KR102493627B1 - Pharmaceutical Composition Comprising Hordenine as Active Ingredient for Preventing or Treating Skin Cancer - Google Patents

Abstract

The present invention relates to a pharmaceutical composition and a cosmetic composition for preventing or treating skin cancer containing Hordenine as an active ingredient.
While the composition according to the invention has excellent ability to inhibit the proliferation and growth of skin cancer cell lines, it does not have the ability to inhibit proliferation and growth in other normal cells of the skin, so it is a pharmaceutical product for external use in various formulations for specifically preventing, treating, or improving skin cancer. And it can be applied to cosmetic products.

Description

Pharmaceutical composition for preventing or treating skin cancer containing hordenine as an active ingredient {Pharmaceutical Composition Comprising Hordenine as Active Ingredient for Preventing or Treating Skin Cancer}

The present invention relates to a pharmaceutical composition and a cosmetic composition for preventing or treating skin cancer containing Hordenine as an active ingredient.

Skin cancer is a malignant tumor of the skin, and representative types include basal cell carcinoma, squamous cell carcinoma, and melanoma.

In general, excessive exposure to UV rays increases the risk of skin cancer. Ultraviolet rays are classified into UV-B and UV-C depending on the wavelength, and UV-B is known to cause basal cell carcinoma and squamous cell carcinoma. These two types account for 80% of all skin cancers. The more lethal UV-C is usually filtered by the ozone layer and rarely reaches the earth's surface, but recent destruction of the ozone layer has increased the number of cases where UV-C reaches the earth's surface unfiltered. UV-C causes melanoma. Melanoma spreads while depositing melanin in each cell, and the prognosis is poor. Melanoma has a higher incidence in the Caucasian population of European descent. On the other hand, lighting fixtures such as halogen lamps or fluorescent lamps also emit UV-C, and melanoma may occur when exposed for a long time.

80% of skin cancers occur in sun-exposed areas such as the face, head, and wrists. In 2002, the US Census of Diseases recorded more than one million skin cancer cases. Most of the cases of skin cancer are basal cell carcinoma or squamous cell carcinoma, and these cancers are relatively treatable. Melanoma accounts for 3% of skin cancer patients, and among melanoma patients, the mortality rate is 14%, which is a dangerous cancer.

For the treatment of such skin cancer, technologies for preventing or treating by inducing apoptosis are being developed. For example, 1,25-dihydroxy-16,22,23-trisdehydro-cholecalciferol derivatives (Republic of Korea Patent No. 10-0240541), a pharmaceutical composition for inhibiting skin hyperplasia and tumor cells , There is a composition for inducing apoptosis (Korean Patent Publication No. 10-2004-0016353) containing a phytosphingosine derivative that induces cell death for the treatment of skin diseases, tumors and cancer.

However, conventional anticancer drugs have the disadvantage of causing side effects due to their toxicity to normal cells. Therefore, there is a demand for the development of improved therapeutic agents without side effects.

Republic of Korea Patent No. 10-0240541 Korean Patent Publication No. 10-2004-0016353

The present inventors, while studying a substance that can specifically express a killing effect only on skin cancer cells without toxicity to normal cells, which is a side effect of conventional anticancer drugs, found that hordenine has the ability to inhibit the proliferation and growth of skin cancer cell lines. While excellent, the present invention was completed by confirming that it could be effectively used for skin cancer treatment because it had no ability to inhibit proliferation and growth in other normal skin cells.

Therefore, the object of the present invention is to contain as an active ingredient a substance capable of minimizing side effects on normal skin cells because it has excellent ability to inhibit the proliferation and growth of skin cancer cell lines, but has no ability to inhibit proliferation and growth in other normal skin cells. To provide a pharmaceutical composition for treating or preventing skin cancer and a cosmetic composition for preventing or improving skin cancer.

In order to achieve the above object, the present invention provides a pharmaceutical composition for treating or preventing skin cancer containing hordenine as an active ingredient.

In addition, the present invention provides a cosmetic composition for preventing or improving skin cancer containing Hordenine as an active ingredient.

While the composition according to the invention has excellent ability to inhibit the proliferation and growth of skin cancer cell lines, it does not have the ability to inhibit proliferation and growth in other normal cells of the skin, so it is a pharmaceutical product for external use in various formulations for specifically preventing, treating, or improving skin cancer. And it can be applied to cosmetic products.

