KR101246682B1 - Composition for whitening of the skin comprising arylthiourea derivatives - Google Patents

Abstract

상기 화학식 I에서,
R₁은 수소, C₁ 내지 C₆ 알킬, C₃ 내지 C₈ 사이클로알킬, 또는 치환기를 갖거나 갖지 않는 페닐을 나타내고, 상기 치환기는 C₁ 내지 C₃ 알킬, 할로겐, 히드록시, 또는 C₁ 내지 C₃ 알콕시이고,
R₂는 수소, 또는 C₁ 내지 C₆ 알킬을 나타내고,
R₃은 수소, C₁ 내지 C₉ 알킬, C₁ 내지 C₆ 알콕시, 히드록시, 할로겐 또는 C₁ 내지 C₆ 알킬아민을 나타내고,
n₁ 및 n₂는 각각 독립적으로 0, 1, 2, 3 또는 4를 나타낸다(단, R_1, R₂, 및 R₃ 가 모두 수소이고, n₁ 및 n₂가 모두 0인 경우를 제외한다). The present invention relates to a whitening composition containing, as an active ingredient, an arylthiourea derivative having the following formula (I), which has a higher inhibition rate of melanin production than N-phenylthiourea:
(I)

In Formula I,
R ₁ represents hydrogen, C ₁ to C ₆ alkyl, C ₃ to C ₈ cycloalkyl, or phenyl with or without substituents, said substituents being C ₁ to C ₃ alkyl, halogen, hydroxy, or C ₁ to C ₃ alkoxy,
R ₂ represents hydrogen or C ₁ to C ₆ alkyl,
R ₃ represents hydrogen, C ₁ to C ₉ alkyl, C ₁ to C ₆ alkoxy, hydroxy, halogen or C ₁ to C ₆ alkylamine,
n ₁ and n ₂ each independently represent 0, 1, 2, 3 or 4, except when R _1, R ₂ , and R ₃ are all hydrogen and n ₁ and n ₂ are both 0 ).

Description

Whitening composition containing arylthiourea derivatives {COMPOSITION FOR WHITENING OF THE SKIN COMPRISING ARYLTHIOUREA DERIVATIVES}

The present invention relates to a whitening composition of an arylthiourea derivative and a whitening composition comprising an arylthiourea derivative as an active ingredient.

The color of human skin is determined by the amount of melanin, carotene and hemoglobin, among which melanin is the most crucial factor. The melanin pigment is a phenolic polymer substance having a complex form of black pigment and protein. It plays a role of blocking ultraviolet rays, and a person who lacks the melanin pigment is very sensitive to sunlight and is likely to burn, and skin cancer is likely to occur at a young age. On the other hand, short-wave UV and carcinogens form harmful free radicals in the skin, and melanin removes these free radicals and plays a useful role in protecting proteins and genes.

Melanin is produced by a complex process from tyrosine by the action of tyrosinase in the pigment cells. The melanin produced at this time is delivered to the skin cells and exhibits a circulating action in which the melanin is lost due to epidermal detachment. This process of melanin production is a naturally occurring phenomenon and does not result in excessive production of melanin in normal skin. However, when the skin responds to external stimuli such as ultraviolet rays, environmental pollution, stress, or the like, melanin is excessively produced and is not discharged out of the skin. It is transferred to keratinocyte and accumulates in the skin surface layer to become stain, freckles, Not only does it cause serious cosmetic problems, it also promotes skin aging and skin cancer.

It is everyone's constant desire to have white, fair skin. If you can suppress the production of melanin pigment, you will get white skin by the whitening effect. In addition, it is possible to treat lesions caused by pigmentation such as blemishes, freckles, and surpluses by preventing excessive melanin pigmentation on the skin or by thinning the color of the melanin pigment already deposited. Kojiic acid (Kojic acid), vitamin C, hydroquinone, arbutin and the like are known as such a representative whitening agent. However, kojic acid is not only effective in inhibiting melanin production in vivo compared to the inhibitory ability of tyrosinase in vitro, but also has a problem in the stability of the formulation, and has recently been reported to cause liver cancer in long-term use. Vitamin C is very unstable and low phase stability in the formulation has a problem that the activity of the active ingredient is lowered over time. Recently, various methods for stabilizing the formulation of vitamin C in capsules or liposomes have been proposed, but no clear stabilization method has yet been proposed. Hydroquinone has excellent whitening effect but is limited to its use as it is defined as a carcinogenic substance. Arbutin is a compound in which sugar is attached to hydroquinone. When sugar is separated by skin enzymes when applied to cosmetics, hydroquinone, a carcinogen, is formed, causing cytotoxicity. Recently, various researches and developments have been made regarding functional whitening agents using herbal medicines such as licorice and mulberry and natural products as the side effects of whitening agents have been released. However, most plant extracts are known to show whitening efficacy only at high concentrations and have little effect at low concentrations. There is not enough verification of the toxicity. Therefore, there is an urgent need to develop a safe alternative whitening agent that is excellent in small amount and has fewer side effects.

