Composition for preventing or treating skin diseases using combination therapy comprising tubulin inhibitor

Abstract

The present disclosure relates to a pharmaceutical composition, a quasi-drug composition, or a cosmetic composition comprising: a tubulin inhibitor; and at least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof as active ingredients for the prevention, alleviation, or treatment of skin diseases. The combination therapy of the present invention can be utilized as an active ingredient simultaneously exhibiting activities such as skin moisturizing, whitening, wrinkle alleviation, acne alleviation, skin barrier strengthening, or skin keratinocyte differentiation, in addition to preventing, treating, or alleviating skin diseases.

Description

Composition for preventing or treating skin diseases using combination therapy containing a tubulin inhibitor

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

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

[0003] In addition, the present invention relates to a cosmetic composition for preventing or improving skin diseases, comprising a tubulin inhibitor and a specific compound as active ingredients.

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

[0005] The primary function of the skin is to act as a barrier, separating the internal environment from the external environment, protecting against aggression from foreign substances and minimizing the loss of moisture and other essential body components to the external space. Filaggrin is particularly important in skin barrier formation due to its fundamental role in the terminal differentiation of the epidermis and its association with skin diseases. First identified as a key structural protein in 1977, filaggrin was given the name filaggrin (filament-aggregating protein) after it was discovered that it acts to aggregate and compact keratin intermediate filaments. This protein is synthesized from the large precursor protein profilaggrin, which is a major component of keratohyalin granules in the granular layer of the epidermis.

[0006] A key component of the skin barrier is the stratum corneum. This layer is the result of differentiation of keratinocytes as they migrate from the basal layer to the granulosum layer. As these cells mature, they express various structural proteins.

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

[0008] That is, filaggrin is converted from the precursor protein profilaggrin to active filaggrin monomer, and forms the stratum corneum by coagulating and compressing keratin filaments to flatten keratinocytes, and the natural moisturizing factor (NMF) produced during the filaggrin processing process contributes to maintaining moisture in the skin and controlling acidic pH.

[0009] Additionally, filaggrin binds intermediate keratin filaments to structural proteins of the cornified epithelium (CE), which gives keratinocytes strong mechanical and chemical resistance. Together with lipids released from lamellar bodies, filaggrin also contributes to the formation of the extracellular lipid matrix of the stratum corneum.

[0010] In particular, filaggrin degradation products UCA (Urocanic acid) and PCA (Pyrroliodone-5-carboxylic acid) help maintain skin moisture, maintain an acidic pH, and protect against UV radiation. UCA also plays a key role in antimicrobial action and immune regulation.

[0011] Accordingly, the inventors of the present invention confirmed that a tubulin inhibitor, when combined with a specific compound, increases the expression of filaggrin in epidermal cells, suppresses inflammatory signals, suppresses photoaging, suppresses collagen decomposition mechanisms, and exhibits a whitening effect, thereby completing the present invention.

[0012] Accordingly, the purpose of the present invention is to provide a pharmaceutical composition or quasi-drug composition for preventing or treating skin diseases. Furthermore, the purpose of the present invention is to provide a cosmetic composition for preventing or improving skin diseases, and a cosmetic composition for moisturizing, whitening, improving wrinkles, alleviating acne, strengthening the skin barrier, or differentiating skin keratinocytes.

[0013] To achieve the above purpose, the present invention provides a pharmaceutical composition for preventing or treating skin diseases, comprising as an active ingredient a tubulin inhibitor; and at least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof.

[0014] In addition, the present invention provides a pharmaceutical composition for preventing, improving or treating a skin disease, a cosmetic composition for preventing or improving a skin disease, and a cosmetic composition for moisturizing, whitening, improving wrinkles, alleviating acne, strengthening the skin barrier or differentiating skin keratinocytes, including the above-described effective ingredient.

[0015] The present invention also provides a method for preventing or treating a skin disease, comprising the step of administering to a subject in need thereof a tubulin inhibitor; and at least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof.

[0016]

[0017] The tubulin inhibitor of the present invention; and the combined use of at least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or a pharmaceutically acceptable salt thereof increases the expression level of filaggrin in epidermal cells, promotes skin differentiation, inhibits the NF-kB signaling process induced by TNF-α, exhibits photoaging inhibition and wrinkle improvement effects, and exhibits a synergistic effect in whitening effects. Accordingly, 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, in addition to preventing, treating, or improving skin diseases.

