Combination therapies containing tubulin inhibitors for the prevention or treatment of skin diseases

Abstract

The present invention relates to a pharmaceutical composition, quasi-drug composition, or cosmetic composition for the prevention, improvement, or treatment of skin diseases, comprising as an active ingredient a tubulin inhibitor and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof. The combination therapy of the present invention can be used as an active ingredient that simultaneously exhibits activities such as skin moisturizing, whitening, wrinkle improvement, acne relief, skin barrier strengthening, or 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 and a specific compound as active ingredients.

[0002] The present invention also relates to an external pharmaceutical composition for preventing or improving skin diseases, containing a tubulin inhibitor and a specific compound as active ingredients.

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

[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 and a specific compound as active ingredients.

Background Art

[0005] The main function of the skin is to act as a barrier separating the internal environment from the external environment, protecting against attacks by external 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 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 of 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 of the present invention have confirmed that when a tubulin inhibitor binds to a specific compound, it increases filaggrin expression in epidermal cells, suppresses inflammatory signals, inhibits photoaging, inhibits collagen degradation mechanisms, and exhibits a whitening effect, thereby 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 as an active ingredient a tubulin inhibitor and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof.

[0014] Furthermore, the present invention provides a quasi-drug composition for the prevention, improvement, or treatment of skin diseases containing the above-mentioned active ingredient, a cosmetic composition for the prevention or improvement of skin diseases, and a cosmetic composition for moisturizing the skin, whitening, improving wrinkles, alleviating acne, strengthening the skin barrier, or differentiating skin keratinocytes.

[0015] Furthermore, the present invention provides a method for preventing or treating a skin disease, comprising the steps of administering to an individual in need a tubulin inhibitor and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof. [Effects of the Invention]

[0016] The combination of the tubulin inhibitor of the present invention with one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or pharmaceutically acceptable salts thereof 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 synergistic effect in skin whitening. Therefore, such combination therapy 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. [Brief explanation of the drawing]

[0017] [Figure 1] This figure shows the changes in filaggrin expression levels following combination therapy including tubulin inhibitors. The results show increased filaggrin expression levels in groups treated with ABT-751 or TN16 alone and in combination therapy. [Figure 2a] This figure shows the photoaging inhibitory effect of combination therapy including tubulin inhibitors. The results show that the expression levels of MMP-1 (Figure 2a), MMP-3 (Figure 2b), and MMP-9 (Figure 2c) decreased in the groups treated with ABT-751 or TN16 alone and in combination therapy after UV ultraviolet irradiation. [Figure 2b] This figure shows the photoaging inhibitory effect of combination therapy including tubulin inhibitors. The results show that the expression levels of MMP-1 (Figure 2a), MMP-3 (Figure 2b), and MMP-9 (Figure 2c) decreased in the groups treated with ABT-751 or TN16 alone and in combination therapy after UV ultraviolet irradiation. [Figure 2c] This figure shows the photoaging inhibitory effect of combination therapy including tubulin inhibitors. The results show that the expression levels of MMP-1 (Figure 2a), MMP-3 (Figure 2b), and MMP-9 (Figure 2c) decreased in the groups treated with ABT-751 or TN16 alone and in combination therapy after UV ultraviolet irradiation. [Figure 3]It is a figure regarding the inhibitory effect on NF-κB signaling. The results show that the inhibitory effect on the NF-κB signaling mechanism increases in the groups treated with ABT-751 or TN16 alone and in the combined treatment group. [Figure 4a] It is a figure regarding the change in the ability of skin cell differentiation by combined treatment containing a tubulin inhibitor. The results show that skin cell differentiation increases in the groups treated with ABT-751 or TN16 alone and in the combined treatment group (Figure 4a: results of fluorescence microscopy, Figure 4b: results of phase contrast microscopy). [Figure 4b] It is a figure regarding the change in the ability of skin cell differentiation by combined treatment containing a tubulin inhibitor. The results show that skin cell differentiation increases in the groups treated with ABT-751 or TN16 alone and in the combined treatment group (Figure 4a: results of fluorescence microscopy, Figure 4b: results of phase contrast microscopy). [Figure 5] It is a figure regarding the whitening effect of suppressing tyrosinase activity. The figure shows that tyrosinase activity is suppressed in the groups treated with ABT-751 or TN16 alone and in the combined treatment group, and the whitening effect further increases in the combined treatment group.

