Composition comprising tubulin inhibitor for preventing or treating skin diseases

Abstract

The present invention relates to a pharmaceutical composition comprising a tubulin inhibitor as an active ingredient for preventing or treating skin diseases. In addition, the present invention relates to a quasi-drug composition or cosmetic composition comprising a tubulin inhibitor as an active ingredient for preventing or alleviating skin diseases. In addition, the present invention relates to a cosmetic composition comprising a tubulin inhibitor as an active ingredient for skin moisturization, whitening, wrinkle alleviation, acne relief, skin barrier strengthening, or skin keratinocyte differentiation. The tubulin inhibitor of the present invention increases the expression level of filaggrin in epidermal cells, promotes skin differentiation, inhibits NF S-kB signaling induced by TNF-α, and exhibits photo-aging inhibition and wrinkle alleviation effects as well as whitening effects. Accordingly, the tubulin inhibitor of the present invention can be utilized as an active ingredient that exhibits activities, such as skin moisturization, whitening, wrinkle alleviation, acne relief, skin barrier strengthening, or skin keratinocyte differentiation, along with the prevention, treatment, or alleviation of skin diseases.

Description

Composition for preventing or treating skin diseases comprising a tubulin inhibitor

[0001] The present invention relates to a pharmaceutical composition for preventing or treating skin diseases, comprising a tubulin inhibitor as an active ingredient.

[0002] In addition, the present invention relates to a pharmaceutical composition for preventing or improving skin diseases, comprising a tubulin inhibitor as an active ingredient.

[0003] In addition, the present invention relates to a cosmetic composition for preventing or improving skin diseases, which comprises a tubulin inhibitor as an active ingredient.

[0004] In addition, the present invention relates to a cosmetic composition for moisturizing the skin, whitening, improving wrinkles, alleviating acne, strengthening the skin barrier, or differentiating skin keratinocytes, which contains a tubulin inhibitor as an active ingredient.

[0005]

[0006] The primary function of the skin is to act as a barrier, separating the internal environment from the external environment, protecting against aggression from foreign substances and minimizing the loss of moisture and other essential body components to the external space. Filaggrin is particularly important in skin barrier formation due to its fundamental role in the terminal differentiation of the epidermis and its association with skin diseases. First identified as a key structural protein in 1977, filaggrin was given the name filaggrin (filament-aggregating protein) after it was discovered that it acts to aggregate and compact keratin intermediate filaments. This protein is synthesized from the large precursor protein profilaggrin, which is a major component of keratohyalin granules in the granular layer of the epidermis.

[0007] A key component of the skin barrier is the stratum corneum. This layer is the result of differentiation of keratinocytes as they migrate from the basal layer to the granulosum layer. As these cells mature, they express various structural proteins.

[0008] Filaggrin is continuously processed by various proteases. This proteolytic degradation releases hygroscopic amino acids and their derivatives, which form the natural moisturizing factor (NMF) and play a role in retaining moisture in the stratum corneum.

[0009] That is, filaggrin is converted from the precursor protein profilaggrin to active filaggrin monomer, and forms the stratum corneum by coagulating and compressing keratin filaments to flatten keratinocytes, and the natural moisturizing factor (NMF) produced during the filaggrin processing process contributes to maintaining moisture in the skin and controlling acidic pH.

[0010] Additionally, filaggrin binds intermediate keratin filaments to structural proteins of the cornified epithelium (CE), which gives keratinocytes strong mechanical and chemical resistance. Together with lipids released from lamellar bodies, filaggrin also contributes to the formation of the extracellular lipid matrix of the stratum corneum.

[0011] In particular, filaggrin degradation products UCA (Urocanic acid) and PCA (Pyrroliodone-5-carboxylic acid) help maintain skin moisture, maintain an acidic pH, and protect against UV radiation. UCA also plays a key role in antimicrobial action and immune regulation.

[0012]

[0013] Accordingly, the present inventors confirmed that a tubulin inhibitor increases filaggrin expression in epidermal cells and promotes epidermal cell differentiation. Furthermore, more surprisingly, they confirmed that the tubulin inhibitor suppresses inflammatory signals, inhibits photoaging, inhibits collagen degradation, and even possesses a whitening effect, thereby completing the present invention.

[0014] Accordingly, the purpose of the present invention is to provide a pharmaceutical composition or quasi-drug composition for preventing or treating skin diseases. Furthermore, the purpose of the present invention is to provide a cosmetic composition for preventing or improving skin diseases, and a cosmetic composition for moisturizing, whitening, improving wrinkles, alleviating acne, strengthening the skin barrier, or differentiating skin keratinocytes.

[0015]

[0016] To achieve the above purpose, the present invention provides a pharmaceutical composition for preventing or treating skin diseases, which comprises a tubulin inhibitor as an active ingredient.

[0017] In addition, the present invention provides a pharmaceutical composition for preventing or improving skin diseases, and a cosmetic composition for preventing or improving skin diseases, which comprises a tubulin inhibitor as an active ingredient.

[0018] In addition, the present invention provides a cosmetic composition for skin moisturizing, whitening, wrinkle improvement, acne relief, skin barrier strengthening, or skin keratinocyte differentiation, which contains a tubulin inhibitor as an active ingredient.