1 is a graph showing the degree of inhibition of cell growth of skin cancer cells by hordenine.
2 is a graph showing the degree of cell growth inhibition of hordenine on skin melanocytes.
3 is a graph showing the degree of inhibition of cell growth of dermal fibroblasts by hordenine.
4 is a graph showing the degree of inhibition of cell growth of keratinocytes by hordenine.
5 is a graph showing the degree of cell growth inhibition of skin cancer cells, skin melanocytes, skin fibroblasts, and skin keratinocytes for each treatment concentration of hordenine.

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

The composition of the present invention contains hordenine as an active ingredient.

The hordenine is found not only in Cactus alkaloids, which are alkaloids belonging to the Cactus family, but also in the genus Cactus and Anhalonium , as well as when barley germinates. Hordenine can be written as N,N-dimethyltyramine and its IUPAC name is 4-(2-dimethylaminoethyl)phenol.

Such hordenine may be obtained from a plant of the genus Cactus Anhalonium , barley, etc. using various known extraction methods, or a commercially available hordenine may be used.

In Experimental Examples 1 to 4 according to the present invention, hordenine showed an apoptotic effect on skin cancer cell malignant melanoma, but had no effect on the death of melanocytes, skin fibroblasts and keratinocytes.

The hordenine presented in the present invention can be applied as a pharmaceutical composition and a cosmetic composition, and when applied to the skin, it exhibits an apoptotic effect on skin cancer cells without affecting the proliferation and death of normal cells, thereby treating skin cancer with minimal side effects. , preventive and ameliorative effects can be obtained.

When formulated as a pharmaceutical composition, the composition of the present invention can be formulated by a method known in the pharmaceutical field, and is mixed with a pharmaceutically acceptable carrier, excipient, etc. to conventional pharmaceutical preparations, such as solutions and syrups. , It can be formulated into capsules, granules, powders, ointments, emulsions, gels, creams, etc., and can be administered by various routes. Preferably, it may be formulated into a liquid, cream, paste, or solid formulation for transdermal administration such as an external solution, emulsion, or ointment.

The pharmaceutical composition of the present invention contains 0.001 to 10% by weight of the hordenine in the total pharmaceutical composition. The pharmaceutical composition thus prepared is used in an effective amount of 0.00001 mg/kg to 400 mg/kg per dose, and the actual dosage of the active ingredient depends on the severity of symptoms, the selected route of administration, the age, sex, weight and health of the subject. It should be understood that the decision must be made in light of several relevant factors, such as condition. Accordingly, the dosage is not intended to limit the scope of the present invention in any way. Administration may be administered once a day to several divided doses.

When formulated as a cosmetic composition, the content of the hordenine is 0.0001 to 10% by weight, preferably 0.01 to 5.0% by weight, based on the total weight of the cosmetic composition. The content of hordenine is preferably equal to or greater than the minimum value so as to achieve a minimal skin cancer improvement or preventive effect, and the content of hordenine is preferably equal to or less than the maximum value in consideration of deterioration in feeling of use due to excessive addition and possibility of application to various formulations. do. At this time, it is preferable that the content of hordenine is appropriately adjusted within the above range according to the content of ingredients contained in the formulation or cosmetic composition.

Ingredients included in the cosmetic composition of the present invention include ingredients commonly used in cosmetic compositions other than hordenine as an active ingredient, such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and conventional adjuvants such as fragrances, and contains a carrier.

The composition of the present invention can be prepared in any formulation commonly produced in the art, for example, softening lotion, astringent lotion, nutrient lotion, nutrient cream, massage cream, essence, eye cream, eye essence, cleansing cream, It can be formulated into cosmetics such as cleansing foam, cleansing water, pack, gel, powder, body lotion, body cream, body oil, and body essence.

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component. can

When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and in particular, in the case of a spray, additional chlorofluorohydrocarbon, propane / May contain a propellant such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizing agent or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butyl glycol oil, fatty acid esters of glycerol, polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystals Star cellulose, aluminum metahydroxide, bentonite, agar or tracanth and the like may be used.

When the formulation of the present invention is surfactant-containing cleansing, as carrier components, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide Ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester may be used.

Hereinafter, a preferred embodiment is presented to aid understanding of the present invention. However, the following examples are only provided to more easily understand the present invention, and the content of the present invention is not limited by the examples.

