을 유산균으로 발효한 것을 특징으로 한다.In the present invention, the fermented product is characterized in that it is fermented with lactic acid bacteria extract of the hole hole.
본 명세서에서 "발효"라 함은 미생물이 자신이 가지고 있는 효소를 이용해 유기물을 분해 시키는 과정을 의미한다. 발효는 본 발명 기술 분야에 알려져 있는 통상의 방법을 선정하거나 그로부터 응용하여 이루어질 수 있으며, 예를 들어 30℃ 내지 45℃, 구체적으로 35℃ 내지 38℃에서 30시간 내지 140시간, 구체적으로 110시간 내지 130시간 동안 이루어질 수 있다. 이와 같은 발효를 통해 정공실 발효물의 피부 미백 효과가 더 높아 질 수 있다.In the present specification, "fermentation" refers to a process in which microorganisms decompose organic matter using an enzyme possessed by them. Fermentation may be performed by selecting or applying a conventional method known in the art of the present invention, for example, 30 to 45° C., specifically 35 to 38° C. for 30 to 140 hours, specifically 110 to Can be done for 130 hours. Through such fermentation, the skin whitening effect of the fermented product of the hole chamber may be higher.
본 발명에 있어서, 정공실 발효물은 정공실 추출물을 미생물로 발효시킨 발효물을 의미한다. 상기 미생물은 유산균(lactic acid bacteria)일 수 있으며, 바람직하게는 락토바실러스(Lactobacillus sp.), 류코노스톡(Leuconostoc sp.) 또는 비피도박테리아(Bifidobacteria sp.)인 것을 특징으로 하나, 이에 제한되는 것은 아니다. In the present invention, the fermented product of the hole chamber means a fermented product obtained by fermenting the extract of the hole chamber with microorganisms. The microorganism may be lactic acid bacteria, preferably Lactobacillus sp., Leuconostoc sp., or Bifidobacteria sp., but is limited thereto. It is not.
본 발명에 있어서, 상기 정공실 추출물 또는 정공실 발효물은 화장료 조성물 총 중량 대비 0.01 내지 90중량% 함유하는 것을 특징으로 한다. 상기의 범위를 벗어나는 경우에는 제형화가 용이하지 않아 바람직하지 않다. In the present invention, the hole chamber extract or hole chamber fermentation product is characterized in that it contains 0.01 to 90% by weight based on the total weight of the cosmetic composition. If it is outside the above range, it is not preferable to formulate it.
본 발명에 있어서, 상기 조성물은 티로시나아제(tyrosinase) 발현 억제 및 멜라닌 생합성을 억제함으로써 피부 미백효과를 나타내는 것을 특징으로 한다. In the present invention, the composition is characterized in that it exhibits a skin whitening effect by inhibiting tyrosinase expression and melanin biosynthesis.
본 발명의 실험예에 따르면, 정공실 추출물은 마가목 줄기와 수피 추출물보다 멜라닌 생성 억제율이 높고, 멜라닌 생성 세포에서 멜라닌의 양을 농도 의존적으로 감소시켰으며, 티로시나아제 발현 또한 농도 의존적으로 억제하는 것을 확인할 수 있다. 이러한 결과로 볼 때, 본 발명의 정공실 추출물을 이용한 발효물도 티로시나아제 및 멜라닌 합성을 억제함으로써 피부 미백 효과를 나타낼 수 있음을 의미한다(실험예3-5 참조). According to the experimental example of the present invention, the extract of Jeonggongsil has a higher inhibition rate of melanogenesis than the extract of rowan stem and bark, concentration-dependently reduced the amount of melanin in melanin-producing cells, and tyrosinase expression is also inhibited in a concentration-dependent manner. I can confirm. From these results, it means that the fermented product using the extract of Jeonggongsil of the present invention can exhibit a skin whitening effect by inhibiting the synthesis of tyrosinase and melanin (see Experimental Example 3-5).
또한, 본 발명의 실험예에 따르면, 정공실 발효물은 발효를 거치지 않은 정공실 추출물보다 더 많이 멜라닌 생성 세포에서 멜라닌의 양을 농도 의존적으로 감소시켰으며, 티로시나아제 발현 또한 농도 의존적으로 억제하는 것을 확인할 수 있다. 이러한 결과로 볼 때, 본 발명의 정공실 발효물이 발효를 하지 않은 정공실 추출물보다 효과적으로 티로시나아제 및 멜라닌 합성을 억제함으로써 더 우수한 피부 미백 효과를 나타낼 수 있음을 의미한다(실험예3-5 참조). In addition, according to the experimental example of the present invention, the fermented hole chamber reduced the amount of melanin in melanin-producing cells in a concentration-dependent manner, and inhibited the expression of tyrosinase in a concentration-dependent manner more than that of the hole chamber extract without fermentation. Can be confirmed. From these results, it means that the fermented product of the present invention can exhibit a better skin whitening effect by inhibiting the synthesis of tyrosinase and melanin more effectively than the non-fermented Jeonggongsil extract (Experimental Example 3-5 Reference).
