KR20070074090A - A composition for inhibiting the c-kit protein containing benzimidazole amine derivates or aminoquinoline derivatives - Google Patents

Abstract

A composition comprising a benzimidazole amine derivative or an aminoquinoline derivative is provided to inhibit signaling of a cell in melanocyte with which a c-kit protein is involved so as to control the function of the melanocyte, thereby providing skin-whitening effect. The composition for inhibiting c-kit protein or skin-whitening comprises 0.0001-10 wt.% of a benzimidazole amine derivative such as 1-(4-methoxyphenyl)-N-(1-naphthalenylmethal)-1H-benzimidazole-5-amine represented by the formula(1) or an aminoquinoline derivative such as 2-amino-3-(3,4-dimethoxyphenyl)-quinoline represented by the formula(2).

Description

C composition for inhibiting the c-kit protein containing benzimidazole amine derivatives or aminoquinoline derivatives

1 is a graph showing the activity inhibitory effect of the concentration-dependent C-kit protein by SU11248.

Figure 2 is 1- (4-methoxyphenyl) -N- (1-naphthalenylmethyl) -1H-benzimidazol-5-amine (1- (4-methoxyphenyl) -N- (1 according to the present invention). -naphthalenylmethyl) -1H-Benzimidazol-5-amine) (hereinafter referred to as “SB-CK-111”) and 2-amino-3- (3,4-dimethoxyphenyl) -quinoline (2-amino-3- (3,4-dimethoxyphenyl) -Quinoline) (hereinafter referred to as "SB-CK-141") is a graph showing the inhibitory effect of C-kit protein activity.

Figure 3 is a photograph showing the induction of C-kit protein activity by SCF (sterm cell factor) and inhibition of C-kit protein activity by SU11248 at the cellular level.

Figure 4 is a photograph showing the inhibitory effect of C-kit protein activity at the cellular level of SB-CK-111 according to the present invention.

Figure 5 is a photograph showing the effect of inhibiting C-kit protein activity at the cellular level of SB-CK-141 according to the present invention.

The present invention relates to a composition for inhibiting C-kit protein or a composition for skin whitening containing any one or more of benzimidazole amine derivatives and aminoquinoline derivatives. In addition to the functions known in the art, by utilizing the activity of inhibiting the activity of the C-kit protein present on the surface of melanocytes, the inclusion of such derivatives can inhibit melanin synthesis and ultimately achieve a skin whitening effect. It relates to a composition.

C-kit is a cell surface protein belonging to the receptor tyrosine kinase type III family and is known as a receptor for stem cell factor (SCF). Studies in white spotting and steel mice show that SCF / c-kit function is important for cell maintenance, hematopoiesis, pigmentation, and reproduction. Play a role.

As a result of the interaction between SCF / c-kit, the C-kit protein has dimerization and self-phosphorylated activity. Subsequently, intracellular signal transduction passes through the Ras-Raf-MAP kinase cascade and finally promotes tyrosinase synthesis by activating the Microphthalmia-Associated Transcription Factor (MITf), and melanosite Induce melanin production.

SCF / c-kit signaling is also associated with UVB-induced pigmentation, which is probably associated with increased SCF expression in keratinocytes and increased C-kit expression in melanocytes, respectively. It is thought to be. Indeed, stimulation with ultraviolet B during cell culture of keratinocytes and melanocytes increases the expression of genes and proteins in SCF and C-kit.

This phenomenon was also found to be significantly increased in the expression of genes and proteins of SCF in the exposed portion compared to the portion not exposed to ultraviolet B when the human epidermis (epidermis) treatment.

The same is true for Brown's Guinea pigs, an excellent model for studying melanosite function. That is, when the inhibitory antibody against C-kit protein was injected into the subepidermal of the experimental animal, it was confirmed that pigmentation induced by ultraviolet B was completely inhibited. Therefore, it can be seen that signaling of SCF / c-kit interaction is ultimately closely related to pigmentation.