Mode for Carrying Out the Invention

[0018] Hereinafter, the present invention will be described in detail.

[0019] The present invention provides a pharmaceutical composition for preventing or treating skin diseases, comprising a tubulin inhibitor and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or pharmaceutically acceptable salts thereof as active ingredients. The present invention also provides a method for preventing, ameliorating, or treating skin diseases, comprising the step of administering to an individual in need thereof a tubulin inhibitor and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof.

[0020] ​

[0021] A tubulin inhibitor (tubulin suppressant) refers 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 within cells and are essential structures in processes such as cell division, cell movement, and intracellular transport.

[0022] Unlike 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 the differentiation of epidermal cells, suppresses the NF-κB signaling process induced by TNF-α, shows the effect of improving photoaging and wrinkles, and shows a whitening effect.

[0023] The tubulin inhibitor in the present invention is not limited in its type and can be, for example, one or more selected from the group consisting of vinca alkaloid, taxane, Cryptophycin 52, Halichondrins, Dolastatins, Hemiasterlins, Combretastatin-A4, 2-Methoxyestradiol, E7010, Plinabuline, Epothilone, and Discodermolide.

[0024] The vinca alkaloid of the present invention can preferably be Vinblastine, Vincristine, Vinorelbine, or Vinflunine.

[0025] Also, the taxane can be Paclitaxel or Docetaxel.

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

[0027] Most preferably, the tubulin inhibitor of the present invention is ABT-751 represented by the following chemical formula I, or TN-16 represented by the following chemical formula. [ka]

[0028] Compound I is compound ABT-751 (N-[2-[(4-Hydroxyphenyl)amino]pyridin-3-yl]-4-methoxybenzenesulfonamide), which is CAS registry number 141430-65-1.

[0029] ABT-751 is a compound that targets microtubules, and it has been reported that this drug has the activity to suppress cell division in certain cancer cells and stop the proliferation of cancer cells.

[0030] Furthermore, compound II is compound 33016-12-5, and is TN-16(3-[1-(phenylamino)ethylidene]-5-(phenylmethyl)-2,4-pyrrolidinedione).

[0031] TN-16 is a novel microtubule inhibitor with antitumor activity, synthesized by modifying tenuazonic acid (3-acetyl-5-sec-butyltetramic acid), a natural antibiotic isolated and characterized from Alternaria tenuis cultures. While TN-16 was found to possess potent anticancer properties in 1967, subsequent research has not uncovered any more potent drugs. Furthermore, in 1983, the mechanism of action of TN-16 was clarified, revealing that it inhibits microtubule formation and acts by binding to sensitive sites.

[0032] The tubulin inhibitor may be derived from an extract, produced biologically, or synthesized chemically, and is not limited by its origin or manufacturing method.

[0033] The present invention is characterized by comprising a tubulin inhibitor and, in combination with the tubulin inhibitor, one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or pharmaceutically acceptable salts thereof.

[0034] In this invention, the term "combination" refers to a use in combination administration of a tubulin inhibitor and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or pharmaceutically acceptable salts thereof, and can be understood as meaning "combined use."

[0035] Specifically, a tubulin inhibitor may be administered in combination with one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof.

[0036] The aforementioned concomitant administration is i) Concomitant administration using a pharmaceutical composition comprising a tubulin inhibitor and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof, either in the form of a mixture or in separate, isolated forms. ii) A pharmaceutical composition containing a tubulin inhibitor may be administered simultaneously, sequentially, or in reverse order in combination with a pharmaceutical composition containing one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof.

