KR20200025836A - Novel material for skin whitening, composition comprising the same and methods for manufacturing the same - Google Patents

Abstract

In the present specification, disclosed are a novel material for whitening, a composition containing the material, and a method for manufacturing the material. The material for whitening according to an aspect of the present invention has no toxicity to skin cells and has an excellent melanin biosynthesis inhibitory effect, thereby being able to be utilized as the composition for whitening or a composition for preventing or alleviating chloasma.

Description

Novel whitening substance, composition containing the substance and manufacturing method thereof {NOVEL MATERIAL FOR SKIN WHITENING, COMPOSITION COMPRISING THE SAME AND METHODS FOR MANUFACTURING THE SAME}

The present invention relates to a novel whitening substance, a composition comprising the substance and a method for producing the substance.

Recently, the public's interest in beauty continues to increase, so the demand and interest for cosmetics is continuously increasing. In particular, due to the tendency to prefer white skin of Asians, research on the whitening cosmetics in the cosmetics market has been constantly competition.

Human skin color is largely determined by various factors such as the amount of melanin, hemoglobin, carotene, and the most important of these is melanin. When melanin is excessively produced in the skin, pigmentation occurs in the skin, and the skin color becomes black or freckles occur. The cause of melanin overproduction is a combination of external environmental changes such as UV exposure, air pollutants, medications, irritant skin preparations, stress, and internal factors such as hormonal imbalance, inflammatory response factors, and cytokines.

Meanwhile, as the cause and mechanism of melanin production have been revealed, a method of combining melanin or cosmetics with substances that directly inhibit melanin production or inhibit the activity of enzymes involved in the melanin production process is generally used. Representative materials used for this purpose include ascorbic acid, kojic acid, arbutin, hydroquinone and various plant extracts. However, these whitening materials have a limited whitening effect or have high skin irritation and thus have limitations in use.

Therefore, there is a need for research and development on a whitening material having excellent skin whitening effect and securing skin stability.

Republic of Korea Patent Publication No. 10-2011-0023435

In one aspect, an object of the present invention is to provide a material for skin whitening having a human safety.

In one aspect, another object of the present invention, to provide a skin whitening material excellent in the whitening effect, and a cosmetic composition and a pharmaceutical composition comprising the same.

In one aspect, another object of the present invention is to provide a method for producing a novel whitening material.

In one aspect, the present invention provides a compound of Formula 1, a derivative thereof, a pharmaceutically acceptable salt thereof, a prodrug thereof, a hydrate thereof, or a solvate thereof:

[Formula 1]

The compound, which is an aspect of the present invention, does not have toxicity to skin cells and has excellent melanin biosynthesis and melanin transmission inhibitory effect, and thus may be used as a whitening composition or a composition for preventing or improving blemishes.

1 is a chemical structure of a compound of formula 1 which is an aspect of the present invention.
2 is a result of HPLC analysis of a compound which is an aspect of the present invention.
Figure 3 is an experimental result of analyzing the cell survival rate in C ₂ C12 cells, which are mouse-derived myoblasts, which is an aspect of the present invention.
4A is an experimental result of comparing and analyzing melanin content in B16F10 cells, a mouse-derived melanoma cell line, which is an aspect of the present invention.
4B is an experimental result of comparing and analyzing melanin content in HEMn-DP cells, which are human-derived epithelial melanocytes, which is an aspect of the present invention.
5 is an experimental result of comparing and analyzing the melanin content in the co-culture environment, which is an aspect of the present invention.
6 is an experimental result of analyzing melanin migration using flow cytometry in a coculture environment, which is an aspect of the present invention.
Figure 7 is a representative photo of the results of the skin brightness improvement human application test, which is an aspect of the present invention.
8 is a result of the improvement rate of skin brightness improvement human application test, which is an aspect of the present invention.
9 is an improvement rate result of the skin melanin improvement human application test, which is an aspect of the present invention.

The terms or words used in the specification and claims are not to be limited to the common or dictionary meanings, and the principle that the inventor can appropriately define the concept of terms in order to best describe his invention. It should be interpreted as meanings and concepts corresponding to the technical idea of the present invention. Therefore, the embodiments described in the specification and the configuration shown in the drawings are only the most preferred embodiments of the present invention and do not represent all of the technical idea of the present invention, various equivalents that may be substituted for them at the time of the present application It should be understood that there may be water and variations. In addition, detailed descriptions of well-known functions and configurations that may unnecessarily obscure the subject matter of the present invention will be omitted. Hereinafter, preferred embodiments of the present invention will be described in detail.

