KR20020049341A - Novel phenyl derivatives, the process for preparing them and the pharmacological composition and the cosmetic composition containing them - Google Patents

Abstract

PURPOSE: Provided are novel phenyl ring derivatives which has tyrosinase inhibiting and lightening effects, a production method thereof, and pharmaceutical and cosmetic composition containing the same. CONSTITUTION: The phenyl ring derivatives have a chemical structure represented by the formula I, in which R1, R2 and R3 are independently hydrogen, halogen or C1-4 alkoxy, X is a carbon atom or nitrogen atom, M is (i) 2-pyridyl, 3-pyridyl or 4-pyridyl, substituted with R4, (ii) 2-puryl or 3-puryl, substituted with R4, (iii) a phenyl ring represented by the formula I-1, in which R4 is a hydrogen, halogen, hydroxy or C1-4 alkoxy group, R5, R6, R7, R8 and R9 are a hydrogen or C1-4 alkoxy group, provided that R1 to R9 may be substituted.

Description

Novel phenyl ring derivatives, preparation methods thereof, and pharmaceutical compositions and cosmetic compositions comprising the same {NOVEL PHENYL DERIVATIVES, THE PROCESS FOR PREPARING THEM AND THE PHARMACOLOGICAL COMPOSITION AND THE COSMETIC COMPOSITION CONTAINING THEM}

Skin melanogenesis is an increase in melanogenesis by melanocytes in melanocytes (melanocytes) within the skin cells, and a large amount of melanin is transferred to keratinocytes. epidermis). Hyperpigmentation of the skin due to blemishes, freckles, skin darkening after skin inflammation, senile pigment spots, etc. can cause not only cosmetic discomfort but also negative psychological effects on the person, leading to discomfort in social activities. Due to the increased demand, cosmetics and pharmaceutical products for whitening are being marketed to prevent and improve them.

The research of whitening agent as a cosmetics has a history of less than 10 years, and the necessity is gradually increasing as skin blackening is recognized as skin aging by ultraviolet rays along with the improvement of living standard in the eastern region which prefers emotionally white skin. Since 1990, arbutin, kojic acid, vitamin C and derivatives thereof have been developed, and whitening cosmetics containing them are commercially available, but the actual clinical effect is not satisfactory.

As a medicine for improving excessive pigmentation, hydroquinone, sulfure, azelaic acid, retinoic acid and the like are used, but they have great irritation and toxicity.

Recently, in order to develop a whitening cosmetics having relatively low toxicity and being natural, it has been found that several herbal medicines are effective for whitening. As an example, Korean Patent Publication No. 1991-8660 discloses a whitening cosmetic material containing a mulberry extract, and Korean Patent Publication No. 1991-11645 discloses a whitening cosmetic containing a extract of Hojang-geun. No. 1993-28783 discloses a whitening composition containing an extract of lettuce extract, and recently, a whitening cosmetic containing mulberryine isolated from mulberry is disclosed in Korean Patent Laid-Open Publication No. 1997-47260.

Early development of the whitening agent focused on the antioxidant function of tyrosine by tyrosinase, the first step in the melanin pigmentation process, and the development of the whitening agent, which is important in screening tyrosinase activity inhibitors in vitro It was a means. In particular, tyrosinase used is derived from mushrooms, and there are many differences in similarity to humans. If the selected whitening agent is applied to humans, there is a problem of skin penetration, cytotoxicity, and formulation stability. The effects are not enough, but arbutin, kojic acid, and vitamin C found in this process are still widely used.

Therefore, in the present invention, on the basis of the tyrosinase inhibitory effect, at the same time, UV protection and SOD-like activity of skin damage prevention function due to reduction of antioxidant melanin, cytotoxicity and melanin production inhibitor using mela-A cell line As a method of measurement, the new problem whitening agent was selected by supplementing the above problems.

The present inventors, while searching for a plant that is expected to have excellent whitening activity from natural resources existing in nature, by separating and purifying the extract fraction of baekryepi, the root of Morus alba L., a Moraceae plant It was confirmed that oxyresveratrol strongly inhibited the activity of tyrosinase (IC ₅₀ = 0.1 µg / ml), which is an enzyme related to whitening, among the obtained Stilbene series compounds.

Morus cortex, Morus alba L., is a root skin removed from the cork layer of Morus alba L., which grows around the village and is cultivated all over the country. It is used in the treatment of the back, clinically known diuretic action, lowering blood pressure, etc. The ingredients known so far are Umbelliferone, Scopoletin, Morusin, Mulberin ) Has been reported.

Et oxy resveratrol is present in the Mori Cortex gods in recent tie or when the inhibitory effect that is better than kojic acid, which is widely used in the cosmetics (Shin. NH, et al .; Biochem. Biophys. Res. Commun. 243 , p801-803, 1998).

However, the amount of oxyresveratrol component is not present in the epidermis, and an effective synthesis method has not yet been established. Currently, the extract of baeksipeun root containing oxyresveratrol is used for the preparation of whitening cosmetics. Oxyresveratrol is not expected to have high cell permeability due to the high polarity due to the hydroxyl group of the phenol ring itself.

Thus, in the present invention, various phenol ring derivatives having lower polarity than oxyresveratrol were synthesized by a simple method and used as an optimal functional cosmetic material.

Accordingly, the present inventors synthesized various types of oxyresveratrol derivatives through various synthesis methods to solve the problems of known oxyresveratrol compounds having tyrosinase inhibitory activity and to obtain compounds having more excellent whitening activity. The present invention was completed by finding out that the novel phenyl ring derivative of the general formula (I) meets the above object.

Accordingly, an object of the present invention is to provide a phenyl ring derivative having a novel chemical structure.

The present invention also aims to provide a novel process for the preparation of compounds of general formula I.

The present invention also aims to provide a pharmaceutical composition comprising a compound of Formula I useful for the treatment of diseases caused by melanin overproduction.

It is also an object of the present invention to provide a whitening cosmetic composition comprising a compound of formula I which inhibits tyrosinase activity and exhibits excellent whitening activity.