더욱이, 본 발명의 정공실 발효물은 티로시나아제 및 DCT(dopachrome tautomerase)의 단백질 발현을 저해함으로써 피부 멜라닌 합성을 억제하는데, 구체적으로는, 정공실 발효물과 정공실 추출물은 멜라닌 생성에 관여하는 유전자인 티로시나아제, DCT의 발현을 모두 감소시키는 것을 확인할 수 있었으며, 정공실 추출물보다 정공실 발효물이 더 효과적으로 티로시나아제 및 DCT 발현을 감소시키는 것을 확인하였다. 이러한 결과로 볼 때, 본 발명의 정공실 발효물은 상기 유전자들의 억제를 통하여 티로시나아제를 억제할 수 있으며 결과적으로 피부 미백 효과를 나타낼 수 있음을 의미한다(실험예 6 참조). Moreover, the hole-sil fermented product of the present invention inhibits skin melanin synthesis by inhibiting the protein expression of tyrosinase and dopachrome tautomerase (DCT). Specifically, the hole-sil fermented product and the hole-silent extract are involved in melanin production. It was confirmed that all of the expressions of genes, tyrosinase and DCT, were reduced, and it was confirmed that the fermented tyrosinase and DCT decreased the expression of the tyrosinase and DCT more effectively than the extract of the jeonggongsil. From these results, it means that the fermented product of the hole chamber of the present invention can suppress tyrosinase through inhibition of the genes, and consequently exhibit a skin whitening effect (see Experimental Example 6).
본 발명에 있어서, 상기 조성물은 스킨로션, 스킨소프너, 스킨토너, 아스트린젠트, 로션, 밀크로션, 모이스쳐 로션, 영양로션, 맛사지크림, 영양크림, 모이스처크림, 핸드크림, 파운데이션, 에센스, 영양에센스, 팩, 비누, 클렌징폼, 클렌징로션, 클렌징크림, 바디로션 및 바디클렌저로 구성된 그룹에서 선택된 하나 이상의 제형인 것을 특징으로 한다. 예를 들면 유연화장수, 수렴화장수, 영양화장수, 영양크림, 마사지크림, 에센스, 아이크림, 아이에센스, 클렌징크림, 클렌징폼, 클렌징워터, 팩, 파우더, 보디로션, 보디크림, 보디오일, 보디에센스 등의 화장료 제형을 가질 수 있으며 또한 로션, 연고, 겔, 크림, 패취 또는 분무제와 같은 경피투과형 제형을 갖는 화장료 조성물 일 수 있다. 본 발명의 화장품 조성물은 담체 이외에 다른 보조제를 포함할 수 있는데, 예를 들면 보존제, 항산화제, 안정화제, 용해화제, 비타민, 안료 및 향료 등을 포함할 수 있다.In the present invention, the composition is a skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, foundation, essence, nutrition essence, pack , Soap, cleansing foam, cleansing lotion, cleansing cream, body lotion, and body cleanser. For example, softening lotion, astringent lotion, nourishing lotion, nourishing cream, massage cream, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, pack, powder, body lotion, body cream, body oil, body essence It may have a cosmetic formulation such as, and may also be a cosmetic composition having a transdermal formulation such as a lotion, ointment, gel, cream, patch or spray. The cosmetic composition of the present invention may contain other adjuvants in addition to the carrier, for example, preservatives, antioxidants, stabilizers, solubilizers, vitamins, pigments and fragrances.
이상 설명한 바와 같이, 정공실 추출물 또는 정공실 추출물을 유산균을 이용하여 발효한 정공실 발효물은 티로시나아제, DCT의 발현을 억제하고 멜라닌 생성세포에서 멜라닌의 생성을 억해함으로써 피부 미백 효과를 나타낸다. As described above, the hallucinogenic fermented product obtained by fermenting the jeonggongsil extract or the jeonggongsil extract using lactic acid bacteria suppresses the expression of tyrosinase and DCT and suppresses the production of melanin in melanogenic cells, thereby exhibiting a skin whitening effect.
도 1 은 마가목 줄기 추출물과 정공실 추출물에 의한 멜라닌 생석 억제 효과를 사람 색소세포(Human epidermal melanocyte)에서 관찰한 결과이다.
