Compositions for the prevention or treatment of skin diseases, containing tubulin inhibitors

Abstract

The present invention relates to a pharmaceutical composition for the prevention or treatment of skin diseases, comprising a tubulin inhibitor as an active ingredient. Furthermore, the present invention relates to a quasi-drug composition or cosmetic composition for the prevention or improvement of skin diseases, comprising a tubulin inhibitor as an active ingredient. Furthermore, the present invention relates to a cosmetic composition for skin moisturizing, whitening, wrinkle improvement, acne relief, skin barrier strengthening, or skin keratinocyte differentiation, comprising a tubulin inhibitor as an active ingredient. The tubulin inhibitor of the present invention increases filaggrin expression in epidermal cells, promotes skin differentiation, suppresses the NF-κB signaling pathway induced by TNF-α, exhibiting photoaging inhibition and wrinkle improvement effects, as well as a whitening effect. Therefore, the tubulin inhibitor of the present invention can be utilized as an active ingredient that simultaneously exhibits activities such as skin moisturizing, whitening, wrinkle improvement, acne relief, skin barrier strengthening, or skin keratinocyte differentiation, along with the prevention, treatment, or improvement of skin diseases.

Description

Technical Field

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

[0002] The present invention also relates to a quasi-drug composition for preventing or improving skin diseases, containing a tubulin inhibitor as an active ingredient.

[0003] The present invention also relates to a cosmetic composition for preventing or improving skin diseases, containing a tubulin inhibitor as an active ingredient.

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

Background Art

[0005] The main function of the skin is to act as a barrier that separates the internal environment from the external environment, protect against attacks by external substances, and minimize 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 final differentiation process of the epidermis and its association with skin diseases. After filaggrin was first identified as an important structural protein in 1977, it was revealed that filaggrin plays a role in aggregating and compressing keratin intermediate filaments, and it was named as an abbreviation for filament-aggregating protein. This protein is synthesized as a large precursor protein, profilaggrin, which is the main component of keratohyalin granules in the granular layer of the epidermis.

[0006] The main component of the skin barrier is the stratum corneum. This stratum corneum is the result of keratinocytes migrating and differentiating from the basal layer to the granulosum layer. These cells express various structural proteins during the maturation process.

[0007] Filaggrin is continuously processed by various proteases. This protein degradation process releases hygroscopic amino acids and their derivatives, which form natural moisturizing factors (NMF) that play a role in retaining moisture in the stratum corneum.

[0008] In other words, filaggrin is converted from its precursor protein, profilaggrin, to the active filaggrin monomer, which aggregates and compresses keratin filaments, flattening the keratinocytes and forming the stratum corneum. The natural moisturizing factors (NMF) produced during the filaggrin processing contribute to the skin's moisture retention and regulation of its acidic pH.

[0009] Furthermore, filaggrin binds intermediate keratin filaments to structural proteins of the keratinized coat (CE), giving keratinocytes strong mechanical and chemical resistance. Along with lipids released from lamellar bodies, filaggrin also contributes to the formation of the extracellular lipid matrix of the stratum corneum.

[0010] In particular, UCA (Urocanic acid) and PCA (Pyrroliodone-5-carboxylic acid), which are breakdown products of filaggrin, retain moisture in the skin, maintain an acidic pH, and protect the skin from UV radiation. UCA also plays an important role in antimicrobial activity and immunomodulation. [Overview of the project] [Problems that the invention aims to solve]

[0011] Therefore, the inventors confirmed that tubulin inhibitors increase filaggrin expression in epidermal cells and promote epidermal cell differentiation. Furthermore, they confirmed that tubulin inhibitors suppress inflammatory signals, inhibit photoaging, inhibit collagen degradation mechanisms, and even possess a skin-whitening effect, thus completing the present invention.

[0012] Therefore, an object of the present invention is to provide pharmaceutical compositions and quasi-drug compositions for the prevention or treatment of skin diseases. Another object of the present invention is to provide cosmetic compositions for the prevention or improvement of skin diseases and cosmetic compositions for moisturizing the skin, whitening, wrinkle improvement, acne relief, strengthening the skin barrier, or differentiation of skin keratinocytes. [Means for solving the problem]

[0013] To achieve the above objective, the present invention provides a pharmaceutical composition for the prevention or treatment of skin diseases, comprising a tubulin inhibitor as an active ingredient.

[0014] Furthermore, the present invention provides a quasi-drug composition for the prevention or improvement of skin diseases, and a cosmetic composition for the prevention or improvement of skin diseases, both containing a tubulin inhibitor as an active ingredient.