[0018]

[0019] Figure 1 shows changes in filaggrin expression levels following combination treatment with a tubulin inhibitor. The results show an increase in filaggrin expression levels in the ABT-751 or TN16 single treatment and combination treatment groups.

[0020] Figure 2 shows the photoaging inhibitory effect according to the combined treatment including a tubulin inhibitor. The results show that the expression levels of MMP-1 (Figure 2a), MMP-3 (Figure 2b), and MMP-9 (Figure 2c) are reduced in the ABT-751 or TN16 single treatment group and the combined treatment group after UV irradiation.

[0021] Figure 3 shows the effect of inhibiting NF-κB signaling. The results show that the NF-κB signaling mechanism inhibition effect is enhanced in the ABT-751 or TN16 single treatment and combination treatment groups.

[0022] Figure 4 shows changes in skin cell differentiation capacity according to combination treatment including a tubulin inhibitor. The results show increased skin cell differentiation in the ABT-751 or TN16 single treatment group and the combination treatment group (Figure 4a: fluorescence microscopy results, Figure 4b: phase contrast microscopy results).

[0023] Figure 5 shows the whitening effect of inhibiting tyrosinase activity. Tyrosinase activity was inhibited in the ABT-751 or TN16 single treatment group and the combination treatment group, and the whitening effect was further enhanced in the combination treatment group.

[0024] Hereinafter, the present invention will be described in detail.

[0025] The present invention relates to a tubulin inhibitor; and

[0026] A pharmaceutical composition for preventing or treating a skin disease is provided, comprising, as an active ingredient, at least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or a pharmaceutically acceptable salt thereof.

[0027] The present invention also provides a method for preventing, improving, or treating a skin disease, comprising the step of administering to a subject in need thereof a tubulin inhibitor; and at least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof.

[0028] Tubulin inhibitors (Tubulin inhibitors) refer to a group of drugs that regulate or inhibit the function of tubulin, a key component of microtubules. Microtubules are part of the cytoskeleton, playing a crucial role within cells and are essential structures for processes such as cell division, cell movement, and intracellular transport.

[0029] In the present invention, unlike the previous functions, the tubulin inhibitor exhibits activity in increasing the expression level of filaggrin in epidermal cells and in skin proliferation according to epidermal cell differentiation, inhibits the NF-kB signaling process induced by TNF-α, exhibits photoaging and wrinkle improvement effects, and exhibits a whitening effect.

[0030] In the present invention, the tubulin inhibitor is not limited in type, but may be, for example, at least one selected from the group consisting of vinca alkaloid, taxane, cryptophycin 52, halichondrins, dolastatins, hemisterlins, combretastatin-A4, 2-methoxyestradiol, E7010, plinabuline, epothilone, and discodermolide.

[0031] The vinca alkaloid of the present invention may preferably be Vinblastine, Vincristine, Vinorelbine, or Vinflunine.

[0032] Additionally, the taxane may be paclitaxel or docetaxel.

[0033] The tubulin inhibitor of the present invention is a concept that includes all of its salts, isomers, derivatives, and analogs that exhibit the same or similar effects.

[0034] 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 II.

[0035] [Chemical Formula I]

[0036]

[0037] [Chemical Formula II]

[0038]

[0039]

[0040] The above compound I is a compound with CAS registry number 141430-65-1, ABT-751 (N-[2-[(4-Hydroxyphenyl)amino]pyridin-3-yl]-4-methoxybenzenesulfonamide).

[0041] ABT-751 is a compound that targets microtubules, and the drug has been reported to exhibit activity in stopping the growth of cancer cells by inhibiting cell division in certain cancer cells.

[0042] In addition, the compound II is a compound with CAS registry number 33016-12-5, TN-16 (3-[1-(phenylamino)ethylidene]-5-(phenylmethyl)-2,4-pyrrolidinedione).

[0043] TN-16 is a novel microtubule inhibitor with antitumor activity. It was synthesized by modifying tenuazonic acid (3-acetyl-5-sec-butyltetramic acid), a natural antibiotic isolated and characterized from cultures of Alternaria tenuis. In 1967, TN-16 was discovered to possess potent anticancer properties, but subsequent studies failed to identify a more potent agent. Furthermore, in 1983, the mechanism of action of TN-16 was revealed to be inhibition of microtubule formation, acting by binding to a sensitive site.

