KR101210471B1 - Competitors of microphthalmia transcription factor and the cosmetic composition for skin whitening comprising the same - Google Patents

Abstract

The present invention relates to a microphthalmia transcription factor (MITF) inhibitor, and more particularly to a cosmetic composition for skin whitening comprising a compound of formula (1) that inhibits the binding of a specific nucleotide sequence called MITF and E-box, The compound of Formula 1 may be usefully used as a cosmetic or whitening agent for skin skin whitening because it has excellent skin whitening activity and no cytotoxicity.
[Formula 1]

Description

Competitors of microphthalmia transcription factor and the cosmetic composition for skin whitening comprising the same}

The present invention relates to a microphthalmia-associated transcription factor (MITF) inducing inhibitor, and more particularly, to search for a MITF inhibitor that inhibits the expression of the enzyme by inhibiting the binding between the gene promoter (E-box) of the pigmentation-related enzyme and MITF. The present invention relates to a MITF inhibitor selected by using a protein chip for use and a cosmetic composition for skin whitening comprising the same.

4- {4-[(4-hydroxy-3-methoxyphenyl) methyl] -3- (hydroxymethyl) oxolan-2-yl} benzene-1,2-diol represented by the following Chemical Formula 1 is a pharma copore database. A single compound selected from compounds expected to inhibit the association of a microphthalmia-associated transcription factor (MITF) with a specific sequence called E-box through computer simulation using the CERIUS II program from:

[Formula 1]

.

.

MITF (microphthalmia-associated transcription factor) is a protein specifically expressed in melanocytes and is an important regulatory factor in melanin production. Skin pigmentation is characterized by tyrosine in melanocytes tyrosinase, tyrp1 (tyrosinase related protein 1), tyrp2 (tyrosinase related protein 2), dopachrom tautomerase (DCT) and dihyduoxyinkole carboxylic acid (DHICA) in Golgi. Occurs by producing eumelanin, which turns black, and pheomelanin, which turns yellow or red by oxidation by pigment forming enzymes such as oxidase (T. Ellenberger et al. Genes Dev ., 8, 970, 1994). The enzymes are specifically expressed in melanocytes, and MITF binds to the E-box (CATGTG) present in the promoter of the pigment-forming enzyme gene such as tyrosinase and the N-terminal transcriptional activation domain (N-terminal) in the E-box. transcription activation domain (TAD) (Sato S et al., Oncogene 14: 3083-3092, 1997), and by promoting the binding of the C-terminal TAD (Takeda K et al. Hum Mol Genet 9: 125-132, 2000) (JM Jung et al., Anal Biochem . , 330 , 251, 2004) induce the expression of pigmentation enzymes resulting in melanin synthesis. Thus, MITF and E-box binding is an important part of regulating the expression of key enzymes involved in pigmentation.

Until now, the development of whitening agents has been mainly focused on finding substances that reduce melanin production by inhibiting the activity of tyrosinase, a basic and most important enzyme in melanin biosynthesis. The developed whitening agents include kojic acid, arbutin, glutathione, vitamin A, vitamin C, etc., but they do not have a satisfactory whitening effect. In addition, the plant extract has a disadvantage that shows a relatively high effect only at high concentrations, the scope of application of the product is very limited.

In order to elucidate the whitening mechanism, there are active researches on ET-1 (endothelin-1), which is important for melanocyte growth and differentiation, PAR-2, which is important for melanocyte delivery, and MITF, which induces melanin formation. MITF-DN and PIAS3 (protein inhibitor of activated STAT3), currently known as inhibitors of MITF and E-box binding, have been reported to inhibit the expression of related enzymes (C. Levy et al. J. Biol . Chem ., 277) . , 1962, 1992) MITF and E-box specific binding inhibitors are believed to be usable as whitening agents for the treatment of skin pigmentation lesions and functional cosmetics. However, the development of the whitening material targeting MITF is inadequate and limited to the intracellular search, which is insufficient to establish a new extracellular search system.