[0015] Furthermore, the present invention provides a cosmetic composition containing a tubulin inhibitor as an active ingredient for moisturizing the skin, whitening, improving wrinkles, alleviating acne, strengthening the skin barrier, or differentiating skin keratinocytes. [Effects of the Invention]

[0016] The tubulin inhibitor of the present invention increases filaggrin expression in epidermal cells, promotes skin differentiation, suppresses the NF-κB signaling pathway induced by TNF-α, exhibits photoaging inhibition and wrinkle improvement effects, and shows a skin whitening effect. Therefore, the tubulin inhibitor of the present invention can be used as an active ingredient that simultaneously exhibits activities such as skin moisturizing, skin whitening, wrinkle improvement, acne relief, skin barrier strengthening, or skin keratinocyte differentiation, in addition to preventing, treating, or improving skin diseases. [Brief explanation of the drawing]

[0017] [Figure 1a] This figure shows the effect of tubulin inhibitors on filaggrin expression in human epidermal cell lines (NHEKs). The results show that filaggrin expression increases with treatment using tubulin inhibitors. [Figure 1b] This figure shows the effect of tubulin inhibitors on filaggrin expression in human epidermal cell lines (NHEKs). The results show that filaggrin expression increases with treatment using tubulin inhibitors. [Figure 2a] This figure shows the effect of tubulin inhibitors on the degree of cell differentiation in human epidermal cell lines (NHEKs). It demonstrates that cell differentiation is promoted after treatment with a tubulin inhibitor. [Figure 2b] This figure shows the effect of tubulin inhibitors on the degree of cell differentiation in human epidermal cell lines (NHEKs). It demonstrates that cell differentiation is promoted after treatment with a tubulin inhibitor. [Figure 2c] This figure shows the effect of tubulin inhibitors on the degree of cell differentiation in human epidermal cell lines (NHEKs). It demonstrates that cell differentiation is promoted after treatment with a tubulin inhibitor. [Figure 3a] This figure shows experimental results to confirm the inhibitory effect of tubulin inhibitors on NF-κB signaling. It shows that NF-κB signaling is suppressed after treatment with a tubulin inhibitor. [Figure 3b]This is a figure regarding the experimental results for confirming the NF-κB signaling inhibition effect of a tubulin inhibitor. After treatment with the tubulin inhibitor, the results show that NF-κB signaling is inhibited. [Figure 4a] This is a figure regarding the effect of reducing the expression levels of MMP-1, MMP-3, and MMP-9 induced by photoaging of a tubulin inhibitor. The figure shows the results that the tubulin inhibitor suppresses the UV-induced MMP expression levels. [Figure 4b] This is a figure regarding the effect of reducing the expression levels of MMP-1, MMP-3, and MMP-9 induced by photoaging of a tubulin inhibitor. The figure shows the results that the tubulin inhibitor suppresses the UV-induced MMP expression levels. [Figure 4c] This is a figure regarding the effect of reducing the expression levels of MMP-1, MMP-3, and MMP-9 induced by photoaging of a tubulin inhibitor. The figure shows the results that the tubulin inhibitor suppresses the UV-induced MMP expression levels. [Figure 4d] This is a figure regarding the effect of reducing the expression levels of MMP-1, MMP-3, and MMP-9 induced by photoaging of a tubulin inhibitor. The figure shows the results that the tubulin inhibitor suppresses the UV-induced MMP expression levels. [Figure 4e] This is a figure regarding the effect of reducing the expression levels of MMP-1, MMP-3, and MMP-9 induced by photoaging of a tubulin inhibitor. The figure shows the results that the tubulin inhibitor suppresses the UV-induced MMP expression levels. [Figure 4f] This is a figure regarding the effect of reducing the expression levels of MMP-1, MMP-3, and MMP-9 induced by photoaging of a tubulin inhibitor. The figure shows the results that the tubulin inhibitor suppresses the UV-induced MMP expression levels. [Figure 5a] This is a figure regarding the tyrosinase activity inhibition and whitening effect of a tubulin inhibitor. The figure shows the results that the tubulin inhibitor inhibits tyrosinase activity and reduces melanin production. [Figure 5b]This is a diagram related to the inhibitory effect of tubulin inhibitors on tyrosinase activity and their whitening effect. It shows the result that tubulin inhibitors suppress tyrosinase activity and reduce melanin production.

Mode for Carrying Out the Invention

[0018] The present invention will be described in detail below.

[0019] The present invention provides a pharmaceutical composition for preventing or treating skin diseases containing a tubulin inhibitor as an active ingredient.

[0020] The present invention also provides a method for preventing or treating skin diseases, which includes the step of administering a therapeutically effective amount of a tubulin inhibitor to an individual who needs it.

[0021] Tubulin inhibitors refer to a group of drugs that regulate or suppress the function of tubulin protein, which is the main component of microtubules. Microtubules are part of the cytoskeleton that play an important role in cells and are essential structures in processes such as cell division, cell movement, and intracellular transport.

[0022] Different from such previous functions, the tubulin inhibitor in the present invention shows activity in increasing the expression level of filaggrin in epidermal cells and skin growth due to epidermal cell differentiation, suppresses the NF-κB signal transduction process induced by TNF-α, shows the effects of suppressing photoaging and improving wrinkles, and shows a whitening effect.

[0023] The tubulin inhibitor in the present invention is not limited in type and may be one or more selected from the group consisting of, for example, vinca alkaloid, taxane, cryptophycin 52, halichondrins, dolastatins, hemiasterlins, combretastatin-A4, 2-methoxyestradiol, E7010, planabuline, epothilone, and discordermolide.