[0044] The above tubulin inhibitor may be derived from an extract, produced biologically, or synthesized chemically, and is not limited in its origin and production method.

[0045] The present invention is characterized in that it further comprises at least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or pharmaceutically acceptable salts thereof, in combination with a tubulin inhibitor.

[0046] In the present invention, the term "combination" refers to the use 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 to have the same meaning as "combined use."

[0047] Specifically, tubulin inhibitors; and one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof may be co-administered.

[0048] The above combination administration

[0049] i) tubulin inhibitors; and, one or more compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof; administered in combination using a pharmaceutical composition comprising the same in the form of a mixture or in separate, separate forms;

[0050] ii) A pharmaceutical composition comprising a tubulin inhibitor and a pharmaceutical composition containing at least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof may be administered concurrently, sequentially, or in reverse order, but is not limited thereto.

[0051] The above combination or composition

[0052] a) administered as a mixture comprising a tubulin inhibitor and at least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof, or

[0053] b) 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 administered in a separate form, but is not limited thereto.

[0054] In the case of a separate form as in the above b), one or more compounds selected from the group consisting of a tubulin inhibitor and calcium, colchicine, tapinarof, fingolimod, tofacitinib, or a pharmaceutically acceptable salt thereof may be formulated together in a separate form; or each may be formulated as a separate formulation so that the separate formulations can be administered simultaneously, separately, sequentially, or in reverse order.

[0055] In the present invention, "combined administration", "combined use", or "using in combination" should be understood not only to mean simultaneous administration, but also to mean an administration form in which one or more compounds selected from the group consisting of a tubulin inhibitor; and calcium, colchicine, tapinarof, fingolimod, tofacitinib, or a pharmaceutically acceptable salt thereof act together on a subject so that each substance can perform a level equivalent to or superior to its original function. Therefore, when the term "combined use" is used herein, it should be understood that it refers to simultaneous, separate, sequential, or reverse administration, and the order is unlimited. When the administration is sequential, reverse, or separate, the order of administration is not particularly limited, but the interval between administrations of two or more components should be such that the beneficial effect of the combination is not lost.

[0056] The preferred dosage of the pharmaceutical composition and concomitant ingredients according to the present invention varies depending on the patient's condition, body weight, severity of the disease, drug form, route of administration, and duration of administration, but can be appropriately selected by those skilled in the art. However, for desirable effects, the dosage may be 0.0001 to 500 mg / kg per day, specifically 0.001 to 500 mg / kg. The dosage may be administered once a day or divided into several doses. However, the scope of the present invention is not limited by the above dosage.

[0057] A composition comprising a combination according to the present invention is for co-administering a tubulin inhibitor; and at least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or a pharmaceutically acceptable salt thereof, wherein the G tubulin inhibitor; and at least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and a pharmaceutically acceptable salt thereof may be formulated into a single formulation.

[0058] The combination or combination composition of the present invention may be provided in the form of a kit. The term "kit" in the present invention refers to a combination for co-administering a tubulin inhibitor; and at least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or a pharmaceutically acceptable salt thereof, and may include a combination or composition according to the present invention for co-administering a tubulin inhibitor. Specifically, the kit according to the present invention comprises: a) a tubulin inhibitor formulated into one formulation and at least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and a pharmaceutically acceptable salt thereof; 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; each of which may be included as an individual formulation, and may additionally include a substance necessary for co-administration of two or more substances, but is not limited thereto.

[0059] The present invention provides a combination therapy in which a tubulin inhibitor and at least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof are used in combination, and a synergistic effect is observed in increasing filaggrin expression in cells, inhibiting photoaging, inhibiting inflammatory signaling mechanisms, differentiating skin cells, and whitening.

[0060] In the present invention, skin disease is a group of diseases caused by structural and functional abnormalities of the skin, and is a concept encompassing abnormal conditions occurring in the skin that can be caused by various causes such as inflammation, infection, allergy, and immune abnormality.