1 is a graph comparing the tyrosinase enzyme inhibitory effects of the compounds of the present invention and the control compounds by concentration,

2 is a spectrum of UV-A and UV-B of Compound 14,

3 is a spectrum of UV-A and UV-B of Compound 15,

4 is a spectrum of UV-A and UV-B of the compound Oxyresveratrol,

5 is a spectrum in UV-A and UV-B of kojic acid,

6 is a graph showing the SOD-like activity (%) for each sample concentration.

The present invention relates to a phenyl ring derivative of the general formula (I).

Formula 1

Where

R _1, R _2, and R ₃ are each independently hydrogen, halogen, C _1-4 alkoxy;

X is a carbon atom or a nitrogen atom;

M is

(Iii) a 2-pyridyl, 3-pyridyl or 4-pyridyl group substituted with R ₄ ;

(Ii) a 2-furyl or 3-furyl group substituted with R ₄ ;

(Iii) a phenyl ring represented by the following general formula (I-1);

Formula 2

Wherein R ₄ is hydrogen, halogen, hydroxy, or a C _1-4 alkoxy group;

The R _5, R _6, R _7, R ₈ and R ₉ groups are hydrogen or a C _1-4 alkoxy group except that neither of the R ₁ to R ₉ substituents are hydrogen.

Preferred compounds in the above general formula I compounds include those in which R _1, R ₂ and R ₃ are hydrogen or a methoxy group, the X substituent is a nitrogen atom and M is a pyridyl, furyl or phenyl ring substituted or substituted with a methoxy group. Further preferred compounds are those wherein R ₁ and R ₃ are hydrogen atoms, R ₂ is a methoxy group, the X substituent is a nitrogen atom and M is a phenyl ring substituted with a methoxy group at the m-position.

The novel compounds of general formula I according to the invention can be prepared as follows.

First, when (a) the X substituent is a carbon atom, the benzyl quaternary phosphine halide derivative of the general formula (IIa) or the phosphonate derivative of the general formula (IIb) below is a benzaldehyde compound of the general formula (III), By Wittig Reaction with 3-furalaldehyde compound or 4-pyridylaldehyde compound of Formula V, phenyl ring derivative compounds of Formula Ia, Formula Ib and Formula Ic can be obtained, respectively.

In addition, (b) when the X substituent is a nitrogen atom, it can be prepared by reacting the benzaldehyde compound of formula (VI) with the phenylamine compound of formula (VII) to obtain a phenyl ring derivative of formula (Id).

In addition, the compound of formula (Ia) obtained in a cis and trans mixture can be selectively obtained by adding a trace amount of iodine and heating in a benzene solvent to obtain a compound of formula (Iat) as a trans.

In the above formula,

R in general formula (IIa) represents an alkyl, cycloalkyl group, phenyl group or the like capable of causing a Wittig reaction; R ′ in formula IIb represents haloalkyl, alkyl, phenyl, alkoxy, phenoxy and the like; X 'represents a halogen atom such as bromine, chlorine, iodine; R ₁ to R ₉ are the same as defined above.

It is well known in the art to obtain a stilbene derivative having a double bond between the benzene rings by reacting the benzyl tertiary phosphine halide derivative of Formula IIa or a benzofonate derivative of IIb with a benzaldehyde compound of Formula III. By Wittig reaction or Horner-Emons reaction, the reaction is carried out for 30 minutes to 24 hours in the presence of a strong base such as sodium hydroxide, potassium hydroxide and the like and a catalyst such as quaternary amine salt. Preferably, 0.1 equivalent of tetrabutylammonium bromide, which is a catalyst, is reacted at 25 ° C. in the presence of 50% aqueous sodium hydroxide solution for one day. This was carried out by reacting a benzyl tertiary phosphine halide derivative of Formula IIa or a phosphonate derivative of Formula IIb with a 3-furalaldehyde compound of Formula IV or a 4-pyridylaldehyde compound of Formula V, respectively. The same applies to the reaction for obtaining the phenyl ring derivative compound of the general formula (Ic).

The reaction for forming an imide bond between two benzene rings by reacting the benzaldehyde compound of Formula VI with the phenylamine compound of Formula X is carried out for 1 hour to 24 hours at a temperature of 80 to 110 ° C. in the presence of a nonpolar solvent. To perform. Preferably, the aldehyde of Formula VI and 1 equivalent of aniline of Formula X are reacted at 110 ° C. for 12 hours using a dean-stark apparatus in the presence of a toluene solvent.

On the other hand, the starting materials of the general formula (IIa) and (IIb) used to prepare the compound of the general formula (Ia) to obtain the secondary alcohol of the general formula (VII) through a reduction reaction to add a suitable reducing agent from the general formula (VI) compound, The halide of the general formula (VII) is obtained through a quaternary phosphine halogenation reaction, followed by addition of a tertiary phosphine or a fifth phosphonate.

Finally, the reduction step of obtaining the trans is carried out for 5 minutes to 2 hours at a temperature of -5 to 30 ° C. in the presence of an ethereal solvent with a reducing agent such as lithium aluminum hydride (LAH). Preferably, 1 equivalent of aldehyde of Formula VI and 1.5 equivalent of LAH are reacted for 1 hour at 0 ° C. in the presence of a tetrahydrofuran and a diethyl ether solvent.

The step of obtaining a halide of the general formula (VII) through the quaternary phosphine halogenation reaction is a tertiary phosphine such as triphenyl phosphine in the presence of a polar solvent such as acetonitrile, chloroform and carbon tetrabromide 25 ~ 70 ℃ The reaction is carried out for 30 minutes to 24 hours under the temperature of. Preferably, 1.1 equivalents of triphenyl phosphine and 1.1 equivalents of carbon tetrahydrobromide are heated to reflux for 2 hours in the presence of an acetonitrile solvent in the alcohol of general formula (VII), and the above-mentioned tertiary phosphine is added to The process for obtaining the starting material is carried out for 1 to 12 hours at 80 to 130 ℃ while heating under reflux in the presence of a non-polar solvent such as benzene, toluene, xylene and the like. Preferably, the halide body of Formula (VII) and 1.5 equivalents of triphenyl phosphine are heated under reflux for 2 hours in the presence of a benzene solvent.

