KR101126818B1 - c-Kit activation inhibitor, skin whitening compound and composition for skin whitening containing the same - Google Patents
c-Kit activation inhibitor, skin whitening compound and composition for skin whitening containing the same Download PDFInfo
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Abstract
본 발명은 c-Kit 활성 저해제, 피부미백제 및 이를 유효성분으로 함유하는 피부미백용 조성물에 관한 것으로서, 본 발명의 c-Kit 활성 저해제는 하기 화학식 1로 표시되는 크리신(Chrysin)이다. 크리신은 멜라닌 생성 및 멜라닌 세포의 분화와 성숙 등에 관여하는 c-Kit의 활성을 저해한다. 따라서, 피부미백제로서 유용하며, 크리신을 유효성분으로 함유하는 화장료 등의 조성물은 기미나 주근깨 개선 등의 피부미백에 매우 효과적으로 사용될 수 있다.The present invention relates to a c-Kit activity inhibitor, a skin whitening agent and a composition for skin whitening containing the same as an active ingredient. The c-Kit activity inhibitor of the present invention is Chrysin represented by the following Chemical Formula 1. Chrysine inhibits the activity of c-Kit involved in melanogenesis and differentiation and maturation of melanocytes. Therefore, it is useful as a skin whitening agent, and compositions such as cosmetics containing chrysin as an active ingredient can be used very effectively for skin whitening such as blemishes and freckles.
크리신, c-Kit, 피부미백 Chrysin, c-Kit, skin whitening
Description
본 발명은 멜라닌 생성 및 멜라닌 세포의 분화와 성숙 등에 관여하는 c-Kit 활성 저해제, 기미나 주근깨 개선 등의 피부미백에 유용한 피부미백제 및 이를 유효성분으로 함유하는 피부미백용 조성물에 관한 것이다.The present invention relates to a skin whitening agent useful for skin whitening, such as c-Kit activity inhibitors involved in melanogenesis and differentiation and maturation of melanocytes, improvement of freckle and freckles, and a composition for skin whitening containing the same as an active ingredient.
c-Kit는 receptor tyrosine kinase (RTK)의 class III에 속하는 수용체로서, 멜라닌 세포의 생존, 증식 및 분화에 관여하는 것으로 알려져 있다.UV에 노출된 피부에서는 멜라닌 세포의 수가 증가하는데, 이 과정에서 c-Kit가 중요한 역할을 한다. K14 (keratin promoter)-steel factor의 형질전환 쥐의 경우, 모낭의 멜라닌 세포를 생성, 분화시키는 'niche'에서 증가된 SCF는 멜라닌 세포를 증식, 분화시켜 niche밖으로 나오게 하는 것이 확인되었고, 태아일 때 c-Kit의 항체인 ACK2를 처리한 쥐는 하얀 털과 하얀 피부를 나타내었다(Nature 416, 854-860, 2002).c-Kit is a class III receptor of receptor tyrosine kinase (RTK) and is known to be involved in the survival, proliferation, and differentiation of melanocytes. In the skin exposed to UV, the number of melanocytes increases. Kit plays an important role. In mice transfected with K14 (keratin promoter) -steel factor, the increased SCF in 'niche', which produces and differentiates melanocytes in hair follicles, has been shown to proliferate and differentiate melanocytes and out of niche. Mice treated with ACK2, a c-Kit antibody, had white hair and white skin (Nature 416, 854-860, 2002).
원래 c-Kit는 미백제의 표적으로 보다는 항암제 개발의 표적으로 더 많이 연구되어져 왔다. Imatinib (Gleevec, STI-571, Novatis, East Hanover, NJ, USA)는 Bcr-Abl kinase를 표적으로 하는 백혈병 등의 항암제로 널리 알려져 있다. 그런데, imatinib를 처방받은 6명의 환자들에게서 피부의 색소 감소현상이 나타났고, 이는 imatinib가 Bcr-Abl 외에도 c-Kit의 활성을 억제하여 나타난 결과이다. 따라서, SCF/c-Kit가 사람의 멜라닌 세포의 성장, 유지에 필수적 역할을 하고 있음을 알 수 있다(Cancer 98, 2483-7, 2003).Originally, c-Kit has been studied more as an anticancer drug target than as a whitening agent. Imatinib (Gleevec, STI-571, Novatis, East Hanover, NJ, USA) is widely known as an anticancer agent such as leukemia targeting Bcr-Abl kinase. However, six patients who were prescribed imatinib showed decreased skin pigmentation, which is a result of imatinib inhibiting c-Kit activity in addition to Bcr-Abl. Therefore, it can be seen that SCF / c-Kit plays an essential role in the growth and maintenance of human melanocytes (Cancer 98, 2483-7, 2003).
