WO2007078034A1

WO2007078034A1 - A composition for inhibiting the c-kit portein containing benzimidazole amine derivatives or aminoquinoline derivatives

Info

Publication number: WO2007078034A1
Application number: PCT/KR2006/001988
Authority: WO
Inventors: Yong-Joo Na; Soo-Mi Ahn; Heung-Soo Baek; Hyun-Jung Shin; Jae-Sung Hwang; Ih-Seop Chang
Original assignee: Amorepacific Corp
Current assignee: Amorepacific Corp
Priority date: 2006-01-06
Filing date: 2006-05-25
Publication date: 2007-07-12
Anticipated expiration: 2008-07-06
Also published as: KR100788161B1; KR20070074090A

Abstract

The invention relates to a composition for inhibiting the c-kit protein or composition for the skin whitening containing at least one of benz imidazole amine derivatives and aminoquinoline derivatives. The composition inhibits a cell signaling in the melanocyte, in which the c-kit protein participates, thereby regulating a function of the melanocyte and ultimately inhibiting the melanogenesis to exhibit the skin whitening effect.

Description

[DESCRIPTION] [Invention Title]

A COMPOSITION FOR INHIBITING THE C-KIT PROTEIN CONTAINING BENZIMIDAZOLE AMINE DERIVATIVES OR AMINOQUINOLINE DERIVATIVES [Technical Field]

<ι> The invention relates to a composition for inhibiting the c-kit protein or composition for the skin whitening containing at least one of benzimidazole amine derivatives and aminoquinoline derivatives, and more particularly to a composition of an external preparation formulation for skin containing the derivatives to inhibit the melanin synthesis and thus to ultimately accomplish the skin whitening effect, using the function of inhibiting the activity of the c-kit protein which is present on a surface of a melanocyte, in addition to the well-known functions of the compounds. [Background Art]

<3> It is known that the c-kit is a cell surface protein belonging to a receptor tyrosine kinase type III family and is a receptor of a stem cell factor (SCF). According to the research on the white spotting and the steel mouse, the SCF/c-kit function plays an important role for the hematopoiesis, pigmentation and reproduction, including the cell maintenance.

<5> As a result of an interaction between the SCF/c-kit, the c-kit protein undergoes the dimerization and has an activity of phosphorylating itself. It then undergoes the Ras-Raf-MAP kinase cascade in the intracellular signal transfer, and activates the Microphthalmia-Associated Transcription Factor (Mitf), thereby accelerating the tyrosinase synthesis and deriving the melanogenesis in the melanocyte.

<7> The SCF/c-kit signal transfer is also associated with the pigmentation that is derived by the ultraviolet B. It is thought that the mechanism thereof may be related to an increase of the SCF expression in the keratinocyte and an increase of the c-kit expression in the melanocyte. In fact, it can be seen that gene and protein expressions of the SCF and the c- kit are increased when the ultraviolet B stimulation is applied during the cell culture of the keratinocyte and melanocyte.

<9> Such phenomena can be also seen when the ultraviolet B treatment is performed for the epidermis of the human. In other words, the gene and protein expressions of the SCF are significantly increased in the portion exposed to the ultraviloet B, as compared to the portions that are not exposed to it . uι> An animal experiment of Brownish guinea pigs, which is a good model of the melanocyte function research, also provides the same result. Specifically, when the inhibitory antibody to the c-kit protein is injected in the subepidermis of the experimental animal, the pigmentation due to the ultraviolet B is completely inhibited. According to these investigations, it can be seen that the signal transfer of the SCF/c-kit interaction is closely related to the pigmentation ultimately.

<i3> In recent years, the customer's interest in the skin whitening has been increasing as the ultraviolet is increased due to the environmental pollution or destruction of the ozone layer. However, as it has been much reported that the chemically synthesized cosmetics, which are produced with the artificial compounds, have caused the side effects and the hypersensitivity, it has been actively performed the attempts to obtain the raw materials of the cosmetics from the natural materials.

<I5> However, the kojic acid, arbutin or derivatives thereof have been unavailable because they cause a safety and stability problem and have little whitening effect when they are mixed with the raw materials of the cosmetics.