In addition, the step of obtaining a single chain isomer of transchain Formula Iat from a mixture of Formula Iac and Iat reacts a halogen compound such as iodine (I ₂ ) with a mixture of Formula Iac and Iat, preferably Formula Iac. 0.01 equivalent of iodine is heated to reflux for 2 hours in the presence of a heptane solvent.

The preparation process for obtaining the target substances of the general formulas Ia, Iat, Ib, and Ic from the starting materials of formula II is described in the methods well known in the art (Zang. J. et al. Org. Chem. 63 , 8125-8132, 1998, Pettit. G., J. Med. Chem. 38, 1666-1672, 1995, and Orsini et al. Carbohydr.Res. 301, 95-109, 1997), and the process for obtaining the target substance of general formula Id from the starting material of formula VI is a method known in the art well known in the art. Takeshi. N., Org. Lett. 7, 985-988, 1999; and Miyano. S., J. Chem. Soc. Perkin Trans. 1.1146, 1976. ], All of which are incorporated herein by reference.

The preparation method for preparing the compounds of formulas (Ia), (Ib), (Ic), (Id) and (Iat) prepared according to the above-described methods is shown in Schemes 1 to 5 below.

Scheme 1 .

Preparation of Formula Ia

Scheme 2.

Preparation of Formula Ib

Scheme 3.

Preparation of Formula Ic

Scheme 4.

Preparation of Formula Id

Scheme 5.

Preparation of Formula Iat

In the above formula,

R, X 'and R ₁ to R ₉ are the same as defined above.

Representative examples of compounds of Formula I prepared according to the methods described above are shown in Table 1 below.

The inventors of the formula 1 compounds tyrosinase inhibitory activity, UV blocking effect test, SOD-like activity test involved in melanin biosynthesis And melan-A cell line (melan-A cell) experiments through the melanin production inhibitory effect experiments, such as the results, it was found that the compound of formula 1 is excellent in whitening activity but low in cytotoxicity.

Therefore, the compound of the present invention having such an effect can be applied to medicines and cosmetics, the amount and method of application may vary depending on the formulation and purpose of use.

For example, the compound of Formula 1 may be mixed with a conventional cosmetic preparation base material to prepare a whitening cosmetic composition.

In the present invention, the compound of Formula 1 alone or in combination with other whitening ingredients is used in an amount of 0.0001% to 30.00% by weight based on the skin cosmetic composition. In addition, the base material for preparing the cosmetic composition is purified water, monohydric or polyhydric alcohol fats and oils, surfactants, and the like on the emulsion, and other additives may include flavoring agents, colorants, preservatives, etc. in the composition. In the solubilized composition, purified water, surfactants, monohydric and polyhydric alcohols, and the like, flavoring agents, colorants, and preservatives are used.

In skin cosmetics, creams and lotions may be prepared in W / O emulsions or O / W emulsions, respectively, and the type of emulsifier used may be anionic form, cationic form, nonionic form, or the like.

In addition, in order to treat diseases associated with the oversynthesis of melanin in humans or animals, the compounds of formula 1 of the present invention may be mixed with a pharmaceutically acceptable carrier as an active ingredient to prepare a composition for preventing and treating melanin-related diseases. have. The pharmaceutical composition may further include conventionally used excipients, disintegrants, sweeteners, lubricants, flavoring agents, etc., tablets, capsules, powders, granules, suspensions, emulsifiers, syrups, It may be formulated in unit dosage forms or in multiple dosage pharmaceutical formulations, such as liquid or parenteral formulations.

Prophylactic and therapeutic compositions for inhibiting melanin synthesis of the present invention can be parenterally or orally administered according to the desired method, the amount of 0.01 to 10g per 1kg body weight, preferably 1 to 5g as an active ingredient per day It may be administered in divided doses of 1 to several times. Dosage levels for a particular patient may vary depending on the patient's weight, age, sex, health condition, diet, time of administration, method of administration, rate of excretion, and severity of disease.

The present invention is illustrated by the following examples and test examples, but is not limited thereto.

Preparation Example 1 Preparation of 3,4,5-trimethoxybenzyl Alcohol

66.5 mg (0.39 mM) of 3,4,5-trimethoxybenzaldehyde was dissolved in 6 ml of an ether: tetrahydrofuran 2: 1 solution, followed by addition of 60 mg of lithium aluminum hydride. The mixture was stirred for 1 hour in the open state and cooled in an ice bath. Saturated aqueous sodium sulfate solution was slowly added until white precipitates were formed, and the resulting white precipitates were filtered and the ether layer in the solvent was separated. The residue obtained by evaporation under reduced pressure was purified by silica gel column chromatography (hexane / ethyl acetate = 2/1) to give 46 mg (60%) of 3,4,5-trimethoxybenzyl alcohol.

Preparation Example 2 Preparation of 3,4,5-trimethoxybenzyl bromide

46 mg (0.23 mM) of 3,4,5-trimethoxybenzyl alcohol obtained in Preparation Example 1, 1.1 equivalents of carbon tetrabromide and 1.1 equivalents of triphenylphosphine were dissolved in 3 ml of acetonitrile and refluxed under nitrogen gas. The residue obtained by evaporation under reduced pressure was purified by silica gel filtration chromatography (dichloromethane _/ methanol = 100/1) to obtain 37.6 mg (63%) of 3,4,5-trimethoxybenzyl bromide.

Preparation Example 3 Preparation of 3,4,5-trimethoxybenzyl triphenylphosphonium bromide

37.6 mg (0.145 mmol) of 3,4,5-trimethoxybenzyl bromide obtained in Preparation Example 2 is dissolved in 2.9 ml of anhydrous benzene, and then 91.9 mg (1.5 equivalents) of triphenylphosphine is added. After refluxing for 2 hours, the mixture was cooled to form a precipitate, and filtered to give 54.5 mg (72%) of 3,4,5-trimethoxybenzyl triphenylphosphonium bromide.

Preparation Example 4 Preparation of 3,5-dimethoxybenzyl triphenylsponium bromide

3 g (13.2 mM) of 3,5-dimethoxybenzyl bromide was dissolved in 44 ml of anhydrous benzene, and then 8.38 g (1.5 equiv) of triphenylphosphine were added. After refluxing for 2 hours, it was cooled to form a precipitate and filtered to give 6.592 g (100%) of 3,5-dimethoxybenzyl triphenylphosphonium bromide.

