Composition for inhibiting, alleviating, ameliorating or treating cutaneous pruritus comprising N-nonyldeoxynojirimycin

Abstract

The present invention provides a pharmaceutical composition for inhibiting, alleviating or treating pruritus cutanea, said pharmaceutical composition comprising N-nonyldeoxynojirimycin, and provides a cosmetic composition for inhibiting, alleviating or improving pruritus cutanea, said cosmetic composition comprising N-nonyldeoxynojirimycin.

Description

Technical Field

[0001] The present invention relates to a pharmaceutical composition for suppressing, alleviating or treating pruritus of the skin, the pharmaceutical composition comprising N-nonyl-deoxynojirimycin, and relates to a cosmetic composition for suppressing, alleviating or improving pruritus of the skin, the cosmetic composition comprising N-nonyl-deoxynojirimycin. Background Art

[0002] Chronic pruritus of the skin can be roughly classified into etiologies related to the skin barrier and etiologies related to abnormal immune responses. Pruritus related to the skin barrier is a symptom that appears widely from infants to adults. It is known that for people with damaged skin barriers, external substances (allergens) easily pass through the stratum corneum, causing the body to become sensitive and thus inducing skin pruritus including atopic dermatitis (Agrawal R and Woodfolk JA, Curr Allergy Asthma Rep, 14:433, 2014). The main cause of pruritus related to the skin barrier is known to originate from the epidermal layer (epidermal keratinocytes) (Kim BE and Leung D, Allergy Asthma Immunol Res, 4:12, 2012; Pastore S et al., Eur J Dermatol, 16:125, 2006). Pruritus related to abnormal immune responses is accompanied by various diseases in addition to atopic dermatitis, and it is difficult to solve with local cosmetic materials.

[0003] The present inventors have disclosed that specific peptide fragments can be usefully used in cosmetic compositions for suppressing the formation of skin wrinkles by promoting the expression of junction proteins by promoting the binding between epidermal cells (Korean Patent No. 10-1817244 and 10-1817250, Korean Patent Application No. 10-2022-0024100, etc.). Summary of the Invention

[0004] (I) Technical Problems to be Solved

[0005] The present inventors conducted various cell-based analytical studies on components derived from natural products, particularly various alkaloid derivatives, to discover components that can inhibit or ameliorate pruritus. As a result, the present inventors found that N-nonyldeoxynojirimycin (NN-DNJ) has excellent inhibitory, alleviating, ameliorating or therapeutic activity against pruritus by inhibiting the secretion of β-hexosaminidase and Prostaglandin 2 (PGE2) in mast cells in a concentration-dependent manner and inhibiting pruritus associated with the skin barrier.

[0006] Accordingly, an object of the present invention is to provide a pharmaceutical composition for inhibiting, alleviating or treating pruritus, which comprises N-nonyldeoxynojirimycin.

[0007] In addition, an object of the present invention is to provide a cosmetic composition for inhibiting, alleviating or ameliorating pruritus, which comprises N-nonyldeoxynojirimycin.

[0008] (II) Technical Solution

[0009] According to one embodiment of the present invention, there is provided a pharmaceutical composition for inhibiting, alleviating or treating pruritus, which comprises N-nonyldeoxynojirimycin.

[0010] According to another embodiment of the present invention, there is provided a cosmetic composition for inhibiting, alleviating or ameliorating pruritus, which comprises N-nonyldeoxynojirimycin.

[0011] In the pharmaceutical composition or cosmetic composition of the present invention, the pruritus may be pruritus caused by one or more diseases or disorders selected from atopic dermatitis, xerosis, contact dermatitis, urticaria, prurigo nodularis, bacterial infection, viral infection, skin parasite infection, lichen simplex chronicus, insect bite and nummular eczema. In a specific embodiment, the pruritus may be pruritus caused by atopic dermatitis or xerosis.

[0012] (III) Advantageous Effects

[0013] According to the present invention, it is disclosed that N-nonyl-deoxynojirimycin significantly inhibits the secretion of β-hexosaminidase and prostaglandin 2 in mast cells in a concentration-dependent manner, and inhibits pruritus related to the skin barrier, and has excellent inhibitory, alleviating, improving or therapeutic activity against skin pruritus. Therefore, N-nonyl-deoxynojirimycin can be usefully applied in a pharmaceutical composition for inhibiting, alleviating or treating skin pruritus, and can be usefully applied in a cosmetic composition for inhibiting, alleviating or improving skin pruritus by effectively inhibiting skin pruritus related to the skin barrier and skin pruritus related to abnormal immune responses. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 The evaluation results showing the inhibitory effects of various alkaloid derivatives on the secretion of β-hexosaminidase induced by PMA / A23187 stimulation in RBL-2H3 cells.