<^|7> In the mean time, as a result of the continuous attempts to find out the whitening active ingredients from the natural substances, it was found the materials showing the tyrosinase activity-inhibiting effect. However, they still have the problems of the stability, the proper effective concentration and the like, which should be solved. [Disclosure] [Technical Problem]

^^|9> In order to solve the problems of the whitening agents, the inventors have verified a variety of compounds so as to develop a material capable of effectively inhibiting the c-kit protein activity present on the melanocyte surface of the skin and thus exhibiting the whitening effect. During the verification, the inventors confirmed that the compounds such as benzimidazole amine derivatives, in particular l-(4-methoxyphenyl)-N-(l- naphthalenylmethyl)-lH-benzimidazole-5-amine (hereinafter, referred to as "SB-CK-111"), and aminoquinol ine derivatives, particularly 2-amino-3-(3,4- dimethoxyphenyO-quinoline (hereinafter, referred to as "SB-CK-141") have an efficacy of inhibiting the c-kit protein activity.

<2i> Accordingly, an object of the invention is to provide a composition for inhibiting the c-kit protein or composition for the skin whitening containing such compounds as effective ingredients. [Technical Solution]

<23> In order to achieve the above object, there is provided a composition for inhibiting the c-kit protein containing at least one of the benzimidazole amine derivatives and the aminoquinol ine derivatives.

<25> In addition, there is provided a composition for the skin whitening containing at least one of the benzimidazole amine derivatives and the aminoquinol ine derivatives.

<27> In one of the compositions, the benzimidazole amine derivative is l-(4- methoxyphenyl)-N-(l-naphthalenylmethyl)-lH-benzimidazole-5^~amine expressed by a following chemistry figure 1.

<29> [chemistry figure 1]

<30>

<3i> In one of the compositions, the aminoquinol ine derivative is 2-amino-3- (3,4-dimethoxyphenyl)-quinoline expressed by a following chemistry figure 2. [chemi stry f igure 2]

<J4>

<35> In the composition for inhibiting the c—kit protein, the composition inhibits an activity of the c-kit protein.

<37> In the composition for inhbiting the c-kit protein, the composition has a use of skin whitening.

<3^ι>> In one of the compositions, at least one of the benzimidazole amine derivative and the aminoquinoline derivative inhibits a cell signaling in the melanocyte, in which the c-kit protein participates, thereby regulating a function of the melanocyte.

<4i> In one of the compositions, at least one of the benzimidazole amine derivative and the aminoquinol ine derivative is contained in an amount of 0.0001-10 wt% for a total weight of the composition.

<43> In one of the compositions, the composition is a cosmetic composition or pharmaceutical composition. [Advantageous Effects]

<45> According to the invention the benzimidazole amine derivative SB-CK-I11 and the aminoquinol ine derivative SB-CK-141 have the effect of inhibiting the c-kit protein activity which is a cell surface protein of the melanocyte. Since it inhibits a cell signaling in the melanocyte, in which the c-kit protein participates, thereby regulating a function of the melanocyte and ultimately inhibiting the melanogenesis to exhibit the skin whitening effect, the composition for inhibiting the c-kit protein or composition for the skin whitening containing them can be usefully used as a cosmetic composition or pharmaceutical composition. [Description of Drawings]

<47> FIG. 1 is a graph showing an inhibiting effect of SU11248 on the c-kit protein activity, which is dependent on the concentration;

<49> FIG. 2 is a graph showing an inhibiting effect of l-(4-methoxyphenyl)- N-(l-naphthalenylmethyl)-lH-benzimidazole-5-amine (hereinafter, referred to as "SB-CK-111") and 2-amino-3-(3,4-dimethoxyphenyl)-quinoline (hereinafter, referred to as "SB-CK-141") of the invention on the c-kit protein activity;

<5i> FIG. 3 is a photograph showing an induction of the c-kit protein activity (i.e. phosphorylation) by a stem cell factor (SCF) and an inhibition of the c-kit protein activity by SU11248 in a cell level;

<53> FIG. 4 is a photograph showing an inhibiting effect of SB-CK-111 of the invention on the c-kit protein activity in a cell level; and