[0037] The aforementioned combination or composition is a) The treatment involves administering a mixture of a tubulin inhibitor and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof, b) The tubulin inhibitor may be administered in an isolated form along with one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or pharmaceutically acceptable salts thereof.

[0038] In the case of the separated forms as described in b) above, the tubulin inhibitor and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or pharmaceutically acceptable salts thereof may be formulated together in separate, separated forms; or they may be formulated in separate formulations, which may be administered simultaneously, individually, sequentially, or in reverse order.

[0039] In this invention, "combined administration," "combined," or "combined" does not merely mean simultaneous administration, but should be understood as a form of administration in which a tubulin inhibitor and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or pharmaceutically acceptable salts thereof act together on an individual, allowing each substance to perform at a level equivalent to or greater than its original function. Therefore, when the term "combined" is used in this application, it should be understood to mean simultaneous, individual, sequential, or reversed administration, without any restriction on the order. When the administration is sequential, reversed, or individual, the order of administration is not particularly restricted, however, the interval between administrations of secondary or higher components should be such that the beneficial effects of the combination are not lost.

[0040] The preferred dosage of the pharmaceutical composition and concomitant components according to the present invention varies depending on the patient's condition, weight, disease severity, drug form, route of administration, and duration, but can be appropriately selected by those skilled in the art. However, for a desirable effect, the dosage is 0.0001 to 500 mg / kg per day, specifically 0.001 to 500 mg / kg. The drug may be administered once a day or in several divided doses. However, the scope of the present invention is not limited by the aforementioned dosage.

[0041] The composition comprising the combination according to the present invention is for the co-administration of a tubulin inhibitor and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or pharmaceutically acceptable salts thereof, and the G tubulin inhibitor and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof may be formulated as a single formulation.

[0042] The combinations or combination compositions of the present invention may be provided in kit form. The term “kit” in the present invention is for the co-administration of a tubulin inhibitor and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or pharmaceutically acceptable salts thereof, and may include the combinations or compositions of the present invention for the co-administration of a tubulin inhibitor and the combinations or combinations of the present invention. Specifically, the kit according to the present invention may include, but is not limited to, a) a tubulin inhibitor formulated in a single formulation and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof, and b) a tubulin inhibitor and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or pharmaceutically acceptable salts thereof, and may further include substances necessary for the co-administration of two or more substances. The present invention has confirmed that when a tubulin inhibitor is used in combination with one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof, a synergistic effect is observed in increasing filaggrin expression from cells, suppressing photoaging, inhibiting inflammatory signaling mechanisms, differentiating skin cells, and whitening, and has thus provided the aforementioned combination therapy.

[0043] 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.

[0044] 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.

[0045] 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.

[0046] Furthermore, the skin conditions caused by photodamage may include lentigines, freckles, hypopigmentation, hyperpigmentation, photodamage due to acute or chronic UV radiation, or photosensitization.

[0047] Furthermore, the skin diseases of the present invention may include squamous cell carcinoma, basal cell carcinoma, benign epithelial tumors, and radiation dermatitis.

[0048] 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.

[0049] In the present invention, the tubulin inhibitor and, One or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof. This can be provided as an active ingredient in pharmaceutical compositions for the prevention or treatment of skin diseases.

[0050] The term "prevention" as used herein may include suppressing the occurrence of disease.

[0051] In this specification, the term “treatment” includes the suppression, mitigation, or elimination of the progression of a disease.

[0052] In this specification, the term "contains as an active ingredient" means that the tubulin inhibitors specified herein and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof are added to an extent that they can exert the effects referred to herein, and includes formulation into various forms by adding various components as adjuncts for purposes such as drug delivery and stabilization.

[0053] The pharmaceutical composition may contain, in a therapeutically effective amount, the tubulin inhibitor and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof.

[0054] In this specification, the term "therapeutic dose" means an amount sufficient to achieve the efficacy or activity of the active ingredient.

[0055] 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.

[0056] 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.

[0057] 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.