[0015] Figure 2 The evaluation results showing the inhibitory effects of treatment with N-nonyl-deoxynojirimycin at different concentrations on the secretion of β-hexosaminidase induced by PMA / A23187 stimulation in RBL-2H3 cells.

[0016] Figure 3 The evaluation results showing the inhibitory effects of treatment with N-nonyl-deoxynojirimycin at different concentrations on the secretion of PGE2 induced by PMA / A23187 stimulation in RBL-2H3 cells.

[0017] Figure 4 The evaluation results showing the inhibitory effects of treatment with N-nonyl-deoxynojirimycin at different concentrations on the secretion of IL-6 induced by TNF-α / IFN-γ stimulation in HaCaT cells.

[0018] Figure 5 The evaluation results showing the inhibitory effects of treatment with N-nonyl-deoxynojirimycin at different concentrations on the secretion of CCL17 induced by TNF-α / IFN-γ stimulation in HaCaT cells.

[0019] Figure 6 The evaluation results showing the inhibitory effects of treatment with N-nonyl-deoxynojirimycin at different concentrations on the secretion of CCL22 induced by TNF-α / IFN-γ stimulation in HaCaT cells. DETAILED DESCRIPTION OF THE INVENTION

[0020] The present invention provides a composition for inhibiting, alleviating, improving or treating skin pruritus, the composition comprising N-nonyl-deoxynojirimycin.

[0021] The present inventors used various natural product-derived components, particularly various alkaloid derivatives, as targets, and screened for substances that inhibit β-hexosaminidase secretion stimulated by PMA / A23187 using mast cells (RBL-2H3 cells), and revealed that N-nonyl-deoxynojirimycin (NN-DNJ) has concentration-dependent inhibitory activity on β-hexosaminidase and prostaglandin 2 secretion. In addition, the present inventors found that N-nonyl-deoxynojirimycin can effectively inhibit pruritus associated with the skin barrier. Therefore, N-nonyl-deoxynojirimycin can be usefully applied in pharmaceutical compositions for inhibiting, alleviating, or treating skin pruritus and cosmetic compositions for inhibiting, alleviating, or improving skin pruritus.

[0022] N-nonyl-deoxynojirimycin (NN-DNJ), as a compound having the structure of the following Chemical Formula 1, is known to have inhibitory activity against glucosidase and to inhibit the breakdown of liver glycogen in vivo.

[0023] <Chemical Formula 1>

[0024]

[0025] In the composition of the present invention, the skin pruritus may be skin pruritus caused by various chronic skin diseases or disorders. For example, the skin pruritus may be skin pruritus caused by one or more diseases or disorders selected from atopic dermatitis, xerosis, contact dermatitis, urticaria, prurigo nodularis, bacterial infection, viral infection, skin parasite infection, lichen simplex chronicus, insect bite, and nummular eczema. In a specific embodiment, the skin pruritus may be skin pruritus caused by atopic dermatitis or xerosis.

[0026] The composition of the present invention may be in the form of a pharmaceutical composition or a cosmetic composition containing N-nonyl-deoxynojirimycin as an active ingredient. For example, the composition of the present invention may be in the form of a pharmaceutical composition for inhibiting, alleviating, or treating skin pruritus, and the composition of the present invention contains N-nonyl-deoxynojirimycin. In addition, the present invention may be in the form of a cosmetic composition containing N-nonyl-deoxynojirimycin for inhibiting, alleviating, or improving skin pruritus.

[0027] The pharmaceutical composition may comprise a pharmaceutically acceptable carrier and can be formulated into parenteral dosage forms such as solutions, suspensions, emulsions, lotions, ointments or lyophilized products according to conventional methods. Preferably, the pharmaceutical composition can be formulated into transdermal dosage forms such as topical solutions, emulsions or ointments. The pharmaceutically acceptable carrier includes aqueous diluents or solvents such as phosphate buffered saline, purified water, sterile water, and non-aqueous diluents or solvents such as propylene glycol, polyethylene glycol, olive oil. In addition, according to needs, the pharmaceutically acceptable carrier may include wetting agents, fragrances, preservatives, etc. The dose of N-nonyl-deoxynojirimycin contained in the pharmaceutical composition can be appropriately selected by those skilled in the art according to the patient's condition, severity of the condition, route of administration and administration time, etc. N-nonyl-deoxynojirimycin can be administered (e.g., applied) to the itchy skin area in an amount of 3 μg to 3 mg per day, and the administration can be carried out once a day or in several divided doses.