Recently, due to the increase of ultraviolet rays due to environmental pollution or destruction of the ozone layer, consumer's interest in whitening is increasing. However, as a large amount of side effects and hypersensitivity reactions to chemical synthetic cosmetics made of artificial compounds have been reported, attempts to obtain raw materials of cosmetics from natural materials are actively underway.

However, kojic acid, arbutin and its derivatives that have been identified so far cannot be used as cosmetic raw materials because of their safety or stability problems or their whitening efficacy is not so high when they are formulated into actual cosmetic ingredients. to be.

On the other hand, studies to find whitening active ingredients in natural products continued to find a substance that inhibits the activity of tyrosinase, but the problem of stability, or the appropriate effective concentration problem remains to be solved.

Thus, the present inventors have been tested by various compounds for the development of a material that exhibits a whitening effect by effectively inhibiting the activity of C-kit protein on the surface of melanocytes in the skin in order to solve the problems of the whitening agent as described above , Benzimidazole amine derivatives, in particular 1- (4-methoxyphenyl) -N- (1-naphthalenylmethyl) -1H-benzimidazole-5-amine (hereinafter referred to as "SB-CK-111") ) And aminoquinoline derivatives, particularly compounds such as 2-amino-3- (3,4-dimethoxyphenyl) -quinoline (hereinafter referred to as “SB-CK-141”) have inhibitory effects on C-kit protein activity Ascertained, the present invention was completed.

Accordingly, it is an object of the present invention to provide a composition for inhibiting C-kit protein or a composition for skin whitening containing the compound as an active ingredient.

In order to achieve the above object, C-kit protein inhibition composition according to the present invention is characterized in that it contains any one or more of benzimidazole amine derivatives and aminoquinoline derivatives.

The composition for skin whitening according to the present invention is characterized by containing any one or more of benzimidazole amine derivatives and aminoquinoline derivatives.

In any one of the above compositions, the benzimidazole amine derivative is 1- (4-methoxyphenyl) -N- (1-naphthalenylmetal) -1H-benzimidazole-5- represented by the following formula (1): Amine (1- (4-methoxyphenyl) -N- (1-naphthalenylmethyl) -1H-Benzimidazol-5-amine).

In any one of the above compositions, the aminoquinoline derivative is 2-amino-3- (3,4-dimethoxyphenyl) -quinoline represented by the following formula (2) 2-amino-3- (3,4-dimethoxyphenyl) -Quinoline).

In the C-kit protein inhibition composition, the composition is characterized by inhibiting the activity of the C-kit protein.

In the C-kit protein inhibition composition, the composition is characterized by having the use of skin whitening.

The composition of any one of the above, wherein at least one of the benzimidazole amine derivative and the aminoquinoline derivative modulates the function of melanocytes by inhibiting the signal transduction of cells in the melanocytes involved in the C-kit protein. It is done.

In any one of the above composition, at least one of the benzimidazole amine derivative and the aminoquinoline derivative is characterized in that it contains in an amount of 0.0001 to 10% by weight based on the total weight of the total composition.

In any one of the above compositions, the composition is characterized in that the cosmetic composition or pharmaceutical composition.

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

Inhibitory effects of the benzimidazole amine derivatives according to the present invention, in particular SB-CK-111 and aminoquinoline derivatives, in particular SB-CK-141, were measured through in vitro experiments and at the cellular level. As a result, the efficacy was confirmed.

Benzimidazole amine derivatives and aminoquinoline derivatives having a whitening function according to the present invention can be added to an external preparation for skin, such as a cosmetic composition or a pharmaceutical composition.

The C-kit protein inhibition composition or skin whitening composition according to the present invention preferably contains at least one of benzimidazole amine derivative and aminoquinoline derivative in an amount of 0.0001 to 10% by weight based on the total weight of the total composition. Do. If the content is less than 0.0001% by weight, the effect can not be expected, because if the content is more than 10.0% by weight may cause difficulties in safety or formulation stability. In consideration of effective effects and stability, and formulation stability, 0.01 to 1.0% by weight is more preferable.