c-Kit는 활성화되면 MAP kinase를 활성화시키고, 다시 helix-loop-helix, leucine zipper 단백질인 Mitf(microphthalmia-associated transcription factor)를 인산화시켜 활성인 상태로 만든다. 활성화된 Mitf는 tyrosinase, Tyrp-2 등의 멜라닌생성 효소의 전사를 활성화시켜 멜라닌 색소를 생성하게 한다. 자외선을 받으면 keratinocyte에서 분비되는 SCF가 c-Kit가 존재하고 있는 전구체 멜라닌 세포를 성숙한 멜라닌 세포로의 분화를 촉진하며, 더불어 멜라닌 합성에 관여하는 효소들의 전사를 촉진함으로서 멜라닌 색소의 합성을 촉진하게 한다.When activated, c-Kit activates MAP kinase and phosphorylates the helix-loop-helix and leucine zipper proteins Mitf (microphthalmia-associated transcription factor), making it active. Activated Mitf activates the transcription of melanogenesis enzymes such as tyrosinase and Tyrp-2 to produce melanin pigments. Upon UV irradiation, SCF secreted by keratinocytes promotes the differentiation of precursor melanocytes containing c-Kit into mature melanocytes, and also promotes the synthesis of melanin pigments by promoting the transcription of enzymes involved in melanin synthesis. .
이와 같이, c-Kit는 자외선에 의한 멜라닌 합성을 촉진하도록 하는 신호전달에 중요하게 관여하고 있다. 따라서, c-Kit에 대한 활성을 저해할 수 있다면, 항암 효과 외에도, 멜라닌 세포의 분화와 성숙을 저해할 수 있다.As such, c-Kit is importantly involved in signal transduction to promote melanin synthesis by ultraviolet light. Therefore, if the activity against c-Kit can be inhibited, in addition to the anticancer effect, it can inhibit the differentiation and maturation of melanocytes.
희고 고운 피부를 갖고자 하는 것은 모든 사람의 한결 같은 소망인데, 멜라닌 합성이 피부 내에서 과도하게 일어나면, 피부 톤을 어둡게 하고, 기미, 주근깨 등을 발생시키기도 한다. 따라서, 피부내의 멜라닌 색소의 합성을 저해시키면, 피부 톤을 밝게 하여 피부미백을 실현할 수 있을 뿐만 아니라, 자외선, 호르몬, 및 유전적인 원인에 기인하여 발생할 수 있는 기미, 주근깨 등의 피부 과색소 침착증 을 개선시킬 수 있다.The desire to have white and fair skin is everyone's constant desire. When too much melanin synthesis occurs in the skin, it can darken the skin tone, and cause blemishes and freckles. Therefore, by inhibiting the synthesis of melanin pigment in the skin, not only can brighten the skin tone to realize skin whitening, but also skin hyperpigmentation such as blemishes and freckles, which may be caused by ultraviolet rays, hormones, and genetic causes. Can be improved.