Example 1 : Preparation of 3,4,5,3 ', 5'-pentamethoxy stilbenes (Compound 1)

6.4 mg (0.038 mM) of 3,4,5-trimethoxybenzyl triphenylphosphonium bromide obtained in Preparation Example 3 and 6.4 mg (1 equivalent of 3,5-dimethoxybenzaldehyde) as a catalyst, 1 mg of tetrabutylammonium bromide After dissolving in 1.28 ml of methylene dichloride, 0.11 ml of 50% aqueous sodium hydroxide solution was slowly added. After stirring overnight, 10 ml hydrous was added and extracted with 20 ml methylenedichloride. The organic layer was dried with magnesium sulfate, and then the residue obtained by evaporation under reduced pressure was purified by silica gel column chromatography to obtain 12 mg (96%) of 3,4,5,3 ', 5'-pentamethoxy stilbene (1 )

¹ H-NMR (CD ₃ COCD ₃ ): δ6.90 (q), 6.67 (s), 6.59 (d), 6.44 (d), 6.38 (d), 6.33 (t), 6.25 (t), 3.85 ( s), 3.80 (s), 3.76 (d), 3.61 (d).

Example 2 Preparation of 3,5,2 ', 4'-Tetramethoxy Stilbene (Compound 2)

Obtained in Preparation Example 4 200 mg (0.407 mM) of 3,5-dimethoxybenzyl triphenylphosphonium bromide and 68 mg (1.1 equiv) of 2,4-dimethoxybenzaldehyde were dissolved in 8.13 ml of methylene dichloride together with 10 mg of tetrabutylammonium bromide as a catalyst. 1.15 ml of 50% aqueous sodium hydroxide solution was slowly added. After stirring overnight, 10 ml aqueous solution was added and extracted with 20 ml methylenedichloride. The organic layer was dried over magnesium sulfate, and the residue obtained by evaporation under reduced pressure was subjected to silica gel column chromatography (CH).₂Cl₂91.3 mg (75%) of 3,5,2 ', 4'-tetramethoxy stilbene (2) Respectively obtained.

Example 3 Preparation of 3,5,3 ', 5'-Tetramethoxy Stilbene (Compound 3)

100 mg (0.203 mM) of 3,5-dimethoxybenzyl triphenylphosphonium bromide obtained in Preparation Example 4 and 33.7 mg (1 equivalent) of 3,5-dimethoxybenzaldehyde were mixed with 5 mg of tetrabutylammonium bromide as a catalyst. After dissolving in 4.06 ml of dichloride, 0.57 ml of 50% aqueous sodium hydroxide solution was slowly added. After stirring overnight, 10 ml brine solution is added and extracted with 20 ml methylenedichloride. The organic layer was evaporated with magnesium sulfate, and the residue obtained by evaporation under reduced pressure was purified by silica gel column chromatography (CH ₂ Cl ₂ ) to obtain 41.2 mg (68%) of 3,5,3 ', 5'-tetramethoxy steel. Ben (3) was obtained.

Example 4 Preparation of 3,5,3 ', 4'-Tetramethoxy Stilbene (Compound 4)

Obtained in Preparation Example 4 200 mg (0.407 mM) of 3,5-dimethoxybenzyl triphenylsporium bromide and 61 mg (0.9 equiv) of 3,4-dimethoxybenzaldehyde were dissolved in 8.13 ml of methylene dichloride together with 10 mg of tetrabutylammonium bromide as a catalyst. 1.15 ml of 50% aqueous sodium hydroxide solution was slowly added. After stirring overnight, 10 ml aqueous solution was added and extracted with 20 ml methylenedichloride. The organic layer was dried over magnesium sulfate, and the residue obtained by evaporation under reduced pressure was subjected to silica gel column chromatography (CH).₂Cl₂Purification to give 80.6 mg (66%) of 3,5,3 ', 4'-tetramethoxy stilbene (4), respectively.

Example 5 Preparation of 3,5,4'-Trimethoxy Stilbene (Compound 5)

Obtained in Preparation Example 4 200 mg (0.407 mM) of 3,5-dimethoxybenzyl triphenylsporium bromide and 44.5 ml (0.9 equiv) of para-anisaldehyde were dissolved in 8.13 ml of methylenedichloride with 10 mg of tetrabutylammonium bromide as a catalyst and then 50% hydroxide. 1.15 ml of aqueous sodium solution was added slowly. After stirring overnight, 10 ml aqueous solution was added and extracted with 20 ml methylenedichloride. The organic layer was dried over magnesium sulfate, and the residue obtained by evaporation under reduced pressure was subjected to silica gel column chromatography (CH).₂Cl₂Purification to give 80.2 mg (73%) of 3,5,4'-trimethoxy stilbene (5), respectively.

Example 6 Preparation of 3,4,5,3 ', 5'-Pentamethoxy Transtilbene (Compound 6)

12 mg (0.037 mmol) of 3,4,5,3 ', 5'-pentamethoxystilbene was dissolved in 2 ml heptane and a small amount of I ₂ was added and refluxed for 2 hours. Purified molecular formula (C ₁₉ O ₅ H ₂₂ ) 3,4,5,3 ', 5'-pentamethoxy transstilbene (6) 12 mg by removing I ₂ by silica gel column chromatography (CH ₂ Cl ₂ ) method (100%) was obtained.

¹ H-NMR (CD ₃ COCD ₃ ): δ 7.00 (s, 1H), 6.96 (s, 1H), 6.74 (s, 2H), 6.67 (d, 2H), 6.40 (t, 1H), 3.92, 3.87, 3.84 (s, 15 H, -OCH ₃ ).

¹³ C-NMR (CDCl ₃ ): δ 161.00, 153.41, 139.22, 132.85, 130.42, 129.14, 106.70, 104.51, 99.99, 60.96, 56.14, 55.36.