도 2 는 정공실 발효물이 정공실 추출물보다 높은 멜라닌 생성 억제효과를 나타냄을 관찰한 결과이다.
도 3 은 정공실 발효물이 정공실 추출물보다 티로시나아제의 발현 억제율이 높음을 관찰한 결과이다.
도 4 은 정공실 발효물이 정공실 추출물보다 DCT의 발현 억제율이 높음을 관찰한 결과이다. 1 is a result of observation in human epidermal melanocytes of melanin formation inhibition effect by rowan stem extract and Jeonggongsil extract.
Figure 2 is a result of observing that the fermented product of the hole chamber exhibits a higher inhibitory effect on melanogenesis than the extract of the hole hole chamber.
3 is a result of observing that the fermented product of Jeonggongsil has a higher inhibition rate of expression of tyrosinase than the extract of Jeonggongsil.
4 is a result of observing that the fermented product of Jeonggongsil has a higher inhibition rate of DCT expression than that of the extract of Jeonggongsil.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하기로 한다. 이들 실시예는 단지 본 발명을 예시하기 위한 것이므로, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지는 않는다.Hereinafter, the present invention will be described in more detail through examples. Since these examples are for illustrative purposes only, the scope of the present invention is not to be construed as being limited by these examples.
비교예Comparative example 1: One: 정공실Precision room 추출물의 제조 Preparation of extract
정공실은 한의원에 약제를 전문적으로 납품하는 동양허브라는 전문업체에서 구입하여 본 발명의 시료로 사용하였다. Jeonggongsil was purchased from a specialized company called Dongyang Herb, which professionally supplies drugs to oriental medicine clinics, and used as a sample of the present invention.
정공실 추출물을 제조하기 위해 100 g의 정공실을 흐르는 물에 세척하여 자연 건조시킨 뒤 파쇄기를 사용하여 열매를 일부 파쇄하고, 착즙기를 사용하여 압착 추출을 실시하였다. 추출된 정공실 추출물은 멸균을 위해 0.2 um 주사기 필터를 사용하여 멸균 처리하였다. In order to prepare the hole chamber extract, 100 g of the hole chamber was washed with running water and dried naturally. Then, a part of the fruit was crushed using a crusher, and compression extraction was performed using a juicer. The extracted hole chamber extract was sterilized using a 0.2 um syringe filter for sterilization.
비교예 2: 마가목 줄기 및 수피 추출물의 제조Comparative Example 2: Preparation of rowan stem and bark extract
마가목 줄기 및 수피는 한의원에 약제를 전문적으로 납품하는 동양허브라는 전문업체에서 구입하여 본 발명의 시료로 사용하였다. Rowan stems and bark were purchased from a specialized company called Dongyang Herb, which professionally supplies drugs to oriental medicine clinics, and used as samples of the present invention.
마가목 추출물을 제조하기 위해 100 g의 마가목 줄기 및 수피를 흐르는 물에 세척하여 자연 건조시킨 뒤 파쇄기를 사용하여 일부 파쇄하고, 착즙기를 사용하여 압착 추출을 실시하였다. 추출된 마가목 추출물은 멸균을 위해 0.2 um 주사기 필터를 사용하여 멸균 처리하였다. In order to prepare rowan extract, 100 g of rowan stem and bark were washed with running water, dried naturally, and partially crushed using a crusher, followed by compression extraction using a juicer. The extracted rowan extract was sterilized using a 0.2 um syringe filter for sterilization.
실시예 1: 정공실 발효물(F-SAF)의 제조Example 1: Preparation of fermented hole chamber (F-SAF)
정공실은 한의원에 약제를 전문적으로 납품하는 동양허브라는 전문업체에서 구입하여 본 발명의 시료로 사용하였다. Jeonggongsil was purchased from a specialized company called Dongyang Herb, which professionally supplies drugs to oriental medicine clinics, and used as a sample of the present invention.
정공실 발효물은 비교예 1(정공실 추출물)을 넣은 액체 배지에 락토바실러스 (Lactobacillus sp. CXHB-4)를 107-109 cell/mL로 접종하고, 5 일(120hr)간 배양을 실시하고, pH 6.5~7.5, 온도 35~38℃, 통기량 1VVM의 조건을 유지하면서 배양하였다. 상기 발효액을 10000 Xg에서 30분간 원심분리하여 고분자 침전물과 균체가 제거된 최종 정공실 발효물을 얻었다.The hole chamber fermentation product was inoculated with Lactobacillus sp. CXHB-4 at 10 7 -10 9 cells/mL in a liquid medium containing Comparative Example 1 (Jeongsil extract), and cultured for 5 days (120 hr). And, while maintaining the conditions of pH 6.5 ~ 7.5, temperature 35 ~ 38 ℃, aeration amount of 1VVM was cultured. The fermentation broth was centrifuged at 10000 Xg for 30 minutes to obtain a final hole chamber fermentation product from which polymer precipitates and cells were removed.