[0061] In the present invention, the skin disease may be at least one selected from the group consisting of, for example, atopic dermatitis, skin thermal damage, pruritus, acne, psoriasis, allergic dermatitis, contact dermatitis, exfoliative dermatitis, seborrheic dermatitis, seborrheic dermatitis, lichen planus, rosacea, pigmentation disorder, hypermelanosis, erythema, wounds, ulcers, bedsores, lupus, skin wrinkle-related diseases, and skin diseases caused by photodamage, and the type thereof is not limited.

[0062] The above skin wrinkle-related diseases may be elastic fibrosis, thinning of the skin, skin atrophy, reduction of collagen fibers and elastic fibers, loss of skin elasticity, dryness, wrinkle formation, or premature skin aging.

[0063] Additionally, the skin diseases caused by the photodamage may be lentigines, freckles, hypopigmentation, hyperpigmentation, photodamage due to acute or chronic UV radiation, or photosensitization.

[0064] Additionally, the skin disease of the present invention may be squamous cell carcinoma, basal cell carcinoma, benign epithelial tumor, and radiation dermatitis.

[0065] In addition, the skin disease of the present invention may be panniculitis, calluses, vitiligo, urticaria, folliculitis, sebaceous keratosis pilaris, eczema, corns, freckles, rashes, athlete's foot, spots, stretch marks, freckles, prickly heat, dry skin, chilblains, boils, keratosis pilaris, scalp dermatitis, or psoriatic arthritis.

[0066] In the present invention, the tubulin inhibitor; and

[0067] At least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof;

[0068] As an active ingredient, it can be provided in a pharmaceutical composition for preventing or treating skin diseases.

[0069] The term “prevention” as used herein may include inhibiting the occurrence of a disease.

[0070] The term “treatment” as used herein includes inhibition, alleviation, or elimination of the development of a disease.

[0071] The term "including as an active ingredient" in this specification means that the tubulin inhibitor of this specification; 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 capable of exhibiting the effect mentioned in this specification, and includes formulation in various forms by adding various components as auxiliary components for drug delivery and stabilization, etc.

[0072] The pharmaceutical composition may comprise a pharmaceutically effective amount of the tubulin inhibitor; and at least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof.

[0073] As used herein, the term “pharmaceutically effective amount” means an amount sufficient to achieve the efficacy or activity of an active ingredient.

[0074] The pharmaceutical composition may additionally comprise a pharmaceutically acceptable diluent or carrier. The diluent may be lactose, corn starch, soybean oil, microcrystalline cellulose, mannitol, or a combination thereof. The carrier may be an excipient, a disintegrant, a binder, a glidant, or a combination thereof. The excipient may be microcrystalline cellulose, lactose, low-substituted hydroxycellulose, or a combination thereof. The disintegrant may be calcium carboxymethylcellulose, sodium starch glycolate, calcium hydrogen phosphate anhydrous, or a combination thereof. The binder may be polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, hydroxypropylcellulose, or a combination thereof. The glidant may be magnesium stearate, silicon dioxide, talc, or a combination thereof.

[0075] The pharmaceutical composition may be formulated as an oral or parenteral dosage form. Oral dosage forms may include granules, powders, liquids, tablets, capsules, dry syrups, or combinations thereof. Parenteral dosage forms may include injections, ointments, aerosols, sprays, patches, and the like, with no particular limitation on the dosage form.

[0076] The dosage of the pharmaceutical composition according to one example is, for example, about 0.0001 to 10000 mg / kg, 0.0001 to 5000 mg / kg, 0.0001 to 1000 mg / kg, 0.0001 to 900 mg / kg, 0.0001 to 800 mg / kg, 0.0001 to 700 mg / kg, 0.0001 to 600 mg / kg, 0.0001 to 500 mg / kg, 0.0001 to 400 mg / kg, 0.0001 to 300 mg / kg, 0.0001 to 200mg / kg, 1 to 1000mg / kg, 1 to 900mg / kg, 1 to 800mg / kg, 1 to 700mg / kg, 1 to 600mg / kg, 1 to 500mg / kg, 1 to 450mg / kg, 1 to 400mg / kg, 1 to 350mg / kg, 1 to 300mg / kg, 1 to 250mg / kg, 10 to 1000mg / kg, 10 to 900mg / kg, 10 to 800mg / kg, 10 to 700mg / kg, 10 to 600mg / kg, 10 to 500mg / kg, 10 to 450mg / kg, 10 to 400mg / kg, 10 to 350mg / kg, 10 to 300mg / kg, 10 to 250mg / kg, 100 to 1000mg / kg, 100 to 900mg / kg, 100 to 800mg / kg, 100 to 700mg / kg, 100 to 600mg / kg, 100 to 500mg / kg, 100 to 450mg / kg, 100 to 400mg / kg, 100 to 350mg / kg, 100 to 300mg / kg, 100 to 250mg / kg, 200 to 1000mg / kg, 200 to 900mg / kg, 200 to 800mg / kg, 200 to 700mg / kg, 200 to 600mg / kg, 200 The dosage may be administered in the range of 500 mg / kg to 500 mg / kg, 200 to 450 mg / kg, 200 to 400 mg / kg, 200 to 350 mg / kg, 200 to 300 mg / kg, or 200 to 250 mg / kg, but is not limited thereto. The frequency of administration of the pharmaceutical composition of the present application is not particularly limited thereto, but may be administered once a day or administered in divided doses several times. The above dosage does not limit the scope of the present application in any way.