MS: 330.37 (M ⁺ )

Example 7 Preparation of 3,5,2 ', 4'-Tetramethoxy Transtilbene (Compound 7)

91.3 mg (0.304 mM) of 3,, 5,2 ', 4'-tetramethoxystilbene is dissolved in 10 ml heptane and a small amount of I ₂ is added and refluxed for 2 hours. 91.3 mg of molecular formula (C ₁₈ O ₄ H ₂₀ ) 3,5,2 ', 4'-tetramethoxy transstilbene compound purified by removing I ₂ by silica gel column chromatography (CH ₂ Cl ₂ ) method (100%) was obtained.

¹ H-NMR (CDCl ₃ ): δ 7.42 (d, 1H), 7.28 (d, 1H), 6.87 (d, 1H), 6.59 (d, 2H), 6.44 (q, 1H), 6.39 (d, 1H), 6.29 (t, 1H), 3.80 (s, 3H, -OCH ₃ ), 3.76 (s, 3H, -OCH ₃ ), 3.76 (s, 6H, -OCH ₃ ).

¹³ C-NMR (CDCl ₃ ): δ 160.88, 160.61, 158.10, 140.37, 127.35, 126.98, 123.85, 119.33, 107.08, 104.99, 104.38, 99.41, 98.50, 97.50, 55.51, 55.40, 55.36, 55.22

MS: 300.34 (M ⁺ )

Example 8 Preparation of 3,5,3 ', 5'-Tetramethoxy Transtilbene (Compound 8)

Dissolve 41.2 mg (0.137 mM) of 3,, 5,3 ', 5'-tetramethoxystilbene in 5 ml heptane, and add trace of I ₂ to reflux overnight. 29.3 mg of molecular formula (C ₁₈ O ₄ H ₂₀ ) 3,5,3 ', 5'-tetramethoxy transstilbene compound purified by removing I ₂ by silica gel column chromatography (CH ₂ Cl ₂ ) method (71%) obtained.

¹ H-NMR (CDCl ₃ ) δ: 6.94 (s, 2H), 6.59 (d, 4H), 6.33 (t, 2H), 3.77 (s, 12H, —OCH ₃ ).

¹³ C-NMR (CDCl ₃ ) δ: 160.99, 139.16, 129.20, 104.65, 100.14, 55.38

MS: 300.34 (M ⁺ )

Example 9 Preparation of 3,5,3 ', 4'-Tetramethoxy Transtilbene (Compound 9)

After dissolving 80.6 mg (0.269 mM) of 3,5,3 ', 4'-tetramethoxy stilbene derivative in 8 ml heptane, a trace amount of iodine was added and refluxed for 6 hours. 50.8 mg (63) of molecular formula (C ₁₈ O ₄ H ₂₀ ) 3,5,3 ', 4'-tetramethoxy transstilbene purified by iodine removal by silica gel column chromatography (CH ₂ Cl ₂ ) %) Was obtained.

¹ H-NMR (CDCl ₃ ) δ: 6.99 (s, 2H), 6.96, 6.94, 6.86, 6.80, 6.77 (d, s, s, s, s, 3H), 6.59 (d, 2H), 6.31 (t , 1H), 3.88 (s, 3H, -OCH ₃ ). 3.83 (s, 3 H, -OCH ₃ ). 3.76 (s, 6H, -OCH ₃ )

¹³ C-NMR (CDCl ₃ ) δ: 160.99, 149.44, 149.07, 139.57, 130.28, 129.00, 126.81, 120.00, 107.9, 106.6, 101.1, 55.4, 55.3

MS: 300.34 (M ⁺ )

Example 10 Preparation of 3,5,4'-Trimethoxy Transtilbene (Compound 10)

After dissolving 80.2 mg (0.297 mM) of 3,5,4'-trimethoxy stilbene derivative in 8 ml heptane, a trace amount of I₂And refluxed for 6 hours. Silica Gel Column Chromatography (CH₂Cl₂) By the way I₂Purified molecular formula by removing (C₁₇O₃H₁₈) 60.2 mg (75%) of 3,5,4'-trimethoxy transstilbene (10) was obtained.

¹ H-NMR (CDCl ₃ ) δ: 7.39 (t, 1H), 7.36 (t, 1H), 6.95 (s, 1H), 6.86 (s, 1H), 6.84 (t, 1H), 6.81 (t, 1H ), 6.57 (d, 2H), 6.31 (t, 1H), 3.76 (s, 6H, -OCH ₃ ), 3.76 (s, 3H, -OCH ₃ ).

¹³ C-NMR (CDCl ₃ ) δ: 160.98, 159.41, 139.71, 129.95, 128.75, 127.79, 126.59, 114.15, 104.35, 99.65, 55.36, 55.33

MS: 270.31 (M ⁺ )

Example 11 3- [2- (3,5-Dimethoxy-phenyl) -vinyl] -furan (Compound 11)

1 g (2.033 mM) of 3,5-dimethoxybenzyl triphenylsporium bromide and 176 μl (1 equivalent) of 3-furalaldehyde (IIIe) were dissolved in 40.7 ml of methylene dichloride together with tetrabutyl ammonium bromide as a catalyst. , 5.3 ml of 50% aqueous sodium hydroxide solution was slowly added. After stirring for 4 hours, 75 ml brine solution was added and extracted with 150 ml methylene dichloride. The organic layer was dried with anhydrous sodium sulfate, and then evaporated under reduced pressure. The residue was purified by silica gel column chromatography (CH ₂ Cl ₂ ) to give 431.8 mg (92%) (6) (C ₁₄ O ₃ H ₁₄ ).

¹ H-NMR (CDCl ₃ ) δ: 7.53 (s, 1H), 7.41 (m, 1H), 6.95 (d, 1H), 6.74 (d, 1H), 6.65 (t, 1H), 6.61 (d, 2H ), 6.38 (t, 1 H), 3.82 (s, 6 H, -OCH ₃ ).