It is an object of the present invention to provide a whitening composition or a whitening cosmetic composition comprising the same, which can produce a safer and superior whitening effect.

Another object of the present invention to provide a composition for the prevention and treatment of skin diseases associated with hyperpigmentation.

In order to achieve the above and other objects, the present invention provides a composition for skin whitening containing an arylthiourea derivative of the formula (I) as an active ingredient:

(I)

In Formula I,

R ₁ represents hydrogen, C ₁ to C ₆ alkyl, C ₃ to C ₈ cycloalkyl, or phenyl with or without substituent X, which substituent C is C ₁ to C ₃ alkyl, halogen, hydroxy, or C ₁ to C ₃ alkoxy,

R ₂ represents hydrogen or C ₁ to C ₆ alkyl,

R ₃ represents hydrogen, C ₁ to C ₉ alkyl, C ₁ to C ₆ alkoxy, hydroxy, halogen or C ₁ to C ₆ alkylamine,

n ₁ and n ₂ each independently represent 0, 1, 2, 3 or 4, provided that R _1, R ₂ , and R ₃ are all hydrogen and n ₁ and n ₂ are all 0 do).

The composition for skin whitening of the present invention is an arylthiourea derivative compound of the formula (I), which is 4-methylphenylthiourea, 3-methylphenylthiourea, 4-ethylphenylthiourea, 4-chlorophenylthiourea, 4-hydroxy Phenylthiourea, 4- tert -butylphenylthiourea, 4-methoxyphenylthiourea, 1,3-diphenylthiourea, 1-phenyl-3- p -tolylthiourea, 1- (4-chlorophenyl) 3-phenylthiourea, 1- (4-methoxyphenyl) -3-phenylthiourea, 1- (4-hydroxyphenyl) -3-phenylthiourea, 1-cyclohexyl-3-phenylthiourea, At least one compound selected from the group consisting of 1-benzyl-3-phenylthiourea, 1-phenethyl-3-phenylthiourea, and 1-phenyl-3- (3-phenylpropyl) thiourea as an active ingredient do.

According to another aspect, the present invention relates to a pharmaceutical composition for preventing and treating hyperpigmentation disorders comprising a composition for skin whitening comprising the arylthiourea derivative compound of Formula (I) as an active ingredient.

In still another aspect, the present invention relates to a whitening cosmetic composition comprising a whitening composition comprising the arylthiourea derivative compound of the formula (I) as an active ingredient.

Hereinafter, the present invention will be described in more detail.

N-phenylthiourea has been known as a representative inhibitor for tyrosinase. These enzymes, along with catechol oxidase, belong to the type-3 copper protein group. The crystal structure of phenylthiourea bound catechol oxidase has been reported [Gerdemann, C .; Eicken, C .; Krebs, B. Acc. Chem. Res. 2002, 35, 183. and Klabunde, T .; Eicken, C .; Sacchettini, J. C .; Krebs, B. Nat. Struct. Biol. 1998, 5, 1084. The sulfur atom of such a compound is bound to both copper ions at the active site of the enzyme [Klabunde, T .; Eicken, C .; Sacchettini, J. C .; Krebs, B. Nat. Struct. Biol. 1998, 5, 1084. In addition to working with double copper centers, van der Waals interactions between residues lining hydrophobic cavities contribute to the high affinity of phenylthiourea for the enzyme [Klabunde, T]. .; Eicken, C .; Sacchettini, J. C .; Krebs, B. Nat. Struct. Biol. 1998, 5, 1084. Although the crystal structure of phenylthiourea bound tyrosinase is unknown, the binding modes of these compounds will be very similar because the active sites of tyrosinase are predominantly conserved and have a hydrophobic cavity similar to Tyr 98.

To the best of the inventors' knowledge, there is no report on the structure activity relationship (SAR) of arylthiourea analogs. Therefore, we assessed the inhibitory activity of a series of arylthiourea derivatives and studied SAR for melanonine formation. General structural formula of the arylthiourea of the present invention is represented by the following general formula (I).

(I)

In Formula I,

R ₁ represents hydrogen, C ₁ to C ₆ alkyl, C ₃ to C ₈ cycloalkyl, or phenyl with or without substituent X, which substituent X is C ₁ to C ₃ alkyl, halogen, hydroxy or C ₁ To C ₃ alkoxy,

R ₁ represents hydrogen or C ₁ to C ₆ alkyl,

R ₃ represents hydrogen, C ₁ to C ₉ alkyl, C ₁ to C ₆ alkoxy, hydroxy, halogen or C ₁ to C ₆ alkylamine,

n ₁ and n ₂ each independently represent 0, 1, 2, 3 or 4, except when R _1, R ₂ , and R ₃ are all hydrogen and n ₁ and n ₂ are both 0 ).

Referring to the method for preparing the arylthiourea derivative of the formula (I) of the present invention are as follows.