[0077] The subject to which the pharmaceutical composition provided herein is administered may be a mammal, including a human, a dog, a cat, a horse, a cow, a pig, a goat, a rabbit, a mouse, a rat, etc., or a cell, tissue, or culture thereof isolated therefrom. In one example, the subject may be a subject (a mammal such as a human) in need of prevention, improvement, and / or treatment of a skin disease as described above, or a cell, tissue, or culture thereof isolated therefrom having a skin disease.

[0078] A pharmaceutical composition according to an example comprises the tubulin inhibitor; And at least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or a pharmaceutically acceptable salt thereof, in an amount of 1 to 80 wt%, 5 to 80 wt%, 5 to 75 wt%, 5 to 70 wt%, 5 to 65 wt%, 50 to 70 wt%, 55 to 65 wt%, 60 to 65 wt%, 10 to 60 wt%, 15 to 60 wt%, 20 to 60 wt%, 1 to 50 wt%, 5 to 50 wt%, 10 to 50 wt%, 15 to 50 wt%, 20 to 50 wt%, 1 to 40 wt%, 5 to 40 wt%, 10 to 40 wt%, 15 to 40 wt%, 20 It may comprise 40 wt%, 1 wt%, 30 wt%, 5 wt%, 10 wt%, 15 wt%, 30 wt%, 20 wt%, 1 wt%, 25 wt%, 5 wt%, 10 wt%, 25 wt%, 15 wt%, 20 wt%, or 23 wt%.

[0079]

[0080] Alternatively, the composition may be a composition for external use in the skin.

[0081] The above skin external preparation may be, but is not limited to, a cream, gel, ointment, skin emulsifier, skin suspension, transdermal patch, drug-containing bandage, lotion, spray, or a combination thereof.

[0082] In the present invention, the external skin preparation may be appropriately mixed with ingredients commonly used in external skin preparations such as cosmetics or medicines, such as aqueous ingredients, oily ingredients, powder ingredients, alcohols, moisturizers, thickeners, ultraviolet absorbers, whitening agents, preservatives, antioxidants, surfactants, fragrances, colorants, various skin nutrients, metal ion sequestrants, sugars, or combinations thereof, as needed.

[0083]

[0084] In addition, the present invention provides a pharmaceutical composition for preventing or treating skin diseases, comprising as an active ingredient at least one compound selected from the group consisting of a tubulin inhibitor; and calcium, colchicine, tapinarof, fingolimod, tofacitinib, or a pharmaceutically acceptable salt thereof.

[0085] In the present invention, quasi-drugs refer to all products other than pharmaceuticals related to the treatment or prevention of diseases. The quasi-drugs above refer to products that are used for the purpose of diagnosing, treating, improving, alleviating, managing, or preventing diseases of humans or animals, and have a milder effect than pharmaceuticals. For example, according to the Pharmaceutical Affairs Act, quasi-drugs are products excluding those used for the purpose of pharmaceuticals, and may include, but are not limited to, fiber and rubber products used for the treatment or prevention of diseases of humans or animals, products that have a mild or no direct effect on the human body, are not instruments or machines, and similar products, and sterilizers and insecticides for preventing infectious diseases.