실험예 1: 정공실 추출물에 의한 인체멜라닌 세포의 멜라닌 생성 억제효과 측정Experimental Example 1: Measurement of the inhibitory effect of melanin production of human melanocytes by extracts from Hallucinations
인체 멜라닌세포(melanocyte)를 이용하여 정공실 추출물(비교예 1)과 마가목 줄기 및 수피 추출물(비교예 2)에 의한 멜라닌 생성 억제효과를 측정하고, 이를 멜라닌 생성을 억제하는 것으로 알려진 H89(10uM)에 의한 멜라닌 생성 억제효과와 비교하였다.H89 (10uM) known to inhibit melanin production by measuring the melanin production inhibitory effect of Jeonggongsil extract (Comparative Example 1) and rowan stem and bark extract (Comparative Example 2) using human melanocytes. It was compared with the inhibitory effect of melanin production.
본 실험은 Cascade Biologics사로부터 구입한 Human epidermal melanocyte (신생아유래) 세포를 역시 같은 회사에서 구입한 Human Melanocyte Growth supplement를 포함한 Medium 254배지에서 배양한다. Human epidermal melanocytes를 6-well plate에 well당 1× 105 개로 접종한 후 5 % CO2, 37℃ 하에서 세포가 well 바닥에 약 80 % 이상 부착될 때까지 배양하였다. 배양 후 배지를 제거하고 시료를 적당 농도로 희석하여 배지에 처리한 후 5% CO2, 37℃ 하에서 이틀에 한 번 배지를 갈아주면서 5일간 배양하였다. 추출물의 농도범위는 세포독성이 없는 10㎍/㎖, 50㎍/㎖로 결정하였다. 처리가 끝난 후 배지를 제거하고 PBS(phosphated buffer saline)로 세척한 후, 트립신으로 처리하여 세포를 회수하였다. 회수된 세포는 hematocytometer를 이용하여 세포수를 측정한 후 각 처리 그룹별 세포수를 동수로 맞추어 5,000 ~ 10,000 rpm으로 10분간 원심 분리한 다음 상등액을 제거하여 세포를 얻었다. 이 세포를 60℃에서 건조한 후, 10% DMSO가 함유된 1M 수산화나트륨액 100㎕를 넣어 60℃ 항온조에서 세포내 멜라닌을 얻었다. 이 액으로 microplate reader로 490㎚에서 흡광도를 측정하여 세포 일정 수당 멜라닌 양을 구하였다. 그 결과를 하기 도 1에 나타내었다. In this experiment, Human epidermal melanocyte (neonatal-derived) cells purchased from Cascade Biologics are cultured in Medium 254 medium containing Human Melanocyte Growth supplement purchased from the same company. Human epidermal melanocytes were inoculated into a 6-well plate at 1×10 5 per well, and then cultured under 5% CO 2 and 37°C until the cells adhered to the bottom of the well by about 80% or more. After cultivation, the medium was removed, the sample was diluted to an appropriate concentration, treated with the medium, and cultured for 5 days while changing the medium once every two days under 5% CO 2 and 37°C. The concentration range of the extract was determined as 10 μg/ml and 50 μg/ml without cytotoxicity. After the treatment was completed, the medium was removed, washed with phosphated buffer saline (PBS), and then treated with trypsin to recover the cells. For the recovered cells, the number of cells was measured using a hematocytometer, the number of cells for each treatment group was adjusted to the same number, centrifuged at 5,000 to 10,000 rpm for 10 minutes, and the supernatant was removed to obtain cells. After drying the cells at 60°C, 100µl of 1M sodium hydroxide solution containing 10% DMSO was added to obtain intracellular melanin in a 60°C incubator. With this solution, absorbance was measured at 490 nm with a microplate reader to determine the amount of melanin per cell. The results are shown in FIG. 1 below.