이에, 종래에는 하이드로퀴논(hydroquinone)이나 아스콜빈산(ascorbic acid), 고지산(kojic acid), 글루타치온(glutathione)같은 티로시나제에 저해 활성을 갖는 물질을 연고나 화장료와 같은 피부 적용 조성물에 배합하여, 기미, 주근깨의 개선 등 피부미백을 실현하고자 하였다. 그러나, 하이드로퀴논은 소정의 미백효과는 인정되지만 피부자극성이 심하여 사용량이 극히 제한된다. 또한, 아스콜빈산은 산화되기 쉬워 이를 배합한 화장료와 같은 조성물은 변색, 변취되는 등의 문제가 발생한다. 한편, 글루타치온, 시스테인 등의 티올계 화합물은 특유의 불쾌한 냄새가 나며 경피흡수에도 문제가 있고, 이들의 배당체 및 유도체들도 극성이 높아 배합 성분으로 사용하기에는 어렵다.Therefore, conventionally, a substance having an inhibitory activity on tyrosinase, such as hydroquinone, ascorbic acid, kojic acid, glutathione, and the like, is formulated into a skin application composition such as an ointment or a cosmetic, We tried to realize skin whitening such as improvement of spots and freckles. However, hydroquinone is recognized to have a predetermined whitening effect but severe skin irritation, so its amount of use is extremely limited. In addition, ascorbic acid tends to be oxidized, and thus a composition such as a cosmetic compound containing the same causes problems such as discoloration and discoloration. On the other hand, thiol-based compounds such as glutathione and cysteine have a characteristic unpleasant odor and have problems with transdermal absorption, and glycosides and derivatives thereof are also highly polar and difficult to use as a blending component.
따라서, 본 발명의 기술적 과제는 멜라닌 합성이나 항암작용 등에 관여하는 c-Kit의 활성을 저해할 수 있는 c-Kit 활성 저해제를 제공하는데 있다.Accordingly, the technical problem of the present invention is to provide a c-Kit activity inhibitor that can inhibit the activity of c-Kit involved in melanin synthesis or anticancer activity.
또한, 본 발명이 이루고자 하는 다른 기술적 과제는 피부에 대한 부작용이 없어 안전하게 사용될 수 있으며, 색소 침착의 저해 효과가 뛰어난 피부미백제 및 이를 함유하는 피부미백용 조성물을 제공하는데 있다.In addition, another technical problem to be achieved by the present invention is to provide a skin whitening agent and a composition for skin whitening having an excellent effect of inhibiting pigmentation can be used safely because there is no side effect on the skin.
상기와 같은 기술적 과제를 해결하기 위하여, 본 발명은 하기 화학식 1로 표시되는 크리신(Chrysin)으로 된 c-Kit 활성 저해제를 제공한다.In order to solve the above technical problem, the present invention provides a c-Kit activity inhibitor of Chrysin (Chrysin) represented by the following formula (1).
<화학식 1><Formula 1>
크리신은 촉규자(Althaea rosea , 접시꽃 씨)로부터 추출하는 것이 바람직하다.Chrysine is preferably extracted from the cucurbita ( Althaea rosea , hollyhock seed).
또한, 본 발명은 상기 화학식 1로 표시되는 크리신(Chrysin)으로 된 피부미백제 및 이를 유효성분으로 함유하는 것을 특징으로 하는 피부미백용 조성물을 제공한다.In another aspect, the present invention provides a skin whitening agent of Chrysin represented by the formula (1) and a composition for skin whitening comprising the same as an active ingredient.
상기 크리신의 함량은 조성물 총 중량을 기준으로 0.000001 내지 10중량%인 것이 바람직한데, 크리신은 스킨, 로션, 크림, 파운데이션, 에센스, 젤, 팩, 폼 클렌징, 비누, 연고 등의 다양한 제형에 첨가되어 피부미백 효과를 나타낼 수 있다.The content of the glycine is based on the total weight of the composition It is preferred that the amount is 0.000001 to 10% by weight, and chrysine may be added to various formulations such as skins, lotions, creams, foundations, essences, gels, packs, foam cleansing, soaps, ointments, and the like to exhibit skin lightening effects.
이하에서 본 발명을 상세하게 설명한다.Hereinafter, the present invention will be described in detail.
본 발명은 상기 화학식 1의 크리신(5,7-dihydroxy-2-phenyl-4H-1-benzopyran-4-one, C15H10O4)으로 된 c-Kit 저해제를 제공한다. 본 발명의 c-Kit 저해제인 크리신(Chrysin)은 촉규자(Althaea rosea , 접시꽃 씨) 및 Oroxylum indicum 식물에 포함된 성분으로 보고되어 있는 물질로써, 항산화 효과가 있다고 알려져 있다. 또한, 크리신의 유도체는 혈당강하효과(KP-1999-0027846), 육모효과(KP-1996-0027845) 등이 알려져 있으나, c-Kit 저해 효과에 대한 보고는 없다.The present invention provides a c-Kit inhibitor of chrysine (5,7-dihydroxy-2-phenyl-4H-1-benzopyran-4-one, C 15 H 10 O 4 ) of Chemical Formula 1. Chrysin, a c-Kit inhibitor of the present invention, has been reported as an ingredient contained in the scavenger ( Althaea rosea , hollyhock seed) and Oroxylum indicum plants, and is known to have an antioxidant effect. In addition, derivatives of chrysine are known to have hypoglycemic effect (KP-1999-0027846), hair growth effect (KP-1996-0027845), etc., but there are no reports on c-Kit inhibitory effect.