[0024] The vinca alkaloid of the present invention may preferably be vinblastine, vincristine, vinorelbine, or vinflunine.

[0025] Furthermore, the taxane may be paclitaxel or docetaxel.

[0026] The tubulin inhibitors of the present invention are a concept that includes all pharmaceutically acceptable salts, isomorphs, or derivatives thereof, as well as analogs that exhibit the same or similar effects.

[0027] In this invention, pharmaceutically acceptable salts mean salts commonly used in the pharmaceutical industry, including, for example, inorganic ion salts made from calcium, potassium, sodium, and magnesium; inorganic salts made from hydrochloric acid, nitric acid, phosphoric acid, bromate, iodic acid, perchloric acid, and sulfuric acid; 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, galactronic acid, glutamic acid, and glutamic acid. Examples include, but are not limited to, organic salts produced from taric acid, glucuronic acid, aspartic acid, ascorbic acid, carboxylic acids, vanillic acid, hydroiodic acid, etc.; sulfonates produced from methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, and naphthalenesulfonic acid, etc.; amino acid salts produced from glycine, arginine, lysine, etc.; and amine salts produced from trimethylamine, triethylamine, ammonia, pyridine, picoline, etc.

[0028] The tubulin inhibitor may be derived from an extract, produced biologically, or synthesized chemically, and is not limited by its origin or manufacturing method.

[0029] In this invention, skin diseases are a group of diseases resulting from structural and functional abnormalities of the skin, and are a comprehensive concept encompassing abnormal conditions originating from the skin that can be triggered by various causes such as inflammation, infection, allergy, and immune abnormalities.

[0030] The skin disease in this invention may be one or more selected from the group consisting of, for example, atopic dermatitis, thermal skin injury, pruritus, acne, psoriasis, allergic dermatitis, contact dermatitis, exfoliative dermatitis, seborrheic dermatitis, seborrheic scalp, lichen planus, rosacea, pigment disorders, melanin hyperplasia, erythema, wounds, ulcers, pressure ulcers, lupus, wrinkle-related skin diseases, and skin diseases caused by photodamage, and the type thereof is not limited.

[0031] The aforementioned wrinkle-related skin conditions may include elastic fibrosis, thinning of the skin, skin atrophy, reduction of collagen and elastic fibers, loss of skin elasticity, dryness, wrinkle formation, or premature skin aging.

[0032] Furthermore, the skin conditions caused by photodamage may include lentigines, freckles, hypopigmentation, hyperpigmentation, acute or chronic UV radiation-induced photodamage, or photosensitization.

[0033] Furthermore, the skin diseases of the present invention may include squamous cell carcinoma, basal cell carcinoma, benign epithelial tumors, and radiation dermatitis.

[0034] Furthermore, the skin disease of the present invention may be panniculitis, calluses, vitiligo, urticaria, folliculitis, sebaceous keratosis pilaris, eczema, corns, melasma, rash, athlete's foot, spots, stretch marks, freckles, heat rash, dry skin, chilblains, suppuration, pox, scalp inflammation, or psoriatic arthritis.

[0035] Furthermore, the present invention may further include, in addition to the tubulin inhibitor, a compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, and tofacitinib.

[0036] The compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, and tofacitinib may be used in combination with or in combination with tubulin inhibitors, and may be administered simultaneously or sequentially.

[0037] The terms calcium, colchicine, tapinarof, fingolimod, or tofacitinib are concepts that include all salts, isomorphs, or derivatives thereof, as well as analogs that exhibit the same or similar effects.

[0038] In the present invention, the tubulin inhibitor can be provided as an active ingredient in a pharmaceutical composition for the prevention or treatment of skin diseases.

[0039] The term "prevention" as used herein may include suppressing the occurrence of disease.

[0040] In this specification, the term “treatment” includes the suppression, mitigation, or elimination of the progression of a disease.

[0041] In this specification, the term "contains as an active ingredient" means that the tubulin inhibitor specified herein is added to an extent that it can exert the effects described herein, and includes formulation into various forms by adding various components as adjuncts for purposes such as drug delivery and stabilization.

[0042] The pharmaceutical composition may contain the tubulin inhibitor in a therapeutically effective amount.

[0043] In this specification, the term "therapeutic dose" means an amount sufficient to achieve the efficacy or activity of the active ingredient.

[0044] The pharmaceutical composition may further comprise a pharmaceutically acceptable diluent or carrier. The diluent may be lactose, corn starch, soybean oil, amorphous cellulose, mannitol, or a combination thereof. The carrier may be an excipient, disintegrant, binder, lubricant, or a combination thereof. The excipient may be microcrystalline cellulose, lactose, low-substituted hydroxycellulose, or a combination thereof. The disintegrant may be carboxymethylcellulose calcium, sodium starch glycolate, anhydrous monohydrogen phosphate, or a combination thereof. The binder may be polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, hydroxypropylcellulose, or a combination thereof. The lubricant may be magnesium stearate, silicon dioxide, talc, or a combination thereof.