In one aspect, the present invention is a compound of Formula 1, a derivative thereof, a pharmaceutically acceptable salt thereof, a prodrug thereof, a hydrate thereof, or a solvate thereof:

[Formula 1]

As used herein, “pharmaceutically acceptable” means that government or equivalent regulatory body approves the use of animals, more specifically humans, by avoiding significant toxic effects when used in conventional medicinal dosages. It means that a person can be approved or approved, listed in a pharmacopoeia, or recognized as another general pharmacopeia.

As used herein, "pharmaceutically acceptable salts" means salts according to one aspect of the invention that are pharmaceutically acceptable and have the desired pharmacological activity of the parent compound. The salt is formed from (1) an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, or the like; Or acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3- (4-hydroxybenzoyl) Benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo [2,2,2] -oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tert Acid addition salts formed with organic acids such as butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid , stearic acid, muconic acid; Or (2) salts formed when the acidic protons present in the parent compound are substituted.

As used herein, "prodrug" refers to a drug that chemically changes a drug to control its physical and chemical properties, and itself does not exhibit physiological activity, but is due to the action of a chemical or enzyme in the body after administration. The drug can be turned into a drug.

As used herein, "hydrate" refers to a compound to which water is bound, and is a broad concept including an inclusion compound having no chemical bonding force between water and the compound.

As used herein, the term "solvate" means a higher order compound produced between molecules or ions of a solute and molecules or ions of a solvent.

The compound, the derivative thereof, the pharmaceutically acceptable salt thereof, the prodrug thereof, the hydrate thereof or the solvate thereof may be prepared by the preparation method described in the specification preparation examples of the present invention.

The compound, the derivative thereof, the pharmaceutically acceptable salt thereof, the prodrug thereof, the hydrate thereof, or the solvate thereof have a remarkably excellent melanin inhibitory effect as compared with the combination of each of tranexamic acid and nicotinic acid. .

In one aspect, the present invention is a whitening composition comprising the compound of Formula 1, its derivatives, its pharmaceutically acceptable salts, its prodrug, its hydrate or its solvate as an active ingredient.

In one aspect, the present invention is a composition for preventing or improving pigmentation comprising the compound of Formula 1, its derivatives, its pharmaceutically acceptable salts, its prodrug, its hydrate or its solvate as an active ingredient.

In one embodiment, the pigmentation may include spots. However, the present invention is not limited thereto, and may include pigmentation, freckles, and the like after inflammation.

In the above aspect, the compound of Formula 1, its derivatives, its pharmaceutically acceptable salts, its prodrugs, its hydrates or solvates thereof, based on the total weight of the composition, 0.1 to 100,000 ppm (w / w). Preferably 0.5 to 70,000ppm (w / w), more preferably 1000 to 70,000ppm (w / w), may be included as 50,000 ppm most preferably. Exceeding the above range is cytotoxic and may be unsuitable for skin application.

In such aspects, the composition may be a pharmaceutical composition or a cosmetic composition.

That is, the cosmetic of the present invention is not particularly limited in the formulation, for example, latex, cream, lotion, essence, pack, gel, powder, makeup base, foundation, lotion, ointment, patch, essence, cleansing foam, cleansing Formulations such as creams, cleansing water, body lotions, body creams, body oils, body essences, shampoos, rinses, body cleansers, soaps, hair dyes, sprays and the like.

The above formulations may contain skin absorption promoting substances to increase the skin lightening effect.

In addition, the cosmetic of the present invention may include one or more selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, sphingolipids and seaweed extract.

The cosmetic of the present invention may be blended with the above essential components, as required, with other components normally blended into the cosmetic.

In addition to fats and oils, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, UV absorbers, preservatives, fungicides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, fragrances, blood circulation accelerators, coolants, It may further include a limiting agent, purified water and the like.

In addition, the compounding component which may be added other than this is not limited to this, Moreover, any of the above components can also be mix | blended within the range which does not impair the objective and effect of this invention.

The pharmaceutical composition, which is an aspect of the present invention, may further include appropriate carriers, excipients, and diluents commonly used in the preparation of pharmaceutical compositions.