Experimental example 1. Hordenine this Effect on proliferation of skin cancer cells

Skin cancer cell melanoma was purchased from Gibco (Life Technologies, Carlsbad, CA, USA). Cells were cultured at 37 °C in DMEM medium (Gibco). The medium was maintained until the attached cells reached 80% confluence and subculture was performed at 80% confluence. Medium was changed every 2 days until confluence. In the subculture, the flask from which the culture medium was removed was washed with phosphate buffer saline (PBS), the cells were dropped with 0.25% trypsin-EDTA (Gibco BRL, Grand Island, NY, USA), and the cell suspension was centrifuged. After that, the number of cells was measured and subcultured three times again in the culture medium.

DMEM medium (Gibco) supplemented with 10% serum was added to a 96-well plate, respectively, and 5 X 10 ³ of skin cancer cells, malignant melanoma, were inoculated, and cultured at 5% CO ₂ and 37 °C to attach the cells to the culture dish. Then, after culturing in a culture solution from which serum was removed for 24 hours, the cells were divided into a negative control group and a hordenine (Sigma Aldrich) treatment group. At this time, 10 μl of hordenine was dissolved by concentration and treated (treatment concentration: 5, 10, 30, 50, and 100 μg/ml).

After the treatment, the cells were cultured for 72 hours at 5% CO ₂ and 37 °C. Thereafter, the MTT reagent was added, and after 4 hours, the medium was removed, 200 μl of DMSO was added to dissolve the formazan crystal, and the absorbance was measured at 570 nm using a microplate reader. The results are shown in Figure 1.

From the results of FIG. 1, it can be seen that apoptosis of skin cancer cells occurs depending on the concentration in the hordenine-treated condition, based on the control group. Apoptosis increased with concentration starting from 10 μg/ml treatment concentration, and almost all cells died when treated with 100 μg/mL treatment.

Experimental example 2. Hordenine this Effects on the proliferation of melanocytes

Human melanocytes were purchased from Gibco (Life Technologies, Carlsbad, CA, USA). Cells were cultured at 37 °C in Medium 254 medium (Gibco). The medium was maintained until the attached cells reached 80% confluence and subculture was performed at 80% confluence. Medium was changed every 2 days until confluence. In the subculture, the flask from which the culture medium was removed was washed with phosphate buffer saline (PBS), the cells were dropped with 0.25% trypsin-EDTA (Gibco BRL, Grand Island, NY, USA), and the cell suspension was centrifuged. After that, the number of cells was measured and subcultured three times again in the culture medium.

Medium 254 medium (Gibco) supplemented with supplements was added to a 96-well plate, and 5 X 10 ³ of human melanocytes were inoculated and cultured under 5% CO ₂ and 37 °C conditions to attach the cells to the culture dish. , After culturing in culture medium without supplement for 24 hours, they were divided into a negative control group and a hordenine (Sigma Aldrich) treatment group. At this time, 10 μl of hordenine was dissolved by concentration and treated (treatment concentration: 5, 10, 30, 50, and 100 μg/ml).

After the treatment, the cells were cultured for 72 hours at 5% CO ₂ and 37 °C. Thereafter, the MTT reagent was added, and after 4 hours, the medium was removed, and 200 μl of DMSO was added to dissolve the formazan crystal, and the absorbance was measured at 570 nm using a microplate reader. The results are shown in Figure 2.

From the results of FIG. 2, it was confirmed that there was no effect on the death of human melanocytes regardless of the concentration in the hordenin-treated condition, based on the control group.

Experimental example 3. Hordenine this Effect on proliferation of fibroblasts

Human dermal fibroblasts were purchased from Gibco (Life Technologies, Carlsbad, CA, USA). Cells were cultured at 37 °C in DMEM medium (Gibco). The medium was maintained until the attached cells reached 80% confluence and subculture was performed at 80% confluence. Medium was changed every 2 days until confluence. In the subculture, the flask from which the culture medium was removed was washed with phosphate buffer saline (PBS), the cells were dropped with 0.25% trypsin-EDTA (Gibco BRL, Grand Island, NY, USA), and the cell suspension was centrifuged. After that, the number of cells was measured and subcultured three times again in the culture medium.

DMEM medium (Gibco) supplemented with 10% serum was added to a 96-well plate, inoculated with 5 X 10 ³ human skin fibroblasts, and cultured under 5% CO ₂ and 37 °C conditions to attach the cells to the culture dish. Next, after culturing in a culture medium from which serum was removed for 24 hours, the cells were divided into a negative control group and a hordenine-treated group (Sigma Aldrich). At this time, 10 μl of hordenine was dissolved by concentration and treated (treatment concentration: 5, 10, 30, 50, and 100 μg/ml).