[0045] The aforementioned pharmaceutical composition may be formulated into oral or parenteral dosage forms. Oral dosage forms may include granules, powders, liquids, tablets, capsules, dried syrups, or combinations thereof. Parenteral dosage forms may include injections, ointments, aerosols, sprays, patches, etc., and the dosage form is not limited.

[0046] For example, the dosage of a pharmaceutical composition for a mammal throughout the day is approximately 0.0001-10000 mg / kg, 0.0001-5000 mg / kg, 0.0001-1000 mg / kg, 0.0001-900 mg / kg, 0.0001-800 mg / kg, 0.0001-700 mg / kg, 0.0001-600 mg / kg, 0.0001-500 mg / kg, 0.0001-400 mg / kg, 0.0001-300 mg / kg, 0 .0001~200mg / kg, 1~1000mg / kg, 1~900mg / kg, 1~800mg / kg, 1~700mg / kg, 1~600mg / kg, 1~500mg / kg, 1~450mg / kg, 1~400 mg / kg, 1~350mg / kg, 1~300mg / kg, 1~250mg / kg, 10~1000mg / kg, 10~900mg / kg, 10~800mg / kg, 10~700mg / kg, 10~600mg / k g, 10~500mg / kg, 10~450mg / kg, 10~400mg / kg, 10~350mg / kg, 10~300mg / kg, 10~250mg / kg, 100~1000mg / kg, 100~900mg / kg, 100~800mg / kg, 100~700mg / kg, 100~600mg / kg, 100~500mg / kg, 100~450mg / kg, 100~400mg / kg, 100~350mg / kg, 100~ The drug can be administered in doses of 300 mg / kg, 100-250 mg / kg, 200-1000 mg / kg, 200-900 mg / kg, 200-800 mg / kg, 200-700 mg / kg, 200-600 mg / kg, 200-500 mg / kg, 200-450 mg / kg, 200-400 mg / kg, 200-350 mg / kg, 200-300 mg / kg, or 200-250 mg / kg, but is not limited thereto. The frequency of administration of the pharmaceutical composition of this application is not particularly limited, but can be administered once a day or in divided doses several times a day. The aforementioned dosages do not limit the scope of this application in any respect.

[0047] The subjects of the pharmaceutical compositions provided herein may be mammals, including humans, dogs, cats, horses, cattle, pigs, goats, rabbits, mice, rats, etc., or cells, tissues, or cultures thereof isolated therefrom. For example, the subjects may be individuals (such as humans) that require the prevention, improvement, and / or treatment of skin diseases as described above, or that have skin diseases, or cells, tissues, or cultures thereof isolated therefrom.

[0048] A pharmaceutical composition, as an example, contains the tubulin inhibitor in amounts of 1-80% by weight, 5-80% by weight, 5-75% by weight, 5-70% by weight, 5-65% by weight, 50-70% by weight, 55-65% by weight, 60-65% by weight, 10-60% by weight, 15-60% by weight, 20-60% by weight, 1-50% by weight, 5-50% by weight, 10-50% by weight, 15-50% by weight, and 20% by weight. It may also contain ~50% by weight, 1~40% by weight, 5~40% by weight, 10~40% by weight, 15~40% by weight, 20~40% by weight, 1~30% by weight, 5~30% by weight, 10~30% by weight, 15~30% by weight, 20~30% by weight, 1~25% by weight, 5~25% by weight, 10~25% by weight, 15~25% by weight, 20~25% by weight, or 23~25% by weight.

[0049] Alternatively, the composition may be a topical skin preparation composition.

[0050] The aforementioned topical skin preparations may be, but are not limited to, creams, gels, ointments, skin emulsifiers, skin suspensions, transdermal patches, drug-containing bandages, lotions, sprays, or combinations thereof.

[0051] In the present invention, the topical skin preparation can be appropriately formulated as needed with ingredients commonly used in topical skin preparations such as cosmetics and pharmaceuticals, such as aqueous components, oily components, powder components, alcohols, humectants, thickeners, UV absorbers, whitening agents, preservatives, antioxidants, surfactants, fragrances, colorants, various skin nutrients, metal ion chelating agents, sugars, or combinations thereof.

[0052] Furthermore, the present invention provides a quasi-drug composition for the prevention or improvement of skin diseases, comprising a tubulin inhibitor as an active ingredient.

[0053] In this invention, "quasi-drug" refers to all products that are not pharmaceuticals related to the treatment or prevention of disease.

[0054] In this invention, "quasi-drug" refers to all products that are not pharmaceuticals and are related to the treatment or prevention of diseases. The term "quasi-drug" means articles used for the purpose of diagnosing, treating, improving, alleviating, managing, or preventing diseases in humans or animals, and which have a milder effect than pharmaceuticals. For example, according to the Pharmaceuticals and Medical Devices Act, a quasi-drug is an article that is not used for pharmaceutical purposes and may include, but is not limited to, textile and rubber products used for the treatment or prevention of diseases in humans and animals, items that have a mild or no direct effect on the human body and are not instruments or machines, similar items, and disinfectants and insecticides for preventing infectious diseases.