[0019]

[0020] The tubulin inhibitor of the present invention increases the expression level of filaggrin in epidermal cells, promotes skin differentiation, inhibits the NF-kB signaling process induced by TNF-α, exhibits photoaging inhibition and wrinkle improvement effects, and exhibits a whitening effect. Accordingly, the tubulin inhibitor of the present invention can be utilized as an effective ingredient that simultaneously exhibits activities such as skin moisturizing, whitening, wrinkle improvement, acne relief, skin barrier strengthening, or skin keratinocyte differentiation, in addition to preventing, treating, or improving skin diseases.

[0021]

[0022] Figure 1 shows the effect of a tubulin inhibitor on filaggrin expression in a human epidermal keratinocyte (NHEK) cell line. The results show that filaggrin expression increases with tubulin inhibitor treatment.

[0023] Figure 2 shows the effect of a tubulin inhibitor on cell differentiation in a human epidermal keratinocyte (NHEK) cell line. The results show that cell differentiation is promoted after treatment with a tubulin inhibitor.

[0024] Figure 3 shows the results of an experiment to confirm the inhibitory effect of a tubulin inhibitor on NF-kB signaling. The results show that NF-kB signaling is inhibited after treatment with a tubulin inhibitor.

[0025] Figure 4 shows the effect of tubulin inhibitors on reducing the expression levels of MMP-1, MMP-3, and MMP-9 induced by photoaging. Tubulin inhibitors show results that suppress UV-induced MMP expression levels.

[0026] Figure 5 shows the inhibition of tyrosinase activity and the whitening effect of a tubulin inhibitor. It shows that a tubulin inhibitor inhibits tyrosinase activity and reduces melanin production.

[0027]

[0028] Hereinafter, the present invention will be described in detail.

[0029]

[0030] The present invention provides a pharmaceutical composition for preventing or treating skin diseases, comprising a tubulin inhibitor as an active ingredient.

[0031] The present invention also provides a method for preventing or treating a skin disease, comprising administering a therapeutically effective amount of a tubulin inhibitor to a subject in need thereof.

[0032] Tubulin inhibitors (Tubulin inhibitors) refer to a group of drugs that regulate or inhibit the function of tubulin, a key component of microtubules. Microtubules are part of the cytoskeleton, playing a crucial role within cells and are essential structures for processes such as cell division, cell movement, and intracellular transport.

[0033] In the present invention, unlike the previous functions, the tubulin inhibitor exhibits activity in increasing the expression of filaggrin in epidermal cells and in skin proliferation according to epidermal cell differentiation, inhibits the NF-kB signaling process induced by TNF-α, exhibits photoaging inhibition and wrinkle improvement effects, and exhibits a whitening effect.

[0034] In the present invention, the tubulin inhibitor is not limited in type, but may be, for example, at least one selected from the group consisting of vinca alkaloid, taxane, cryptophycin 52, halichondrins, dolastatins, hemisterlins, combretastatin-A4, 2-methoxyestradiol, E7010, plinabulin, epothilone, and discodermolide.

[0035] The vinca alkaloid of the present invention may preferably be Vinblastine, Vincristine, Vinorelbine, or Vinflunine.

[0036] Additionally, the taxane may be paclitaxel or docetaxel.

[0037] The tubulin inhibitor of the present invention is a concept that includes all pharmaceutically acceptable salts, isomers, derivatives, and analogs that exhibit the same or similar effects.

[0038] In the invention, pharmaceutically acceptable salts refer to salts commonly used in the pharmaceutical industry, and examples thereof include inorganic ion salts manufactured with calcium, potassium, sodium and magnesium, inorganic acid salts manufactured with hydrochloric acid, nitric acid, phosphoric acid, hydrobromic acid, iodic acid, perchloric acid and sulfuric acid, organic acid salts manufactured with acetic acid, trifluoroacetic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid and hydroiodic acid, sulfonic acid salts manufactured with methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid and naphthalenesulfonic acid, glycine, arginine, Examples thereof include, but are not limited to, amino acid salts manufactured from lysine, etc., and amine salts manufactured from trimethylamine, triethylamine, ammonia, pyridine, picoline, etc.

[0039] The above tubulin inhibitor may be derived from an extract, produced biologically, or synthesized chemically, and is not limited in its origin and production method.

[0040] In the present invention, skin disease is a group of diseases caused by structural and functional abnormalities of the skin, and is a concept encompassing abnormal conditions occurring in the skin that can be caused by various causes such as inflammation, infection, allergy, and immune abnormality.

[0041] In the present invention, the skin disease may be at least one selected from the group consisting of, for example, atopic dermatitis, skin thermal damage, pruritus, acne, psoriasis, allergic dermatitis, contact dermatitis, exfoliative dermatitis, seborrheic dermatitis, seborrheic dermatitis, lichen planus, rosacea, pigmentation disorder, hypermelanosis, erythema, wounds, ulcers, bedsores, lupus, skin wrinkle-related diseases, and skin diseases caused by photodamage, and the type thereof is not limited.