도 1에 나타난 바와 같이, 인체 멜라닌 세포에서 비교예 1인 정공실 추출물 처리에 의한 멜라닌 양이 농도 의존적으로 감소됨을 확인하였다. 반면에 마가목 줄기 및 수피 추출물인 비교예 2의 경우, 멜라닌의 생성억제 효과가 있지만, 비교예 1인 정공실 추출물 비해 억제율이 낮음을 확인할수있었다. 그러므로, 정공실 추출물이 인체 세포에도 미백 효능을 나타내고 있음을 확인하였다. As shown in FIG. 1, it was confirmed that the amount of melanin was decreased in a concentration-dependent manner by the treatment of the extract of the hole in Comparative Example 1 in human melanocytes. On the other hand, in the case of Comparative Example 2, which is a rowan stem and bark extract, it was confirmed that there was an effect of inhibiting the production of melanin, but the inhibition rate was lower than that of the extract of Jeonggongsil of Comparative Example 1. Therefore, it was confirmed that the extract of Jeonggongsil also exhibited whitening efficacy in human cells.
실험예 2: 정공실 추출물의 Liver X Receptor Response Element(LXRE) 프로모터 활성 촉진 효과Experimental Example 2: Effect of Promoting Liver X Receptor Response Element (LXRE) Promoter Activity of Jeonggongsil Extract
멜라닌 양을 감소시키는 정공실 추출물의 억제 메카니즘을 규명하기 위한 실험을 수행하였다. Liver X receptor(LXR)가 활성화 되면, 멜라닌 생성이 억제된다고 보고되어 있어 LXR 단백질에 의해 활성화된 LXRE 프로모터를 이용하여, 정공실 추출물이 LXRE 프로모터 리포터(LXRE promoter luciferase reporter)를 활성화 시킬 수 있는지의 여부를 조사하였다. LXRE reporter DNA를 murine melanoma(B-16 F10) 세포에 superfect를 이용하여 삽입함으로써, 형질전환을 유도하였고, 형질전환된지 24시간 후에 추출물을 처리하였다. 16시간이 경과된 후에 세포를 수집하고, Luminometer (Berthold, Germany)를 이용하여 450㎚에서 luminescence를 측정하였다. (Planta medica 2005; 71:338-343) 양성대조군으로 LXR의 활성화제로 알려진 TO901317을 사용하였다. 그 결과를 하기 표 1에 나타내었다.An experiment was conducted to determine the mechanism of inhibition of the extract of Jeonggongsil that reduces the amount of melanin. It is reported that when Liver X receptor (LXR) is activated, melanin production is reported to be inhibited. Whether or not the hole chamber extract can activate the LXRE promoter luciferase reporter using the LXRE promoter activated by the LXR protein. Was investigated. Transformation was induced by inserting LXRE reporter DNA into murine melanoma (B-16 F10) cells using a superfect, and the extract was treated 24 hours after transformation. After 16 hours elapsed, cells were collected, and luminescence was measured at 450 nm using a Luminometer (Berthold, Germany). (Planta medica 2005; 71:338-343) TO901317, known as an activator of LXR, was used as a positive control. The results are shown in Table 1 below.
표 1에서 나타나는 바와 같이, 마가목이라도 부위에 따라 그 효능효과가 다름을 관찰할 수 있었는데, 마가목의 줄기 및 수피 추출물인 비교예 2의 경우 LXR 단백질의 활성화 효과가 없는 반면, 마가목의 열매인 정공실 추출물, 비교예 1의 경우 LXR 단백질을 농도 의존적으로 활성화시킴을 관찰하였다. 그러므로, 본 실험을 통해 정공실이 뛰어난 미백 효능을 가지고 있음을 증명하였다.As shown in Table 1, it could be observed that even rowan trees had different efficacy effects depending on the part.In the case of Comparative Example 2, which is an extract of rowan stem and bark, there was no activating effect of LXR protein, whereas Jeonggongsil, a fruit of rowan tree In the case of the extract, Comparative Example 1, it was observed that the LXR protein was activated in a concentration-dependent manner. Therefore, through this experiment, it was proved that the hole chamber has excellent whitening effect.
실험예 3: 정공실 발효물에 의한 색소 세포의 멜라닌 생성 억제효과 측정Experimental Example 3: Measurement of the melanin production inhibitory effect of pigment cells by fermentation in the hole chamber
쥐의 색소세포(B16 melanoma cells)를 이용하여 비교예 1 및 실시예 1에 의한 멜라닌 생성 억제효과를 측정하였으며, 이를 멜라닌 생성을 억제하는 것으로 알려진 알부틴(100 ppm)에 의한 멜라닌 생성 억제효과와 비교하였다.The melanin production inhibitory effect according to Comparative Example 1 and Example 1 was measured using mouse pigment cells (B16 melanoma cells), and this was compared with the melanin production inhibitory effect by arbutin (100 ppm), which is known to inhibit melanogenesis. I did.