The arylthiourea derivatives of the present invention can be prepared from benzoyl isothiocyanates and commercially available amines according to Scheme 1 below. That is, benzoyl chloride is treated with ammonium thiocyanate while refluxing in acetonitrile for about 30 minutes to produce the intermediate benzoyl isothiocyanate. Then, the obtained benzoyl isothiocyanate was treated with amine while refluxing for about 30 minutes to obtain an intermediate (5), and then hydrolyzed by hot 5% NaOH for 20 minutes to arylthiourea derivative as a desired compound. To prepare.

Scheme 1

In Scheme 1, R ₃ represents a substituent set forth in Table 1, wherein (i) is NH ₄ SCN, reflux, 15 minutes conditions, (ii) R-NH ₂ , reflux for 30 minutes and (iii ) 5% NaOH, warm for 20 minutes.

Alternatively, arylthiourea derivatives of the present invention can be prepared from benzoyl isothiocyanate and commercially available amines according to Scheme 2 below. That is, after dissolving the alkylamine in benzene, treating with carbon disulfide and triethylamine, filtering the obtained solution to obtain a solid, then dissolving in chloroform, and then adding a mixture solution of triethylamine and ethyl chlorocarbonate. From the solution, isothiocyanate was obtained and the isothiocyanate was reacted with an amine compound to obtain a desired arylthiourea derivative compound.

Scheme 2

In Scheme 2, R ₁ and R ₂ are substituents shown in Table 1 below, R ₃ is hydrogen, (iv) carbon disulfide and triethylamine treatment, (v) ethylchlorocarbonate and triethylamine In the solution of the mixed solution of (vi), the reaction proceeds in acetonitrile for 2 hours under stirring conditions.

It is preferable that the arylthiourea derivative compound of this invention is contained by 0.0001-5 weight% with respect to the total weight of a composition.

The pharmaceutical composition of the present invention is used for the purpose of preventing or treating hypermelanin pigmentation such as blemishes, freckles, and senile plaques, and includes various pharmaceutically acceptable carriers, excipients, and / or additives. It may be formulated in a dosage form.

Representative examples include transdermal preparations such as creams, ointments, gels, lotions, solutions or patches.

Formulation of the cosmetic composition of the present invention can be selected arbitrarily, conventional cosmetic formulations essence, whitening cream, lotion, emulsion, pack, general cosmetics, skin milk, cream, serum, beauty soap, softening cosmetics, medicinal cosmetics, hair tonic It can be prepared in various forms, such as systemic cleaners and oil gels.

Cosmetic composition of the present invention further comprises at least one additive selected from the group consisting of oil, water, surfactants, moisturizers, thickeners, chelating agents, pigments, preservatives, and fragrances to produce a cosmetic composition, characterized in that Can be.

The composition for skin whitening in the present invention has a very excellent effect on melanin production inhibitory effect than the known N-phenylthiourea (Compound 1a) as shown in Table 1 below.

Hereinafter, the present invention will be described by the following examples. These Examples are for explaining the present invention, and the present invention is not limited to these Examples.

Example 1: Preparation of the following compound 1a (phenylthiourea)

To 0.012 mol of ammonium chloride solution in acetone, 0.01 mol of benzoyl chloride was added over 5 minutes, and the mixture was heated to reflux for 15 minutes. The reaction mixture was cooled to room temperature and an amount of aniline in acetone was added dropwise over about 15 minutes. Then, the reaction mixture was again refluxed for 30 minutes and then poured on ice. The resulting solid was collected and washed with water. The collected solid was added to the preheated aqueous sodium hydroxide solution and stirred for 20 minutes. The resulting mixture was poured into 5% hydrochloric acid. The reaction was treated with sodium carbonate to remove benzoic acid. The crude product was collected, dried and purified by flash column chromatography to afford the title compound (reaction mechanism see Scheme 1 above).

Yield 70%; Yellow solid; mp 159-161 ° C .; IR (neat); 3412, 3172, 1655, 1637, 1588; ¹ H NMR (CDCl ₃ ): δ 6.15 (bs, 2H, NH ₂ ), 7.25 (d, 2H, Ar-H, J = 7.6 Hz), 7.33 (t, 1H, Ar-H, J = 7.6 Hz) 7.45 (t, 2H, Ar-H, J = 7.6 Hz), 8.31 (bs, 1H, NH).

Example 2: Preparation of the following compound 1b (4-methylphenylthiourea)

Yield 80%; Yellow solid; mp 160-162 ° C .; IR (neat); 3435, 3159, 1614, 1582, 1560; ¹ H NMR (CDCl ₃ ): δ 2.36 (s, 3H, CH ₃ ), 6.02 (bs, 2H, NH ₂ ), 7.12 (d, 2H, Ar-H, J = 8.0 Hz), 7.24 (d, 2H , Ar-H, J = 8.0 Hz), 7.82 (bs, 1H, NH).