The pharmaceutical composition comprising the compound of formula 1 according to the present invention may be in any form suitable for pharmaceutical preparations, including external preparations such as injections, ointments, gels, creams, patches or sprays, etc. according to a conventional method. Can be used.

The dosage of the preparation varies depending on the age, sex, weight, symptoms, and method of administration of the subject, but it is preferable to continue the application for 1 to 5 times at 1.0 to 3.0 ml per day.

In one aspect, the present invention provides a method of preparing a compound of Formula 1, a derivative thereof, a pharmaceutically acceptable salt thereof, a prodrug thereof, a hydrate thereof, or a solvate thereof, wherein the method comprises: trinexamic acid and fluorenylme; Reacting the oxycarbonyl protecting group-containing material to produce fluorenylmethoxycarbonyl-capped tranexamic acid; Reacting the fluorenylmethoxycarbonyl protected tranexamic acid (TXA) with N, N-diisopropylethylamine to prepare tranexamic acid resin; And reacting the tranexamic acid resin with nicotinic acid (NA) to prepare a compound of Formula 1.

[Formula 1]

Specifically, the fluorenylmethoxycarbonyl protecting group-containing material may include N- (9-fluorenylmethoxycarbonyloxy) succinimide (Fmoc-Osu).

Hereinafter, the present invention will be described in more detail with reference to the following examples and comparative examples. However, the following examples are provided only for the purpose of illustration in order to facilitate understanding of the present invention, the scope and scope of the present invention is not limited thereto.

Preparation Example 1 Preparation of Compound of Formula 1

Hereinafter, the compound of Formula 1 of the present invention was named Nicotinoyl Tranexamic acid (NA-TXA). The manufacturing process proceeded in three steps.

Preparation Example 1-1 Preparation of Fmoc-TXA

62.99 g of H-TXA-OH was dissolved in 400 ml of 0.75NK ₂ CO ₃ , and then 300 ml of ACN (acetonitrile) was added thereto, followed by stirring for 20 minutes. 129.5 g of Fmoc-OSu and 140 ml of ACN were added to the reaction solution in an ice-bath, followed by reaction for about 2 hours, followed by distillation under reduced pressure. After distillation under reduced pressure, the sample was titrated to pH 3 using 500 mL of 1N-HCl, washed with 500 mL of tertiary distilled water, and then washed again with 500 mL of ether. The washed reaction solution was solidified by distillation under reduced pressure, and dried to synthesize Fmoc-TXA in powder form.

Preparation Example 1-2 Preparation of TXA-Resin

38 g of CTC (chlorotrityl chloride) resin and 300 mL of DCM (dichloromethane) were added to a 4L reactor, followed by stirring for 20 minutes. After washing with 300 ml of DCM, 32 g of Fmoc-TXA-OH was dissolved in 100 mL of DMF, and 30 mL of DIPEA (N, N-Diisopropylethylamine) was added and reacted. After confirming the reaction progress by the kaiser test, the reaction was washed with DMF 200ml. 150 mL of DCM, 15 mL of MeOH, and 10 mL of DIPEA were added thereto, stirred for 1 hour, washed with 200 mL of DMF, and stirred for 1 hour with 20% de-blacking solution to remove Fmoc. Then, washed three times with 200 mL of DMF and 200 mL with DCM.

Preparation Example 1-3 Synthesis of NA-TXA

Dissolve NA 10g and HOBt 10g with DMF 300mL in synthesized TXA-Resin, dissolve HBTU 50g with DMF 150mL, add 20mL of DIPEA, wash 3 times with DMF 200mL and wash with DCM 200mL to wash NA-TXA-Resin. Synthesized. 50 mL of TFA and 200 mL of DCM were added to the reactor in which the reaction was completed and reacted for 30 minutes. The solution was collected, concentrated, solidified with ACN, filtered and dried.

The yield of synthesized NA-TXA was 12.92 g and the yield was 98.55%, and the purity was 98.38%.

Example 1 Identification and Purity Analysis of the Compound of Formula 1

The synthesized NA-TXA was analyzed by high performance liquid chromatography (HPLC) on a Waters XBridge C18 column. The analytical conditions were analyzed by linear concentration gradient of distilled water solution A containing 0.1% trifluoroacetic acid and acetonitrile solution B containing 0.1% trifluoroacetic acid from 0% to 100% at a rate of 1 mL / min. It was.