본 발명자들은 멜라닌 생성 및 멜라닌 세포의 분화와 성숙 등에 관여하는 c-Kit의 활성을 저해하는 물질을 스크리닝(screening)하기 위하여 천연에서 자생하는 동식물 등을 대상으로 연구를 거듭한 결과, 크리신이 매우 강력한 c-Kit의 활성 저해능을 나타냄을 밝혀내고 본 발명을 완성하게 되었다.The present inventors have conducted studies on native plants and animals for screening substances that inhibit c-Kit activity involved in melanogenesis and differentiation and maturation of melanocytes. The present invention was completed by revealing the activity of c-Kit.
즉, 크리신은 c-Kit에 대한 활성을 저해하므로, 항암 효과 외에도 멜라닌 세포의 분화와 성숙 및 멜라닌 합성에 관여하는 효소의 합성도 저해하는 다기능 피부미백제로 사용이 가능하다. 특히, 크리신과 같은 c-Kit 활성 저해제는 멜라닌 합성의 신호전달 초기에 작용함으로서 소량으로도 효과적인 피부미백 효과를 달성할 수 있다.In other words, since chrysine inhibits the activity of c-Kit, it can be used as a multifunctional skin whitening agent that inhibits the synthesis of enzymes involved in melanocyte differentiation and maturation and melanin synthesis in addition to anti-cancer effects. In particular, c-Kit activity inhibitors, such as chrysine, act early in the signaling of melanin synthesis to achieve effective skin whitening effects in small amounts.
크리신을 촉규자로부터 추출하는 방법을 예시하면 다음과 같으며, 추출 대상이나 추출방법이 이에 한정되는 것은 아니다.Illustrating a method of extracting chrysine from the culprit is as follows, but the extraction target or extraction method is not limited thereto.
생약재로 시판되는 촉규자(Althaea rosea , 접시꽃 씨)를 구입하여 잘게 분쇄하고, 분쇄물 건조 중량에 대하여 5-20부피의 메탄올로 환류 냉각기가 달린 추출기에서 50-100℃로 1-5시간 가열하여 추출한다. 이 결과물을 여과포로 여과한 후 잔사를 같은 방법으로 1회이상씩 더 추출한다. 추출액을 합하여 감압농축한 후 동결건조나 분무 건조하여 건조추출물을 얻는다. 제조된 촉규자의 메탄올 추출물로부터 유효성분을 확인하기 위해 메탄올 추출물을 10배의 물에 현탁시킨후 에칠아세테이트로 용매 분획 후 건조하여 촉규자 비극성 분획을 얻는다. 얻어진 비극성 분획은 실리카겔 컬럼 크로마토그라피를 용리액으로서 에칠아세테이트, 메탄올 혼합용매로 수행하여 유효성분을 분리하고, 얻은 유효성분은 분취용 HPLC사용하여 동일 조건으로 수행하여 순수하게 정제한다. 정제된 유효성분은 핵자기공명스펙트럼(NMR) 및 질량분석(Mass)을 통하여 확인할 수 있다.A commercially available herb ( Althaea rosea ) was purchased as a medicinal herb and pulverized.Then, the dried powder was heated at 50-100 ° C. for 1-5 hours in an extractor equipped with a reflux condenser with 5-20 volumes of methanol. Extract. The resultant is filtered with a filter cloth, and the residue is extracted one more time in the same manner. The extracts were combined and concentrated under reduced pressure, followed by freeze drying or spray drying to obtain a dry extract. In order to identify the active ingredient from the methanol extract of the prepared catalyst, the methanol extract was suspended in 10 times of water, and the solvent fraction was then dried with ethyl acetate to obtain a nonpolar fraction. The obtained non-polar fraction was subjected to silica gel column chromatography as an eluent as an ethylacetate and methanol mixed solvent to separate the active ingredient, and the obtained active ingredient was purified under the same conditions using preparative HPLC. Purified active ingredient can be confirmed by nuclear magnetic resonance spectrum (NMR) and mass spectrometry (Mass).