[0086] The type or formulation of the pharmaceutical composition of the present invention is not particularly limited, but may preferably be a disinfectant, a shower foam, a gargle, a wet tissue, a detergent soap, a hand wash, a humidifier filler, a mask, an ointment, or a filter filler.

[0087] When the composition of the present invention is included in an over-the-counter drug for skin moisturizing purposes, the composition may be used as is or together with other over-the-counter drug ingredients, and may be used appropriately according to a conventional method. The mixing amount of the active ingredients may be appropriately determined depending on the intended use, and the over-the-counter drug composition according to the present invention may contain 0.01 to 20 wt% of the tubulin inhibitor; and at least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or pharmaceutically acceptable salts thereof, based on the total weight of the composition.

[0088]

[0089] In addition, the present invention provides a cosmetic composition for preventing or improving skin diseases, comprising as an active ingredient at least one compound selected from the group consisting of a tubulin inhibitor and calcium, colchicine, tapinarof, fingolimod, tofacitinib, or a pharmaceutically acceptable salt thereof.

[0090] The term "improvement" herein may mean any action that at least reduces the severity of a symptom, for example, a parameter associated with alleviating or treating a condition.

[0091] The above tubulin inhibitor; And at least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or a pharmaceutically acceptable salt thereof is present in an amount of 0.0001 to 80 wt%, 0.01 to 70 wt%, 0.01 to 60 wt%, 0.01 to 50 wt%, 0.01 to 40 wt%, 0.01 to 30 wt%, 0.1 to 70 wt%, 0.1 to 60 wt%, 0.1 to 50 wt%, 0.1 to 40 wt%, 0.1 to 30 wt%, 0.5 to 70 wt%, 0.5 to 60 wt%, 0.5 to 50 wt%, 0.5 to 40 wt%, 0.5 to 30 wt%, 1 to It may be present in an amount of, but is not limited to, 70 wt%, 1 to 60 wt%, 1 to 50 wt%, 1 to 40 wt%, or 1 to 30 wt%, for example, 30 wt%.

[0092] The above cosmetic composition is not particularly limited to a specific formulation, and the formulation may be appropriately selected depending on the intended purpose. The cosmetic composition may, for example, have a solubilized formulation, an emulsified formulation, or a dispersed formulation. The cosmetic composition may have a cosmetic formulation of an emollient toner, a nourishing toner, a massage cream, a nourishing cream, an essence, a pack, a gel, an ampoule, or a skin-adhesive type, but is not limited thereto.

[0093] The above cosmetic composition may additionally include ingredients commonly used in cosmetic compositions in addition to the effective ingredients disclosed herein, and may include, for example, conventional auxiliary agents and carriers such as antioxidants, stabilizers, solubilizers, surfactants, dispersants, emulsifiers, preservatives, vitamins, pigments, and fragrances.

[0094]

[0095] In addition, the present invention provides a cosmetic composition for skin moisturizing, whitening, wrinkle improvement, acne relief, skin barrier strengthening, or skin keratinocyte differentiation, comprising as an active ingredient at least one compound selected from the group consisting of a tubulin inhibitor; and calcium, colchicine, tapinarof, fingolimod, tofacitinib, or a pharmaceutically acceptable salt thereof.

[0096] As used herein, the term “skin moisturizing” may mean any action that maintains skin moisture or prevents moisture loss.

[0097] The term "whitening" as used herein may mean improving the brightness of skin tone or alleviating pigmentation by inhibiting melanin production.

[0098] The term "wrinkle improvement" as used herein may mean increasing skin elasticity, reducing the depth of wrinkles, or increasing the smoothness of the skin surface.

[0099] The term "acne relief" as used herein may mean suppressing inflammation, redness, or excessive sebum production of acne, or promoting healing of acne lesions.

[0100] In this specification, the term “strengthening the skin barrier” may mean any action that enhances the function of the skin barrier, which is located at the outermost part of the skin and prevents moisture and nutrient loss.

[0101] The term "skin keratinocyte differentiation" as used herein may mean promoting the process of keratinocytes maturing to form the stratum corneum, or thereby strengthening the function of the skin barrier.

[0102]

[0103] Another aspect of the present invention provides a method for preventing, improving, or treating a skin disease, comprising administering to a subject in need thereof an effective amount of the tubulin inhibitor; and at least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or a pharmaceutically acceptable salt thereof.

[0104] The term "effective amount" means an amount effective enough to produce the effect mentioned above.