FIG. 2 shows the results of analyzing the synthetically prepared NA-TXA at 216 nm wavelength using HPLC. A major peak appeared at 10.27 minutes of RT (retention time), and the corresponding peak area was 98.48%. -TXA purity is 98.48%.

In addition, the synthesized NA-TXA confirmed that the molecular weight is 263.105 (M + H ⁺ : CALC. 263.30) by MALDI-TOF mass spectrometer.

Example 2 Cytotoxicity Evaluation of NA-TXA

C2C12 cells (ATCC CRL-1772), a mouse-derived myoblast, were purchased from ATCC and contained Dulbeccos's Modified Eagle's containing 10% heat-inactivated fetal bovine serum (FBS, hyclone), 50 U / ml penicillin, and 100 ug / ml streptomycin. Cytotoxicity was evaluated in Medium (DMEM, Gibco) at 37 ℃, 5% CO ₂ conditions. Powdered NA-TXA was diluted to 0.5%, 1%, 2%, 5%, 10% w / v using DMEM medium (serum-free) and dispensed in 96 well plates at 3x10 ⁴ (100 uL / well). Incubated for 24 hours. After that, the cells were incubated for 24 hours by replacing with serum-free medium diluted with NA-TXA. After incubation, wash twice with PBS, dispense 200 μL / well of MTT solution (0.5 mg / mL, serum-free medium), incubate for 4 hours, remove medium, add 200 uL / well of DMSO solution, and absorbance at 570 nm. Was measured. It was determined that the cell viability was potentially cytotoxic when the cell viability was reduced to less than 70% of the sample-free group, and the cell viability was calculated according to the following equation and is shown in FIG. 3. Experimental results were verified by performing t-test (paired t-test) using SPSS (Ver. 19) statistical program.

[Equation 1]

Cell viability (%) = (O.D. at 570 nm of the sample addition group) / (O.D. at 570 nm of the sample addition group) x 100

In the results of FIG. 3, cell viability of 99.77 ± 9.52%, 97.58 ± 11.73%, 89.43 ± 9.50%, 98.02 ± 8.55% and 19.10 ± 1.72% was observed at 0.5%, 1%, 2%, 5%, and 10% concentrations. It became. NA-TXA is considered to be non-cytotoxic at concentrations below 5% since it is determined to be potentially cytotoxic if cell viability is reduced to less than 70% of the sample-free group.

Example 3 Evaluation of Whitening Efficacy of NA-TXA in Melanocytes

The whitening efficacy was evaluated using B16F10 cells (ATCC CRL-6475), a mouse-derived melanoma cell line, and Human epidermal melanocytes (neonatal, darkly pigmented donor, HEMn-DP, Gibco C20225PB). High glucose and low glucose DMEM (Hyclone) containing 10% heat-inactivated FBS, 50 U / ml penicillin, 100 ug / ml streptomycin for B16F10 cells, 50 U / ml penicillin, 100 for HEMn-DP cells Medium 254 (Gibco) containing ug / ml streptomycin and Human Melanocyte Growth Supplement-2 (HMGS-2, Gibco) was incubated at 37 ° C and 5% CO ₂ for melanin content analysis. B16F10 cells and HEMn-DP cells were inoculated in ⁵ mm 10 ⁵ cells in a 60 mm culture dish and incubated for 24 hours, followed by the production of α-melanocyte stimulation hormone (α-MSH, Sigma-aldrich M4135) to induce melanin synthesis. After treatment with nM, each material (NA, TXA, NA + TXA, NA-TXA) was treated to a final concentration of 1 mM and incubated for 48 hours and 72 hours, respectively. The cultured cells were washed with PBS, harvested, and the pellets were completely dried at 85 ° C., and 200 ul 1 M NaOH (containing 10% DMSO) was added to lyse the cells while heating at 85 ° C. for 30 minutes. Transfer 150 ul of the dissolved solution to each well of the 96-well plate, measure the melanin by measuring the absorbance at 405 nm, and quantify the protein (Bradford quantitative) to calculate the amount of melanin relative to the total protein in the cell. 4 is shown.