[0042] The above skin wrinkle-related diseases may be elastic fibrosis, thinning of the skin, skin atrophy, reduction of collagen fibers and elastic fibers, loss of skin elasticity, dryness, wrinkle formation, or premature skin aging.

[0043] Additionally, the skin diseases caused by the photodamage may be lentigines, freckles, hypopigmentation, hyperpigmentation, photodamage due to acute or chronic UV radiation, or photosensitization.

[0044] Additionally, the skin disease of the present invention may be squamous cell carcinoma, basal cell carcinoma, benign epithelial tumor, and radiation dermatitis.

[0045] In addition, the skin disease of the present invention may be panniculitis, calluses, vitiligo, urticaria, folliculitis, sebaceous keratosis pilaris, eczema, corns, freckles, rashes, athlete's foot, spots, stretch marks, freckles, prickly heat, dry skin, chilblains, boils, keratosis pilaris, scalp dermatitis, or psoriatic arthritis.

[0046]

[0047] In addition, the present invention may further include a compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, and tofacitinib, in addition to the above tubulin inhibitor.

[0048] A compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, and tofacitinib may be used in combination or complex with a tubulin inhibitor, and administration may be simultaneous or sequential.

[0049] The above calcium, colchicine, tapinarof, fingolimod, or tofacitinib is a concept that includes all of its salts, isomers, or derivatives, and analogues that exhibit the same or similar effects.

[0050]

[0051] In the present invention, the tubulin inhibitor can be provided as an active ingredient in a pharmaceutical composition for preventing or treating skin diseases.

[0052] The term “prevention” as used herein may include inhibiting the occurrence of a disease.

[0053] The term “treatment” as used herein includes inhibition, alleviation, or elimination of the development of a disease.

[0054] The term "included as an active ingredient" in this specification means that the tubulin inhibitor of this specification is added to an extent that it can exhibit the effect mentioned in this specification, and includes the formulation in various forms by adding various components as auxiliary components for drug delivery and stabilization, etc.

[0055] The pharmaceutical composition may contain the tubulin inhibitor in a pharmaceutically effective amount.

[0056] As used herein, the term “pharmaceutically effective amount” means an amount sufficient to achieve the efficacy or activity of an active ingredient.

[0057] The pharmaceutical composition may additionally comprise a pharmaceutically acceptable diluent or carrier. The diluent may be lactose, corn starch, soybean oil, microcrystalline cellulose, mannitol, or a combination thereof. The carrier may be an excipient, a disintegrant, a binder, a glidant, or a combination thereof. The excipient may be microcrystalline cellulose, lactose, low-substituted hydroxycellulose, or a combination thereof. The disintegrant may be calcium carboxymethylcellulose, sodium starch glycolate, calcium hydrogen phosphate anhydrous, or a combination thereof. The binder may be polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, hydroxypropylcellulose, or a combination thereof. The glidant may be magnesium stearate, silicon dioxide, talc, or a combination thereof.

[0058] The pharmaceutical composition may be formulated as an oral or parenteral dosage form. Oral dosage forms may include granules, powders, liquids, tablets, capsules, dry syrups, or combinations thereof. Parenteral dosage forms may include injections, ointments, aerosols, sprays, patches, and the like, with no particular limitation on the dosage form.

[0059] The dosage of the pharmaceutical composition according to one example is, for example, about 0.0001 to 10000 mg / kg, 0.0001 to 5000 mg / kg, 0.0001 to 1000 mg / kg, 0.0001 to 900 mg / kg, 0.0001 to 800 mg / kg, 0.0001 to 700 mg / kg, 0.0001 to 600 mg / kg, 0.0001 to 500 mg / kg, 0.0001 to 400 mg / kg, 0.0001 to 300 mg / kg, 0.0001 to 200mg / kg, 1 to 1000mg / kg, 1 to 900mg / kg, 1 to 800mg / kg, 1 to 700mg / kg, 1 to 600mg / kg, 1 to 500mg / kg, 1 to 450mg / kg, 1 to 400mg / kg, 1 to 350mg / kg, 1 to 300mg / kg, 1 to 250mg / kg, 10 to 1000mg / kg, 10 to 900mg / kg, 10 to 800mg / kg, 10 to 700mg / kg, 10 to 600mg / kg, 10 to 500mg / kg, 10 to 450mg / kg, 10 to 400mg / kg, 10 to 350mg / kg, 10 to 300mg / kg, 10 to 250mg / kg, 100 to 1000mg / kg, 100 to 900mg / kg, 100 to 800mg / kg, 100 to 700mg / kg, 100 to 600mg / kg, 100 to 500mg / kg, 100 to 450mg / kg, 100 to 400mg / kg, 100 to 350mg / kg, 100 to 300mg / kg, 100 to 250mg / kg, 200 to 1000mg / kg, 200 to 900mg / kg, 200 to 800mg / kg, 200 to 700mg / kg, 200 to 600mg / kg, 200 The dosage may be administered in the range of 500 mg / kg to 500 mg / kg, 200 to 450 mg / kg, 200 to 400 mg / kg, 200 to 350 mg / kg, 200 to 300 mg / kg, or 200 to 250 mg / kg, but is not limited thereto. The frequency of administration of the pharmaceutical composition of the present application is not particularly limited thereto, but may be administered once a day or administered in divided doses several times. The above dosage does not limit the scope of the present application in any way.