After the treatment, the cells were cultured for 72 hours at 5% CO ₂ and 37 °C. Thereafter, the MTT reagent was added, and after 4 hours, the medium was removed, and 200 μl of DMSO was added to dissolve the formazan crystal, and the absorbance was measured at 570 nm using a microplate reader. The results are shown in Figure 3.

From the results of FIG. 3 , it was confirmed that the apoptosis of human fibroblasts was not affected regardless of the concentration in the hordenine-treated condition, based on the control group.

Experimental example 4. Hordenine this Effects on proliferation of keratinocytes

Human epidermal keratinocytes were purchased from CELLnTEC Advanced Cell Systems. Cells were cultured at 37 °C in CnT-Prime medium (CELLnTEC). The medium was maintained until the attached cells reached 80% confluence and subculture was performed at 80% confluence. Medium was changed every 2 days until confluence. In the subculture, the flask from which the culture medium was removed was washed with phosphate buffer saline (PBS), the cells were dropped with 0.25% trypsin-EDTA (Gibco BRL, Grand Island, NY, USA), and the cell suspension was centrifuged. After that, the number of cells was measured and subcultured three times again in the culture medium.

Put CnT-Prime medium (CELLnTEC) in a 96-well plate, inoculate 5 X 10 ³ of human dermal keratinocytes, incubate under 5% CO ₂ and 37 °C conditions to attach the cells to the culture dish, and then for 24 hours After culturing in CnT-Prime Basal Medium culture medium, it was divided into a negative control group and a hordenine (Sigma Aldrich) treatment group. At this time, 10 μl of hordenine was dissolved by concentration and treated (treatment concentration: 5, 10, 30, 50, and 100 μg/ml).

After the treatment, the cells were cultured for 72 hours at 5% CO ₂ and 37 °C. Thereafter, the MTT reagent was added, and after 4 hours, the medium was removed, and 200 μl of DMSO was added to dissolve the formazan crystal, and the absorbance was measured at 570 nm using a microplate reader. The results are shown in FIG. 4 .

From the results of FIG. 4 , it was confirmed that the apoptosis of human keratinocytes was not affected regardless of the concentration in the hordenin-treated condition, when the control group was used as a standard.

5 is a graph showing the degree of cell growth inhibition of skin cancer cells, skin melanocytes, skin fibroblasts, and skin keratinocytes for each treatment concentration of hordenine.

As shown in FIG. 5, hordenine exhibits a cell death effect specifically for skin cancer cells, and it can be seen that there is no ability to inhibit proliferation and growth of skin normal cells such as skin melanocytes, skin fibroblasts, and skin keratinocytes. there is.

Hereinafter, preferred formulation examples are presented to aid understanding of the present invention. However, the following formulation examples are only provided to more easily understand the present invention, and the content of the present invention is not limited by the formulation examples.

formulation example 1: Soft lotion (skin lotion)

As shown in Table 1 below, softening lotion containing hordenine was prepared according to a conventional method.

ingredient Content (% by weight) Hordenine 0.0001 glycerin 5.0 1,3-butylene glycol 3.0 PEG 1500 1.0 allantoin 0.1 DL-panthenol 0.3 EDTA-2Na 0.02 Benzophenone-9 0.04 sodium hyaluronate 5.0 ethanol 10.0 Octyldodeces-16 0.2 polysorbate 20 0.2 incense 0.02 Distilled water balance Sum 100

formulation example 2: Astringent lotion

As shown in Table 2 below, an astringent lotion containing hordenine was prepared according to a conventional method.

ingredient Content (% by weight) Hordenine 0.0001 glycerin 2.0 1,3-butylene glycol 2.0 allantoin 0.2 DL-panthenol 0.2 EDTA-2Na 0.02 Benzophenone-9 0.04 sodium hyaluronate 3.0 ethanol 15.0 polysorbate 20 0.3 Witch Hagel Extract 2.0 citric acid a very small amount incense 0.02 Distilled water balance Sum 100

formulation example 3: Nutrition lotion

As shown in Table 3 below, nutritional lotion containing hordenine was prepared according to a conventional method.