본 실험은 murine melanoma(B16 F10) 세포를 10% FBS를 포함하는 DMEM에 현탁시켜 well당 1 × 105 cells 으로 6-well plate에 접종하여 well 바닥에 부착될때까지 배양한다. 멜라닌 생성 유도를 위하여 0.1uMα-MSH를 처리하고, 이후 시료를 농도별로 첨가한 후 3일간 배양하였다. 물질 처리의 농도범위는 0.1%, 1% 로 설정하였다. 멜라닌 생성능 실험의 양성대조군으로 알부틴 100 ppm 을 처리하였다. 배양한 세포는 phosphate buffered saline(PBS)으로 씻고 트립신(trypsin)으로 회수하였다. 회수한 세포를 혈구계(hematocytometer)로 계수하여 각 처리 그룹별로 1× 106 cells/mL 으로 동일한 세포수가 되도록 모아 1000rpm, 10분간 원심분리하여 세포만을 얻었다. 회수된 세포를 60℃에서 1시간 건조시켜 10% DMSO가 포함된 1M NaOH용액 100 ㎕을 넣어 세포내의 멜라닌을 얻었다. 이 멜라닌 액을 Microplate reader로 490nm에서 흡광도를 측정하여 멜라닌 생성 저해율을 평가하였다. 그 결과를 하기 도 2에 나타내었다. In this experiment, murine melanoma (B16 F10) cells were suspended in DMEM containing 10% FBS, inoculated into a 6-well plate at 1 × 10 5 cells per well, and cultured until adhered to the bottom of the well. In order to induce melanin production, 0.1uMα-MSH was treated, and then samples were added for each concentration and cultured for 3 days. The concentration range of the material treatment was set to 0.1% and 1%. 100 ppm of arbutin was treated as a positive control for the melanin production ability experiment. The cultured cells were washed with phosphate buffered saline (PBS) and recovered with trypsin. The recovered cells were counted with a hematocytometer, collected so as to have the same number of cells at 1×10 6 cells/mL for each treatment group, and centrifuged at 1000 rpm for 10 minutes to obtain only cells. The recovered cells were dried at 60° C. for 1 hour, and 100 μl of 1M NaOH solution containing 10% DMSO was added to obtain intracellular melanin. The melanin production inhibition rate was evaluated by measuring the absorbance at 490 nm with a Microplate reader. The results are shown in FIG. 2 below.
도 2에서 나타나는 바와 같이, 물질 처리에 의해 멜라닌 양이 농도 의존적으로 감소함을 관찰하였다. 특히, 실시예 1의 경우, 알부틴에 비해 우수한 미백효능을 보였다. 그러므로, 본 실험을 통해 정공실 발효물이 가장 뛰어난 미백 효능을 가지고 있음을 증명하였다.As shown in FIG. 2, it was observed that the amount of melanin decreased in a concentration-dependent manner by the material treatment. In particular, in the case of Example 1, compared to arbutin showed excellent whitening effect. Therefore, through this experiment, it was proved that the fermented product of Jeonggongsil has the most excellent whitening effect.
실험예 4: 정공실 발효물에 의한 세포내 티로시나아제 유전자 발현 측정Experimental Example 4: Measurement of intracellular tyrosinase gene expression by fermentation of the hole chamber
쥐의 색소세포(B16 melanoma cells)를 이용하여 비교예 1 및 실시예 1에 의한 멜라닌 생성에 중요한 효소인 티로시나아제의 발현 억제효과를 확인하였다. The effect of inhibiting the expression of tyrosinase, an enzyme important for melanin production according to Comparative Examples 1 and 1, was confirmed using mouse pigment cells (B16 melanoma cells).