[0055] The type and dosage form of the quasi-drug composition of the present invention are not particularly limited, but preferably include disinfectant cleansers, shower foams, mouthwashes, wet wipes, detergents, soaps, hand washes, humidifier fillers, masks, ointments, or filter fillers.

[0056] When the composition of the present invention is included in a quasi-drug for moisturizing the skin, the composition can be used as is or in combination with other quasi-drug ingredients, and can be used appropriately according to conventional methods. The amount of active ingredients mixed can be appropriately determined according to the purpose of use, and the quasi-drug composition according to the present invention may contain the tubulin inhibitor in an amount of 0.01 to 20% by weight relative to the total weight of the composition.

[0057] Furthermore, the present invention provides a cosmetic composition for the prevention or improvement of skin diseases, comprising a tubulin inhibitor as an active ingredient.

[0058] In this specification, the term “improvement” may mean any action that alleviates or treats a condition, such as any action that reduces the severity of the symptoms.

[0059] The tubulin inhibitor may be present in amounts of 0.0001 to 80% by weight, 0.01 to 70% by weight, 0.01 to 60% by weight, 0.01 to 50% by weight, 0.01 to 40% by weight, 0.01 to 30% by weight, 0.1 to 70% by weight, 0.1 to 60% by weight, 0.1 to 50% by weight, 0.1 to 40% by weight, 0.1 to 30% by weight, 0.5 to 70% by weight, 0.5 to 60% by weight, 0.5 to 50% by weight, 0.5 to 40% by weight, 0.5 to 30% by weight, 1 to 70% by weight, 1 to 60% by weight, 1 to 50% by weight, 1 to 40% by weight, or 1 to 30%, for example, 30% by weight, relative to the total weight of the cosmetic composition, but is not limited thereto.

[0060] The cosmetic composition is not particularly limited to a specific dosage form, and the dosage form can be appropriately selected according to the purpose. The cosmetic composition may, for example, have a solubilizing dosage form, an emulsifying dosage form, or a dispersing dosage form. The cosmetic composition may, but is not limited to, a softening lotion, a nourishing lotion, a massage cream, a nourishing cream, an essence, a pack, a gel, an ampoule, or a skin-applied cosmetic dosage form.

[0061] In addition to the active ingredients disclosed herein, the cosmetic composition may further contain ingredients commonly used in cosmetic compositions, such as common auxiliary agents and carriers including antioxidants, stabilizers, solubilizers, surfactants, dispersants, emulsifiers, preservatives, vitamins, pigments, and fragrances.

[0062] Furthermore, the present invention provides a cosmetic composition containing a tubulin inhibitor as an active ingredient for moisturizing the skin, whitening, improving wrinkles, alleviating acne, strengthening the skin barrier, or differentiating skin keratinocytes.

[0063] In this specification, the term "skin moisturizing" may mean any action that retains moisture in the skin or prevents moisture loss.

[0064] In this specification, the term "whitening" may mean improving the brightness of skin tone or reducing hyperpigmentation by suppressing melanin production.

[0065] In this specification, the term “wrinkle improvement” may mean an improvement in skin elasticity, a reduction in the depth of wrinkles, or an improvement in the smoothness of the skin surface.

[0066] In this specification, the term “acne relief” may mean suppressing acne inflammation, redness, or excessive sebum production, or promoting the healing of acne lesions.

[0067] In this specification, the term "skin barrier enhancement" may mean all actions that improve the function of the skin barrier, which is located on the outermost layer of the skin and prevents the loss of moisture and nutrients.

[0068] In this specification, the term “keratinocyte differentiation” may mean the process by which keratinocytes mature and form the stratum corneum, or thereby enhance the function of the skin barrier.

[0069] Another aspect of the present invention provides a method for preventing, improving, or treating a skin disease, comprising the step of administering an effective amount of the tubulin inhibitor to an individual in need.

[0070] The term "effective quantity" refers to a quantity that is effective enough to produce the effects mentioned above.

[0071] The individual may be a mammal, such as a human, cattle, horse, pig, dog, sheep, goat, or cat. The individual may be one that requires preventive, ameliorative, or therapeutic effects for a skin disease.

[0072] In this specification, the term “administer” is interchangeable with “introduce” and “implant,” and may mean the placement of a particular composition into an individual by a method or route that results in at least partial localization of the composition to a desired site.

[0073] Administration may be by methods known in the art. The method of administration, including the route and number of doses, can be appropriately selected by a skilled technician. Administration may be directly administered to the individual by any means, such as intravenous, intramuscular, oral, transdermal, mucosal, intranasal, oral, intratracheal, or subcutaneous administration. The administration may be systemic or topical. The administration may include topical application to the skin.