In the results of FIG. 4A, melanin content increased by 55% with α-MSH treatment in B16F10 cells, and melanin content decreased by 25% in the TXA treated group. In addition, the group treated with NA + TXA decreased by 18%. On the other hand, melanin content was reduced by 34% in the group treated with NA-TXA.

In the results of FIG. 4B, the melanin content increased by 56% with α-MSH treatment, the melanin content decreased by 12% in the TXA treated group, and decreased by 35% in the NA + TXA treated group. On the other hand, melanin content was reduced by 49% in the NA-TXA treated group.

Example 4 Evaluation of Whitening Efficacy of NA-TXA in Co-Culture Environment

Human epidermal keratinocyte (neonatal, HEKn, Gibco C0015C) cells and HEMn-DP cells, human-derived epidermal keratinocytes, 50 U / ml penicillin, 100 ug / ml streptomycin and Human Keratinocyte Growth Supplement (HKGS, Gibco), HMGS-2 Melanin content analysis was performed to evaluate the whitening efficacy at 37 ° C and 5% CO2 co-culture with EpiLife (Gibco) and Medium 254 (Gibco) containing (Gibco), respectively. HEKn cells were aliquoted in 5 × 10 5 cells / 60 mm dish and incubated for 24 hours, after which the culture medium was removed and washed once with PBS. Then 1 mL of fresh PBS was added and 5 mJ of UVB was irradiated using a UVB irradation chamber (UV-Testkammer BS-04, Opsytec, Germany). Existing PBS was removed quickly and replaced with fresh mixed medium (keratinocyte medium: melanocyte medium = 1: 1), and HEMn-DP cells were dispensed in 5 × 105 cell / 60 mm dishes. After the addition of each material to a final concentration of 1 mM and incubated for 72 hours in a CO2 incubator. The cultured cells were analyzed for melanin content in the same manner as in [Example 0], and the results are shown in FIG. 5.

As a result, the UVB irradiation resulted in a 20% increase in melanin content, a 26% decrease in melanin content in the TXA and NA treated groups, and a 47% decrease in the NA + TXA treated groups. On the other hand, melanin content was reduced by 60% in the NA-TXA treated group.

In order to confirm the ability of NA-TXA to inhibit the melanosome migration into keratinocytes, analysis was performed using flow cytometry. HEKn cells were dispensed in 5 × 105 cell / 60 mm dishes and incubated for 24 hours. Melanocytes were then diluted with PBS to a final concentration of 5 uM for CFDA-SE staining and mixed with HEMn-DP cells and incubated for 10 min at RT. Then, to stop the reaction of CFDA-SE, 4-5 times the volume of cold medium (10% FBS) was added and incubated for 5 minutes on ice, and then washed three times with medium. After removing the culture medium of HEKn cells after the incubation and washing with PBS, 5 mJ UVB was irradiated with 1 mL of fresh PBS. The PBS was removed quickly, and the HEMn-DP cells reacted with CFDA-SE were previously dispensed in a dish of 2.5 × 105, and each material was treated to a final concentration of 1 mM and incubated for 72 hours. After washing with PBS and using a cell lifter cells were collected and centrifuged for 3 minutes at 800 rpm. After removing the supernatant and proceeding with fixation and permeability, the primary antibody (pan-cytokeratin, abcam) and the secondary antibody (Goat Anti-Mouse IgG H & L, Alexa Fluor® 647) are reacted in turn and flow cytometry (FACS Calibur, BD) is shown in Figure 6 the results. The UVB irradiation increased the rate of melanosome transfer to keratinocytes to 21.36%, and the inhibition rate was calculated by substituting the decrease of melanosome migration according to test substance treatment in the following formula.

[Formula 2]

% Inhibition = [1- (Melanosome ratio of each substance treatment group / Melanosome ratio of control group)] * 100

PBS PBS NA TXA NA + TXA NA-TXA UVB irradiation (5 mJ) - + + + + + Keratinocyte Containing Melanosome (%) 5.25 21.36 14.82 18.5 10.71 5.02 Inhibition Rate (%) - - 30.6 14.4 49.9 76.5

As a result, the amount of keratinocytes containing melanosomes increased to 21.36% in the group irradiated with UVB, but 30.6% with NA, 14.4% with TXA, and 49.9% with NA + TXA. It was. On the other hand, after treatment with NA-TXA after UVB irradiation it was confirmed that the inhibition rate of 76.5%.