The cosmetic composition according to the present invention, in addition to any one or more of the benzimidazole amine derivative and the aminoquinoline derivative described above, preferably give a synergistic effect to the main effect within the range that does not impair the main effect of the present invention. It may also contain other ingredients that may be.

Any one or more of benzimidazole amine derivatives and aminoquinoline derivatives according to the present invention are suitably prepared and applied as they are in the cosmetic formulation, but can also be used as a general external preparation.

The cosmetic composition of the present invention can be used in various ways, such as cosmetics for c-kit protein inhibition or skin whitening effect. Examples of products to which the present composition can be added include cosmetics such as various creams, lotions, skins, and the like, cleansing agents, face washes, soaps, and essences.

The cosmetic to which the composition containing any one or more of the benzimidazole amine derivative and the aminoquinoline derivative of the present invention is added may take the form of a solution, an emulsion, a viscous mixture, or the like.

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, beauty liquid, 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.

In the cosmetic composition of each formulation, in addition to any one or more of the benzimidazole amine derivatives and aminoquinoline derivatives described above, other components can be appropriately selected and blended by those skilled in the art without difficulty according to the formulation or use purpose of other cosmetics.

The formulations may contain a skin absorption promoting substance to increase the C-kit protein inhibitory effect and skin whitening effect.

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

In addition to the said essential component, you may mix | blend the cosmetics of this invention with the other components normally mix | blended with cosmetics as needed.

Other components that may be added include fats and oils, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, fungicides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, flavorings, Blood circulation accelerators, cooling agents, restriction agents, 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 be mix | blended within the range which does not impair the objective and effect of this invention.

The pharmaceutical composition according to the present invention may be administered orally, parenterally, rectally, topically, transdermally, intravenously, intramuscularly, intraperitoneally, subcutaneously, and the most preferred route of administration is transdermal administration.

In addition, the dosage of the active ingredient will vary depending on the age, sex and weight of the subject to be treated, the specific disease or pathology to be treated, the severity of the disease or pathology, the route of administration and the judgment of the prescriber. Dosage determination based on these factors is within the level of skill in the art and generally dosages range from 0.001 mg / kg / day to approximately 2000 mg / kg / day. Preferred dosages are from 0.5 mg / kg / day to 2.5 mg / kg / day. Alternatively, it may be applied for 1 to 5 times per day at 1.0 to 3.0 ml per day to continue for more than 1 month.

On the other hand, depending on the intended dosage form, the pharmaceutical composition may be in solid, semisolid or liquid dosage form. Examples of dosage forms include but are not limited to tablets, pills, capsules, suppositories, small bags, granules, powders, creams, lotions, ointments, gels, patches, sprays, plasters, liquid solutions, suspensions, dispersions, emulsions, syrups, and the like. It is not limited. The active ingredient may be encapsulated in liposomes, microparticles or microcapsules. However, the most preferred formulations are formulations for transdermal administration such as creams, lotions, ointments, gels, patches, sprays, liquid solutions, suspensions, dispersions or emulsions.

Solid compositions such as tablets, pills, granules and the like in the formulations may be conveniently coated and typically compositions for intravenous administration are solutions in sterile isotonic aqueous buffer and include local anesthetics to relieve pain at the injection site.

In addition, if desired, the medicament may also contain small amounts of nontoxic auxiliary substances, such as wetting agents, emulsifiers, pH buffers, and the like, examples of which include, but are not limited to, sodium acetate, sorbitan monolaurate, triethanolamine and triethanolamine oleate. It is not limited.

In addition, the pharmaceutical compositions of the present invention may further comprise excipients, carriers and diluents such as stabilizers, antioxidants, binders, colorants, flavors, preservatives and thickeners.

Hereinafter, the present invention will be described in more detail with reference to the following test examples, but the present invention is not limited only to these examples.