크리신은 조추출물의 형태로 또는 정제한 순수 화합물의 형태로 스킨, 로션, 크림, 파운데이션, 에센스, 젤, 팩, 폼 클렌징, 비누 등의 화장료나 연고와 같은 다양한 조성물에 유효량 첨가되어 피부미백 효과를 나타낼 수 있다. 피부미백 효과와 경제성을 고려할 때, 크리신의 첨가량은 조성물 총 중량을 기준으로 0.000001 내지 10중량%인 것이 바람직한데, 더욱 바람직하게는 0.0001 내지 1.0중량%이다.Chrysine can be added to various compositions such as skin, lotions, creams, foundations, essences, gels, packs, foam cleansings, soaps, etc. in the form of crude extracts or in the form of purified pure compounds. Can be represented. In consideration of the skin lightening effect and economical efficiency, the amount of chrysine added is preferably 0.000001 to 10% by weight based on the total weight of the composition, more preferably 0.0001 to 1.0% by weight.
이하, 본 발명을 구체적으로 설명하기 위해 실시예를 들어 상세하게 설명하기로 한다. 그러나, 본 발명에 따른 실시예들은 여러 가지 다른 형태로 변형될 수 있으며, 본 발명의 범위가 아래에서 상술하는 실시예들에 한정되는 것으로 해석 되어서는 안된다. 본 발명의 실시예들은 당업계에서 평균적인 지식을 가진 자에게 본 발명을 보다 완전하게 설명하기 위해서 제공되는 것이다.BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to examples. However, embodiments according to the present invention can be modified in many different forms, the scope of the present invention should not be construed as limited to the embodiments described below. Embodiments of the invention are provided to more fully describe the present invention to those skilled in the art.
실시예 1Example 1
생약재로 시판되는 건조된 촉규자를 구입하여 잘게 분쇄하고, 분쇄물 10kg에 대하여 100L 메탄올로 환류 냉각기가 달린 추출기에서 70℃로 3시간 가열하여 추출하였다. 추출물을 여과포로 여과한 후 잔사를 같은 방법으로 1회 이상 더 추출하였다. 추출액을 합하여 감압농축하므로서 건조 추출물 1.1kg을 얻었다.A dried cucumber commercially available as a herbal medicine was purchased and ground finely, and extracted by heating at 70 ° C. for 3 hours in an extractor equipped with a reflux condenser with 100 L methanol for 10 kg of the ground product. The extract was filtered with a filter cloth and the residue was extracted one more time in the same manner. The combined extracts were concentrated under reduced pressure to give 1.1 kg of dry extract.
실시예 2Example 2
실시예 1에서 제조된 촉규자 메탄올 추출물 1kg을 10L의 물에 현탁시킨 후, 에칠아세테이트 10L씩을 가해 3회 용매분획하여 에칠아세테이트 분획을 얻고, 이를 감압농축하여 건조물 60g을 얻었다.After 1 kg of the methanol extract prepared in Example 1 was suspended in 10 L of water, 10 L of ethyl acetate was added thereto to obtain a solvent fraction three times, and the ethyl acetate fraction was concentrated under reduced pressure to obtain 60 g of a dried product.
실시예 3Example 3
실시예 2에서 제조된 촉규자 에칠아세테이트 가용분획 50g을 실리카겔 컬럼 크로마토그라피를 용리액으로서 에칠아세테이트, 메탄올 혼합용매로 수행하여 유효분획을 얻고, 이 분획을 분취용 HPLC를 수행하여 유효성분 60mg을 얻었다. 분리한 유효성분은 핵자기공명스펙트럼(NMR) 및 질량분석(Mass)을 통하여 아래 구조의 크리신으로 확인하였다.50 g of the soluble fraction of the eluent acetate prepared in Example 2 was subjected to silica gel column chromatography as an eluent to obtain an effective fraction, and an effective fraction was obtained by performing preparative HPLC to obtain 60 mg of the active component. The separated active ingredient was identified as Krycine having the following structure through nuclear magnetic resonance spectrum (NMR) and mass spectrometry (Mass).