[0058] The subjects to whom the pharmaceutical compositions provided herein are administered 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 subject may be an individual (such as a human mammal) that requires the prevention, improvement, and / or treatment of a skin disease as described above, or that has a skin disease, or cells, tissues, or cultures thereof isolated therefrom.

[0059] An example of a pharmaceutical composition is the tubulin inhibitor and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or pharmaceutically acceptable salts thereof, 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% It may be included in amounts of 60% by weight, 1-50% by weight, 5-50% by weight, 10-50% by weight, 15-50% by weight, 20-50% 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.

[0060] Alternatively, the composition may be a topical skin preparation composition.

[0061] 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.

[0062] 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.

[0063] Furthermore, the present invention provides a quasi-drug composition for the prevention or treatment of skin diseases, comprising a tubulin inhibitor and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or pharmaceutically acceptable salts thereof, as active ingredients.

[0064] 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.

[0065] 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. When the composition of the present invention is included in a quasi-drug for skin moisturizing purposes, 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 and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or pharmaceutically acceptable salts thereof, in an amount of 0.01 to 20% by weight of the total weight of the composition.

[0066] Furthermore, the present invention provides a cosmetic composition for the prevention or improvement of skin diseases, comprising a tubulin inhibitor and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or pharmaceutically acceptable salts thereof, as active ingredients.

[0067] 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.

[0068] The tubulin inhibitor and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or pharmaceutically acceptable salts thereof are present in amounts of 0.0001-80% by weight, 0.01-70% by weight, 0.01-60% by weight, 0.01-50% by weight, 0.01-40% by weight, and 0.01-3% by weight relative to the total weight of the cosmetic composition. It may, but is not limited to, be included in amounts of 0% by weight, 0.1–70% by weight, 0.1–60% by weight, 0.1–50% by weight, 0.1–40% by weight, 0.1–30% by weight, 0.5–70% by weight, 0.5–60% by weight, 0.5–50% by weight, 0.5–40% by weight, 0.5–30% by weight, 1–70% by weight, 1–60% by weight, 1–50% by weight, 1–40% by weight, or 1–30%, for example, 30% by weight.

[0069] 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.

[0070] 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.

[0071] Furthermore, the present invention provides a cosmetic composition for moisturizing the skin, whitening, improving wrinkles, alleviating acne, strengthening the skin barrier, or differentiating skin keratinocytes, comprising a tubulin inhibitor and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or pharmaceutically acceptable salts thereof, as active ingredients.

[0072] In this specification, the term "skin moisturizing" may mean any action that retains moisture in the skin or prevents moisture loss.

[0073] In this specification, the term "whitening" may mean improving the brightness of skin tone or reducing hyperpigmentation by suppressing melanin production.

[0074] 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.

[0075] In this specification, the term “acne relief” may mean suppressing acne inflammation, redness, or excessive sebum production, or promoting the healing of acne lesions.

[0076] 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.

[0077] 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.

[0078] Another aspect of the present invention provides a method for preventing, improving, or treating a skin disease, comprising the step of administering to an individual in need an effective amount of the tubulin inhibitor and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or pharmaceutically acceptable salts thereof.

[0079] The term "effective quantity" refers to a quantity that is effective enough to produce the effects mentioned above.

[0080] 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.

[0081] 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.

[0082] 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.

[0083] 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.

[0084] Another embodiment provides uses for the tubulin inhibitor and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof, for use in manufacturing compositions for the prevention, improvement, or treatment of skin diseases.

[0085] Another embodiment provides uses for the tubulin inhibitor and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof, for use in manufacturing agents for the prevention, improvement, or treatment of skin diseases.

[0086] Another embodiment provides a food composition for the prevention or improvement of skin diseases, comprising a tubulin inhibitor and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof, as active ingredients. The food composition may include a health functional food.

[0087] 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.

[0088] 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.

[0089] 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.