[0028] The cosmetic composition may be in the form of a functional cosmetic composition containing N-nonyl-deoxynojirimycin as an active ingredient. The cosmetic composition can be made into various forms according to conventional cosmetic manufacturing methods. For example, the cosmetic composition can be made into forms such as makeup products, lotions, creams, emulsions containing the peptide, which can be used after dilution with conventional cleansing liquids, astringents and moisturizing liquids. In addition, the cosmetic composition may include conventional adjuvants such as stabilizers, solubilizers, vitamins, pigments and fragrances commonly used in the field of cosmetic compositions. In the cosmetic composition, the N-nonyl-deoxynojirimycin may contain an effective amount for inhibiting, alleviating or improving skin pruritus. For example, the content of the N-nonyl-deoxynojirimycin may be 0.001-10% by weight based on the total weight of the composition.

[0029] Hereinafter, the present invention will be described in detail by way of examples. However, these examples are only for illustrating the present invention, and the scope of the present invention is not limited to these examples.

[0030] Example 1: Cultivation of mast cells (RBL-2H3 cells)

[0031] The mast cells (RBL-2H3 cells) were purchased from the American Type Culture Collection (ATCC, USA). The RBL-2H3 cells were cultured in Dulbecco's Modified Eagle's medium (high glucose, Hyclone, USA) containing 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin in an incubator at 37 °C with 5% CO2.

[0032] Example 2: Evaluation of the inhibitory effect on β-hexosaminidase secretion in RBL-2H3 cells

[0033] To evaluate the inhibitory effect on the degranulation of RBL-2H3 cells using various compounds as objects, the secretion amount of β-hexosaminidase was measured. Specifically, the cultured RBL-2H3 cells were seeded at 2×10 5Cells / well were seeded at a density into each well of a 24-well plate, and DMEM medium containing 10% FBS and 1% penicillin / streptomycin was added for culture. After 24 hours, the existing medium was removed, and the cells were washed twice with extracellular buffer (5 mM KCl, 119 mM NaCl, 25 mM PIPES, 0.4 mM MgCl2, 1 mM CaCl2, 40 mM NaOH, 5.6 mM glucose, 0.1% BSA, pH 7.2), and then the medium was replaced with extracellular buffer. The test substances [1-Deoxynojirimycin, 1-Deoxymannojirimycin, Miglitol, 1-Deoxy-L-idonojirimycin, Deoxygalactonojirimycin, nojirimycin-1-sulfonic acid, NN-DNJ, N-butyl-deoxynojirimycin (Miglustat), Voglibose, Isofagomine D-Tartrate, Swainsonine, Deoxyfuconojirimycin, Castanospermine, N-methyl-1-deoxynojirimycin] were each added at 20 μM and cultured in an incubator at 37 °C with 5% CO2 for 1 hour, and then treated with 50 nM Phorbol 12-myristate 13-acetate (PMA) and 1 μM A23187 (Merck, Germany) as a calcium ionophore, and further cultured for 30 minutes. The plate was placed in an ice bath at 4 °C for 10 minutes to terminate the degranulation reaction, and then the supernatant was mixed in equal proportion with substrate buffer (1 mM 4-p-nitrophenyl N-acetyl-β-D-glucosamine, 0.05 M sodium citrate, pH 4.5) and reacted in an incubator at 37 °C with 5% CO2 for 1 hour.After the reaction was terminated by adding the Stop solution (0.1 M carbonate buffer, pH 10), the absorbance was measured at 405 nm using a microplate reader. The results are shown in. Figure 1 in. From Figure 1 the results, it was confirmed that, compared with other alkaloid analogs, N-nonyl-deoxynojirimycin showed a significantly superior effect of inhibiting degranulation of RBL-2H3 cells by significantly inhibiting the secretion of β-hexosaminidase.

[0034] Example 3: Evaluation of the inhibitory effect of N-nonyl-deoxynojirimycin on the secretion of β-hexosaminidase in RBL-2H3 cells according to the treatment concentration

[0035] The amount of β-hexosaminidase secreted was measured by the same method as in Example 2, except that RBL-2H3 cells were treated with different concentrations (1 - 200 μM) of N-nonyl-deoxynojirimycin. The results are as Figure 2 shown. From Figure 2 the results, it was confirmed that N-nonyl-deoxynojirimycin inhibited the secretion of β-hexosaminidase in a concentration-dependent manner.