So far known methods for detecting specific binding inhibitors of MITF and E-box include gel mobility-shift analysis (C. Jansen et al., Biochem . J. 246: 227-232, 1987; K. Ruscher et al., J. Biotechnol . 78: 163-170, 2000), Southwestern blotting (B. Bowen et al., Nucleic Acids Res . 8: 120, 1980; WK Miskimins et al., Proc . Natl. Acad . Sci . USA 82: 6741-6744, 1985), ELISA (Y. Choo et al., Nucleic Acids Res . 21, 1993), reporter constructs in yeast assay (SD Hanes et al., Science 251: 426-430, 1991). Although these methods have been performed, there have been problems such as the use of radioisotopes, the limited number and time of conducting experiments, and the complex experimental methods.

The present inventors have reported a MITF inhibitor selected using a microphthalmia-associated transcription factor (MITF) and a protein chip for detecting specific binding inhibitors of E-box, and a cosmetic composition for skin whitening comprising the same (Korea Patent Registration) 10-0872821-00-00). In the present invention, 27 candidates prepared by mimicking the binding site of MITF and E-box as a factor that inhibits expression by binding to the promoters of the MITF and tyrosinase gene were synthesized, and three compounds showing the whitening effect were identified. I did.

Therefore, the present inventors are further studying the substance that can inhibit the binding site of MITF and E-box, and the substance that can inhibit the binding site of MITF and E-box from Pharmacopore database. The compounds selected using the CERIUS II program were found to be safe and have a very good whitening effect, thus completing the present invention.

It is an object of the present invention to provide a cosmetic composition for skin whitening, a melanin inhibitor or a pharmaceutical composition for skin whitening.

In order to achieve the above object, the present invention provides 4- {4-[(4-hydroxy-3-methoxyphenyl) methyl] -3- (hydroxymethyl) oxolan-2-yl} benzene- as a MITF inhibitor. It provides a cosmetic composition for skin whitening comprising 0.005 to 50% by weight of 1,2-diol as an active ingredient.

The present invention also provides a melanin production inhibitor containing the MITF inhibitor as an active ingredient.

Furthermore, the present invention provides a pharmaceutical composition for skin skin whitening containing the MITF inhibitor as an active ingredient.

Single compound for skin skin whitening according to the present invention, 4- {4-[(4-hydroxy-3-methoxyphenyl) methyl] -3- (hydroxymethyl) oxolan-2-yl} benzene-1, 2-diol inhibits the binding of MITF and E-box to inhibit the expression of tyrosinase (pigmentase) protein, which not only inhibits melanin production but also shows little cytotoxicity. It can be usefully used.

1 is a diagram measuring the extent to which the compound of formula 1 inhibits the binding between MITF and E-box according to an embodiment of the present invention using an EMSA test.
FIG. 2 is a diagram illustrating the effect of the compound of Formula 1 according to an embodiment of the present invention on the production of tyrosinase protein, expressed by combining MITF and E-box, using Western blot.

Prior to the detailed description of the present invention, the 'MITF inhibitor' used in the present specification is' 4- {4-[(4-hydroxy-3-methoxyphenyl) methyl] -3- (hydroxy) represented by Formula 1 below. Methyl) oxolan-2-yl} benzene-1,2-diol '.

Hereinafter, the present invention will be described in detail.

The present invention provides 4- {4-[(4-hydroxy-3-methoxyphenyl) methyl] -3- (hydroxymethyl) oxolan-2-yl} benzene-1 represented by the following general formula (1) as a MITF inhibitor: Provided is a cosmetic composition for skin whitening comprising, 2-diol as an active ingredient.

The compound of Formula 1 is selected from 10 candidates obtained from the Pharmacopore database using a CERIUS II program to inhibit the binding site of MITF and E-box. Got it.

