KR100817662B1 - c-Kit activation inhibitor, skin whitening compound and composition for skin whitening containing the same - Google Patents

Abstract

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 selected from the group consisting of compounds represented by Formulas 1 to 9 Flavone) derivative. These flavone derivatives inhibit the activity of c-Kit involved in melanogenesis and differentiation and maturation of melanocytes. Therefore, the above-described flavone derivatives are useful as skin whitening agents, and compositions such as cosmetics containing these flavone derivatives as active ingredients can be used very effectively for skin whitening such as blemishes and freckles.

Flavone derivatives, c-Kit, skin whitening

Description

c-Kit activation inhibitor, skin whitening compound and composition for skin whitening containing the same}

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-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 skin exposed to UV, the number of melanocytes increases, and c-Kit plays an important role in this process. 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).

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).

On the other hand, SCF, a ligand of c-kit, is overexpressed in the lesion area of melasma (Korean Journal of Dermatology 2005; 43 (8): 1046 ~ 1052), and UVB-Melanosis by UV rays Overexpression has been reported in pathologic blackening of pathological skin such as, senile spots (Lentigo senilis), dermatofibroma and Caffe aure macule (Pigment Cell Research 17: 96-110. 2004).

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. .

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.

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 blemishes, freckles, senile spots (Lentigo senilis) that can be caused by ultraviolet rays, hormones, and genetic causes Skin dermatitis, such as dermatofibroma and caffe aure macule, can improve skin whitening.

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.

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.

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.

In order to solve the above technical problem, the present invention provides a c-Kit activity inhibitor of the flavone (Flavone) derivative selected from the group consisting of compounds represented by the following formula (1) to (9).

The present invention also provides a skin whitening agent of a flavone derivative selected from the group consisting of the compounds represented by Formulas 1 to 9, and a composition for skin whitening containing any one or more of these as an active ingredient.

In the composition for skin whitening, the content of the flavone derivative is preferably 0.000001 to 10% by weight based on the total weight of the composition, such flavone derivatives are skin, lotion, cream, foundation, essence, gel, pack, foam cleansing, It can be added to various formulations, such as soaps, ointments, to exhibit skin lightening effects.

Hereinafter, the present invention will be described in detail.

The present invention is 6-Methoxyflavone of Formula 1 (6-Methoxy-2-phenyl-4H-chromen-4-one, C ₁₆ H ₁₂ O _{3 ,} CAS No. 26964-24-9), 3, 7-dihydroxyflavone (C ₁₅ H ₁₀ O _4, CAS No. 492-00-2), 3,6-Dihydroxyflavone (C ₁₅ H ₁₀ O _4, CAS No. 08238-41-1) of Formula 3, Formula 3-hydroxy-6-methylflavone (6-Methyl-3-hydroxyflavone, C ₁₆ H ₁₂ O _{3 ,} CAS No. 6971-18-2) of 4, 3,3'-dihydroxyflavone (C ₁₅ H) ₁₀ O _{4 ,} CAS No. 55977-09-8), 6,2 ', 3'-trihydroxyflavone of Formula 6 (C ₁₅ H ₁₀ O _5, CAS No. 108238-47-7), 6 of Formula 7 , 4'-dimethoxy-3-hydroxyflavone (C ₁₇ H ₁₄ O _5, CAS No. 93176-02-4), 4'-hydroxy-β-naphthoflavone (C ₁₉ H ₁₂ O _3, CAS No. 98166-72-4) and a flavone derivative of 3,7,3'-trihydroxyflavone (C ₁₅ H ₁₀ O ₅ ) of Chemical Formula 9 provides a c-Kit inhibitor.

The present inventors have conducted studies on substances that inhibit the activity of c-Kit involved in melanogenesis, differentiation and maturation of melanocytes, and found that the above-described flavone derivatives exhibit very potent inhibitory activity of c-Kit. The present invention has been completed.

That is, the above-described flavone derivatives inhibit the activity against c-Kit, and thus can be used as a multifunctional skin whitening agent that inhibits the synthesis of enzymes involved in the differentiation and maturation of melanocytes and the synthesis of melanin in addition to the anticancer effect. In particular, c-Kit activity inhibitors, such as the flavone derivatives described above, can act at the early stage of signaling of melanin synthesis to achieve an effective skin whitening effect in small amounts.

The flavone derivatives of the above formulas (1) to (3) can be easily purchased on the market, either individually or by mixing two or more kinds, such as skin, lotion, cream, foundation, essence, gel, pack, foam cleansing, soap, etc. An effective amount can be added to various compositions such as ointments and skin whitening effects. In consideration of the skin lightening effect and economical efficiency, the amount of the flavone derivatives added is preferably 0.000001 to 10% by weight, more preferably 0.0001 to 1.0% by weight based on the total weight of the composition.

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. The embodiments of the present invention are provided to more completely explain the present invention to those skilled in the art.

Example

Commercially available flavone derivative compounds of Formulas 1 to 9 were used and used. Each flavone derivative compound was brought to a final concentration of 1 uM, and c-Kit RTK was added to each well of an ATP and 384-well plate, followed by primary 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.