[0074] In the present invention, administration may involve administering a composition according to one specific example at a dose of 0.1 mg to 1,000 mg per individual 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 to 100 mg, 10 mg to 50 mg, or 10 mg to 25 mg. However, the dosage can be prescribed in various ways depending on factors such as the formulation method, administration method, patient's age, weight, sex, condition, diet, administration time, administration route, excretion rate, and response sensitivity, and a person skilled in the art can appropriately adjust the dosage considering these factors. The number of administrations can be once a day or two or more times a day within the range of clinically acceptable side effects, and it can be administered to one or more sites, and the total number of administration days in a single treatment can be from 1 to 30 days, administered daily or at intervals of 2 to 5 days. If necessary, the same treatment can be repeated after an appropriate period. For non-human animals, the dosage can be the same as for humans per kg, or the dosage can be converted using, for example, the volume ratio of organs (such as the heart) of the target animal and humans (e.g., average value) and administered.

[0075] Another embodiment provides the use of the tubulin inhibitor for use in manufacturing compositions for the prevention, improvement, or treatment of skin diseases.

[0076] Another embodiment provides the use of the tubulin inhibitor for use in manufacturing agents for the prevention, improvement, or treatment of skin diseases.

[0077] Another embodiment provides a food composition for preventing or improving skin diseases, comprising a tubulin inhibitor as an active ingredient.

[0078] A food composition according to one embodiment of the present invention may be in liquid or solid form, and may be in the form of tablets, capsules, soft capsules, pills, granules, beverages (drinks), diet bars, chocolates, caramel, or confectionery, and the form is not particularly limited. In addition to the active ingredients, the food composition of the present invention may optionally contain excipients, sugars, flavorings, colorings, oils and fats, proteins, etc.

[0079] Repetitive content has been omitted in consideration of the complexity of this specification, and terms not otherwise defined herein have the meanings commonly used in the art to which this invention pertains.

[0080] The present invention will be described in detail below with reference to examples. However, the following examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following examples.

[0081] [Experimental Example 1] Strain The normal human epidermal keratinocyte (NHEK) cell line was obtained from ATCC and used as a human embryonic transcutaneous cell line. The cells were obtained through primary culture and were used to confirm intracellular biochemical changes induced by tubulin inhibitors under normal, not diseased, conditions. Human embryonic kidney cells (HEKs) were obtained from ATCC and used. They were used to confirm intracellular biochemical changes induced by tubulin inhibitors under normal conditions, not disease conditions.

[0082] [Experimental Example 2] 1.1 Measurement of filaggrin expression using cell-based promoter activity measurement Human epidermal cell lines (NHEK) were treated with a filaggrin promoter-linked luciferase luminescence gene using the Fugene4K reagent (promega). After 24 hours, each tubulin inhibitor was administered at a concentration of 1.0 μM. 24 hours after drug treatment, filaggrin expression levels were measured using a luciferase luminescence intensity measurement kit (promega). The control group was administered the same dose of DMSO as the treated compounds.

[0083] 1.2 Observation of cell differentiation degree measurement using phase-contrast microscopy After treating normal human epidermal keratinocytes (NHEKs) with each tubulin inhibitor at a concentration of 1.0 μM, the degree of cell differentiation was observed daily using a phase-contrast microscope.

[0084] The cells were photographed using a phase-contrast microscope camera on day 5 of cell differentiation, and the results are shown in Figure 2.

[0085] [Experimental Example 3] Confirmation of suppression of inflammatory signaling mechanisms using cell-based promoter activity measurement. NF-κB, a DNA transcription factor, is present in the cell matrix and exists in a form bound to IκB, which suppresses NF-κB. NF-κB is involved in both congenital and acquired immune responses and is a representative pro-inflammatory cytokine exhibited in many inflammatory diseases.

[0086] The NF-κB mechanism involves a group of proteins (protein family) involved in regulating inflammatory responses, immune system modulation, apoptosis, cell proliferation, and epithelial differentiation. They regulate the expression of various genes and form the central axis of intracellular signaling pathways.

[0087] Human embryonic kidney cells (HEKs) were given a luciferase luminescence gene with an NF-κB promoter using the Fugene4K reagent (Promega). Six hours later, TNF-α was used to induce inflammation and NF-κB signaling. Eighteen hours later, each tubulin inhibitor was treated at a concentration of 1.0 μM.

[0088] 24 hours after drug treatment, the inhibitory effect on the NF-κB signaling mechanism associated with each tubulin inhibitor was measured using a luciferase luminescence intensity measurement kit (Promega). Furthermore, it was confirmed whether the inhibitory effect on the NF-κB signaling mechanism was increased when tubulin inhibitors were combined with 2 mM CaCl2, 1 μM colchicine, 1 μM tapinarof, 1 μM fingolimod, or 1 μM tofacitinib compared to the group treated with the tubulin inhibitor alone.

[0089] [Experimental Example 4] Inhibitory effect of tubulin inhibitors on the mechanism of collagen reduction Photoaging refers to the skin aging phenomenon caused by exposure to ultraviolet rays from the sun. This is a form of aging distinct from biological aging (senescence or biological aging), which is caused by a gradual decline in biological functions, and generally acts as a major cause of wrinkles and dryness, elastic fibrosis, and hyperpigmentation.