분자식) C15H10O4 Molecular Formula) C 15 H 10 O 4
분자량) 254.2446Molecular weight) 254.2446
융점) 290 ~ 292℃Melting point) 290 ~ 292 ℃
UV(λmax, nm)(MeOH) 250, 269, 314UV (λmax, nm) (MeOH) 250, 269, 314
1H-NMR(300MHz, acetone-d6) σ 6.49(d,1H,J=2.1Hz), 6.58(s,1H), 6.65(d,1H,J=2.1Hz), 7.58(m,3H), 8.02(m,2H), 9.63(brs.2H)1H-NMR (300MHz, acetone-d6) σ 6.49 (d, 1H, J = 2.1Hz), 6.58 (s, 1H), 6.65 (d, 1H, J = 2.1Hz), 7.58 (m, 3H), 8.02 (m, 2H), 9.63 (brs.2H)
상기 실시예 1-3에 따라 얻어진 각 시료를 실시예 1(메탄올 추출물) 및 실시예 2(에칠아세테이트 가용분획)의 농도는 100ug/ml, 실시예 3(Chrysin)은 최종농 도가 14.8uM가 되도록 하여 c-Kit RTK을 ATP와 384-well plate의 각 웰에 첨가하여 상온에서 1차 반응시킨 후, 기질로서 비오티닐화된 폴리[글루타민:티로신](4:1){(biotinylated-poly[Glu:Tyr] (4:1))을 첨가하여 2차 효소 반응을 진행시켰다.Each sample obtained in accordance with Example 1-3, the concentration of Examples 1 (methanol extract) and Example 2 (Ethyl Acetate soluble fraction) is 100ug / ml, Example 3 (Chrysin) so that the final concentration is 14.8uM. C-Kit RTK was added to each well of an ATP and 384-well plate, followed by a first reaction at room temperature, followed by biotinylated poly [glutamine: tyrosine] (4: 1) {(biotinylated-poly [Glu]. : Tyr] (4: 1)) was added to proceed with the secondary enzyme reaction.
기질들과 결합하도록 스트렙타비딘(Streptavidin)이 피복된 증여체 비드(donor bead)와 항체(P-Tyr-100)가 결합된 수용체 비드(acceptor bead)가 포함된 캡쳐 완충액(capture buffer)을 첨가하여 3차 반응시켰다. 기질의 인산화 정도는 퓨전 마이크로플레이트(FusionTM microplate) 분석장치를 이용하여 알파스크린 신호를 측정함으로써 결정한다. 또한 기존에 c-Kit의 저해제로 알려진 Tyrphostin A51 를 이용하여 저해효과를 비교하였다.Add capture buffer containing donor bead coated with streptavidin and acceptor bead bound to antibody (P-Tyr-100) to bind substrates To the third reaction. The degree of phosphorylation of the substrate is determined by measuring the alpha screen signal using a Fusion ™ microplate analyzer. In addition, the inhibitory effect was compared using Tyrphostin A51, which was previously known as an inhibitor of c-Kit.
알파스크린 신호를 정확하게 측정하기 위하여 각각의 시료에 대하여 3회 실시하였으며 c-Kit에 대한 저해율을 구하기 위해서 양성 대조군에는 시료대신 DMSO를 가하였고, 음성 대조군에는 시료 대신 DMSO를 가하고 효소대신 완충액을 가하였다.In order to accurately measure the alpha screen signal, the test was performed three times for each sample, and DMSO was added to the positive control instead of the sample, and DMSO was added to the negative control and the buffer was added instead of the enzyme to determine the inhibition rate for c-Kit. .
감지된 signal로부터 c-Kit에 대한 활성 저해율(%inhibition)은 다음의 식에 의하여 계산하고 결과를 표 1에 정리하였다.The activity inhibition rate (% inhibition) for c-Kit from the detected signal was calculated by the following equation and the results are summarized in Table 1.