Example 3: Preparation of the following compound 1c (4-ethylphenylthiourea)

Yield 80%; Pale yellow solid; mp 125-127 ° C .; IR (neat); 3436, 3160, 1738, 1616, 1581; ¹ H NMR (CDCl ₃ ): δ 1.23 (t, 3H, CH ₃ , J = 7.6 Hz), 2.66 (q, 2H, CH ₂ , J = 7.6 Hz), 6.16 (bs, 2H, NH ₂ ), 7.15 (d, 2H, Ar-H, J = 8.4 Hz), 7.25 (d, 2H, Ar-H, J = 8.0 Hz), 8.18 (bs, 1H, NH).

Example 4: Compound 1d (4- tert -Butylphenylthiourea)

Yield 89%; White solid; mp 154-156 ° C; IR (neat): 3435, 3160, 1734, 1615, 1580 cm ⁻¹ ; ¹ H NMR (CDCl ₃ ): δ 1.30 (s, 9H, C (CH ₃ ) ₃ ), 6.10 (bs, 2H, NH ₂ ), 7.12 (d, J = 8.8 Hz, 2H, Ar-H), 7.44 (d, J = 8.8 Hz, 2H, Ar-H), 8.00 (bs, 1H, NH).

Example 5: Preparation of the following compound 1e (4-chlorophenylthiourea)

Yield 80%; Pale yellow solid; mp 170-172 ° C .; IR (neat): 3412, 3160, 3156, 1621, 1581 cm ⁻¹ ; ¹ H NMR (CDCl ₃ ): δ 6.04 (bs, 2H, NH ₂ ), 7.20 (d, 2H, Ar-H, J = 8.8 Hz), 7.42 (d, 2H, Ar-H, J = 8.8 Hz) , 7.90 (bs, 1 H, NH).

Example 6: Preparation of the following compound 1f (4-hydroxyphenylthiourea)

Yield 68%; Red solid; mp 199-201 ° C; IR (neat): 3399, 3303, 3185, 1650, 1548 cm ⁻¹ ; ¹ H NMR (DMSO): δ 6.72 (d, 2H, Ar-H, J = 8.0 Hz), 7.06 (d, 2H, Ar-H, J = 7.6 Hz), 9.1 (bs, 1H, NH), 9.28 (bs, 1 H, OH).

Example 7: Preparation of the following compound 1g (4-methoxyphenylthiourea)

Yield 70%; White solid; mp 155-156 ° C; IR (neat): 3422, 3330, 3145, 1645, 1589, 1446 cm ⁻¹ ; ¹ H NMR (CDCl ₃ ): δ 3.85 (s, 3H, OCH ₃ ), 6.88 (d, 2H, Ar-H, J = 7.6 Hz), 7.22 (bs, 2H, NH ₂ ), 7.40 (d, 2H , Ar-H, J = 7.6 Hz), 9.10 (bs, 1H, NH).

Example 8: Preparation of the following compound 1h (3-methylphenylthiourea)

Yield 80%; White solid; mp 151-153 ° C .; IR (neat): 3421, 3161, 1621, 1538, 1458 cm ⁻¹ ; ¹ H NMR (CDCl ₃ ): δ 2.30 (s, 3H, Ar-CH ₃ ), 6.32 (bs, 2H, NH ₂ ), 7.11 (d, 2H, Ar-H, J = 7.6 Hz), 7.29-7.33 (m, 2H, Ar-H), 8.45 (bs, 1H, NH).

Example 9: Preparation of the following compound 9a (1,3-diphenylthiourea)

After dissolving 0.1 mol of aniline in a minimum amount of benzene, 6 ml of carbon disulfide and 14 ml of triethylamine were treated in an ice bath, and then the solution was filtered to give a solid, which was dissolved in 75 ml of chloroform. A mixture solution of 14 ml of triethylamine and 10.2 ml of ethyl chlorocarbonate was added dropwise and stirred for 1 hour. The chloroform solution was washed sequentially with 3N hydrochloric acid and water, and then dried over sodium sulfate. Then, the obtained organic phase was evaporated under reduced pressure to obtain a crude solid, and then recrystallized from ethanol to obtain isothiocyanate (compound of 8a). The isothiocyanate was dissolved in a minimum amount of acetonitrile (2 mL) and stirred while dropping into a solution containing 0.1 mL of aniline in acetonitrile (10 mL). The resulting solid was filtered, dried and purified by column chromatography to give 1,3-diphenylthiourea (Compound 9a) (see Reaction Scheme 2 above for reaction mechanism).

Yield 96%; White solid; mp 157-159 ^o C; IR (neat); 3200, 2850, 1475, 1455, 1190 cm ⁻¹ ; ¹ H NMR (CDCl ₃ ): δ 6.07 (bs, 1H, NH), 7.22 (m, 2H, Ar-H), 7.35 (m, 4H, Ar-H), 7.59 (m, 4H, Ar-H) , 7.82 (bs, 1 H, NH). HRMS calcd for C ₁₃ H ₁₂ N ₂ S m / z 228.0721, found 228.0724.

Example 10 The following compound 9b (1-phenyl-3- p -Tolylthiourea)

Using the reaction conditions described in Example 9, Compound 9b was obtained from p -toluidine (Compound 7b).