[0090] [Example 1] Confirmation of filaggrin expression levels by combined treatment including a tubulin inhibitor Human epidermal cell lines (NHEK) were inserted with a filaggrin promoter-tagged luciferase luminescence gene using Fugene4K reagent (Promega), and then treated with ABT-751 or TN16 at a concentration of 1.0 μM 24 hours later. In addition, 2 mM CaCl2, 1 μM colchicine, 1 μM tapinarof, 1 μM fingolimod, and 1 μM tofacitinib were treated either alone or in combination with ABT-751 or TN16.

[0091] 24 hours after drug treatment, the amount of filaggrin expressed after treatment with each compound was measured using a luciferase luminescence measurement kit (Promega), and the results are shown in Figure 1.

[0092] As can be seen in Figure 1, filaggrin expression levels were confirmed to be increased in the ABT-751 or TN16 treated groups compared to the untreated control group.

[0093] Furthermore, we were able to confirm that filaggrin expression levels were significantly increased in the ABT-751 or TN16 treatment groups treated in combination with calcium or colchicine, fingolimod, tapinarov, or tofacitinib compared to the ABT-751 or TN16 monotherapy groups.

[0094] [Example 2] Inhibitory effect of combination treatment including a tubulin inhibitor 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.

[0095] 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.

[0096] 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.

[0097] Human NHEK cells were coated with either ABT-751 or TN-16, and photoaging (particularly wrinkle improvement) was induced by UV irradiation 30 minutes later. Alternatively, cells were treated with 2 mM CaCl2, 1 μM colchicine, 1 μM tapinarof, 1 μM fingolimod, or 1 μM tofacitinib in combination with ABT-751 or TN16, followed by UV irradiation. For comparison and validation, a negative control group (no treatment) and a positive control group (irradiated only with UV-A) were included in the experiment.

[0098] 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.

[0099] Specifically, the photoaging-inhibiting effect of ABT-751 or TN-16 was measured through changes in Matrix Metalloproteinase (MMP) protein expression levels, as described above. That is, MMP protein expression levels were measured in a control group irradiated with ultraviolet light only, and in experimental groups that were treated with ABT-751 or TN-16, respectively, followed by ultraviolet light irradiation, to determine the degree to which MMP protein expression was suppressed.

[0100] The results are shown in Figure 2.

[0101] 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.

[0102] As can be seen in Figure 2, when ABT-751 or TN-16 were applied to the skin, respectively, the expression levels of MMP-1, MMP-3, and MMP-9 were all reduced to approximately 50% or less compared to the control group that was irradiated with ultraviolet light alone.

[0103] Furthermore, we verified that in experimental groups treated with 2 mM CaCl2, 1 μM colchicine, 1 μM tapinarof, 1 μM fingolimod, and 1 μM tofacitinib in combination with ABT-751 or TN16, MMP protein expression was significantly suppressed compared to experimental groups treated with ABT-751 or TN-16 alone. This allowed us to confirm an overall increase in the suppression of photoaging by UV ultraviolet light.

[0104] [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.

[0105] 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.

[0106] 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, the cells were treated with either ABT-751 or TN16 at a concentration of 1.0 μM. In addition, 2 mM CaCl2, 1 μM colchicine, 1 μM tapinarof, 1 μM fingolimod, or 1 μM tofacitinib were used for treatment alone or in combination with ABT-751 or TN16. Twenty-four hours after drug treatment, the inhibitory effect of each compound on the NF-κB signaling mechanism was measured using a luciferase luminescence level measurement kit (Promega), and the results are shown in Figure 3.

[0107] As shown in Figure 3, we confirmed that treatment with ABT-751 or TN-16 at a concentration of 1.0 μM suppressed the NF-κB signaling pathway. Furthermore, in groups treated with ABT-751 or TN16 in combination with 2 mM CaCl2, 1 μM colchicine, 1 μM tapinarof, 1 μM fingolimod, and 1 μM tofacitinib, we confirmed a significantly increased inhibitory effect on the NF-κB signaling mechanism compared to groups treated with ABT-751 or TN16 alone.