<55> FIG. 5 is a photograph showing an inhibiting effect of SB-CK-141 on the c-kit protein activity in a cell level. [Best Mode]

<57> The benzimidazole amine derivatives and the aminoquinoline derivatives, having a skin whitening function, of the invention can be added to a composition of an external preparation formulation for skin, such as cosmetic composition or pharmaceutical composition. o9> The composition for inhibiting the c-kit protein or composition for the skin whitening according to the invention preferably contains at least one of the benzimidazole amine derivatives and the aminoquinoline derivatives in an amount of 0.0001-10 wt% for a total weight of the composition. When the content is less than 0.0001 wt%, the effects thereof cannot be expected, and when the content is more than 10 wt%, it can cause the safety or formulation stability problem. If the effective effects, the safety and the formulation stability are considered, the content is more preferably 0.01-1.0 wt%.

<6i> The cosmetic composition of the invention may further contain other ingredients preferably capable of providing a synergy effect to the main effect, within a range of not deteriorating the main effect of the invention, in addition to at least one of the benzimidazole amine derivatives and the aminoquinoline derivatives.

<63> At least one of the benzimidazole amine derivatives and the aminoquino11ne derivatives of the invention is preferably prepared in a form of the cosmetic formulation and applied as it is However, they can be used as a typical external preparation

6s> The cosmetic composition of the invention can be used in the cosmetics for inhibiting the c-kit protein or the skin whitening effect The product to which the composition of the invention can be added includes, for example, the cosmetics such as various creams, lotions and the like, cleansing agent, washing agent, soap, cosmetic solution, etc

«)7> The cosmetics to which the composition of the invention containing at least one of the benzimidazole amine derivatives and the aminoquinoline derivatives is added may take a form of solution, emulsion, viscous mixture

_^69> In other words, the cosmetics of the invention are not particularly limited with regard to the formulations thereof. For example, the formulations may include emulsion, cream, toilet water, essence, pack, gel, powder makeup base foundation, lotion, ointment, patch, cosmetic solution, cleansing foam, cleansing cream, cleansing water, body lotion, body cream, body oil, body essence, shampoo rinse, body cleansing agent, soap, hair-dye, spray and the like

-7i> In the cosmetic composition of the respective formulations, the other ingredients except at the least of the benzimidazole amine derivatives and the aminoquinoline derivatives may be arbitrarily mixed by the one skilled in the art without any difficulty depending on the formulations or use of the cosmetic compositions

<73> The above pharmaceutical preparation may contain a material of enhancing the absorption so as to improve the effect of inhibiting the c-kit protein and the skin whitening.

<?">> In addition the cosmetics of the invention may contain any ingredient selected from a group consisting of water-soluble vitamin, oil-soluble vitamin, polymer peptide polymer polysaccharide, sphingolipid and seaweed extracts <77> The cosmetics of the invention may contain another ingredients, which are mixed in the typical cosmetics, as required, in addition to the essential ingredients,

<7')> The other ingredients which can be added may include oil and fat ingredient, moisturizing agent, emollient agent, surfactant, organic and inorganic pigments, organic powder, ultraviolet absorbent, antiseptic, fungicide, antioxidant, plant extract, pH adjuster, alcohol, colorant, flavour, blood circulation accelerant, frigidity agent, anhydrotics, purified water and the like.

<8i> In addition, the ingredients that can be added are not limited to the above ingredients and any ingredient may be mixed within a range of not deteriorating the object and the effect of the invention.

<83> The pharmaceutical composition of the invention may be administrated in oral, non-oral, rectal, local, percutaneous, intravenous, intramuscular, intraperitoneal, subcutaneous administrations. The percutaneous administration is the most preferable.

<85> In addition, the dosage of the active ingredient is different depending on ages, sexes and weights of the patient to be treated, specific diseases or pathological state to be treated, seriousness of the disease or pathological state, administration route, prescriber's decision and the like. The dosage decision based on such factors is within a level of the one skilled in the art. In general, the dosage is about 0.001 mg/kg/day to 2000 mg/kg/day. The preferred dosage is 0.5 mg/kg/day to 2,5 mg/kg/day. Alternatively, 1.0 to 3.0 ml/day may be applied one to five times per day for one month or more.