<Test Example 1> Inhibitory effect of c-kit protein activity of SB-CK-111, a benzimidazole amine derivative, and SB-CK-141, an aminoquinoline derivative in an in vitro experiment

<1-1> Evaluation of C-Kit Protein Activity and Inhibitory Effect of C-Kit Protein Activity by SU11248 in In Vitro Experiments

In this Test Example 1-1, the seed kit using SU11248 (Sugen, USA), which binds to the seed kit protein and inhibits the activity by specifically attaching the seed kit protein, to the system verification used for the activity evaluation The effect of inhibiting protein activity was observed.

This experiment is consistent with the method described in the Panvera Z-lyte Kinase Assay Kit Tyr 4 peptide, but the actual reaction temperature or time, Concentrations, ATP concentrations, etc. were conducted under optimized conditions.

First, 2 × ATP was dispensed in 5ul portions into a 384 well dish for fluorescence measurement. Subsequently, SU11248 made of 150 nM, 50 nM, 15 nM, 5 nM, 1.5 nM, and 0.5 nM, respectively, was treated using a 96-pin dish. To ensure the accuracy of the reaction, each sample was paired and the group without SU11248 / the group without the C-kit protein / phospho-peptide 0% group / phospho-peptide Control experiments of the 100% group were also conducted simultaneously. Thereafter, 5 μl of the solution containing the seed kit protein and the substrate peptide (peptide) was dispensed, followed by reaction at 37 ° C. for 45 minutes. After the reaction was completed, the development solution was dispensed by 5ul each for 40 minutes at room temperature, and finally, the stop solution was dispensed by 5ul each to complete the experiment. In this case, the final concentration of each material used in the reaction corresponds to ATP 500uM, 1.5 ng of Kinase protein, 2 uM peptide, SU11248 corresponds to the final concentration of 30nM, 10nM, 3nM, 1nM, 0.3nM, 0.1nM do.

On the other hand, by measuring the fluorescence value of the reaction was completed samples to derive the inhibition rate of SU11248 on the C-kit protein activity. The activation of C-kit protein can be measured by reading two fluorescence values using light activated through 405 nm and 535 nm filters, and calculating the ratio between them.

As a result, as shown in FIG. 1, it was observed that the activity of the C-kit protein was inhibited in a concentration-dependent manner of SU11248.

<1-2> C-Kit Protein Inhibitory Effect of Benzimidazole Derivatives SB-CK-111 and Aminoquinoline Derivative SB-CK-141 in In Vitro Experiments

From the results of the C-kit protein activity evaluation system verification result of Test Example 1-1, it was confirmed that the C-kit protein activity measurement and the inhibition of C-kit protein activity by a specific inhibitor can be evaluated. Based on this, an experiment was conducted to evaluate whether the benzimidazole derivative SB-CK-111 and the aminoquinoline derivative SB-CK-141 can inhibit the activity of C-kit protein under the same reaction conditions as in Test Example 1-1. Was performed.

The reaction conditions used in this experiment were the same as those of Test Example 1-1, but the final concentrations of the benzimidazole derivative SB-CK-111 and the aminoquinoline derivative SB-CK-141 were used to show the inhibitory effect. Corresponding to 100uM, 30uM, 10uM, 3uM, 1uM, 0.3uM, 0.1uM, respectively. Inhibition rate derivation method is also described in Test Example 1-1, by calculating the value obtained by the fluorescence meter after the reaction is completed, the degree of inhibition of C-kit protein activity is shown in Figure 2.

As a result, as can be seen in Figure 2, the concentration of the benzimidazole derivative SB-CK-111 and aminoquinoline derivative SB-CK-141 according to the present invention was confirmed that the activity of the c-kit protein is inhibited It was.