본 실험은 murine melanoma(B-16 F1) 세포를 10%의 FBS (fetal bovine serum)가 함유된 DMEM 배지로 6-well plate에 well 당 5× 105 개로 접종한 후 5% CO2, 37℃ 하에서 세포가 well 바닥에 약 80 % 이상 부착될 때까지 배양하였다. 멜라닌 생성 유도를 위하여, 0.1 uM α-MSH를 처리하고, 이후 시료를 1% 첨가한 후 24시간 배양하였다. 양성대조군으로 알부틴 100 ppm 을 처리하였다. 배양한 세포는 phosphate buffered saline으로 씻고 1 ㎖ Trizol을 넣어 회수하였다. 0.2㎖ Chloroform(Trizol의 1/5 비율)을 넣어 voltexing 하여 4℃에서 14000rpm, 15 분간 원심분리로 상층액을 얻었다. 동 상층액에 0.5 ㎖ isopropanol 넣고 10분간 반응시킨 후, 14000 rpm, 15 분간 원심분리하여 침전물을 얻었다. 상층액을 제거하고 200 ㎕ 100% EtOH를 이용하여 2회 씻어준다. 이후 침전물을 상온에서 완전히 말려 DEPC water 20~30 ㎕ 에 녹였다. DEPC에 녹인 RNA를 정량하여 각 그룹 당 RNA 동량을 사용하여 cDNA를 합성하였다. 합성된 cDNA를 타겟으로 시아닌 염료인 SYBR GREEN master mix 를 사용하여 실시간 중합효소 연쇄반응 (PCR)을 실시하여 최종적으로 tyrosinase의 유전자 발현 정도를 평가하였다. 그 결과를 하기 도 3에 나타내었다. In this experiment, murine melanoma (B-16 F1) cells were inoculated in a 6-well plate with DMEM medium containing 10% FBS (fetal bovine serum) at 5×10 5 per well, 5% CO 2 , 37°C. The cells were cultured until about 80% or more adhered to the bottom of the well. In order to induce melanin production, 0.1 uM α-MSH was treated, and then 1% of the sample was added, followed by incubation for 24 hours. 100 ppm of arbutin was treated as a positive control. The cultured cells were washed with phosphate buffered saline, and 1 ml of Trizol was added to recover them. 0.2ml Chloroform (1/5 ratio of Trizol) was added and voltexed to obtain a supernatant by centrifugation at 14000rpm at 4℃ for 15 minutes. 0.5 ml of isopropanol was added to the supernatant and reacted for 10 minutes, followed by centrifugation at 14000 rpm for 15 minutes to obtain a precipitate. Remove the supernatant and wash twice with 200
도 3에서 나타나는 바와 같이, 물질 처리에 의해 티로시나아제의 발현이 감소함을 관찰하였다. 실시예 1의 경우, 비교예1에 비해 우수한 발현 억제 효능을 보였다. 그러므로, 본 실험을 통해, 정공실 발효물이 정공실 추출물보다 뛰어난 미백 효능을 가지고 있음을 증명하였다.As shown in Figure 3, it was observed that the expression of tyrosinase decreased by the treatment of the substance. In the case of Example 1, compared to Comparative Example 1, it showed superior expression inhibitory effect. Therefore, through this experiment, it was proved that the fermented product of Jeonggongsil has superior whitening effect than the extract of Jeonggongsil.
실험예 5: 정공실 발효물에 의한 세포내 DCT 유전자 발현 측정Experimental Example 5: Measurement of DCT gene expression in cells by fermentation in the hole chamber
쥐의 색소세포(B16 melanoma cells)를 이용하여 비교예 1 및 실시예 1에 의한 멜라닌 생성기작에서 검은 유멜라닌 합성을 조절하는 인자인 DCT (dopachrome tautomerase)의 발현 억제효과를 확인하였다. The effect of inhibiting the expression of DCT (dopachrome tautomerase), a factor that regulates black eumelanin synthesis, was confirmed in the melanin production mechanism according to Comparative Examples 1 and 1 using mouse pigment cells (B16 melanoma cells).
본 실험은 murine melanoma(B-16 F1) 세포를 10%의 FBS (fetal bovine serum)가 함유된 DMEM 배지로 6-well plate에 well 당 5×105 개로 접종한 후 5% CO2, 37℃ 하에서 세포가 well 바닥에 약 80 % 이상 부착될 때까지 배양하였다. 멜라닌 생성 유도를 위하여, 0.1 uM α-MSH를 처리하고, 이후 시료를 1% 첨가한 후 24시간 배양하였다. 양성대조군으로 알부틴 100 ppm 을 처리하였다. 배양한 세포는 phosphate buffered saline으로 씻고 1 ㎖ Trizol을 넣어 회수하였다. 0.2 ㎖ Chloroform(Trizol의 1/5 비율)을 넣어 voltexing 하여 4℃에서 14000 rpm, 15 분간 원심분리로 상층액을 얻었다. 동 상층액에 0.5 ㎖ isopropanol 넣고 10분간 반응시킨 후, 14000 rpm, 15 분간 원심분리하여 침전물을 얻었다. 상층액을 제거하고 200 ㎕ 100% EtOH를 이용하여 2회 씻어준다. 이후 침전물을 상온에서 완전히 말려 DEPC water 20~30 ㎕ 에 녹였다. DEPC에 녹인 RNA를 정량하여 각 그룹 당 RNA 동량을 사용하여 cDNA를 합성하였다. 합성된 cDNA를 타겟으로 시아닌 염료인 SYBR GREEN master mix 를 사용하여 실시간 중합효소 연쇄반응 (PCR)을 실시하여 최종적으로 MITF의 유전자 발현 정도를 평가하였다. 그 결과를 하기 도 4에 나타내었다. In this experiment, murine melanoma (B-16 F1) cells were inoculated in a 6-well plate with DMEM medium containing 10% FBS (fetal bovine serum) at 5×10 5 per well, 5% CO 2 , 37°C. The cells were cultured until about 80% or more adhered to the bottom of the well. In order to induce melanin production, 0.1 uM α-MSH was treated, and then 1% of the sample was added, followed by incubation for 24 hours. 