[0060]

[0061] The subject of administration of the pharmaceutical composition provided herein may be a mammal including a human, a dog, a cat, a horse, a cow, a pig, a goat, a rabbit, a mouse, a rat, etc., or a cell, tissue, or culture thereof isolated therefrom. In one example, the subject may be a subject (a mammal such as a human) that requires prevention, improvement, and / or treatment of a skin disease as described above, or a cell, tissue, or culture thereof isolated therefrom.

[0062] According to an example, a pharmaceutical composition comprises the tubulin inhibitor in an amount of 1 to 80 wt%, 5 to 80 wt%, 5 to 75 wt%, 5 to 70 wt%, 5 to 65 wt%, 50 to 70 wt%, 55 to 65 wt%, 60 to 65 wt%, 10 to 60 wt%, 15 to 60 wt%, 20 to 60 wt%, 1 to 50 wt%, 5 to 50 wt%, 10 to 50 wt%, 15 to 50 wt%, 20 to 50 wt%, 1 to 40 wt%, 5 to 40 wt%, 10 to 40 wt%, 15 to 40 wt%, 20 to 40 wt%, 1 to 30 wt%, 5 to 30 wt%, 10 to It may comprise 30 wt%, 15 to 30 wt%, 20 to 30 wt%, 1 to 25 wt%, 5 to 25 wt%, 10 to 25 wt%, 15 to 25 wt%, 20 to 25 wt%, or 23 to 25 wt%.

[0063]

[0064] Alternatively, the composition may be a composition for external use in the skin.

[0065] The above skin external preparation may be, but is not limited to, a cream, gel, ointment, skin emulsifier, skin suspension, transdermal patch, drug-containing bandage, lotion, spray, or a combination thereof.

[0066] In the present invention, the external skin preparation may be appropriately mixed with ingredients commonly used in external skin preparations such as cosmetics or medicines, such as aqueous ingredients, oily ingredients, powder ingredients, alcohols, moisturizers, thickeners, ultraviolet absorbers, whitening agents, preservatives, antioxidants, surfactants, fragrances, colorants, various skin nutrients, metal ion sequestrants, sugars, or combinations thereof, as needed.

[0067]

[0068] In addition, the present invention provides a pharmaceutical composition for preventing or improving skin diseases, which comprises a tubulin inhibitor as an active ingredient.

[0069] In the present invention, a quasi-drug refers to any product other than a drug related to the treatment or prevention of a disease.

[0070] In the present invention, quasi-drugs refer to all products other than pharmaceuticals related to the treatment or prevention of diseases. The quasi-drugs above refer to products that are used for the purpose of diagnosing, treating, improving, alleviating, managing, or preventing diseases of humans or animals, and have a milder effect than pharmaceuticals. For example, according to the Pharmaceutical Affairs Act, quasi-drugs are products excluding those used for the purpose of pharmaceuticals, and may include, but are not limited to, fiber and rubber products used for the treatment or prevention of diseases of humans or animals, products that have a mild or no direct effect on the human body, are not instruments or machines, and similar products, and sterilizers and insecticides for preventing infectious diseases.

[0071] The type or formulation of the pharmaceutical composition of the present invention is not particularly limited, but may preferably be a disinfectant, a shower foam, a gargle, a wet tissue, a detergent soap, a hand wash, a humidifier filler, a mask, an ointment, or a filter filler.

[0072] When the composition of the present invention is included in an over-the-counter drug for skin moisturizing purposes, the composition may be used as is or in combination with other over-the-counter drug ingredients, and may be used appropriately according to conventional methods. The mixing amount of the active ingredients may be appropriately determined depending on the intended use, and the over-the-counter drug composition according to the present invention may contain the tubulin inhibitor in an amount of 0.01 to 20 wt% based on the total weight of the composition.

[0073]

[0074] In addition, the present invention provides a cosmetic composition for preventing or improving skin diseases, which comprises a tubulin inhibitor as an active ingredient.

[0075] The term "improvement" herein may mean any action that at least reduces the severity of a symptom, for example, a parameter associated with alleviating or treating a condition.

[0076] The tubulin inhibitor is present in an amount of 0.0001 to 80 wt%, 0.01 to 70 wt%, 0.01 to 60 wt%, 0.01 to 50 wt%, 0.01 to 40 wt%, 0.01 to 30 wt%, 0.1 to 70 wt%, 0.1 to 60 wt%, 0.1 to 50 wt%, 0.1 to 40 wt%, 0.1 to 30 wt%, 0.5 to 70 wt%, 0.5 to 60 wt%, 0.5 to 50 wt%, 0.5 to 40 wt%, 0.5 to 30 wt%, 1 to 70 wt%, 1 to 60 wt%, 1 to 50 wt%, 1 to 40 wt%, or 1 to 30%, based on the total weight of the cosmetic composition. For example, it may be included at 30 wt%, but is not limited thereto.