Preparation Example 2 Preparation of NA-TXA Ampoule and Control Ampoule

To prepare the NA-TXA Ampoule and control Ampoule in the amount described in Table 2 below. Specifically, glycerol, sodium hyuronate, hydroxyethyl cellulose, and nittin noyl tranxamic acid were dissolved and mixed in purified water at 70 ° C., cooled to 30 ° C., and the remaining raw materials were added and mixed to prepare two Ampoules in Table 2. Was prepared.

Raw material name NA-TXA Ampoule Contrast Ampoule Purified water 78.15% 83.15% Arginine 6% 6% glycerin 5% 5% Nicotine Oil Trinexamic Acid 5% - 1,2-hexanediol 2% 2% ethanol 2% 2% Fiji-26-Bootes-26 1.1% 1.1% Caprylyl Glycol 0.5% 0.5% Sodium hyaluronate 0.1% 0.1% Hydroxyethyl cellulose 0.1% 0.1% Spices 0.05% 0.05%

Example 5 Confirmation of Skin Brightness Improvement Effect of NA-TXA Ampoule

The test subjects were 6 in their 30s, 13 in their 40s, and 2 in their 50s. A total of 21 subjects were used. Prohibited all, medical treatment, massage and other procedures were also prohibited. NA-TXA Ampoule (control group, right facial part) and control Ampoule (control group, left facial part) provided to the subjects for evaluation were used for 4 weeks in the morning and evening after washing the face with appropriate amount (dropper 2-3 drops). In case of using the product, the test substance is completely absorbed before use. In order to confirm the effect of improving skin brightness, the skin L * value was measured by using a spectrophotometer before, 2 weeks, and 4 weeks (total 3 times) before applying the test substance to the left and right face parts of the test subjects. The test site was measured three times and the average value is shown in Table 3 and a representative photograph is shown in FIG. 7. Higher readings mean better skin brightness. The significance of the test material before and after use was checked using the SPSS (Ver. 25) program (P <0.05, Repeated measures ANOVA), and post-test was performed with Bonferroni. Skin brightness improvement (%) compared to before use was calculated by substituting the following formula and the results are shown in FIG. 8.

[Equation 3]

% Improvement = (Measurement after use-Measurement before use) / Measurement before use x 100 (%)

Before use 2 weeks later 4 weeks later Test group Average 61.56 63.46 ^** 64.61 ^** Standard Deviation 3.44 3.03 2.93 Control Average 62.48 63.06 63.76 ^* Standard Deviation 3.31 3.14 2.73

^* P <0.05 , ^** P <0.01 , Number of subjects: 21, Unit: L ^*

As a result, in the subjects using the test group (NA-TXA Ampoule), the skin brightness improvement rate was 3.09% after 2 weeks and 4.95% after 4 weeks, and the brightness change after 2 weeks and 4 weeks. There was a statistically significant difference (P <0.01). On the other hand, the control group (control Ampoule) showed a skin improvement of 0.93% after 2 weeks and 2.05% after 4 weeks, and only the brightness change after 4 weeks showed a statistically significant difference. P <0.05) showed a lower rate of improvement than the test group.

Example 6 Confirmation of Skin Melanin Improvement Effect of NA-TXA after Artificial Pigment Deposition

The test subjects were 1 in 20s, 5 in 30s, 10 in 40s, and 6 in 50s (a total of 20 females and 2 males). The use of therapies, cosmetics and quasi-drugs for the purpose of improving the skin is prohibited, and medical treatment and massage procedures are also prohibited. Minimal Erythema Dose (MED) was determined by observing the irradiation site for 16-24 hours after UV (UVA + B) irradiation using the Solar Simulator on the subject's forearm area. In consideration of peace, up to three MEDs were identified. The subjects were allowed to absorb the appropriate amount (2 drops) of skin after using morning and evening after washing the test group (NA-TXA Ampoule) and control group (control Ampoule) from one week after the end of UV irradiation (blackening confirmation). In order to compare the melanin change, melanin (N, melanon) of the test site was measured before, after 2 weeks, after 4 weeks, and after 8 weeks using the test substance. After measuring five times of each test site, three average values except the maximum and minimum values were obtained and shown in Table 4. Lower readings indicate improved skin melanin. The significance of the test material before and after use was checked using the SPSS (Ver. 25) program (P <0.05, Repeated measures ANOVA), and post-test was performed with Bonferroni. Skin brightness improvement (%) compared to before use was calculated using the same calculation formula as in Example 5. (Fig. 9).