The compound of formula 1 as a MITF inhibitor inhibits the binding between the MITF and the oligo-DNA including the E-box. The compound of Formula 1 can bind the oligo-DNA but modifies the binding reaction site between the MITF and the oligo-DNA. Inhibits melanogenesis by disabling the role of transcription factors (see Table 1 and Table 2). In addition, the compound of Formula 1 was confirmed to have little toxicity in cytotoxicity experiments (see Table 3 and Table 4).

MITF inhibitor represented by the compound of Formula 1 according to the present invention can be effectively used in the cosmetic composition for skin whitening because it inhibits excessive melanin production as described above.

In the composition of the present invention, the MITF inhibitor is preferably contained in an amount of 0.005 to 50% by weight based on the total weight of the composition.

The composition of the present invention may contain one or more active ingredients exhibiting the same or similar functions in addition to the MITF inhibitor.

Cosmetics prepared with a cosmetic composition containing the MITF inhibitor of the present invention as an active ingredient can be prepared in the form of a general emulsion formulation and solubilized formulation. Cosmetics of emulsified form include nutrition lotion, cream, essence, etc., and cosmetics of solubilized form have softening longevity. In addition to cosmetics containing the MITF inhibitor of the present invention, by containing a dermatologically acceptable medium or base can be prepared in the form of adjuvants that can be applied topically or systemically applied commonly used in the field of dermatology.

Suitable cosmetic formulations include, for example, emulsions, suspensions, microemulsions, microcapsules, microgranules or ionics (liposomes), nonionics obtained by dispersing an oil phase in a solution, gel, solid or pasty anhydrous product, aqueous phase. It may be provided in the form of a vesicle dispersant, in the form of a cream, skin, lotion, powder, ointment, spray or conceal stick. It may also be prepared in the form of a foam or in the form of an aerosol composition further containing a compressed propellant.

In addition, the cosmetic composition of the present invention, in addition to MITF inhibitors, fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water Commonly used in ionic or nonionic emulsifiers, fillers, metal ion sequestrants and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or cosmetics It may contain adjuvants conventionally used in the cosmetic or dermatology field, such as any other ingredient. And the above ingredients may be introduced in amounts generally used in the field of dermatology.

Specific formulations of the cosmetic composition of the present invention include skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisturizing lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, essence, nutrition essence, pack, Formulations such as soaps, shampoos, cleansing foams, cleansing lotions, cleansing creams, body lotions, body cleansers, emulsions, press powders, loose powders, eye shadows and the like.

The present invention also provides a melanogenesis inhibitor containing the composition as an active ingredient.

As described above, since the composition has a melanin production inhibitory effect similar to arbutin, which is well known as a melanin formation inhibitor (see Table 1 and Table 2), the MITF inhibitor of the present invention may be provided as a melanin production inhibitor.

As a specific formulation of the melanogenesis inhibitor of the present invention, syrups, tablets, capsules, troches, liquids or suspensions may be used, but is not limited thereto.

In addition, the present invention provides a pharmaceutical composition for skin skin whitening containing the composition as an active ingredient.

Pharmaceutical compositions of the present invention may be formulated for oral administration, for example, oral administration such as tablets, powders, capsules, troches, solutions, suspensions; Or it can be formulated into various preparations such as ointment. Pharmaceutical formulations prepared using conventional carriers can be administered orally or parenterally, for example, for dermal or topical application. In addition, the dosage of the active ingredient is different depending on the age, condition, etc. of the patient, but in general, 10 to 500, preferably 50 to 300 mg per day to be administered to adults, the judgment of the doctor or pharmacist Depending on the time interval can be administered several times a day, preferably once to six divided doses.

Hereinafter, the present invention will be described in more detail with reference to examples. However, the following examples are merely to illustrate the present invention, but the content of the present invention is not limited thereto.

< Example  1> MITF  -E- box  Interfering MITF  Selection of Inhibitor Candidates

Candidates were obtained from the Pharmacopore database using the CERIUS II program to inhibit the binding sites of MITF and E-box. One compound was selected from ten candidate materials obtained as described above, and represented by the following Chemical Formula 1.