[0105] The subject may be a mammal, such as a human, cow, horse, pig, dog, sheep, goat, or cat. The subject may be an object in need of a preventive, ameliorating, or therapeutic effect for a skin disease.

[0106] As used herein, the term "administering" is used interchangeably with "introducing" and "implanting" and may mean placement of a composition according to one embodiment into a subject by a method or route that results in at least partial localization of the composition to a desired site.

[0107] Administration may be by any method known in the art. The route of administration, frequency of administration, and other administration methods can be appropriately selected by those skilled in the art. Administration may be administered directly to a subject by any means, including intravenous, intramuscular, oral, transdermal, mucosal, intranasal, buccal, intratracheal, or subcutaneous administration. Administration may be systemic or local. Administration may include application to the skin.

[0108] In the present invention, administration is 0.1 mg to 1,000 mg of the composition according to one specific example per subject per day, for example, 0.1 mg to 500 mg, 0.1 mg to 100 mg, 0.1 mg to 50 mg, 0.1 mg to 25 mg, 1 mg to 1,000 mg, 1 mg to 500 mg, 1 mg to 100 mg, 1 mg to 50 mg, 1 mg to 25 mg, 5 mg to 1,000 mg, 5 mg to 500 mg, 5 mg to 100 mg, 5 mg to 50 mg, 5 mg to 25 mg, 10 mg to 1,000 mg, 10 mg to 500 mg, 10 mg The dosage may be administered in the range of 100 mg to 100 mg, 10 mg to 50 mg, or 10 mg to 25 mg. However, the dosage may be prescribed in various ways depending on factors such as the formulation method, administration method, patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and response sensitivity, and a person skilled in the art can appropriately adjust the dosage by considering these factors. The number of administrations may be once a day or twice or more within the range of clinically acceptable side effects, and the administration site may be administered in one or more sites, and the total number of administration days may be from 1 to 30 days per treatment at intervals of 2 to 5 days daily or every day. If necessary, the same treatment may be repeated after an appropriate period. For animals other than humans, the same dosage per kg as for humans may be administered, or the dosage may be converted to the above dosage based on the volume ratio (e.g., average value) of the organs (heart, etc.) of the target animal and humans.

[0109] Another aspect provides the use of the tubulin inhibitor; and at least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof, for use in preparing a composition for preventing, improving, or treating a skin disease.

[0110] Another aspect provides the use of the tubulin inhibitor; and at least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof, for use in the manufacture of a medicament for preventing, improving, or treating a skin disease.

[0111] Another aspect provides a food composition for preventing or improving skin diseases, comprising as an active ingredient at least one compound selected from the group consisting of a tubulin inhibitor; and calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof. The food composition may include a health functional food.

[0112] A food composition according to one embodiment of the present invention may be in the form of a liquid or solid formulation, and may be in the form of a tablet, capsule, soft capsule, pill, granule, beverage (drink), diet bar, chocolate, caramel formulation, or confectionery formulation, and the formulation is not particularly limited. In addition to the above-mentioned effective ingredient, the food composition of the present invention may contain excipients, sugars, flavorings, colorings, fats, proteins, etc. as needed.

[0113]

[0114] Duplicate contents are omitted in consideration of the complexity of this specification, and terms not otherwise defined in this specification have meanings commonly used in the technical field to which the present invention belongs.

[0115] Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are only illustrative of the present invention, and the content of the present invention is not limited to the following examples.

[0116]

[0117] [Example 1] Confirmation of filaggrin expression level according to combined treatment including tubulin inhibitor

[0118] After inserting a luciferase gene with a filaggrin promoter into a human epidermal keratinocyte (NHEK) cell line using Fugene4K reagent (Promega), the cells were treated with 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 alone or in combination with ABT-751 or TN16.

[0119] 24 hours after drug treatment, the level of filaggrin expression according to each compound treatment was measured using a kit (Promega) that measures the level of luciferase luminescence, and the results are shown in Figure 1.

[0120] As can be seen in Figure 1, it was confirmed that the expression level of filaggrin increased in the ABT-751 or TN16 treatment group compared to the untreated control group.

[0121] Additionally, it was confirmed that the expression of filaggrin was significantly increased in the ABT-751 or TN16 treatment group combined with calcium or colchicine, fingolimod, tapinarop, or tofacitinib compared to the ABT-751 or TN16 treatment group alone.