<Test Example 2> Inhibitory effect of c-kit protein activity of benzimidazole derivative SB-CK-111 and aminoquinoline derivative SB-CK-141 at the cellular level

<2-1> Evaluation of C-Kit Protein Activity at the Cell Level and Inhibitory Effect of C-Kit Protein Activity by SU11248

In this Test Example 2-1, a seed using SU11248 (Sugen, USA), which binds to the C-kit and inhibits its activity in combination with the C-kit protein alone activity at the cellular level, for validation of the system used for activity evaluation. -Kit protein activity inhibitory effect was observed.

At this time, the cell line used was HM3KO (supplied by Dr. Y Funasaka- Kobe University school of medicine), a melanosite derived from humans, and the culture was performed in MEM medium containing 10% placental serum, 37 ° C, and 5% CO _2. Under the conditions of. Cultured HM3KO cells were detached with 0.25% Trypsin-EDTA, laid back on 6-well plates with the same number (2.4 × 10 ⁵ cells / well) and left for 24 hours. Thereafter, all of the medium was removed, and then replaced with MEM medium containing 1% fetal placental serum, followed by incubation for another 24 hours. On the third day, SU11248 was treated with a final concentration of 50 nM after again changing to MEM medium with 1% placental serum. After 24 hours, the stem cell factor (SCF), which is specifically attached to the seed kit to induce its activity on HM3KO cells, was added at a final concentration of 50ng / ml for 15 minutes to induce the activity of the seed kit protein. It was. At this time, by using the group treated with nothing (denoted by UN) / the SCF-treated group (denoted SCF) / SU11248 pre-treated and SCF treated group (denoted SU11248), the increase in the activity of C-kit protein alone and The effect of C-kit protein deactivation by SU11248 was compared. The results are shown in Figure 3 below.

As can be seen in Figure 3, it was observed that the activity of C-kit protein was inhibited by SU11248.

<2-2> C-Kit Protein Inhibitory Effects of Benzimidazole Derivatives SB-CK-111 and Aminoquinoline Derivatives SB-CK-141

From the results of Test Example 2-1, it was confirmed that the measurement of C-kit protein activity at the cellular level and the inhibition of activity by a specific inhibitor can be evaluated at the cellular level. Based on this, an experiment was conducted to evaluate whether the benzimidazole derivative SB-CK-111 and the aminoquinoline derivative SB-CK-141 can inhibit the activity of C-kit protein under the same reaction conditions as in Test Example 2-1. Was performed.

The reaction conditions used in this experiment were the same as those of Test Example 2-1, but the final concentrations of the benzimidazole derivative SB-CK-111 and the aminoquinoline derivative SB-CK-141 used to show the inhibitory effect were It corresponds to 500, 50, 5 nM. The final concentration of the positive control SU11248 (Sugen, USA) was 50 nM, which was the same as in Test Example 2-1. The results are shown in FIGS. 4 and 5.

As a result, as can be seen in Figures 4 and 5, the benzimidazole derivative SB-CK-111 and the aminoquinoline derivative SB-CK-141 concentration-dependently effective the activity of the seed kit protein Inhibition was confirmed, showing a superior effect than the positive control SU11248.

Examples of the formulation of the composition will be described below, but the present invention is not intended to be limited thereto, but is intended to be described in detail.

<Formulation Example 1: Nutritious Longevity (Milk Skin Lotion)>

Nutrients according to the present invention was prepared in the composition of Table 1.

ingredient Content (% by weight) Purified water To 100 glycerin 8.0 Butylene glycol 4.0 Hyaluronic acid extract 5.0 Beta Glucan 7.0 Carbomer 0.1 SB-CK-111 0.05 Caprylic Capric Triglycerides 8.0 Squalane 5.0 Cetearyl Glucoside 1.5 Sorbitan stearate 0.4 Cetearyl Alcohol 1.0 antiseptic Quantity incense Quantity Pigment Quantity Triethanolamine 0.1

<Formulation Example 2: Nutrition Cream>

Nutritional cream according to the invention was prepared in the composition of Table 2 below.