Yield 94%; White solid; mp 146-148 ^o C; IR (neat): 3200, 2850, 1470, 1457, 1320, 1280 cm ⁻¹ ; ¹ H NMR (CDCl ₃ ): δ 2.48 (s, 3H, CH ₃ ), 6.02 (bs, 1H, NH), 7.09 (d, J = 8.0 Hz, 2H, Ar-H), 7.21 (d, J = 8.4 Hz, 1H), 7.25-7.46 (m, 4H, Ar-H), 7.59 (t, J = 8.4 Hz, 2H, Ar-H), 7.79 (bs, 1H, NH). HRMS calcd for C ₁₄ H ₁₄ N ₂ S m / z 242.0878, found 242.0873.

Example 11: Preparation of the following compound 9c (1- (4-chlorophenyl) -3-phenylthiourea)

Using the reaction conditions described in Example 9, compound 9c was obtained from p -chloroaniline (compound 7c).

Yield 94%; White solid; mp 153-155 ^o C; IR (neat): 3200, 3020, 2980, 1620, 1500, 1475 cm ⁻¹ ; ¹ H NMR (CDCl ₃ ): δ 5.99 (bs, 1H, NH), 7.18 (d, J = 8.0 Hz, 2H), 7.27 (t, J = 8.4 Hz, 1H, Ar-H), 7.44 (t, J = 8.4 Hz, 2H, Ar-H). 7.82 (bs, 1 H, NH). HRMS calcd for C ₁₃ H ₁₁ ClN ₂ S m / z 262.0331, found 262.0327.

Example 12: Preparation of the following compound 9d (1- (4-methoxyphenyl) -3-phenylthiourea)

Using the reaction conditions described in Example 9 above, compound 9d was obtained from p -anisidine (compound 7d).

Yield 92%; White solid; mp 163-165 ^o C; IR (neat); 3200, 3173, 3020, 2920, 1620, 1580, 1480, 1450, 1325 cm ⁻¹ ; ¹ H NMR (CDCl ₃ ): δ 3.85 (s, 1H, OCH ₃ ), 6.02 (bs, 1H, NH), 6.99 (d, J = 7.8 Hz, 2H, Ar-H), 7.20 (m, 3H, Ar-H), 7.36 (t, J = 8.0 Hz, 2H, Ar-H), 7.59 (m, 2H, Ar-H), 7.84 (bs, 1H, NH). HRMS calcd for C ₁₄ H ₁₄ N ₂ OS m / z 258.0827, found 258.0823.

Example 13: Preparation of the following compound 9e (1- (4-hydroxyphenyl) -3-phenylthiourea)

Using the reaction conditions described in Example 9 above, Compound 9e was obtained from p -hydroxyaniline (Compound 7e).

Yield 87%; White solid; mp 191-193 ^o C; IR (neat): 3221, 3175, 2840, 1660, 1610, 1510, 1455, 1405 cm ⁻¹ ; ¹ H NMR (CDCl ₃ ): δ 5.98 (bs, 1H, NH), 6.88 (d, J = 8.0 Hz, 1H, Ar-H), 6.99 (d, J = 8.0 Hz, 2H, Ar-H), 7.21 (t, J = 8.0 Hz, 1H, Ar-H), 7.40-7.56 (m, 4H, Ar-H), 7.78 (bs, 1H, NH), 10.9 (s, 1H, OH). HRMS calcd for C ₁₃ H ₁₂ N ₂ OS m / z 244.0670, found 244.0667.

Example 14: Preparation of the following compound 9f (1-cyclohexyl-3-phenylthiourea)

Using the reaction conditions described in Example 9, compound 9f was obtained from cyclohexylamine (compound 7f).

Yield 93%; White solid; mp 149-151 ^o C; IR (neat): 3203, 3164, 2854, 1470, 1320, 1285, 1240 cm ⁻¹ ; ¹ H NMR (CDCl ₃ ): δ 1.07-2.07 (m, 10H, C ₆ H ₁₁ ), 3.68 (m, 1H), 5.91 (bs, 1H, NH), 7.13 (d, J = 8.0 Hz, 2H, Ar-H), 7.27 (t, J = 7.6 Hz, 1H, Ar-H), 7.45 (t, J = 8.0 Hz, 1H, Ar-H), 7.89 (bs, 1H, NH). HRMS calcd for C ₁₃ H ₁₈ N ₂ S m / z 234.1191, found 234.1195.

Example 15: Preparation of the following compound 9g (1-benzyl-3-phenylthiourea)

Using the reaction conditions described in Example 9 above, 9 g of the compound was obtained from benzylamine (compound 7 g).

Yield 96%; White solid; mp154-156 ^o C; IR (neat): 3200, 3167, 2927, 2750, 1620, 1609, 1500, 1457, 1430 cm ⁻¹ ; ¹ H NMR (CDCl ₃ ): δ 5.01 (s, 2H, NCH ₂ Ph), 5.99 (bs, 1H, NH), 7.06-7.18 (m, 5H, Ar-H), 7.23 (t, J = 8.0 Hz , 2H, Ar-H), 7.40 (t, J = 8.0 Hz, 1H, Ar-H), 7.44 (t, J = 8.0 Hz, 2H, Ar-H), 7.85 (bs, 1H, NH). HRMS calcd for C ₁₄ H ₁₄ N ₂ S m / z 242.0878, found 242.0873.