[Formula 1]

4- {4-[(4-hydroxy-3-methoxyphenyl) methyl] -3- (hydroxymethyl) oxolan-2-yl} benzene-1,2-diol

< Example  2> Cultivation of B16F10 Cells

B16F10 murine melanoma cell line (B16F10 murine melanoma cell; hereinafter, B16F10; Suwon Forestry Research Institute), which is commonly used for screening whitening materials, is a melanin-producing cell line. ), 37 ° C, 5 in sterile DMEM medium (Dulbecco's modified Eagle's medium; GIBCO, USA) with 1% (v / v) of penicillin (Invitrogen, USA) -streptomycin (Invitrogen, USA) Cultured in an incubator in% CO ₂ conditions. B16F10 cell cultures were passaged each time the 100 π dish was 90% full and passages were no more than 30 times.

< Example  3> Melan -a( melan -a) culture of cells

As melanin-producing cell lines, melan-a cells (Amorepacific), which are generally used for screening of whitening substances, were treated with 10% (v / v) FBS (fetal bovine serum; Sigma, USA), penicillin; Invitrogen, USA) -incubator at 37 ° C and 10% CO ₂ in sterile RPMI medium (Rosewel Park Memorial Institute; manufacturer, country of origin) with 1% (v / v) of streptomycin (Invitrogen, USA) Incubated at. Melan-A cell cultures were passaged each time the 100 π dish was 90% full and passages were no more than 30 times.

< Comparative example  1> MITF  Selection of compounds with no inhibitory effect

In order to compare the MITF inhibitory effect of the compound of Formula 1 according to Example 1, the compound of Formula 2 was selected from among the 10 candidates selected in Example 1 that did not inhibit the binding between MITF and E-box.

3- {2,5,7-Triazatricyclo [6.4.0.0 (2,6)] dodeca-1 (12), 6,8,10-tetraen-5-yl} propanoic acid

< Experimental Example  1> Evaluation of the degree of inhibition of melanogenesis

< Experimental Example  1-1> Evaluation of Inhibition of Melanin Production in B16F10 Cells

To determine the effect of the compound of formula 1 selected by the method of Example 1 on the inhibition of melanin production in B16F10 cells. Melanin content determination method is Hosoi method (Hosoi et al. , CANCER) RESEARCH 45: 1474-1478, 1985).

Specifically, B16F10 cells were incubated for 24 hours in the same medium and conditions as in Example 2 in 12-well plates at a concentration of 6 × 10 ⁴ cells / well, and then treated with DMSO in the untreated group and whitening material in the comparative group. Arbutin (manufacturer, country of manufacture) known as was treated with 200 ppm, the treatment group was treated with a compound of Formula 1 at 5, 15, 30 μM. In addition, after incubating for 48 hours, trypsin containing EDTA was treated to recover B16F10 cells. The cells were washed twice with PBS and then clouded in 200 ml of 1N NaOH containing 10% DMSO, followed by agitation at 90 ° C. for 1 hour, followed by cooling to allow sufficient melanin to dissolve, followed by ELISA (enzyme-linked immunosorbent assay). Absorbance was measured at a wavelength of 405 nm using a reader (Molecular Device, USA), and melanin production was calculated using Equation 1 below.

Using Equation 1, the results of calculating the melanin production in the untreated group, the comparative group and the treated group are shown in Table 1 below.

Sample Name Treated with B16F10 Cells Melanin Production (%) Untreated group DMSO 100 Comparison   Arbutin (200 ppm) 51.1
Treated group
  Compound of Formula 1 (5 μM) 83.5   Compound of Formula 1 (15 μM) 71.8   Compound of Formula 1 (30 μM) 57.5

As shown in Table 1, when the compound of Formula 1 according to the present invention was treated with 5, 15, and 30 μM, melanin production was inhibited in a concentration-dependent manner, and when treated with 30 μM, melanin synthesis inhibitory effect was similar to that of arbutin. It could be confirmed that it represents.