[0036] Example 4: Evaluation of the inhibitory effect of the treatment of N-nonyl-deoxynojirimycin on the secretion of PGE2 in RBL-2H3 cells

[0037] To evaluate the inhibitory effect of the treatment of N-nonyl-deoxynojirimycin on the secretion of prostaglandin 2 (PGE2) by RBL-2H3 cells, an enzyme-linked immunosorbent assay (ELISA) was performed using a PGE2 ELISA kit (R&D Systems, USA). Specifically, the cultured RBL-2H3 cells were seeded into each well of a 6-well plate at a density of 1×10 6 cells / well and cultured by the same method as in Example 2. After 24 hours, the existing medium was removed, washed with PBS, and then DMEM medium without FBS was added. The cells were treated with different concentrations (1 - 40 μM) of N-nonyl-deoxynojirimycin and cultured in a 37 °C, 5% CO2 incubator for 1 hour, then treated with 50 nM PMA and 1 μM A23187 and further cultured for 30 minutes. The cell culture supernatant was collected in a 1.5 ml tube, centrifuged at 2000 relative centrifugal force (rcf), and then the supernatant was used for ELISA analysis using a PGE2 ELISA kit (R&D Systems, USA). After the reaction was terminated, the absorbance at 450 nm was measured using a microplate reader. The results are shown in Figure 3 in. From Figure 3The results confirmed that N-nonyldeoxynojirimycin inhibited the secretion of PGE2 in a concentration-dependent manner.

[0038] Example 5: Culture of epidermal cells (HaCaT cells)

[0039] Epidermal cells (HaCaT cells) were purchased from Thermo Fisher Scientific (USA). HaCaT cells were cultured in DMEM medium containing 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin at 37°C in a 5% CO2 incubator.

[0040] Example 6: Evaluation of the inhibitory effect of N-nonyldeoxynojirimycin treatment on IL-6, CCL17, and CCL22 secretion in HaCaT cells

[0041] To evaluate the inhibitory effect on IL-6, CCL17, and CCL22 secretion induced by TNF-α and IFN-γ stimulation in epidermal HaCaT cells, a cell model for skin barrier-related pruritus, IL-6 ELISA kit (Abcam, UK), CCL17 ELISA kit (R&D Systems, USA), and CCL22 ELISA kit (R&D Systems, USA) were used. HaCaT cells were plated at 1×10 6 The density of cells / well was inoculated into each well of a 6-well plate with a cell culture-specific coating, and DMEM medium containing 10% FBS and 1% antibiotics (penicillin / streptomycin) was added for culture. After 24 hours, the existing culture medium was removed, washed with PBS, and then DMEM medium without FBS was added, treated with different concentrations of N-nonyldeoxynojirimycin, and cultured in an incubator at 37°C and 5% CO2 for 1 hour, and then treated with 10ng / ml of TNF-α and IFN-γ, respectively, and further cultured for 24 hours. The cell culture fluid was collected in a 1.5ml test tube, centrifuged at 2000rcf, and the supernatant was used for ELISA analysis. After termination of the reaction, the absorbance at 450nm was measured using a microplate reader. The results are shown in Figure 4 、 Figure 5 and Figure 6 In. By Figure 4 、 Figure 5 and Figure 6 The results confirmed that N-nonyldeoxynojirimycin inhibited the secretion of IL-6, CCL17, and CCL22 from epidermal cells induced by TNF-α / IFN-γ stimulation of pruritus in a concentration-dependent manner.

[0042] This invention was carried out with the support of the Gyeonggi Regional Research and Cooperation Center (GRRC) (Advanced Center for Special Resources in Northeast Gyeonggi - Development of Health Functional Foods of Functional Kimchi and Tae Myung Cheong Beverage).

Claims

1. A pharmaceutical composition for inhibiting, alleviating or treating pruritus, comprising N-nonyl-deoxynojirimycin.

2. The pharmaceutical composition according to claim 1, wherein The pruritus is pruritus caused by one or more diseases or disorders selected from atopic dermatitis, xerosis, contact dermatitis, urticaria, prurigo nodularis, bacterial infection, viral infection, skin parasite infection, lichen simplex chronicus, insect bite and nummular eczema.

3. The pharmaceutical composition according to claim 1, wherein The pruritus is pruritus caused by atopic dermatitis or xerosis.

4. A cosmetic composition for inhibiting, alleviating or improving pruritus, comprising N-nonyl-deoxynojirimycin.

5. The cosmetic composition according to claim 4, characterized in that, The pruritus is pruritus caused by one or more diseases or disorders selected from atopic dermatitis, xerosis, contact dermatitis, urticaria, prurigo nodularis, bacterial infection, viral infection, skin parasite infection, lichen simplex chronicus, insect bite and nummular eczema.

6. The cosmetic composition according to claim 4, characterized in that, The pruritus is pruritus caused by atopic dermatitis or xerosis.

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