100 ppm of arbutin was treated as a positive control. The cultured cells were washed with phosphate buffered saline, and 1 ml of Trizol was added to recover them. 0.2 ml Chloroform (1/5 ratio of Trizol) was added and voltexed to obtain a supernatant by centrifugation at 14000 rpm for 15 minutes at 4°C. 0.5 ml of isopropanol was added to the supernatant and reacted for 10 minutes, followed by centrifugation at 14000 rpm for 15 minutes to obtain a precipitate. Remove the supernatant and wash twice with 200
도 4에서 나타나는 바와 같이, 물질 처리에 의해 DCT 전사인자의 발현이 감소함을 관찰하였다. 특히, 실시예 1의 경우, 비교예 1에 비해 우수한 발현 억제 효능을 보였다. 그러므로, 본 실험을 통해, 정공실 발효물이 정공실 추출물보다 뛰어난 미백 효능을 가지고 있음을 증명하였다.As shown in Figure 4, it was observed that the expression of the DCT transcription factor decreased by the treatment of the substance. In particular, in the case of Example 1, compared to Comparative Example 1, it showed excellent expression inhibition effect. Therefore, through this experiment, it was proved that the fermented product of Jeonggongsil has superior whitening effect than the extract of Jeonggongsil.
Claims (7)
A cosmetic composition for skin whitening containing an extract of Jeonggongsil or fermented product of Jeonggongsil as an active ingredient.
The cosmetic composition of claim 1, wherein the hole chamber extract is extracted with water, a lower alcohol having 1 to 4 carbon atoms, glycerin, ethyl acetate, butylene glycol, propylene glycol, dichloromethane, or nucleic acid.
The cosmetic composition for skin whitening according to claim 1, wherein the fermented product is obtained by fermenting the extract of the hole hole with lactic acid bacteria.
The cosmetic composition for skin whitening according to claim 3, wherein the lactic acid bacteria are Lactobacillus sp., Leuconostoc sp., or Bifidobacteria sp.
The cosmetic composition for skin whitening according to claim 1, wherein the hole chamber extract or the hole chamber fermentation product contains 0.01 to 90% by weight based on the total weight of the cosmetic composition.
The cosmetic composition for skin whitening according to claim 1, wherein the hole extract or fermented hole hole exhibits a skin whitening effect by inhibiting tyrosinase expression and melanin biosynthesis.
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CN110478309A (en) * | 2019-09-27 | 2019-11-22 | 无锡宾西利悦科技有限公司 | A kind of composition of the shining face of tender skin |
KR102633132B1 (en) | 2021-05-21 | 2024-02-05 | (주)메디톡스 | Composition Comprising Extracellular Vesicles Derived from Yeast, and methods related thereto |
FR3123657B1 (en) * | 2021-06-08 | 2023-05-26 | Soredec Soc De Recherches Et Detudes Cosmetologiques | PROTEIN HYDROLYZATE DERIVED FROM ROMAN BERRIES, COSMETIC COMPOSITION CONTAINING SUCH A HYDROLYZATE AND USE FOR MAINTAINING SKIN HYDRATION |
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KR100535259B1 (en) * | 2003-07-11 | 2005-12-09 | 주식회사 태평양 | Phagocytosis inhibitor composition containing extract of galenical medicines |
KR100563548B1 (en) | 2004-05-18 | 2006-03-27 | 한불화장품주식회사 | A cosmetic composition containing an extract of sorbus commixta |
KR101265955B1 (en) * | 2011-09-07 | 2013-05-22 | 한불화장품주식회사 | Cosmetic composition using microcapsules containing fermented herb medicine extracts |
KR101725623B1 (en) * | 2015-03-20 | 2017-04-12 | 안동대학교 산학협력단 | Pharmaceutical composition comprising the organic solvent fraction of sorbus commixta fruit as an effective component for prevention or treatment of thrombosis and health functional food comprising the same |
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