< Experimental Example  1-2> Melan -a( melan -a) Evaluation of the degree of inhibition of cell melanogenesis

Except for using Melan-A cells instead of B16F10 cells, the effect of the selected compound of Formula 1 on melanin-producing inhibition in Melan-A cells was confirmed by the same method as Experimental Example 1-1. Table 2 shows.

Name of sample processed on melan-A cells Melanin Production (%) Untreated group DMSO 100 Comparison   Arbutin (200 ppm) 29.4
Treated group
  Compound of Formula 1 (5 μM) 89.3   Compound of Formula 1 (15 μM) 90.3   Compound of Formula 1 (30 μM) 58.5

As shown in Table 2, when the compound of Formula 1 according to the present invention was treated with 5, 15 and 30 μM, the melanin production was inhibited in a concentration-dependent manner. Could. This indicates that the compound of formula 1 according to the present invention has an effect of inhibiting melanin production.

< Experimental Example  2> cytotoxicity test

Cytotoxicity experiment was conducted to determine whether the compound of Formula 1 selected in Example 1 can be used as a cosmetic composition for skin whitening.

Specifically, B16F10 cells and melan-A cells were incubated for 24 hours in the same medium and conditions as in Example 2 in 96-well plates at concentrations of 2.5 × 10 ³ cells / well, respectively, and then treated with DMSO. In the comparison group, arbutin (arbutin, manufacturer, country of manufacture) known as a whitening material was treated with 200 ppm, and the treatment group was treated with 5, 15, 30 μM of the compound of formula 1. Further, after 48 hours of incubation, MTT (3- (4,5-dimethyl-2-thiazolyl) -2,5-diphenyl-2H-tetrazolium bromide; Sigma, USA) dissolved in PBS (phosphate-buffered saline) 100 μl of solution (5 mg / ml) was injected. After 4 hours of incubation, the medium was removed and formazan formed by adding 100 μl of DMSO was dissolved in an enzyme-linked immunosorbent assay (ELISA) microplate reader [MDS (Molecular Device, USA)]. Absorbance was measured at 540 nm using. Cell viability was calculated using Equation 2 below.

The results of calculating the viability of B16F10 cells and melan-A cells in the untreated group, the comparative group and the treated group using Equation 2 are shown in Tables 3 and 4 below.

Sample Name Treated with B16F10 Cells Cell viability (%) Untreated group DMSO 100 Comparison   Arbutin (200 ppm) 90.1
Treated group
  Compound of Formula 1 (5 μM) 107.4   Compound of Formula 1 (15 μM) 97.4   Compound of Formula 1 (30 μM) 110.9

Name of sample processed on melan-A cells Cell viability (%) Untreated group DMSO 100 Comparison   Arbutin (200 ppm) 88.4
Treated group
  Compound of Formula 1 (5 μM) 104.6   Compound of Formula 1 (15 μM) 88.2   Compound of Formula 1 (30 μM) 84.4

As shown in Table 3 and Table 4, high cell viability was obtained even when the compound of Formula 1 was treated to cells at a high concentration of 30 μM. This shows cell viability similar to that of the untreated group without any treatment, indicating that the compound of Formula 1 has little cytotoxicity.

< Experimental Example  3> MITF And E- box  Binding inhibition assessment

In order to confirm whether the compound of Formula 1 according to the present invention has a whitening activity by inhibiting the binding of MITF and E-box, an electrophoretic mobility shift assay (EMSA) test, which can confirm the interaction between DNA and protein through chemiluminescence, is performed It was. The experiment was carried out in the use of ^{LightShif ® Chemiluminesecent EMSA Kit (Pierce,} IL, USA).

Specifically, the biotin-labeled oligo-DNA containing E-box (CATGTG) capable of binding to MITF and the MBPek-MITF produced and separated by inserting a vector into the E-coli were added without DMSO. A treatment group sample was prepared, and a comparative group was prepared by adding 1 mM of the compound of Formula 2, and treated with a sample containing 0.5 mM, 1 mM, 2 mM, and 3 mM of the compound of Formula 1, respectively, and 2 μg of MITF antibody. A sample was prepared, and the results of the comparative analysis by the EMSA test method are shown in FIG. 1. Samples 1 to 7 carried out in the EMSA test of Figure 1 are as follows.