MS: 230.25 (M ⁺ )

Example 12 4- [2- (3,5-dimethoxyphenyl) -vinyl] -pyridine (Compound 12)

1 g (2.033 mmol) of the 3,5-dimethoxybenzyl triphenylsporium bromide and 4-pyridine-carboxaldehyde were dissolved in 40.7 ml of methylene dichloride together with tetrabutyl ammonium bromide as a catalyst, and then 50% aqueous sodium hydroxide solution. 5.3 ml was added slowly. After stirring for 4 hours, 75 ml brine solution was added and extracted with 150 ml methylenedichloride. The organic layer was removed with anhydrous sodium sulfate, and the residue obtained by evaporation under reduced pressure was purified by silica gel column chromatography (CH ₂ Cl ₂ / MeOH = 20: 1) to obtain a molecular formula (C ₁₅ O ₂ NH ₁₅ ) 4- [2- 490.5 mg (100%) of (3,5-dimethoxyphenyl) -vinyl] -pyridine (12) were obtained.

¹ H-NMR (CDCl ₃ ) δ: 8.49 (d, 1H), 8.39 (d, 1H), 3.76 (s, 3H, —OCH ₃ ), 3.58 (s, 3H, —OCH ₃ ).

MS: 241.28 (M ⁺ )

Example 13 : (3,5-dimethoxybenzylidene)-(2,4-dimethoxy-phenyl) amine (Compound 13)

1 g (6 mM) of 3,5-dimethoxybenzaldehyde and 858 μl (1 equivalent) of 2,4-dimethoxyaniline were dissolved in 12 ml toluene. It was heated to reflux under a Dean-Stark apparatus overnight. The residue obtained by concentrating under reduced pressure was dissolved in 30 ml of methanol, and then cooled to -20 ° C to recrystallize the molecular formula (C ₁₇ NO ₄ H ₂₁ ) (3,5-dimethoxybenzylidene)-(2,4-dimethoxy-phenyl ) 440.6 mg (27%) of an amine compound (13) were obtained.

¹ H-NMR (CDCl ₃ ) δ: 8.42 (s, 1H). 7.26 (d, 2H), 7.03 (s, 1H), 6.55 (m, 2H), 6.50 (d, 1H), 3.87 (s, 3H, -OCH ₃ ), 3.84 (s, 6H, -OCH ₃ ), 3.82 (s, 3 H, -OCH ₃ ). ¹³ C-NMR (CDCl ₃ ) δ: 159.5, 138.6, 134.8, 120.7, 115.1, 107.1, 107.0, 116.2, 104.3, 104.1, 103.8, 99.4, 99.3, 55.8, 55.7, 55.5, 55.4

MS: 301.33 (M ⁺ )

Example 14 (4-methoxybenzylidene)-(3-methoxyphenyl) amine (Compound 14)

269 μl (2.2 mM) of p-anisaldehyde and 247 μl (1 equivalent) of m-anisidine were dissolved in 11 ml toluene. Reflux heating under a Dean Stark apparatus overnight. The residue obtained by concentrating under reduced pressure was dissolved in 10 ml of methanol, and then cooled to -20 ° C to recrystallize the (4-methoxybenzylidene)-(3-methoxyphenyl) amine compound of molecular formula (C ₁₅ NO ₂ H ₁₅ ) (14 ) 53 mg (10%) were obtained.

¹ H-NMR (CDCl ₃ ) δ: 8.39 (s, 1H), 7.86 (t, 1H), 7.83 (t, 1H), 7.28 (m, 1H), 7.00 (t, 1H), 6.97 (t, 1H ), 6.79 (t, 1H), 6.76 (m, 2H), 3.88 (s, 3H, -OCH ₃ ), 3.84 (s, 3H, -OCH ₃ ).

¹³ C-NMR (CDCl ₃ ) δ: 159.8, 132.0, 130.6, 129.8, 129.1, 114.2, 112.9, 111.5, 107.9, 106.6, 101.1, 55.4, 55.3

MS: 241.28 (M ⁺ )

Example 15 (4-methoxybenzylidene) -phenylamine (Compound 15)

1.2 ml (10 mM) of p-anisaldehyde and 911 μl (1 equiv) of aniline were dissolved in 50 ml methanol. After refluxing overnight, the mixture was cooled to −20 ° C. to obtain 27 mg (1.3%) of the recrystallized molecular formula (C ₁₄ NOH ₁₃ ) (4-methoxybenzylidene) -phenylamine compound (15).

¹ H-NMR (CDCl ₃ ) δ: 8.38 (s, 1H), 7.87 (t, 1H), 7.84 (d, 1H), 6.59 (d, 2H), 6.44 (q, 1H), 6.39 (d, 1H ), 6.29 (t, 1H), 3.80 (s, 3H, -OCH ₃ ), 3.76 (s, 3H, -OCH ₃ ), 3.76 (s, 6H, -OCH ₃ ).

¹³ C-NMR (CDCl ₃ ) δ: 159.7, 132.0, 130.5, 129.3, 129.1, 125.5, 120.9, 115.0, 114.2, 55.4

MS: 211.25 (M ⁺ )

By the following experiments, it was confirmed whether the phenol ring derivatives of the present invention effectively inhibit the tyrosinase and effective for whitening.

Reference Example 1 : Target Samples and Reagents

Tyrosinase (product number: T7755) was obtained from Sigma, a mushroom extract, L-dopa (product number: D9628), xanthine oxidase (product number: X1875). ), TPA (phobol 12- Myristate 13- Acetate) (product number: P3135), N-1-naphthyl ethylene diamine (product number: N9125), etc. were also purchased from Sigma, resveratrol (product number: R5010), koji acid (Product number: K3125), hydroxylamine hydrochloride (product number: H6025-5) and sulfanic acid (product number: 48150-5), hypoxanthine (product number: H6120-0) and all reagents used in organic synthesis. It was purchased from Aldrich.

Penicillin-Streptomycin (Product No .: 15140-122), FBS (Product No .: 26140-079), RPMI1640 (Product No .: 31800-014) are manufactured by Gibco, and Teddy (Product No .: 3R1902) is Teddy. It was purchased from Tedia and used for the experiment.

Reference Example 1 : Instrument Analysis

A tyrosinase activity test, SOD-like activity test, and Melan-A test method used a spectrophotometer (precision microplate reader, E09090 model Molecular Devices), and UV-absorbance was measured using the Hewlett Packard 8453 model (Hewlett packard). It was. Gemini-2000 model (Gemini Co., Ltd.) was used to obtain the material structure, and ¹ H-NMR of 300 MHz and ¹³ C-NMR of 75 MHz were obtained, and Mass Spectrum was used as GC-mass instrument of 2000 amu / sec. Packard) was used.