<87> In the mean time, the pharmaceutical composition may be administrated in a solid, semi-solid or liquid form depending on the intended administration types. The administration types may include, but not limited to, tablet, pill, capsule, suppository, small bag, granule, powder, cream, lotion, ointment, gel, patch, spary, sticking plaster, liquid solution, suspensions, dispersions, emulsions, syrup and the like. The active ingredient may be encapsulated in liposome, fine particles or microcapsules. However, the most preferred formulation is the formulation for the percutaneous administration, such as cream, lotion, ointment, gel, patch, spray, liquid solution, suspensions, dispersions or emulsions.

<89> Among the formulations, the solid composition such as tablet, pill and granule may be conveniently coated. Typically, the composition for the intravenous administration is the solution in sterilized isotonic aqueous buffer solution and includes the local anesthetic so as to alleviate the pain in the syringed portion.

<9i> Additionally, the pharmaceutical preparation may contain a small amount of a non-toxic adjuvant material, such as wetting agent, emulsifying agent, pH buffer agent and the like. The non-toxic adjuvant material may include, but not limited to, sodium acetate, sorbitan monolaurate, triethanolamine and triethanolamine oleate.

<9i> Further, the pharmaceutical composition of the invention may further contain stabilizer, antioxidant, binding agent, colorant, flavouring, excipient such as antiseptic and thickening agent, carrier and diluent. [Mode for Invention]

<95> Hereinafter, the invention will be more specifically described with reference to following experimental examples. However, it should be noted that the invention is not limited thereto.

<97> The inhibiting effects of the benzimidazole amine derivatives, particularly SB-CK-111 and the aminoquinoline derivatives, particularly SB- CK-141 on the c-kit protein activity were measured through the experiment in the test tube and the experiment in the cell level and thus the efficacy thereof was verified.

<99>

<ιoo> Experimental example l: The inhibiting effect of the benzimidazole amine derivative SB-CK-Il1 and the aminoquinoline derivative SB-CK-141 on the c-kit protein activity in the experiment in the test tube>

<102> <!-!> Evaluation of the c-kit protein activity and inhibiting effect of the SU11248 on the c-kit protein activity in the experiment in the test tube <i04> In the experimental example 1-1, it was observed the activity of the single c-kit protein and the effect of inhibiting the c-kit protein activity using SU11248 (Sugen, USA) which sticks specifically to the c-kit protein and thus inhibits the activity thereof, so as to verify a system used for the activity evaluation.

<i06> This experimental method was the same as that suggested in the Panvera Z-lyteTM Kinase assay kit Tyr 4 peptide, but was performed in the optimized conditions with regard to the actual reaction temperature and time, the concentration of the used c-kit, the concentration of ATP and the like.

<i()8> First, 2X ATP was divided to the 384 well dishes for the fluorescent measurement by 5 μH.. Then, SU11248 prepared with 150 nM, 50 nM, 15 nM, 5 nM, 1.5 nM and 0.5 nM were treated with 96 pin dish. To eusure the correctness of the reaction, each sample was prepared with pairs and the control tests of a group having no SU11248/a group to which the c-kit protein was not added/a group of phospho-pept ide 0%/a group of phospho-peptide 100% were performed at the same time. After that, the solution in which the c-kit protein and the substrate peptide therefore were mixed, was applied by 5 μl and then the reaction was carried out at 37⁰C for 45 minutes. After the reaction was completed, the development solution was applied by 5 μi, the reaction was conducted at room temperature for 40 minutes, and then the stop solution was applied by 5 μl, thereby completing the reaction. At this time, the final concentrations of the respective materials used for the reaction were ATP 500 U M, the c-kit (kinase) protein 1.5 ng and the peptide 2 μM, and the final concentrations of SU11248 were 30 nM, 10 nM, 3 nM, 1 nM, 0.3 nM and 0.1 nM.

<Hϋ> In the mean time, it was measured the fluorescent values for the sample having completed the reaction so as to derive the inhibiting rate of SU11248 for the c-kit protein activity. The activation degree of the c-kit protein was measured by reading two fluorescent values using the fluorescence light activated with 405 nm and 535 nm filters and calculating the ratio between them.