Sample 1.No Treatment Group: DMSO

Sample 2. Comparative Group: Compound 2 (1 mM)

Sample 3. Treatment Group: Compound of Formula 1 (0.5 mM)

Sample 4. Treatment Group: Compound of Formula 1 (1 mM)

Sample 5. Treatment Group: Compound of Formula 1 (2 mM)

Sample 6. Treatment Group: Compound of Formula 1 (3 mM)

Sample 7. Control: MITF Antibody (2 μg)

1 is a diagram measuring the extent to which the compound of formula 1 inhibits the binding between MITF and E-box using EMSA test.

As shown in FIG. 1, the band generated by the binding between the MITF and the E-box is generated one space below the band generated by the reaction between the MITF and the MITF-antibody (see Sample 7). It was confirmed that the band is generated by the combination between the MITF and the E-box. In addition, as the concentration of Formula 1 increases, the thickness of the band generated by the coupling between the MITF and the E-box becomes thinner and disappears (see Samples 3 to 6). By directly inhibiting the binding of the box, the compound of formula 1 shows an excellent effect as a MITF inhibitor.

< Experimental Example  4> Tyrosinase Protein Expression level  Measure

To determine more clearly whether the compound of Formula 1 according to the present invention has a whitening activity by inhibiting the binding of MITF and E-box, a tyrosinase (pigmentase) protein expressed as the MITF and E-box are combined Measurement was performed by western blot.

Specifically, DMSO (untreated group); Arbutin (comparative group); And B16F10 cells treated with the compound of Formula 1 (treatment group) at concentrations of 5, 15, and 30 μM were subjected to lysis buffer [40 mM Tris (pH 8.0), 120 mM NaCl, 0.5% Nonidet P-40. (NP-40), 0.1 mM sodium orthovanadate, 2 mg / ml aprotinin, 2 mg / ml leupeptin, and 100 mg / ml phenylmethylsulfonyl fluoride (PMSF)]. Next, the protein was quantified using protein activity detection reagents (Pierce, IL, U.S.A.) and the same amount of protein was heated for 10 minutes. Total protein was separated by size by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), transferred to PVDF membrane, and tyrosinase protein by chemiluminescence meter (Amersham Life Science, IL, USA). Expression was measured.

In the above experiment, the expression level of MITF was slightly changed while the expression level of tyrosinase protein was rapidly decreased as the concentration of the compound of Formula 1 was increased, and the results are shown in FIG. 2.

Figure 2 is a measure of the degree of inhibition of tyrosinase protein production by the compound of formula 1 using Western blot.

As shown in FIG. 2, as the concentration of the compound of Formula 1 increases, the amount of tyrosinase protein produced by combining MITF and E-box is markedly reduced, and the amount of MITF remaining without binding to E-box is increased. A small amount could be confirmed. This indicates that the compound of formula 1 directly inhibits the binding of MITF and E-box, and the compound of formula 1 is a tyrosinase (pigmenting enzyme) as a MITF inhibitor as compared to arbutin (comparative group), which is commonly known as a whitening agent. ) Has an excellent effect on inhibiting the production of protein.

Examples of formulations for the composition of the present invention are illustrated below.

< Formulation example  1> whitening containing compound of formula 1 as active ingredient Cosmetic  Produce

<1-1> Manufacture of flexible lotion

Formulation example of the flexible lotion containing Formula 1 as an active ingredient was prepared as shown in Table 5.