In addition, tyrosinase inhibitory activity involved in melanin biosynthesis is described by Aroca et al. P., et al .; J. Biol. Chem. 268 (34) , p25650-25655, 1993 and Tomita et al., Tomita. K., et al .; J. Antibiot. 43 (12) , 1601-1604, 1990, reference was made to the activity assay, and the UV blocking effect and Superoxide Dismutase-like activity assay were described by Jacobson et al., Jacobson et al., Et al. al .; J. Infect. Immun. 62 (9) , p4085-4086, 1994], Oyanagui. Y., Anal. Biochem. 142 , 290-296, 1984 and Matsuda et al., Biol. Pharm. Bull. 18 (8) , 1148-1150, 1995., and a method for verifying the effect of inhibiting melanogenesis using melan-A cells is described in Bennett et al., Int. J. Cancer. 39, 414-418. 1987.].

Test Example 1

Confirmation of inhibitory effect on the whitening enzyme tyrosinase

Test samples dissolved in 80 μl of 25 mM L-dopa 40 μl, 67 mM phosphate buffer (PH 6.8) and 40 μl methanol to investigate the inhibitory effect of the test substance on tyrosinase, an enzyme associated with melanogenesis. μM, 100 μM, 500 μM, 1 mM) was added to a 96-well microplate and 40 μl of mushroom extract tyrosinase (125 U / ml) was added. After incubation at 37 ° C. for 20 minutes, the amount of dopa chromium produced was measured at 492 nm using a spectrophotometer (precision microplate reader, Molecular Devices E09090), and 600 μM L-dopa 90 μl, 50 mM phosphoric acid. Test sample (500 μM) dissolved in 1800 μl of buffer (PH 6.8) and 200 μl of methanol is added to the tube and 900 μl of mushroom tyrosinase (60 U / ml) is added. After 20 minutes of incubation at 37 ° C., UV absorption was measured at 200 nm to 600 nm (Matsuda et al., Biol. Pharm. Bull. 19 (1) , 153-156, 1996). Based on the following formula, the tyrosinase inhibitory effect was determined based on the control group to which no sample was added.

From the tyrosinase inhibition rate of the dilution solution of each sample obtained by the above formula, the dose of each sample (IC ₅₀ = 50% inhibition concentration) that reduced the tyrosinase activity by 50% was determined.

The mushroom extract tyrosinase inhibitory activity at 100 μM concentration of each compound is shown in Table 2. (Each number is the average of three experiments.)

Tyrosinase Inhibitory Activity of Phenyl Ring Derivatives sample Absorbance % Inhibition sample Absorbance % Inhibition 6 0.463 11.0 13 0.454 12.7 7 0.463 11.0 14 0.310 40.0 8 0.470 9.6 15 0.393 24.4 9 0.447 14.0 Kojisan 0.421 19.0 10 0.468 10.0 Resveratrol 0.388 25.4 11 0.445 14.4 Oxyresveratrol 0.146 71.9 12 0.463 11.0 Control 0.520 -

The inhibition rate by concentration of the substances showing similar or higher activity than kojic acid, the double reference material, is shown in Table 3 and FIG. 1.

Inhibitory Effect of Tyrosinase Enzyme on the Concentration Gradient of Samples sample 2 μM 20 μM 100 μM 200 μM IC ₅₀ 14 10.4 16.7 40.0 46.4 198.2 15 5.5 5.8 24.4 26.3 > 200 Kojisan 6.6 5.2 19 39.9 264.3 Resveratrol 8.5 10.4 25.4 24 > 200 Oxyresveratrol 16.5 35.5 71.9 76.6 53.7

In addition, the results of analyzing the absorption of UV-A (270nm -290nm) and UV-B (350-370nm) of 33.4 μM phenyl ring derivative samples including the compounds of the present invention and oxyresveratrol, resveratrol and kojic acid , Compound Nos. 14 and 15 exhibited characteristic absorption peaks in UV-A. (See FIGS. 2-5)

Test Example 2: Superoxide Dismutase (SOD) -Like Active Enzyme Experiment

0.2 ml of Reagent A solution consisting of 0.5 mM hypoxanthine (product number: H6120-0), 10 mM hydroxylamine hydrochloride (product number: H6025-5) dissolved in water, 0.77 ml of EDTA-phosphate The buffer (PH 8.2) and 32 μl of sample dissolved in methanol were mixed well and incubated at 37 ° C. for 10 minutes. To this was added 0.2 ml of 5 mU / ml xanthine oxidase solution (product number: X1875) and incubated at 37 ° C. for 30 minutes, followed by 20 μM of N-naphthylethylene diamine (product number: N9125), 2 After dissolving mM sulfanic acid (product number: 48150-5) in water and adding 2 ml of reagent B prepared by adding 16.7% acetic acid, the solution was left at room temperature for 30 minutes and absorbance was measured at 492 nm.

As a result of the experiment, compound 14 and oxyresveratrol and resveratrol showed equivalent or similar activity with kojic acid at 50 μM (see Table 4 and FIG. 6).

Test Example 3: Melan-A Cell Experiment sample OD (absorbance) SOD Activity (%) 10 μm 50 μm Control 10 μm 50 μm 14 0.238 0.229 0.282 15.6 18.8 Oxyresveratrol 0.200 0.186 0.208 3.9 10.6 Resveratrol 0.199 0.168 0.204 2.5 17.6 Kojisan 0.230 0.221 0.283 18.8 21.9

(1) cell culture

Melan-A cell line in RPMI1640 medium (31800-104) with 10% fetal placental serum (Fetal Bovine Serum, Article no .: 26140-079) and 200 nm Phorbol 12-Myristate 13-Acetate, Product number: P3135) incubated in the conditions. 10 ml of the culture is placed in a 100π tissue culture dish and about 5 × 10 ⁵ cells are seeded. After 3-4 days at 37 ° C. and 5% CO ₂ , the cells were inoculated at 10 ⁵ cells / well in 24 well plates and cultured for 24 hours.