Example 16: Preparation of the following compound 9h (1-phenethyl-3-phenylthiourea)

Using the reaction conditions described in Example 9, compound 9h was obtained from phenylethylamine (compound 7h).

Yield 93%; White solid; mp 112-114 ^o C; IR (neat): 3258, 2925, 2680, 1667, 1605, 1454, 1327 cm ⁻¹ ; ¹ H NMR (CDCl ₃ ): δ 2.78 (t, J = 5.6 Hz, 2H, CH ₂ Ph), 3.87 (t, J = 6.0 Hz, 2H, NCH ₂ ), 5.99 (bs, 1H, NH), 7.09 -7.18 (m, 5H, Ar-H), 7.26 (t, J = 7.6 Hz, 2H, Ar-H), 7.37-7.49 (m, 3H, Ar-H), 7.87 (bs, 1H, NH). HRMS calcd for C ₁₅ H ₁₆ N ₂ S m / z 256.1034, found 256.1031.

Example 17: Preparation of the following compound 9i (1-phenyl-3- (3-phenylpropyl) thiourea)

Using the reaction conditions described in Example 9, compound 9i was obtained from phenylpropylamine (compound 7i).

Yield 97%; White solid; mp 130-132 ^o C; IR (neat); 3252, 3020, 2980, 2857, 1620, 1475, 1370, 1320 cm ⁻¹ ; ¹ H NMR (CDCl ₃ ): δ 1.90-1.96 (m, 2H, CH _2- ), 2.63 (t, J = 6.0 Hz, 2H, CH ₂ Ph), 3.69 (t, J = 6.4 Hz, 2H, NCH _2- ), 5.98 (bs, 1H, NH), 7.04-7.16 (m, 5H, Ar-H), 7.26 (t, J = 8.0 Hz, 2H, Ar-H), 7.32 (t, J = 8.0 Hz , 1H, 7.52 (t, J = 8.0 Hz, 2H, Ar-H), 7.79 (bs, 1H, NH) HRMS calcd for C ₁₆ H ₁₈ N ₂ S m / z 270.1191, found 270.1187.

Experimental Example

Measurement of melanogenesis inhibitory effect through cAMP / protein kinase A

Cell line melanoma B16 with alpha-melanosite stimulating hormone produces melanin through signal transduction of cAMP / protein kinase A (Rasmussen, N., et al. , Neuroendocrinol. Lett., 20: 265-282 , 1999; Scott, MC, et al., J. Cell. Sci. 115: 2349-55, 2002). All synthesized derivatives described above were tested for their inhibitory activity on melanin formation in melanoma B16 cell lines as shown in Table 1 below. The reason why the tyrosinase itself is not used in this experimental example is that these inhibitors have the purpose of reducing melanin formation. Melanin production by comparing the amount of melanin produced after simultaneously treating the compounds prepared in the above embodiments with alpha-melanosite stimulating hormone and the amount of melanin produced when the compounds of the examples were not treated Inhibitory effect was calculated.

Where

Mc is the melanin production amount of the control group which added only alpha-melanosite stimulating hormone;

Mo is the amount of melanin produced in cell lines cultured without the addition of alpha-melanosite stimulating hormone,

Mi represents the melanin production amount of the test group to which the compound (1a-1h, 9a-9i) and alpha-melanosite stimulating hormone obtained through the Example were added.

More specifically, 2,500 cell lines of melanoma B16 were dispensed per well of a 96-well plate, and then cultured in DMEM medium containing 10% placental serum at 37 ° C. and 5% CO ₂ for 24 hours. After incubation, each of the alpha-melanosite stimulating hormone (final concentration 1 nM) and the derivatives (1a-1h, 9a-9i) prepared in the examples were subjected to various sample concentrations (10 μM, 5 μM, 3 μM, 1 μM, 0.5 μM). , Or 0.05 μM) simultaneously and further incubated for 3 days, and then the amount of melanin released into the medium was quantified by measuring absorbance at a wavelength of 405 nm. All the compounds according to the present invention did not inhibit the growth of B16 cells at the measured concentrations of this example was confirmed that there is no cytotoxicity.

Cell line melanoma B16 cultured in DMEM medium containing 10% fetal placental serum without addition of alpha-melanosite stimulating hormone released melanin at 59 ± 1 μg / ml and alpha- in the same medium. When melanocyte stimulating hormone was added, melanin was released at 156 ± 2 μg / ml.