[0077] The above cosmetic composition is not particularly limited to a specific formulation, and the formulation may be appropriately selected depending on the intended purpose. The cosmetic composition may, for example, have a solubilized formulation, an emulsified formulation, or a dispersed formulation. The cosmetic composition may have a cosmetic formulation of an emollient toner, a nourishing toner, a massage cream, a nourishing cream, an essence, a pack, a gel, an ampoule, or a skin-adhesive type, but is not limited thereto.

[0078] The above cosmetic composition may additionally include ingredients commonly used in cosmetic compositions in addition to the effective ingredients disclosed herein, and may include, for example, conventional auxiliary agents and carriers such as antioxidants, stabilizers, solubilizers, surfactants, dispersants, emulsifiers, preservatives, vitamins, pigments, and fragrances.

[0079]

[0080] In addition, the present invention provides a cosmetic composition for skin moisturizing, whitening, wrinkle improvement, acne relief, skin barrier strengthening, or skin keratinocyte differentiation, which contains a tubulin inhibitor as an active ingredient.

[0081] As used herein, the term “skin moisturizing” may mean any action that maintains skin moisture or prevents moisture loss.

[0082] The term "whitening" as used herein may mean improving the brightness of skin tone or alleviating pigmentation by inhibiting melanin production.

[0083] The term "wrinkle improvement" as used herein may mean increasing skin elasticity, reducing the depth of wrinkles, or increasing the smoothness of the skin surface.

[0084] The term "acne relief" as used herein may mean suppressing inflammation, redness, or excessive sebum production of acne, or promoting healing of acne lesions.

[0085] In this specification, the term “strengthening the skin barrier” may mean any action that enhances the function of the skin barrier, which is located at the outermost part of the skin and prevents moisture and nutrient loss.

[0086] The term "skin keratinocyte differentiation" as used herein may mean promoting the process of keratinocytes maturing to form the stratum corneum, or thereby strengthening the function of the skin barrier.

[0087]

[0088] Another aspect of the present invention provides a method for preventing, improving, or treating a skin disease comprising administering an effective amount of the tubulin inhibitor to a subject in need thereof.

[0089] The term "effective amount" means an amount effective enough to produce the effect mentioned above.

[0090] The subject may be a mammal, such as a human, cow, horse, pig, dog, sheep, goat, or cat. The subject may be an object in need of a preventive, ameliorating, or therapeutic effect for a skin disease.

[0091] As used herein, the term "administering" is used interchangeably with "introducing" and "implanting" and may mean placement of a composition according to one embodiment into a subject by a method or route that results in at least partial localization of the composition to a desired site.

[0092] Administration may be by any method known in the art. The route of administration, frequency of administration, and other administration methods can be appropriately selected by those skilled in the art. Administration may be administered directly to a subject by any means, including intravenous, intramuscular, oral, transdermal, mucosal, intranasal, buccal, intratracheal, or subcutaneous administration. Administration may be systemic or local. Administration may include application to the skin.

[0093] In the present invention, administration is 0.1 mg to 1,000 mg of the composition according to one specific example per subject per day, for example, 0.1 mg to 500 mg, 0.1 mg to 100 mg, 0.1 mg to 50 mg, 0.1 mg to 25 mg, 1 mg to 1,000 mg, 1 mg to 500 mg, 1 mg to 100 mg, 1 mg to 50 mg, 1 mg to 25 mg, 5 mg to 1,000 mg, 5 mg to 500 mg, 5 mg to 100 mg, 5 mg to 50 mg, 5 mg to 25 mg, 10 mg to 1,000 mg, 10 mg to 500 mg, 10 mg The dosage may be administered in the range of 100 mg to 100 mg, 10 mg to 50 mg, or 10 mg to 25 mg. However, the dosage may be prescribed in various ways depending on factors such as the formulation method, administration method, patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and response sensitivity, and a person skilled in the art can appropriately adjust the dosage by considering these factors. The number of administrations may be once a day or twice or more within the range of clinically acceptable side effects, and the administration site may be administered in one or more sites, and the total number of administration days may be from 1 to 30 days per treatment at intervals of 2 to 5 days daily or every day. If necessary, the same treatment may be repeated after an appropriate period. For animals other than humans, the same dosage per kg as for humans may be administered, or the dosage may be converted to the above dosage based on the volume ratio (e.g., average value) of the organs (heart, etc.) of the target animal and humans.

[0094] Another aspect provides the use of the tubulin inhibitor for preparing a composition for preventing, improving or treating a skin disease.

[0095] Another aspect provides the use of the tubulin inhibitor for use in the manufacture of a medicament for preventing, improving or treating a skin disease.

[0096] Another aspect provides a food composition for preventing or improving skin diseases, comprising a tubulin inhibitor as an active ingredient.

[0097] A food composition according to one embodiment of the present invention may be in the form of a liquid or solid formulation, and may be in the form of a tablet, capsule, soft capsule, pill, granule, beverage (drink), diet bar, chocolate, caramel formulation, or confectionery formulation, and the formulation is not particularly limited. In addition to the above-mentioned effective ingredient, the food composition of the present invention may contain excipients, sugars, flavorings, colorings, fats, proteins, etc. as needed.