[0090] The sun emits two types of ultraviolet radiation: UV-A and UV-B. UV-A has wavelengths of 320-400 nm, while UV-B has wavelengths of 290-320 nm. Collagen is a prime example of tissue damaged by this type of UV radiation. Collagen is the main component of connective tissue, primarily found in bones and skin, and is broadly classified into five types. Type 1 collagen is the most abundant type found in the connective tissue of skin, followed by Type 3.

[0091] Matrix metalloproteinases (MMPs) are proteolytic enzymes that break down collagen and other components of connective tissue, and their expression increases when exposed to UV ultraviolet light. Therefore, when the skin is exposed to UV light, the expression of these Matrix Metalloproteinases (MMPs) increases, destroying collagen in the dermis and accelerating photoaging, such as wrinkles and hyperpigmentation.

[0092] Human skin cells (NHEK) were treated with either a tubulin inhibitor or a control group for 6 hours, then irradiated with UB ultraviolet light for 30 minutes. 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™ Reverse Transcription kit (Promega), and the expression levels of MMP-1, MMP-3, and MMP-9 present in the skin cells were measured quantitatively using PCR (qPCR) with a Real-Time PCR Detection System (Bio-Rad, CFX96).

[0093] [Experiment Example 5] Confirmation of skin whitening function using tyrosinase activity measurement To confirm whether the tubulin inhibitor of the present invention has a skin-whitening function, tyrosinase activity was measured. Tyrosinase is an enzyme that oxidizes tyrosine in the presence of oxygen to produce melanin, and inhibiting tyrosinase activity suppresses melanin production. When tyrosine is oxidized by tyrosinase, it takes on an orange color, but when tyrosinase activity is inhibited, the oxidation reaction of tyrosine by tyrosinase decreases, so the color gradually fades and becomes transparent depending on the degree of inhibition of tyrosinase activity. The absorbance value that decreases at this time was measured at 490 nm to evaluate the degree of tyrosinase inhibition.

[0094] [Example 1] Filaggrin expression level and cell differentiation in human epidermal cell lines following treatment with a tubulin inhibitor. Human epidermal cell lines (NHEKs) in culture were treated with a tubulin inhibitor, and filaggrin expression levels were measured by cell-based reporter assay and RT-PCR.

[0095] Each tubulin inhibitor was treated with NHEK at a concentration of 1.0 μM. To compare filaggrin expression and cell differentiation, a control group was treated with NHEK at the same dose as the treated drug. After 24 hours, filaggrin expression and cell differentiation were observed.

[0096] The results are shown in Figures 1 and 2.

[0097] As can be seen in Figure 1a, filaggrin expression levels increased in the tubulin inhibitor-treated group. Furthermore, as can be seen in Figure 2a, a significant increase in cell differentiation was confirmed in the tubulin inhibitor-treated group.

[0098] Furthermore, to confirm the combined effects of tubulin inhibitors, when each tubulin inhibitor was combined with 2 mM CaCl2, 1 μM colchicine, 1 μM tapinarof, 1 μM fingolimod, or 1 μM tofacitinib, it was confirmed that filaggrin expression (Figures 1a-1b) and cell differentiation potential (Figures 2b-2c) were increased compared to the experimental group treated with the tubulin inhibitor alone.

[0099] [Example 2] Confirmation of the suppression of inflammation by tubulin inhibitors Through a cell-based reporter assay, we confirmed that treating human epidermal cell lines (NHEK) with TNF-α to induce NF-κB activation, followed by treatment with a tubulin inhibitor at a concentration of 1.0 μM, suppressed the NF-κB signaling pathway. The results are shown in Figure 3a.

[0100] As can be seen in Figure 3a, treatment with the tubulin inhibitor according to the present invention suppressed NF-κB signaling compared to a control group treated with TNF-α alone. TPCA1 was used as a positive control, a well-known NF-κB signaling inhibitor.

[0101] In addition, when tubulin inhibitors were used in combination with 2 mM CaCl2, 1 μM colchicine, 1 μM tapinarof, 1 μM fingolimod, and 1 μM tofacitinib, the inhibitory effect on the NF-κB signaling mechanism was increased compared to the experimental group treated with the tubulin inhibitor alone (Figure 3a-b).

[0102] [Example 3] Confirmation of the suppression of collagen reduction mechanism by a tubulin inhibitor. Human NHEK cells were coated with a tubulin inhibitor, and photoaging (particularly wrinkle improvement) was induced by UV irradiation 30 minutes later. To compare and validate the tubulin inhibitors, a negative control group (untreated) and a positive control group (irradiated only with UV-A) were included in the experiment.

[0103] After the application and UV ultraviolet experiments were completed, 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 the skin tissue were measured using quantitative PCR (qPCR) with a Real-time PCR machine.

[0104] The photoaging-inhibiting effect of tubulin inhibitors was measured by the change in Matrix Metalloproteinase (MMP) protein expression levels described above. Specifically, MMP protein expression levels were measured in a control group irradiated with ultraviolet light only, an experimental group irradiated with ultraviolet light after applying the tubulin inhibitor, and a group treated with the tubulin inhibitor in combination with 2 mM CaCl2, 1 μM colchicine, 1 μM tapinarof, 1 μM fingolimod, and 1 μM tofacitinib. The degree to which MMP protein expression was suppressed by the application of the tubulin inhibitor was measured.