ingredient Content (% by weight) Hordenine 0.0001 Glyceryl Stearate SE 1.5 Cetearyl Alcohol 1.5 lanolin 1.5 Polysorbate 60 1.3 Sorbitan Stearate 0.5 hydrogenated vegetable oil 1.0 mineral oil 5.0 squalane 3.0 trioctanoin 2.0 dimethicone 0.8 Tocopherol Acetate 0.5 carboxyvinyl polymer 0.12 glycerin 5.0 1,3-butylene glycol 3.0 sodium hyaluronate 5.0 triethanolamine 0.12 incense 0.01 Distilled water balance Sum 100

formulation example 4: Nutrition Cream

As shown in Table 4 below, a nutritious cream containing hordenine was prepared according to a conventional method.

ingredient Content (% by weight) Hordenine 0.0001 Pro-type glycerin monostearate 2.0 Cetearyl Alcohol 2.2 stearic acid 1.5 beeswax 1.0 Polysorbate 60 1.5 Sorbitan Stearate 0.6 hydrogenated vegetable oil 1.0 squalane 3.0 mineral oil 5.0 trioctanoin 5.0 dimethicone 1.0 sodium magnesium silicate 0.1 glycerin 5.0 betaine 3.0 triethanolamine 1.0 sodium hyaluronate 4.0 incense 0.01 Distilled water balance Sum 100

formulation example 5: massage cream

As shown in Table 5 below, a massage cream containing hordenine was prepared according to a conventional method.

ingredient Content (% by weight) Hordenine 0.0001 Pro-type glycerin monostearate 1.5 Cetearyl Alcohol 1.5 stearic acid 1.0 Polysorbate 60 1.5 Sorbitan Stearate 0.6 isostearyl isostearate 5.0 squalane 5.0 mineral oil 35 dimethicone 0.5 Hydroxyethyl Cellulose 0.12 glycerin 6.0 triethanolamine 0.7 incense 0.01 Distilled water balance Sum 100

formulation example 6 : Essence

As shown in Table 6 below, an essence containing hordenine was prepared according to a conventional method.

ingredient Content (% by weight) Hordenine 0.0001 glycerin 10 betaine 5.0 PEG 1500 2.0 allantoin 0.1 DL-panthenol 0.3 EDTA-2Na 0.02 Benzophenone-9 0.04 Hydroxyethyl Cellulose 0.1 sodium hyaluronate 8.0 carboxyvinyl polymer 0.2 triethanolamine 0.18 Octyldodecanol 0.3 Octyldodeces-16 0.4 ethanol 6.0 incense 0.02 Distilled water balance Sum 100

formulation example 7: Pack

As shown in Table 7 below, a pack containing hordenine was prepared according to a conventional method.

ingredient Content (% by weight) Hordenine 0.0001 polyvinyl alcohol 15.0 cellulose gum 0.15 glycerin 3.0 PEG 1500 2.0 cyclodextrin 0.15 DL-panthenol 0.4 allantoin 0.1 Monoammonium Glycyrrhizinate 0.3 nicotinamide 0.5 ethanol 6.0 PEG 40 hydrogenated castor oil 0.3 incense 0.01 Distilled water balance Sum 100

formulation example 8: ointment

As shown in Table 8 below, an ointment containing hordenine was prepared according to a conventional method.

ingredient Content (% by weight) Hordenine 0.0001 Purified water balance Cetostearyl Alcohol 3.0 hard liquid paraffin 5.0 Tocopherol Acetate 2.0 Ceteareth-20 3.0 panthenol 0.5 Methyl paraoxybenzoate Appropriate amount Paraoxybenzoic Acid Profile Appropriate amount

Claims (8)

A pharmaceutical composition for treating or preventing malignant melanoma skin cancer containing Hordenine as an active ingredient. The pharmaceutical composition according to claim 1, wherein the hordenine is contained in an amount of 0.01 to 10% by weight based on the total weight of the pharmaceutical composition. The method of claim 1, wherein the pharmaceutical composition
A pharmaceutical composition characterized in that it is liquid, cream, paste or solid. delete A cosmetic composition for preventing or improving malignant melanoma skin cancer containing hordenine as an active ingredient. The cosmetic composition according to claim 5, wherein the hordenine is contained in an amount of 0.01 to 10% by weight based on the total weight of the cosmetic composition. The method of claim 5, wherein the cosmetic composition
It is characterized in that it is any one formulation selected from the group consisting of softening lotion, astringent lotion, nutrient lotion, nutrient cream, massage cream, essence, pack, body lotion, body cream, body oil, body essence, gel, emulsion and body cleanser The cosmetic composition to be. delete

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