In the following, the process and results and the detailed description to the present invention based on Table 1 are presented.

compound R ₁ R ₂ R ₃ n ₁ n ₂ 10 μM
Inhibition rate at 3 μM
Inhibition rate at IC ₅₀ value (μM) Clog P 1a H H 4-H 0 0 30 - 54.4 0.745 ¹⁾ 1b H H 4-CH ₃ 0 0 83 - 5.5 1.244 ¹⁾ 1c H H 4-CH ₂ CH ₃ 0 0 88 - 4.9 1.213 ¹⁾ 1d H H 4-C (CH ₃ ) ₃ 0 0 95 - 3.7 2.571 ¹⁾ 1e H H 4-Cl 0 0 84 - 4 1.764 ¹⁾ 1f H H 4-OH 0 0 70 - 6.3 0.073 ¹⁾ 1 g H H 4-OCH ₃ 0 0 80 - 5.6 0.847 ¹⁾ 1h H H 3-CH ₃ 0 0 78 - 5 1.244 ¹⁾ 9a -Ph H H 0 0 88 40 4.5 2.170 ²⁾ 9b -Ph (4-CH ₃ ) H H 0 0 79 24 6.3 2.669 ²⁾ 9c -Ph (4-Cl) H H 0 0 > 100 47 3.4 3.185 ²⁾ 9d -Ph (4-OCH ₃ ) H H 0 0 59 39 8.2 2.272 ²⁾ 9e -Ph (4-OH) H H 0 0 24 10 20.7 1.503 ²⁾ 9f -cyclohexyl H H 0 0 > 100 62 2.4 2.192 ²⁾ 9g -Ph H H One 0 91 47 4.7 2.649 ²⁾ 9h -Ph H H 2 0 > 100 87 2.2 2.978 ²⁾ 9i -Ph H H 3 0 > 100 89 1.8 3.357 ²⁾ Kojic acid 261 - Arbutin 151 - ^{1) The} ClogP value is calculated by Chemdraw9.0.
^{2) The} ClogP value is calculated by Chemdraw11.0.

As can be seen from Table 1, the result of 50% inhibition concentration of kojic acid and arbutin in the same experimental model was 261μM and 151μM, respectively, which indicates that the arylthiourea derivative according to the present invention has excellent inhibitory efficacy. Phenylthiourea (Compound 1a) showed good inhibition rate for melanin formation (30% inhibition at 10 μM, IC ₅₀ = 54.4 μM, Clog P = 0.745). To investigate the role of substituents on the phenyl ring of phenylthiourea, the inventors introduced various substituents at different positions of the phenyl ring. The p-methyl substituted analog (Compound 1b) showed much better inhibition (83% inhibition at 10 μM, IC ₅₀ = 5.5 μM, Clog P = 1.244) than the phenylthiourea (Compound 1a) described above. Testing with increasing volume at this location showed 4-ethylphenylthiourea (Compound 1c, 88% inhibition at 10 μM, IC ₅₀ = 4.9 μM, Clog P = 1.773), 4-tert-butylphenylthiourea (Compound 1d , 95% inhibition at 10 μM, IC ₅₀ = 3.7 μM, Clog P = 2.571) and 4-chlorophenylthiourea (Compound 1e, 84% inhibition at 10 μM, IC ₅₀ = 4.0 μM, Clog P = 1.764) Showed. To investigate the effects of hydrophobic or predominantly electron donor groups, we prepared hydroxyl analogs and methoxy compounds. These 4-hydroxy analogs (Compound 1f, 70% inhibition at 10 μM, IC ₅₀ = 6.3 μM, Clog P = 0.073) and 4-methoxy compounds (1 g of Compound, 80% inhibition at 10 μM, IC ₅₀ = 5.6 μM, Clog P = 0.847) also showed significantly higher inhibition rate than the phenylthiourea (Compound 1a).

In addition, as shown in Table 1, when the phenyl group is bonded to both amine groups of the thiourea, it showed an excellent inhibitory effect on melanin formation than phenylthiourea (Compound 1a). That is, a phenyl group or a phenyl group R ₁ (compound 9a, 88% inhibition at _{10μM, IC 50 = 4.5μM, Clog} P = 2.170) of the aryl thiourea derivatives of the formula I, wherein the R ₁ group has a substituent, p - Tolyl (79% inhibition at compound 9b, 10 μM, IC ₅₀ = 6.3 μM, Clog P = 2.669), chlorophenyl (compound 9c,> 100% inhibition at 10 μM, IC ₅₀ = 3.4 μM, Clog P = 3.185), or In the case of methoxyphenyl (Compound 9d, 59% inhibition at 10 μM, IC ₅₀ = 8.2 μM, Clog P = 2.272), it showed excellent melanin formation inhibitory effect.

In addition, compounds having an alkyl group in the R ₁ group of the arylthiourea derivative compound of Formula I are bonded to the nitrogen atom of the amine, that is, 1-benzyl-3-phenylthiourea (9 g, 91% inhibition rate at 10 μM) , IC ₅₀ = 4.7 μM, Clog P = 2.649), 1-phenethyl-3-phenylthiourea (9 h, 87% inhibition at 10 μM, IC ₅₀ = 2.2 μM, Clog P = 2.978), 1-phenyl-3- (3-phenylpropyl) thiourea (9i, 89% inhibition at 10 μM, IC ₅₀ = 1.8 μM, Clog P = 3.357) and compounds in which the R ₁ group is cycloalkyl, such as 1-cyclohexyl-3-phenylthio Urea (9f, greater than 100% inhibition at 10 μM, IC ₅₀ = 2.4 μM, Clog P = 2.191) also showed excellent inhibitory efficacy against melanin formation.