[0108] [Example 4] Confirmation of skin cell differentiation ability by combination treatment including a tubulin inhibitor To confirm the efficacy of combination therapy including tubulin inhibitors against NHEK in human epidermal cells, cells were observed after drug treatment.

[0109] We prepared groups of cells treated with 1.0 μM ABT-751 or TN16, groups treated with 2 mM CaCl2, 1 μM colchicine, 1 μM tapinarof, 1 μM fingolimod, or 1 μM tofacitinib individually, and groups treated with CaCl2, colchicine, tapinarof, fingolimod, or tofacitinib together with ABT-751 or TN16. The degree of differentiation was measured 5 days after drug treatment.

[0110] In the groups treated with calcium and colchicine, skin cells were fluorescently stained with Keratin-14 antibody and imaged using a fluorescence microscope. Keratin-10 protein is a well-known marker of skin cell differentiation. The results obtained using a fluorescence microscope are shown in Figure 4a.

[0111] Furthermore, in the groups treated with tapinarov, fingolimod, or tofacitinib, skin cells were imaged using a phase-contrast microscope, and the results are shown in Figure 4b.

[0112] As shown in Figure 4a, we confirmed that cell differentiation was increased in the experimental groups treated with ABT-751 or TN16 compared to the negative control group.

[0113] Furthermore, as shown in Figures 4a and 4b, we were able to confirm that cell differentiation was significantly increased in the experimental groups treated with calcium, colchicine, tapinarov, fingolimod, tofacitinib, and ABT-751 or TN16 in combination, compared to the experimental groups treated with ABT-751 or TN16 alone.

[0114] [Example 5] Confirmation of skin whitening function using tyrosinase activity measurement To confirm whether the combination therapy containing the tubulin inhibitor of the present invention exhibits 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, and 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. In the tyrosinase activity inhibition experiment, L-tyrosine 2 mM, which was the matrix, was treated with ABT-751 or TN16 at a concentration of 1.0 μM.

[0115] In addition, subjects were treated with 2 mM CaCl2, 1 μM colchicine, 1 μM tapinarof, 1 μM fingolimod, or 1 μM tofacitinib alone, or in combination with ABT-751 or TN16. Kojic acid, known to inhibit tyrosinase activity, was used as a comparison group.

[0116] After measuring the absorbance at 490 nm, the tyrosinase enzyme was added again and the reaction was carried out at 37°C. Furthermore, after measuring the absorbance at 490 nm, the absorbance value was calculated to confirm the degree to which each compound inhibited the tyrosinase activity. The results are shown in Figure 5.

[0117] As shown in Figure 5, the degree of inhibition of tyrosinase activity was significantly increased in the experimental groups treated with ABT-751 or TN16 in combination with one of the following: calcium, colchicine, tapinarov, fingolimod, or tofacitinib, compared to the groups treated with ABT-751 or TN16 alone. This confirms that combination treatment including tubulin inhibitors can suppress melanin production and exert a skin-whitening effect.

Claims

1. Tubulin inhibitors, A pharmaceutical composition for the prevention or treatment of skin diseases, comprising as an active ingredient one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof.

2. The composition according to claim 1, wherein the tubulin inhibitor is a compound represented by the following chemical formula I or chemical formula II, or a pharmaceutically acceptable salt thereof. 【Chemistry 1】

3. 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.

4. The composition according to claim 3, 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.

5. The composition according to claim 3, 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.

6. 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.

7. 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.

8. Tubulin inhibitors, A quasi-drug composition for the prevention or improvement of skin diseases, comprising as an active ingredient one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof.

9. Tubulin inhibitors, A cosmetic composition for the prevention or improvement of skin diseases, comprising as an active ingredient one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or pharmaceutically acceptable salts thereof.

10. Tubulin inhibitors, A cosmetic composition for moisturizing, whitening, wrinkle improvement, acne relief, strengthening the skin barrier, or differentiation of skin keratinocytes, comprising as an active ingredient one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof.

11. Tubulin inhibitors, A method for preventing or treating a skin disease, comprising the step of administering to an individual in need one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof.

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