Before use 2 weeks later 4 weeks later 8 weeks later Test group Average 115.85 105.78 ^* 103.19 ^* 97.60 ^* Deviation 21.06 12.46 13.47 14.35 Control Average 112.87 108.46 104.70 ^* 100.43 ^* Deviation 19.38 15.89 13.75 15.76

^* P <0.05 , number of subjects: 22, unit: AU

As a result, the subjects using the test group (NA-TXA Ampoule) showed skin melanin improvement rate of 8.69% after 2 weeks, 10.93% after 4 weeks, and 15.75% after 8 weeks, compared to before 2 weeks. After 4 weeks, the skin melanin index change after 8 weeks showed a statistically significant difference (P <0.05). On the other hand, the control group (control Ampoule) showed 3.91% improvement in skin brightness after 2 weeks, 7.24% after 4 weeks, and 11.02% after 8 weeks. There was a statistically significant difference (P <0.05) but lower than the test group.

Hereinafter, the formulation example of the composition according to the present invention, but the pharmaceutical composition and cosmetic composition is applicable to various formulations, which is intended to explain in detail only, not intended to limit the present invention.

Formulation Example 1 Soap

Ingredient content(%) Compound of Formula 1 50000ppm maintain Quantity Sodium hydroxide Quantity Sodium chloride Quantity Spices Quantity Purified water Remaining amount

[Formulation Example 2] Lotion

Ingredient content(%) Compound of Formula 1 50000ppm L-ascorbic acid-2-phosphate magnesium salt 1.00 Water Soluble Collagen (1% Aqueous Solution) 1.00 Sodium citrate 0.10 Citric acid 0.05 Licorice Extract 0.20 1,3-butylene glycol 3.00 Purified water Remaining amount

Formulation Example 3 Cream

Ingredient content(%) Compound of Formula 1 50000ppm Polyethylene Glycol Monostearate 2.00 Self-emulsifying glycerin monostearate 5.00 Cetyl alcohol 4.00 Squalene 6.00 Glyceryl tri2-ethylhexanoate 6.00 Sphingolipid 1.00 1.3-butylene glycol 7.00 Purified water Remaining amount

Formulation Example 4 Ointment

Ingredient content(%) Compound of Formula 1 50000ppm Polyvinyl alcohol 13.00 L-ascorbic acid-2-phosphate magnesium salt 1.00 Lauroylhydroxyproline 1.00 Soluble collagen (1% aqueous solution) 2.00 1,3-butylene glycol 3.00 ethanol 5.00 Purified water Remaining amount

Formulation Example 5 Preparation of Serum

Ingredient content(%) Compound of Formula 1 50000ppm Hydroxyethylene Cellulose (2% aqueous solution) 12.00 Xanthan gum (2% aqueous solution) 2.00 1,3-butylene glycol 6.00 Dark glycerin 4.00 Sodium hyaluronate (1% aqueous solution) 2.00 Purified water Remaining amount

Claims (6)

A compound of Formula 1, a derivative thereof, a pharmaceutically acceptable salt thereof, a prodrug thereof, a hydrate thereof, or a solvate thereof:
[Formula 1]

. A whitening composition comprising the compound of claim 1, a derivative thereof, a pharmaceutically acceptable salt thereof, a prodrug thereof, a hydrate thereof, or a solvate thereof as an active ingredient. A composition for preventing or improving pigmentation, comprising the compound of claim 1, a derivative thereof, a pharmaceutically acceptable salt thereof, a prodrug thereof, a hydrate thereof, or a solvate thereof as an active ingredient. The method of claim 3,
The pigmentation is a composition for preventing or improving pigmentation containing a blemish. The method according to any one of claims 2 to 4,
The composition comprises a compound of Formula 1, a derivative thereof, a pharmaceutically acceptable salt thereof, a prodrug thereof, a hydrate thereof, or a solvate thereof in an amount of 0.1 to 100,000 ppm (w / w) based on the total weight of the composition. , Composition. The method according to any one of claims 2 to 4,
The composition is a pharmaceutical composition or cosmetic composition.

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