[0105] The results are shown in Figures 4a to 4f.

[0106] MMP-1 and MMP-9 are enzymes that promote the degradation of type 1 and type 3 collagen, which are mainly expressed in skin tissue, while MMP-3 is a regulatory enzyme that degrades type 1 collagen and adjusts changes in MMP-1 expression levels. These MMP proteins have a very important impact on photoaging.

[0107] As can be seen in Figures 4a, 4c, and 4e, when tubulin inhibitors were applied to the skin, the expression levels of MMP-1, MMP-3, and MMP-9 decreased by approximately 50% compared to the control group irradiated only with ultraviolet light. Thus, it was verified that the application of tubulin inhibitors suppresses the expression of Matrix Metalloproteinases (MMP) proteins, and an overall effect of suppressing photoaging by UV ultraviolet light was confirmed. The effect of each tubulin inhibitor on suppressing the expression levels of MMP-1, MMP-3, and MMP-9, which were elevated by UV-B irradiation, was measured.

[0108] In addition, when tubulin inhibitors were used in combination with 2 mM CaCl2, 1 μM colchicine, 1 μM tapinarof, 1 μM fingolimod, and 1 μM tofacitinib, the suppression of MMP-1, MMP-3, and MMP-9 expression levels was increased compared to the experimental group treated with tubulin inhibitors alone (Figures 4a to 4f).

[0109] [Example 4] Verification of the effect of tubulin inhibitors on improving pigmentation Through tyrosinase activity inhibition experiments, each tubulin inhibitor was treated with a 2mM L-tyrosine matrix at a concentration of 1.0 μM, and the absorbance was measured at 490 nm. Then, tyrosinase enzyme was added again and the reaction was carried out at 37°C. After measuring the absorbance at 490 nm, the absorbance value was calculated to confirm the degree to which each tubulin inhibitor inhibited tyrosinase activity. Kojic acid, known to inhibit tyrosinase activity, was used as a comparison group. This allowed us to determine the degree to which each tubulin inhibitor suppressed melanin production. The results are shown in Figures 5a and 5b.

[0110] As shown in Figure 5a, each tubulin inhibitor has the activity to suppress melanin production through inhibition of tyrosinase activity.

[0111] Furthermore, as can be seen in Figures 5a to 5b, when tubulin inhibitors were used in combination with 2 mM CaCl2, 1 μM colchicine, 1 μM tapinarof, 1 μM fingolimod, and 1 μM tofacitinib, the inhibitory effect on tyrosinase activity was increased compared to the experimental group treated with the tubulin inhibitor alone.

Claims

1. A pharmaceutical composition for the prevention or treatment of skin diseases, comprising a tubulin inhibitor as an active ingredient.

2. The composition according to claim 2, wherein the tubulin inhibitor is selected from one or more of the group consisting of vinca alkaloid, taxane, cryptophycin 52, halichondrin, dolastatin, hemiasterlin, combretastatin-A4, 2-methoxyestradiol, E7010, prinabuline, epothilon, discodermold, and pharmaceutically acceptable salts of the said compounds.

3. The composition according to claim 2, wherein the vinca alkaloid is one or more selected from the group consisting of vinblastine, vincristine, vinorelbine, vinflunine, and pharmaceutically acceptable salts of the said compound.

4. The composition according to claim 2, wherein the taxane is selected from one or more of the group consisting of paclitaxel, docetaxel, and pharmaceutically acceptable salts of the said compound.

5. The composition according to claim 1, wherein the skin disease is one or more selected from the group consisting of atopic dermatitis, thermal skin injury, pruritus, acne, psoriasis, allergic dermatitis, contact dermatitis, exfoliative dermatitis, seborrheic dermatitis, seborrheic scalp, lichen planus, rosacea, pigment disorders, melanin hyperplasia, erythema, wounds, ulcers, pressure ulcers, lupus, wrinkle-related skin diseases, and skin diseases caused by photodamage.

6. The composition according to claim 5, wherein the wrinkle-related skin disease is one or more selected from the group consisting of elastic 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. The composition according to claim 5, wherein the skin disease caused by photodamage is one or more selected from the group consisting of lentigines, freckles, hypopigmentation, hyperpigmentation, photodamage due to acute or chronic UV radiation, and photosensitization.

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

9. The composition according to claim 1, wherein the skin disease is one or more selected from the group consisting of panniculitis, calluses, vitiligo, urticaria, folliculitis, sebaceous keratosis pilaris, eczema, corns, melasma, rash, athlete's foot, spots, stretch marks, freckles, heat rash, dry skin, chilblains, suppuration, pox, scalp inflammation, and psoriatic arthritis.

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

11. A quasi-drug composition for the prevention or improvement of skin diseases, comprising a tubulin inhibitor as an active ingredient.

12. A cosmetic composition for the prevention or improvement of skin diseases, comprising a tubulin inhibitor as an active ingredient.

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

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

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