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 a c-Kit inhibitor.

In order to accurately measure the alpha screen signal, three samples were performed for each sample, and DMSO was added to the positive control instead of the sample, DMSO was added to the negative control and buffer was added to the negative control to determine the inhibition rate for c-Kit. .

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.

% Inhibition = (mean value of sample-mean value of negative control) / (mean value of positive control-mean value of negative control) X 100

sample % Inhibition 6-Methoxyflavone (Formula 1) 45 3,7-dihydroxyflavone (Formula 2) 51 3,6-Dihydroxyflavone (Formula 3) 44 3-hydroxy-6-methylflavone (Formula 4) 51 3,3'-dihydroxyflavone (Formula 5) 70 6,2'3'-trihydroxyflavone (Formula 6) 37 6,4'-dimethoxy-3-hydroxyflavone (Formula 7) 56 4'-hydroxy-β-naphthoflavone (Formula 8) 47 3,7,3'-trihydroxyflavone (Formula 9) 65 Tyrphostin A51 14.8uM 80

As can be seen from the results of Table 1, it can be seen that the flavone derivative compounds of Formulas 1 to 9 have a strong inhibitory effect of c-Kit activity at lower concentrations compared to Tyrphostin A51, which is known as an inhibitor of c-Kit activity.

Some of the above-mentioned flavone derivative compounds were added to the cosmetic cream in the amounts and ingredients shown in Table 2 below to evaluate the whitening effect.

Component (weight,%) Preparation Example 1 1 Preparation Example 2 Preparation Example 3 Comparative Example 1 6-Methoxyflavone (Formula 1) 0.5 - - - 3,3'-dihydroxyflavone (Formula 5) - 0.5 - - 6,4'-dimethoxy-3-hydroxyflavone (Formula 7) - - 0.5 - Stearic acid 15.0 15.0 Cetanol 1.0 1.0 Potassium hydroxide 0.7 0.7 glycerin 5.0 5.0 Propylene glycol 3.0 3.0 antiseptic Quantity Quantity incense Quantity Quantity Purified water to 100 to 100

In order to verify the skin whitening effect on the cream prepared by the above-described composition, the pigmentation inhibition and blemish improvement experiment was performed as follows.

Pigmentation Inhibitory Effect Experiment

Selected healthy men and women 20 people by putting fine aluminum foil and thin portions of both arms have a diameter of 7mm size hole 6 two rows, were the amount of light of 60mJ / cm ² and at a distance using ORIEL solar simulator 1000W 10cm from the arm . The irradiation site was washed well with 70% ethanol aqueous solution before irradiation. The bases prepared according to Preparation Examples 1-3 and Comparative Example 1 were applied twice a day from three days before the irradiation to the third week after the irradiation.

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.

Experimental substance Effective (persons) No difference (persons) Preparation Example 1 14 6 Preparation Example 2 10 10 Preparation Example 3 11 9

As can be seen from the results in Table 3, the flavone derivative-containing cream of Preparation Examples 1-3 showed a whitening effect on at least 10 of 20 subjects, and did not show any side effects in the skin.

Blemish improvement effect experiment

Ten healthy women with blemishes were selected and the base prepared according to Preparation Example 1 was applied twice a day to the blemish site of the face for three weeks.

 After completion of the experiment, the degree of pigmentation by the expert was visually determined, and the degree of improvement of the blemishes was determined by subjective judgment of the test subject, and the evaluation of whether the cream of Preparation Example 1 improved the blemishes was performed in two stages: effective and no effect. The results are shown in Table 4.

Effective (persons) No difference (persons) Expert judgment 6 4 Subjective Judgment 7 3

As can be seen from the results of Table 4, the cream containing 6-Methoxyflavon prepared according to Preparation Example 1 showed a blemish-improving effect on at least six of 10 subjects, did not show any action.

As such, the flavone derivatives of the invention described above are effective c-Kit activity inhibitors. In particular, the aforementioned flavone derivatives have no side effects on the skin, and inhibit melanin production and differentiation and maturation of melanocytes, so compositions such as cosmetics containing them as an active ingredient are used for blemishes, freckles, senile spots (Lentigo senilis), Dermatofibroma, Caffe aure macule, etc. It can be used very effectively to improve skin hyperpigmentation and realize skin whitening.

Claims (5)

delete Skin whitening agent selected from the group consisting of compounds represented by the formula (1), (4) and (8). < Formula 1>

< Formula 4>

< Formula 8>

A skin whitening composition comprising any one or more selected from the group consisting of compounds represented by Formulas 1, 4 and 8 as an active ingredient. < Formula 1>

< Formula 4>

< Formula 8>

According to claim 3, wherein the content of the active ingredient is a skin whitening composition, characterized in that 0.000001 to 10% by weight based on the total weight of the composition. According to claim 3, wherein the skin whitening composition is skin, lotion, cream, foundation, essence, gel, pack, foam cleansing, soap and ointment is a composition for skin whitening, characterized in that formed in any one formulation .