<ιi2> As a result, as shown in Fig. 1, the activity of the c-kit protein was inhibited SU11248 dose-dependent Iy.

< I 14>

<ii5> <l-2> The inhibiting effect of the benzimidazole derivative SB-CK-IU and the aminoquinoline derivative SB-CK-141 on the c^~kit protein activity in the experiment in the test tube

<ii7> It was confirmed that it was possible to measure the activity of the c- kit protein and to evaluate the inhibition of the c-kit protein activity by the specific inhibitors from the verification result of the system for evaluating the c-kit protein activity in the experimental example 1-1. Based on this, it was performed a test of evaluating whether the benzimidazole derivative SB-CK-I11 and the aminoquinoline derivative SB-CK-141 could inhibit the c-kit protein activity under the same reaction conditions as those of the experimental example 1-1.

<ii9> Although the reaction conditions used in this test were the same as those of the experimental example 1-1, the final concentrations of the benzimidazole derivative SB-CK-I11 and the aminoquinoline derivative SB-CK- 141, which were used to examine the activity inhibiting effect, were respectively 100 μM, 30 μM, 10 μM, 3 uM, 1 μM, 0.3 μM and 0.1 μM. Regarding the inhibiting rate, as described in the experimental example 1-1, the degree of the inhibition of the c-kit protein activity was shown in Fig. 2 based on the values obtained with the device for measuring the fluorescent after the reaction was completed.

<i2i> As a result, as can be seen in Fig. 2, the activity of the c-kit protein was inhibited dose dependent manner of the benzimidazole amine derivative SB-CK-I11 and the aminoquinoline derivative SB-CK-141, respectively.

<I23>

<ι24> <Experimental example 2' The inhibiting effect of the benzimidazole amine derivative SB-CK-I11 and the aminoquinoline derivative SB-CK-141 on the c-kit protein activity in the cell level>

<i26> <2-l> Evaluation on the c-kit protein activity and the inhibiting effect of SU11248 on the c-kit protein activity in the cell level

<(28> In the experimental example 2-1, it was observed the activity of the single c-kit protein and the effect of inhibiting the c-kit protein activity using SU11248 (Sugen, USA) which sticks specifically to the c-kit protein and thus inhibits the activity thereof, in the cell level, so as to verify a system used for the activity evaluation.

<i30> At this time, the used cell lines were HM3K0 (which was supplied from Dr. Y Funasaka-Kobe University School of Medicine) which was derived from the human, and the culture was carried out in the MEM culture media to which 10% fetal bovine serum was added, under the conditions of 37°C and 5% C(V The cultured HM3K0 cells were separated with 0.25% trypsin-EDTA, again provided to the 6-well plates in the same amount (2.4X10 cells/well) and then left as they were for 24 hours. After that, all of the culture media were removed, and replaced with the MEM culture media to which 1% fetal bovine serum was added and then again cultured for 24 hours. On the third day, after the media were again replaced with fresh MEM culture media to which 1% fetal bovine serum was added, SU11248 was applied with the final concentration of 50 nM. Then, after 24 hours, the stem cell factor (SCF), which binds specifically to the c-kit protein and thus induces the activation thereof, was applied to the HM3K0 cells with the final concentration of 50 ng/mt for 15 minutes, thereby inducing the activation of the c-kit protein. At this time, it was compared the increase of the activity of the c-kit protein and the inhibiting effect of SU11248 on the c-kit protein activity by using a group which was treated with nothing (notated as UN)/a group which was treated with SCF only (notated as SCF)/a group which was pre-treated with SU11248 and treated with SCF (notated as SU11248) . The result is shown in Fig. 3,

<i32> As can be seen in Fig. 3, it was observed that the activity of the c- kit protein was inhibited by SU11248.

<I34> <i35> <2-2> The inhibiting effect of the benzimidazole derivative SB-CK-I11 and the aminoαuinoline derivative SB-CK-141 on the c^~kit protein activity in the eel 1 level

<!37> It was confirmed that it was possible to measure the c-kit protein activity and to evaluate the inhibition of the c-kit protein activity by the specific inhibitors in the cell level, from the result of the experimental example 2-1. Based on this, it was performed a test of evaluating whether the benzimidazole derivative SB-CK-I11 and the aminoquinol ine derivative SB- CK-141 could inhibit the c-kit protein activity under the same reaction conditions as those of the experimental example 2-1.