Formulation Example 1

Preparation of nourishing cream

A nutritive cream containing one of Compounds 1b to 1h and 9a to 9i as an active ingredient in the composition shown in Table 2 was prepared according to a conventional method.

Ingredients Addition amount (% by weight) One of the compounds 1b to 1h and 9a to 9i; 0.5 Liquid paraffin 5.0 Squalane 10.0 glycerin 10.0 beeswax 5.0 Glyceryl stearate 2.0 Stearic acid 3.0 Polysorbate 60 1.5 Sorbitan sesquistearate 0.3 Carboxy polymer 0.3 Preservative, fragrance, pigment, pH adjuster Suitable amount Purified water up to 100

Formulation Example 2

Preparation of Soft Cosmetics

In the composition as shown in Table 3 below, the flexible longevity containing one of compounds 1b to 1h and 9a to 9i as an active ingredient was prepared according to a conventional method.

Ingredients Addition amount (% by weight) One of the compounds 1b to 1h and 9a to 9i; 0.2 Tocopheryl acetate 5.0 ethanol 10.0 glycerin 5.0 1,3-butyl glycol 5.0 Carboxy polymer 0.3 Preservative, fragrance, pigment, pH adjuster Suitable amount Purified water up to 100

Formulation Example 3

Pack

A pack containing one of Compounds 1b to 1h and 9a to 9i as an active ingredient in the composition shown in Table 4 was prepared according to a conventional method.

Ingredients Addition amount (% by weight) One of the compounds 1b to 1h and 9a to 9i; 1.0 Polyvinyl alcohol 15.0 ethanol 10.0 glycerin 5.0 Polyoxyethylene oleylethyl 1.0 Preservative, fragrance, pigment, pH adjuster Suitable amount Purified water up to 100

Formulation Example 4

Ointment preparation

To the composition as shown in Table 5, ointments containing one of compounds 1b to 1h, and 9a to 9i as an active ingredient were prepared according to a conventional method.

Ingredients Addition amount (% by weight) One of the compounds 1b to 1h and 9a to 9i; 2.0 Diethyl sebacate 8.0 Prepayment 5.0 Polyoxyethylene oleyl ether phosphate 6.0 Paraoxybenzoic Acid Ester Suitable amount vaseline up to 100

Skin irritation test

In a closed patch test method, 20 healthy men and women were checked for the skin irritation degree of the nourishing cream of Formulation Example 1 for 24 hours.

Claims (7)

A skin whitening composition containing the arylthiourea derivative of the formula (I) as an active ingredient:
(I)

In Formula I,
R ₁ represents C ₃ to C ₈ cycloalkyl, or phenyl with or without substituent X, which substituent C is C ₁ to C ₃ alkyl, halogen, hydroxy or C ₁ to C ₃ alkoxy,
R ₂ represents hydrogen or C ₁ to C ₆ alkyl,
R ₃ represents hydrogen, C ₁ to C ₉ alkyl, C ₁ to C ₆ alkoxy, hydroxy, halogen or C ₁ to C ₆ alkylamine,
n ₁ and n ₂ each independently represent 0, 1, 2, 3 or 4, except when R _1, R ₂ , and R ₃ are all hydrogen and n ₁ and n ₂ are both 0 ). The method of claim 1,
The arylthiourea derivative compound of Formula (I),
1,3-diphenylthiourea, 1-phenyl-3- para -tolylthiourea, 1- (4-chlorophenyl) -3-phenylthiourea, 1- (4-methoxyphenyl) -3-phenylthio Urea, 1- (4-hydroxyphenyl) -3-phenylthiourea, 1-cyclohexyl-3-phenylthiourea, 1-benzyl-3-phenylthiourea, 1-phenethyl-3-phenylthiourea, And 1-phenyl-3- (3-phenylpropyl) thiourea; at least one compound selected from the group consisting of: A pharmaceutical composition for the prevention and treatment of skin hyperpigmented skin diseases, comprising the composition for skin whitening according to claim 1. The method of claim 3, wherein
Pharmaceutical composition for the prevention and treatment of skin hyperpigmentation disease, characterized in that the formulation of the composition is a cream, ointment, gel, lotion, or patch. A whitening cosmetic composition comprising a composition for skin whitening according to claim 1 as an active ingredient. 6. The method of claim 5,
The whitening cosmetic composition is an essence, whitening cream, lotion, emulsion, pack, general cosmetics, skin milk, cream, serum, cosmetic soap, softening cosmetics, hair tonic, systemic cleaner or oil gel, characterized in that the whitening cosmetic composition. The method according to claim 6,
The whitening cosmetic composition further comprises one or more additives selected from the group consisting of oils, water, surfactants, moisturizers, thickeners, chelating agents, pigments, preservatives and fragrances.