[0098]

[0099] Duplicate contents are omitted in consideration of the complexity of this specification, and terms not otherwise defined in this specification have meanings commonly used in the technical field to which the present invention belongs.

[0100]

[0101] Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are only illustrative of the present invention, and the content of the present invention is not limited to the following examples.

[0102]

[0103] [Experimental Example 1] Strain

[0104] Normal Human Epidermal Keratinocyte (NHEK) cell line is a human fetal keratinocyte cell line obtained from ATCC. These cells were obtained through primary culture and used to examine intracellular biochemical changes induced by tubulin inhibitors under normal, non-pathogenic conditions.

[0105] Human embryonic kidney cells (HEK) were obtained from ATCC and used to examine intracellular biochemical changes induced by tubulin inhibitors under normal, non-disease-specific conditions.

[0106]

[0107] [Experimental Example 2]

[0108] 1.1 Measurement of filaggrin expression using cell-based promoter activity measurement

[0109] After inserting a luciferase gene with a filaggrin promoter into a human epidermal keratinocyte (NHEK) cell line using Fugene4K reagent (Promega), the cells were treated with each tubulin inhibitor at a concentration of 1.0 μM 24 hours later. 24 hours after drug treatment, the level of filaggrin expression in response to each tubulin inhibitor treatment was measured using a kit for measuring the level of luciferase luminescence (Promega). DMSO at the same volume as the treated compound was used as a control.

[0110]

[0111] 1.2 Observation of cell differentiation using phase contrast microscopy

[0112] After treating human epidermal cell lines (Normal Human Epidermal Keratinocytes; NHEK) with each tubulin inhibitor at a concentration of 1.0 uM, cell differentiation was observed daily using a phase contrast microscope.

[0113] On the 5th day of cell differentiation, images were taken using a phase contrast microscope camera, and the results are shown in Figure 2.

[0114]

[0115] [Experimental Example 3] Confirmation of Inflammatory Signaling Mechanism Inhibition Using Cell-Based Promoter Activity Measurement

[0116] NF-κB, a DNA transcription factor, exists in the cytosol and exists in a bound form with IκB, which inhibits NF-κB. NF-κB is involved in innate and adaptive immune responses and is a representative pro-inflammatory cytokine that appears in many inflammatory diseases.

[0117] The NF-κB mechanism is a protein family involved in inflammatory response regulation, immune modulation, apoptosis, cell proliferation, and epithelial differentiation. It regulates the expression of various genes and forms the central axis of the intracellular signal transmission system.

[0118] After inserting a luciferase gene with the NFkb promoter into human embryonic kidney cells (HEK) using Fugene4K reagent (Promega), inflammation was induced by treatment with TNF-α 6 hours later, and Nfkb signaling was induced. After 18 hours, each tubulin inhibitor was treated at a concentration of 1.0 μM.

[0119] After 24 hours of drug treatment, the NfkB signaling pathway inhibition effect according to each tubulin inhibitor treatment was measured using a kit (Promega) that measures the degree of luciferase emission. In addition, when 2 mM CaCl2, 1 μM colchicine, 1 μM tapinarof, 1 μM fingolimod, and 1 μM tofacitinib were co-treated with tubulin inhibitors, it was confirmed whether the NfkB signaling pathway inhibition effect was enhanced compared to the experimental group treated with tubulin inhibitors alone.

[0120]

[0121] [Experimental Example 4] Inhibitory effect of tubulin inhibitor on collagen reduction mechanism

[0122] Photoaging refers to the skin aging process caused by exposure to the sun's ultraviolet rays. It is distinct from senescence (or biological aging), which is caused by the gradual decline in bodily functions. It is generally the primary cause of wrinkles, dryness, loss of elasticity, and pigmentation.

[0123] The sun emits two types of ultraviolet rays: UV-A and UV-B, with wavelengths ranging from 320 to 400 nm for UV-A and 290 to 320 nm for UV-B. Collagen is a prime example of tissue damage caused by this UV radiation. Collagen is a major component of connective tissue, primarily found in bone and skin, and is broadly classified into five types. Type 1 collagen is the most abundant type found in skin connective tissue, followed by Type 3.

[0124] Matrix Metalloproteinases (MMPs) are proteolytic enzymes that break down collagen, a major component of human connective tissue. Their expression levels are increased by UV rays. Therefore, when skin is exposed to UV rays, the expression of these matrix metalloproteinases (MMPs) increases, destroying collagen in the dermal layer and contributing to photoaging, including wrinkles and pigmentation.

[0125] Six hours after application of tubulin inhibitors or control groups to human skin cells (NHEK), the cells were exposed to UB UV light for 30 minutes. Intracellular RNA was extracted from the skin cells using the RNeasy mini kit (Qiagen). The extracted RNA was reverse transcribed into cDNA using the ImProm-II TM Reverse Transcription kit (Promega). The expression levels of MMP-1, MMP-3, and MMP-9 in the skin cells were measured by quantitative PCR (qPCR) using a Real-Time PCR Detection System (Bio-Rad, CFX96).