Raw material Content (parts by weight) Formula 1 10.00 1,3-butylene glycol 1.00 Disodium iodide 0.05 Allantoin 0.10 Dipotassium glycyrrhizate 0.05 Citric acid 0.01 Sodium citrate 0.02 Glycereth-26 1.00 Arbutin 2.00 Hydrogenated castor oil 1.00 ethanol 30.00 Preservative a very small amount coloring agent a very small amount Flavoring agent a very small amount Purified water Balance

<1-2> Preparation of nourishing cream

Formulation of nutrition cream containing a compound of Formula 1 was prepared as shown in Table 6.

Raw material Content (parts by weight) The compound of formula (1) 10.0 1,3-butylene glycol 7.0 glycerin 1.0 D-panthenol 0.1 Plant extract 3.2 Magnesium aluminum silicate 0.3 PEG-40 stearate 1.2 Stearic acid 2.0 Polysorbate 60 1.5 Chin type glyceryl stearate 2.0 Sorbitan sesquioleate 1.5 Cetearyl Alcohol 3.0 Mineral oil 4.0 Squalane 3.8 Carlyric / capric triglyceride 2.8 vegetable oil 1.8 Dimethicone 0.4 Dipotassium glycyrrhizate a very small amount Allantoin a very small amount Sodium hyaruronate a very small amount Tocopheryl acetate Suitable amount Triethanolamine Suitable amount Preservative Suitable amount Flavoring agent Suitable amount Purified water Balance

< Formulation example  2> Preparation of a melanin inhibitor containing the compound of formula 1 as an active ingredient

<2-1> Preparation of Syrup

Syrup containing the compound of Formula 1 as an active ingredient was prepared as shown in Table 7.

Ingredient Content (parts by weight) The compound of formula (1) 2 saccharin 0.8 Party 25.4 glycerin 8 Spices 0.04 ethanol 4 Sorbic acid 0.4 Distilled water 60

<2-2> Preparation of Tablet

Tablets containing the compound of formula 1 as an active ingredient were prepared as shown in Table 8.

Ingredient Content (parts by weight) The compound of formula (1) 250 Lactose 175.9 Potato starch 180 Colloidal silicic acid 32 10% gelatin solution Suitable amount Potato starch 160 talc 50 Magnesium stearate 5

250 parts by weight of the compound of Formula 1, 175.9 parts by weight of lactose, 180 parts by weight of potato starch and 32 parts by weight of colloidal silicic acid were mixed. 10% gelatin solution was added to the mixture, followed by pulverization and passed through a 14-mesh sieve. This was dried, to which 160 parts by weight of potato powder, 50 parts by weight of talc and 5 parts by weight of magnesium stearate were added, and the resulting mixture was purified.

< Formulation Example 3> Preparation of a pharmaceutical formulation containing the compound of formula 1 as an active ingredient

<3-1> Sanje  Produce

2 g of a compound of formula 1

1 g lactose

The above components were mixed and packed in airtight bags to prepare powders.

<3-2> Preparation of Tablet

100 mg of compound of Formula 1

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

<3-3> Preparation of Capsule

100 mg of compound of Formula 1

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, the capsule was prepared by filling in gelatin capsules according to the conventional method for producing a capsule.

Claims (3)

4- {4-[(4-hydroxy-3-methoxyphenyl) methyl] -3- (hydroxymethyl) oxolan-2-yl represented by the following formula (1) as a microphthalmia-associated transcription factor (MITF) inhibitor } Cosmetic composition for skin whitening comprising 0.005 to 50% by weight of benzene-1,2-diol as an active ingredient.
[Formula 1]

The method of claim 1,
4- {4-[(4-hydroxy-3-methoxyphenyl) methyl] -3- (hydroxymethyl) oxolan-2-yl} benzene-1,2-diol represented by Chemical Formula 1 is A skin cosmetic composition for inhibiting melanin production.
4- {4-[(4-hydroxy-3-methoxyphenyl) methyl] -3- (hydroxymethyl) oxolane represented by Chemical Formula 1 of claim 1 as a microphthalmia-associated transcription factor (MITF) inhibitor 2-yl} benzene-1,2-diol A pharmaceutical composition for skin whitening comprising 0.005 to 50% by weight as an active ingredient.

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