10 μl of the sample was treated for 3 days (solvent: 50% propylene glycol, 30% ethanol, 20% water) while 990 μl of the culture medium was changed daily, and then cultured for 1 day.

(2) Determination of melanin content

After removing the medium, washed with PBS and immediately added 1 ml of 1 N sodium hydroxide to dissolve the melanin and the absorbance was measured at 400 (450) nm.

(3) measuring cell viability

The medium is removed and washed with phosphate buffer saline. 200 μl of crystal violet (CV 0.1%, 10% ethanol, remaining PBS) reagent is added and incubated at room temperature for 5 minutes, followed by washing twice with water. After adding 1 ml of ethanol and shaking for 10 minutes at room temperature, the absorbance was measured at 590 nm.

As a result of the experiment, as shown in Table 5, Compound 14 was found to inhibit melanin production with a relatively low toxicity at 100ppm, and oxyresveratrol and resveratrol having a hydroxy (-OH) group have relatively high cytotoxicity. Indicated.

Activity of Detected Compounds by Melan-A Cell Line Melanin production Cell viability compound Concentration (ppm) OD (absorbance) Control Melanin (%) OD (absorbance) Control Survival rate (%) 14 100 0.074 0.128 57.8 1.112 1.559 71.3 10 0.129 0.128 100.8 1.475 1.559 94.6 One 0.120 0.128 93.8 1.590 1.559 102.0 Resveratrol 100 0.006 0.099 6.1 0.084 1.548 5.4 10 0.047 0.099 47.5 0.544 1.548 35.1 One 0.091 0.099 91.9 1.211 1.548 78.2 Kojisan 100 0.091 0.099 91.9 1.345 1.595 84.3 10 0.095 0.099 96.0 1.573 1.595 98.6 One 0.106 0.099 107.1 1.541 1.595 96.6 Oxyresveratrol 100 0.046 0.177 26.0 0.325 1.933 16.8 10 0.156 0.177 88.1 1.824 1.933 94.4 One 0.168 0.177 94.9 2.020 1.933 104.5

The present invention is Tyrosinase inhibitory activity, UV blocking effect test, SOD-like activity test involved in melanin biosynthesis And through melan-A cell (melan-A cell) melanin production inhibitory effect experiments, such as excellent whitening activity, low cytotoxicity, and provides a phenyl ring compounds of the general formula 1 of a novel chemical structure that suppresses melanin production do.

Claims (14)

Phenyl ring derivative of the general formula (I) Formula 1 Where R _1, R _2, and R ₃ are each independently hydrogen, halogen, C _1-4 alkoxy; X is a carbon atom or a nitrogen atom; M is (Iii) a 2-pyridyl, 3-pyridyl or 4-pyridyl group substituted with R ₄ ; (Ii) a 2-furyl or 3-furyl group substituted with R ₄ ; (Iii) a phenyl ring represented by the following general formula (I-1); Formula 2 Wherein R ₄ is hydrogen, halogen, hydroxy, or a C _1-4 alkoxy group; The R _5, R _6, R _7, R ₈ and R ₉ groups are hydrogen or a C _1-4 alkoxy group except that neither of the R ₁ to R ₉ substituents are hydrogen. The method of claim 1, Phenyl ring derivative of the general formula (Ia) Formula 6 Where R ₁ to R ₃ and R ₅ to R ₉ are as defined in claim 1. The method of claim 2, Phenyl ring derivative of the general formula Iac Formula 7 Where R ₁ to R ₃ and R ₅ to R ₉ are as defined in claim 1. The method of claim 2, Phenyl ring derivatives of the general formula Iat: Formula 8 Where R ₁ to R ₃ and R ₅ to R ₉ are as defined in claim 1. The method of claim 1, Phenyl ring derivative of the general formula (Ib) Formula 10 Where R ₁ to R ₄ are as defined in claim 1. The method of claim 1, Phenyl ring derivative of the general formula (Ic) Formula 12 Where R ₁ to R ₄ are as defined in claim 1. The method of claim 1, Phenyl ring derivatives of the general formula Id: Formula 15 Where R ₁ to R ₃ and R ₅ to R ₉ are the same as defined in claim 1. A process for preparing a compound of Formula Ia by subjecting a compound of Formula IIa or Formula IIb to a Wittig reaction in the presence of a strong base with the following Formula III compound: Formula 3 Formula 4 Formula 5 Formula 6 In the above formula, R in general formula (IIa) represents an alkyl, cycloalkyl group, phenyl group or the like capable of causing a Wittig reaction; X 'represents a halogen atom such as bromine, chlorine, iodine; R ′ in formula IIb represents haloalkyl, alkyl, phenyl, alkoxy, phenoxy and the like; R ₁ to R ₃ and R ₅ to R ₉ are the same as defined in claim 1. A method of selectively preparing the compound of Formula Iat by heating the compound of Formula Ia in the presence of iodine: Formula 6 Formula 8 Wherein R ₁ to R ₃ and R ₅ to R ₉ are the same as defined in claim 1. A process for preparing the following general formula (Ib) compound by subjecting the compound of formula (IIa) or (IIb) according to claim 8 to Witig in the presence of the following general formula (IV) compound and strong base: Formula 9 Formula 10 Where R ₁ to R ₄ are as defined in claim 1. A process for preparing the following general formula (Ic) compound by subjecting the compound of formula (IIa) or (IIb) according to claim 8 to Witig in the presence of a compound (V) with a strong base: Formula 11 Formula 12 Where R ₁ to R ₄ are as defined in claim 1. A method of preparing the compound of formula Id by subjecting the compound of formula VI to an imidization reaction with the compound of formula Formula 13 Formula 14 Formula 15 Where R ₁ to R ₃ and R ₅ to R ₉ are as defined in claim 1. A pharmaceutical composition for treating diseases caused by melanogenesis, comprising the compound of formula I according to any one of claims 1 to 7 as an active ingredient and a pharmaceutically acceptable carrier. A whitening cosmetic composition comprising the general formula I compound according to any one of claims 1 to 7 as an active ingredient and a cosmetically acceptable base.