[0126]

[0127] [Test Example 5] Confirmation of whitening function using tyrosinase activity measurement

[0128] To confirm whether the tubulin inhibitor of the present invention has a whitening function, tyrosinase activity was measured. Tyrosinase is an enzyme that produces melanin by oxidizing tyrosine in the presence of oxygen, and inhibition of tyrosinase activity suppresses melanin production. Tyrosine is oxidized by tyrosinase, resulting in an orange color. However, when tyrosinase activity is inhibited, the oxidation reaction of tyrosine through tyrosinase is reduced, so the color gradually becomes lighter and transparent depending on the degree of tyrosinase inhibition. The degree of tyrosinase inhibition was evaluated by measuring the decrease in absorbance value at 490 nm.

[0129]

[0130] [Example 1] Filaggrin expression and cell differentiation in human epidermal cell lines treated with tubulin inhibitors

[0131] The expression level of filaggrin was measured using cell-based reporter assay and RT-PCR by treating cultured human epidermal keratinocyte (NHEK) cell lines with a tubulin inhibitor.

[0132] Each tubulin inhibitor was treated with NHEK at a concentration of 1.0 μM, and for comparison of filaggrin expression and cell differentiation, DMSO at the same dose as the treatment drug was treated as a control group, and after 24 hours of treatment, the amount of filaggrin expression and the degree of cell differentiation were observed.

[0133] The results are shown in Figures 1 and 2.

[0134] As can be seen in Figure 1a, the expression level of filaggrin increased in the tubulin inhibitor-treated group. In addition, as can be seen in Figure 2a, the degree of cell differentiation was significantly increased in the tubulin inhibitor-treated group.

[0135] In addition, in order to confirm the effect of combined use of tubulin inhibitors, when each tubulin inhibitor was combined with 2 mM CaCl2, 1 μM colchicine, 1 μM tapinarof, 1 μM fingolimod, and 1 μM tofacitinib, it was confirmed that the expression of filaggrin (Fig. 1a to 1b) and cell differentiation ability (Fig. 2b to 2c) increased compared to the experimental group treated with the tubulin inhibitor alone.

[0136]

[0137]

[0138] [Example 2] Confirmation of the inhibition of inflammatory mechanisms by tubulin inhibitors.

[0139] Through a cell-based reporter assay, it was confirmed that NF-kB activation was induced by treating human epidermal cell lines (NHEK) with TNF-α, and then the NF-kB signaling process was inhibited by treating with a tubulin inhibitor at a concentration of 1.0 μM. The results are shown in Fig. 3a.

[0140] As can be seen in Fig. 3a, treatment with a tubulin inhibitor according to the present invention inhibited NF-kB signal transduction compared to the control group treated with only TNF-α. TPCA1, a well-known NF-kB signal transduction inhibitor, was used as a positive control.

[0141] In addition, when 2 mM CaCl2, 1 μM colchicine, 1 μM tapinarof, 1 μM fingolimod, 1 μM tofacitinib, and a tubulin inhibitor were co-treated, it was confirmed that the NfkB signaling pathway inhibition effect was enhanced compared to the experimental group treated with a tubulin inhibitor alone (Figures 3a to b).

[0142]

[0143] [Example 3] Confirmation of the inhibition mechanism of collagen reduction by tubulin inhibitors.

[0144] A tubulin inhibitor was applied to human NHEK cells, and after 30 minutes, UV irradiation was performed to induce photoaging (particularly, wrinkle improvement). To compare and verify the tubulin inhibitor, a negative control group with no treatment and a positive control group irradiated only with UV-A were included in the experiment.

[0145] After application and UV exposure experiments, RNA was extracted from skin cells. The extracted RNA was reverse transcribed into cDNA, and the amounts of MMP-1, MMP-3, and MMP-9 present in skin tissue were measured using quantitative PCR (qPCR) using a real-time PCR machine.

[0146] The photoaging inhibitory effect of tubulin inhibitors was measured by measuring the change in the expression level of Matrix Metalloproteinases (MMP) proteins described above. That is, the expression level of MMP proteins was measured in each of the control group irradiated with UV rays only, the experimental group irradiated with UV rays after applying the tubulin inhibitor, and the group treated with a combination of the tubulin inhibitor and 2 mM CaCl2, 1 μM colchicine, 1 μM tapinarof, 1 μM fingolimod, and 1 μM tofacitinib, to determine the degree to which the expression level of MMP proteins was suppressed by application of the tubulin inhibitor.

[0147]

[0148] The results are shown in Figures 4a to 4f.

[0149] MMP-1 and MMP-9 are enzymes that promote the decomposition of type 1 and type 3 collagen, which are mainly expressed in skin tissue, and MMP-3 is a regulatory enzyme that decomposes type 1 collagen and controls changes in the expression level of MMP-1, and is an MMP protein that has a very important effect on photoaging.

[0150] As can be confirmed in Fig. 4a, Fig. 4c, and Fig. 4e, when the tubulin inhibitor was applied to the skin, the expression levels of MMP-1, MMP-3, and MMP-9 were all reduced by approximately 50% compared to the control group that was only irradiated with UV rays. In this way, it was verified that the application of the tubulin inhibitor suppressed the expression levels of Matrix Metalloproteinases (MMP), and the overall effect of suppressing photoaging by UV rays was confirmed. The effect of suppressing the expression levels of MMP-1, MMP-3, and MMP-9, which were increased by UV-B irradiation, was measured according to the treatment with each tubulin inhibitor.