ingredient Content (% by weight) Purified water To 100 glycerin 3.0 Butylene glycol 3.0 Liquid paraffin 7.0 Beta Glucan 7.0 Carbomer 0.1 SB-CK-141 3.0 Caprylic Capric Triglycerides 3.0 Squalane 5.0 Cetearyl Glucoside 1.5 Sorbitan stearate 0.4 Polysorbate 60 1.2 antiseptic Quantity incense Quantity Pigment Quantity Triethanolamine 0.1

<Formulation Example 3: Massage Cream>

Massage cream according to the invention was prepared in the composition of Table 3 below.

ingredient Content (% by weight) Purified water To 100 glycerin 8.0 Butylene glycol 4.0 Liquid paraffin 45.0 Beta Glucan 7.0 Carbomer 0.1 SB-CK-141 1.0 Caprylic Capric Triglycerides 3.0 Beeswax 4.0 Cetearyl Glucoside 1.5 Sesqui oleic acid sorbitan 0.9 Vaseline 3.0 antiseptic Quantity incense Quantity Pigment Quantity paraffin 1.5

<Formulation Example 4: Pack>

A pack according to the invention was prepared with the composition of Table 4 below>

ingredient Content (% by weight) Purified water To 100 glycerin 4.0 Polyvinyl alcohol 15.0 Hyaluronic acid extract 5.0 Beta Glucan 7.0 Allantoin 0.1 SB-CK-111 0.5 Nonyl Phenyl Ether 0.4 Polysorbate 60 1.2 antiseptic Quantity incense Quantity Pigment Quantity ethanol 6.0

<Formulation Example 5: Ointment among the external preparations for skin>

Ointment in the external preparation for skin according to the present invention was prepared in the composition of Table 5.

ingredient Content (% by weight) Purified water To 100 glycerin 8.0 Butylene glycol 4.0 Liquid paraffin 15.0 Beta Glucan 7.0 Carbomer 0.1 SB-CK-141 1.0 Caprylic Capric Triglycerides 3.0 Squalane 1.0 Cetearyl Glucoside 1.5 Sorbitan stearate 0.4 Cetearyl Alcohol 1.0 antiseptic Quantity incense Quantity Pigment Quantity Beeswax 4.0

As described above, the benzimidazole derivative SB-CK-111 and the aminoquinoline derivative SB-CK-141 inhibit the activity of C-kit protein, which is a cell surface protein of melanocytes. Thereby, it inhibits the signal transduction of the cells in the melanocytes involved in the C-kit protein, thereby regulating the function of melanocytes, and ultimately inhibiting melanin production, thus exhibiting skin whitening effect. -Kit protein inhibition composition or skin whitening composition can be usefully used as a cosmetic composition or a pharmaceutical composition.

Claims (9)

A composition for inhibiting c-kit protein containing any one or more of benzimidazole amine derivatives and aminoquinoline derivatives. A composition for skin whitening containing any one or more of benzimidazole amine derivatives and aminoquinoline derivatives. The benzimidazole amine derivative according to claim 1 or 2, wherein 1- (4-methoxyphenyl) -N- (1-naphthalenylmetal) -1H-benzimidazole- 5-amine composition. [Formula 1]

The composition of claim 1 or 2, wherein the aminoquinoline derivative is 2-amino-3- (3,4-dimethoxyphenyl) -quinoline represented by the following formula (2). [Formula 2]

The composition for inhibiting C-kit protein according to claim 1, wherein the composition inhibits the activity of C-kit protein. The composition of claim 1, wherein the composition has the use of skin whitening. The method according to claim 1, wherein at least one of the benzimidazole amine derivative and the aminoquinoline derivative modulates the function of melanocytes by inhibiting signal transduction of cells in the melanocytes involved with the C-kit protein. Characterized in that the composition. The composition according to claim 1 or 2, wherein at least one of the benzimidazole amine derivative and the aminoquinoline derivative is contained in an amount of 0.0001 to 10% by weight based on the total weight of the total composition. The composition of claim 1 or 2, wherein the composition is a cosmetic composition or a pharmaceutical composition.

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