저해율(%) = (시료의 평균값 음성대조군의 평균값)/(양성대조군의 평균값 음성대조군의 평균값) X 100% Inhibition = (average value of sample negative control) / (average value of positive control)
표 1의 결과에서 볼 수 있듯이, 크리신을 함유하는 조추출물인 실시예 1-2의 시료는 상당한 c-Kit에 대한 저해율을 나타내었으며, 정제한 크리신인 실시예 3의 시료는 c-Kit의 활성 저해제로 알려진 Tyrphostin A51과 비교할 때 동등 수준의 c-Kit 활성의 억제능이 있음을 알 수 있다.As can be seen from the results of Table 1, the sample of Example 1-2, which is a crude extract containing chrysine, showed a significant inhibition rate for c-Kit, and the sample of Example 3, which was purified chrysine, showed the activity of c-Kit. Compared with Tyrphostin A51, known as an inhibitor, it can be seen that there is an equivalent level of inhibitory effect of c-Kit activity.
크리신을 화장료에 첨가시 미백효과를 평가하기 위하여, 하기 표 2에 기재된 성분과 함량으로 크림을 제조하였다.In order to evaluate the whitening effect when the creamin was added to the cosmetic, a cream was prepared with the ingredients and contents shown in Table 2 below.
전술한 조성으로 제조한 크림에 대한 색소침착 저해효과를 검증하기 위해 사용한 방법은 다음과 같다. 건강한 남여 20명을 선정하여 양팔의 하박부에 직경 15mm크기의 구멍이 2개씩 2줄로 파인 알루미늄 호일을 붙이고, 팔에서 10cm 떨어진 거리에서 ORIEL solar simulator 1000W를 사용하여 60mJ/cm2의 광량을 조사하였다. 조사전에 70% 에탄올 수용액으로 조사 부위를 잘 세척하였다. 조사하기 3일전부터 조사 후 3주째까지 1일 2회씩 제조예 1 및 비교예 1에 따라 제조된 기제를 한 쌍으로 같은 줄에 도포하였다.The method used to verify the pigmentation inhibitory effect on the cream prepared with the above composition is as follows. 20 healthy males and females were placed in two lower rows of aluminum foil with two 15 mm diameter holes in the lower part of both arms, and 60mJ / cm 2 light was irradiated using ORIEL solar simulator 1000W at a distance of 10cm from the arm. . The irradiation site was washed well with 70% ethanol aqueous solution before irradiation. The bases prepared according to Preparation Example 1 and Comparative Example 1 were applied twice a day from 3 days before irradiation to 3 weeks after irradiation, in pairs, on the same row.
각각에 대하여 제조예와 비교예의 색소침착도를 육안으로 판정하고, 제조예가 비교예에 비해 색소침착을 억제한 정도를 효과있음, 차이없음의 2단계로 평가하였으며, 그 결과는 표 3과 같다.For each, the degree of pigmentation of the preparation example and the comparative example was visually determined, and the degree of suppression of the pigmentation of the preparation example compared to the comparative example was evaluated in two stages of effective and no difference, and the results are shown in Table 3.
위의 표 3의 결과에서 볼 수 있듯이, 제조예 1의 크리신 함유 크림은 피시험자 20명 중에서 11명에 대하여 미백 효과를 나타내었으며, 피부내에서 어떤 부작용도 나타나지 않았다.As can be seen from the results in Table 3, the creamin-containing cream of Preparation Example 1 showed a whitening effect on 11 out of 20 subjects, and did not show any side effects in the skin.
이와 같이, 크리신은 효과적인 c-Kit 활성 저해제이다. 특히, 크리신은 피부에 대한 부작용이 없으며, 멜라닌 생성 및 멜라닌 세포의 분화와 성숙을 저해하므 로, 크리신을 유효성분으로 함유하는 화장료 등의 조성물은 기미나 주근깨 개선 등의 피부미백에 매우 효과적으로 사용될 수 있다.As such, chrysine is an effective c-Kit activity inhibitor. In particular, since there is no side effect on the skin and inhibits melanin production and differentiation and maturation of melanocytes, compositions such as cosmetics containing chrysine as an active ingredient can be used very effectively for skin whitening such as blemishes and freckles. have.
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