[0151] In addition, when 2 mM CaCl2, 1 μM colchicine, 1 μM tapinarof, 1 μM fingolimod, 1 μM tofacitinib, and a tubulin inhibitor were combined, it was confirmed that the inhibitory effect on the expression of MMP-1, MMP-3, and MMP-9 was increased compared to the experimental group treated with the tubulin inhibitor alone (Fig. 4a to Fig. 4f).

[0152]

[0153] [Example 4] Verification of the pigmentation improvement effect of a tubulin inhibitor.

[0154] In a tyrosinase activity inhibition experiment, 1.0 μM concentration of each tubulin inhibitor was treated with 2 mM of the substrate L-tyrosine, and the absorbance was measured at 490 nm. Then, tyrosinase enzyme was added again and the reaction was performed at 37ºC. After measuring the absorbance at 490 nm again, the absorbance value was calculated to determine the degree to which each tubulin inhibitor inhibits tyrosinase activity. Kojic acid, known to inhibit tyrosinase activity, was used as a control group. Through this, the degree to which each tubulin inhibitor inhibits melanin production can be determined. The results are shown in Figs. 5a and 5b.

[0155] As shown in Fig. 5a, each tubulin inhibitor has the activity of inhibiting melanin production through inhibition of tyrosinase activity.

[0156] In addition, as can be confirmed in FIGS. 5a and 5b, when 2 mM CaCl2, 1 μM colchicine, 1 μM tapinarof, 1 μM fingolimod, 1 μM tofacitinib, and a tubulin inhibitor were treated together, the effect of inhibiting tyrosinase activity was confirmed to be increased compared to the experimental group treated with the tubulin inhibitor alone.

Claims

1. A pharmaceutical composition for preventing or treating skin diseases, comprising a tubulin inhibitor as an active ingredient.

2. A composition according to claim 2, wherein the tubulin inhibitor is at least one selected from the group consisting of vinca alkaloid, taxane, cryptophycin 52, Halichondrins, Dolastatins, Hemiasterlins, Combretastatin-A4, 2-Methoxyestradiol, E7010, Plinabuline, Epothilone, Discodermolide, and pharmaceutically acceptable salts of the above compounds.

3. A composition according to claim 2, wherein the vinca alkaloid is at least one selected from the group consisting of Vinblastine, Vincristine, Vinorelbine, Vinflunine, and pharmaceutically acceptable salts of the above compounds.

4. A composition according to claim 2, wherein the taxane is at least one selected from the group consisting of paclitaxel, docetaxel, and pharmaceutically acceptable salts of the compounds.

5. A composition according to claim 1, wherein the skin disease is at least one selected from the group consisting of atopic dermatitis, skin thermal damage, pruritus, acne, psoriasis, allergic dermatitis, contact dermatitis, exfoliative dermatitis, seborrheic dermatitis, seborrheic dermatitis, lichen planus, rosacea, pigmentation disorders, hypermelanosis, erythema, wounds, ulcers, bedsores, lupus, skin wrinkle-related diseases, and skin diseases caused by photodamage.

6. A composition according to claim 5, wherein the skin wrinkle-related disease is at least one selected from the group consisting of elasticity fibrosis, thinning of the skin, skin atrophy, reduction of collagen fibers and elastic fibers, loss of skin elasticity, dryness, wrinkle formation, and premature skin aging.

7. A composition according to claim 5, wherein the skin disease caused by photodamage is at least one selected from the group consisting of lentigines, freckles, hypopigmentation, hyperpigmentation, photodamage caused by acute or chronic UV radiation, and photosensitization.

8. A composition according to claim 1, wherein the skin disease is at least one selected from the group consisting of squamous cell carcinoma, basal cell carcinoma, benign epithelial tumor, and radiation dermatitis.

9. A composition according to claim 1, wherein the skin disease is at least one selected from the group consisting of panniculitis, calluses, vitiligo, urticaria, folliculitis, sebaceous keratosis pilaris, eczema, styes, freckles, blemishes, rashes, athlete's foot, spots, stretch marks, prickly heat, dry skin, chilblains, suppuration, keratoses, scalp dermatitis, and psoriatic arthritis.

10. A composition according to claim 1, wherein the composition further comprises a compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, and tofacitinib.

11. A pharmaceutical composition for preventing or improving skin diseases containing a tubulin inhibitor as an active ingredient.

12. A cosmetic composition for preventing or improving skin diseases, comprising a tubulin inhibitor as an active ingredient.

13. A cosmetic composition for moisturizing the skin, whitening, improving wrinkles, alleviating acne, strengthening the skin barrier, or differentiating skin keratinocytes, containing a tubulin inhibitor as an active ingredient.

14. A method for preventing or treating a skin disease, comprising the step of administering a therapeutically effective amount of a tubulin inhibitor to a subject in need thereof.

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