<i39> Although the reaction conditions used in this test were the same as those of the experimental example 2-1, the final concentrations of the benzimidazole derivative SB-CK-I11 and the aminoquinol ine derivative SB-CK- 141, which were used to examine the activity inhibiting effect, were 500 nM, 50 nM and 5 nM. The final concentration of SU11248 (Sugen, USA), which was a positive control group, was 50 nM which was the same as those of the experimental example 2-1. The result is shown in Figs. 4 and 5.

<i4i> As a result, as can be seen in Figs. 4 and 5, the benzimidazole amine derivative SB-CK-111 and the aminoquinol ine derivative SB-CK-141 effectively inhibited the c-kit protein activity dependently on the concentrations thereof and exhibited the equivalent effect to SU11248 which was the positive control group.

<i43> Hereinafter, although it will be described formulation examples of the above composition, it should be noted that they are provided to illustrate the invention specifically, not to limit it.

<145> <Formulation example 1: Nutritional toilet water (milk skin lotion)>

<i46> The nutrtional toilet water of the invention was prepared with the following composition shown in Table 1.

<i48> [Table 1] < 149>

< | S0> <Formulation example 2: Nutritional cream>

< ! 5 1 > The nutrtional cream of the invention was prepared with the following composition shown in Table 2.

< 153> [Table 2] < 1 54>

<I56> The massage cream of the invention was prepared with the following composition shown in Table 3.

<158> [Table 3] <I59>

<16ϋ> <Forinulat ion example 4: Pack>

< I 61> The pack of the invention was prepared with the following composition shown in Table 4.

<I63> [Table 4]

< 164>

<I65> <Formulation example 5: Ointment of skin external preparation> <I66> The ointment of the skin external preparation of the invention was prepared with the following composition shown in Table 5.

<.I68> [Table 5] 1 6⁽)->

< 1 70>

[Industrial Applicability]

< 1 7 I > According to the invention, the benzimidazole amine derivative SB-CK- 111 and the aminoquinoline derivative SB-CK-141 have the effect of inhibiting the c-kit protein activity which is a cell surface protein of the melanocyte. Since it inhibits a cell signaling in the melanocyte, in which the c-kit protein participates, thereby regulating a function of the melanocyte and ultimately inhibiting the melanogenesis to exhibit the skin whitening effect, the composition for inhibiting the c-kit protein or composition for the skin whitening containing them can be usefully used as a cosmetic composition or pharmaceutical composition.

<173>

Claims

[CLAIMS] [Claim 1]

A composition for inhibiting the c-kit protein containing at least one of benzimidazole amine derivatives and aminoquinoline derivatives.

[Claim 2]

A composition for skin whitening containing at least one of benzimidazole amine derivatives and aminoquinoline derivatives.

[Claim 3]

The composition according to claim 1 or 2, wherein the benzimidazole amine derivative is l-(4-methoxyphenyl)-N-(l-naphthalenylraethyl)-lH- benzimidazole-5-amine expressed by the following chemistry figure 1. [chemistry figure 1]

[Claim 4]

The composition according to claim 1 or 2, wherein the aminoquinoline derivative is 2-amino-3-(3,4-dimethoxyphenyl)-quinoline expressed by the following chemistry figure 2. [chemistry figure 2]

[Claim 5]

The composition according to claim 1, wherein the composition inhibits the c-kit protein activity. [Claim 6]

The composition according to claim 1, wherein the composition has a use of skin whitening. [Claim 7]

The composition according to claim 1 or 2, wherein at least one of the benzimidazole amine derivative and the aminoquinoline derivative inhibits a cell signaling in the melanocyte, in which the c-kit protein participates, thereby regulating a function of the melanocyte. [Claim 8]

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 0.0001-10 wt% for a total weight of the composition. [Claim 9]

The composition according to claim 1 or 2, wherein the composition is a cosmetic composition or pharmaceutical composition.