[0122]

[0123] [Example 2] Inhibition of collagen reduction mechanism by combined treatment including tubulin inhibitor

[0124] Photoaging refers to the skin aging process caused by exposure to the sun's ultraviolet rays. It is distinct from senescence (or biological aging), which is caused by the gradual decline in bodily functions. It is generally the primary cause of wrinkles, dryness, loss of elasticity, and pigmentation.

[0125] The sun emits two types of ultraviolet rays: UV-A and UV-B, with wavelengths ranging from 320 to 400 nm for UV-A and 290 to 320 nm for UV-B. Collagen is a prime example of tissue damage caused by this UV radiation. Collagen is a major component of connective tissue, primarily found in bone and skin, and is broadly classified into five types. Type 1 collagen is the most abundant type found in skin connective tissue, followed by Type 3.

[0126] Matrix Metalloproteinases (MMPs) are proteolytic enzymes that break down collagen, a major component of human connective tissue. Their expression levels are increased by UV rays. Therefore, when skin is exposed to UV rays, the expression of these matrix metalloproteinases (MMPs) increases, destroying collagen in the dermal layer and contributing to photoaging, including wrinkles and pigmentation.

[0127] ABT-751 or TN-16 were applied to human NHEK cells, respectively, and UV irradiation was performed 30 minutes later to induce photoaging (especially wrinkle improvement). In addition, 2 mM CaCl2, 1 μM colchicine, 1 μM tapinarof, 1 μM fingolimod, and 1 μM tofacitinib were co-treated with ABT-751 or TN16, and then UV irradiation was performed. For comparison and verification, a negative control group without any treatment and a positive control group irradiated only with UV-A were included in the experiment.

[0128] After application and UV exposure experiments, RNA was extracted from skin cells. The extracted RNA was reverse transcribed into cDNA, and the amounts of MMP-1, MMP-3, and MMP-9 present in skin tissue were measured using quantitative PCR (qPCR) using a real-time PCR machine.

[0129] Specifically, the photoaging inhibitory effect of ABT-751 or TN-16 was measured through changes in the expression level of Matrix Metalloproteinases (MMP) proteins described above. That is, the expression level of MMP proteins was measured in each of the control group that was irradiated with UV rays only and the experimental group that was irradiated with UV rays after applying ABT-751 or TN-16, respectively, to determine the degree of inhibition of MMP protein expression.

[0130] The results are shown in Fig. 2.

[0131] MMP-1 and MMP-9 are enzymes that promote the decomposition of type 1 and type 3 collagen, which are mainly expressed in skin tissue, and MMP-3 is a regulatory enzyme that decomposes type 1 collagen and controls changes in the expression level of MMP-1, and is an MMP protein that has a very important effect on photoaging.

[0132] As can be seen in Fig. 2, when ABT-751 or TN-16 was applied to the skin, the expression levels of MMP-1, MMP-3, and MMP-9 were mostly reduced by less than 50% compared to the control group that was exposed to only UV rays.

[0133] In addition, it was verified that in the experimental group 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, the MMP protein expression level was significantly suppressed compared to the experimental group treated with ABT-751 or TN-16 alone, and thus, the overall synergistic effect of suppressing photoaging by UV ultraviolet rays could be confirmed.

[0134]

[0135] [Example 3] Confirmation of Inflammatory Signaling Mechanism Inhibition Using Cell-Based Promoter Activity Measurement

[0136] NF-κB, a DNA transcription factor, exists in the cytosol and exists in a bound form with IκB, which inhibits NF-κB. NF-κB is involved in innate and adaptive immune responses and is a representative pro-inflammatory cytokine that appears in many inflammatory diseases.

[0137] The NF-κB mechanism is a protein family involved in inflammatory response regulation, immune modulation, apoptosis, cell proliferation, and epithelial differentiation. It regulates the expression of various genes and forms the central axis of the intracellular signal transmission system.

[0138] Human embryonic kidney cells (HEK) were transfected with a luciferase gene driven by the NFkb promoter using Fugene4K reagent (Promega). Six hours later, TNF-α was treated to induce inflammation and NFkb signaling. After 18 hours, ABT-751 or TN16 was treated at a concentration of 1.0 μM. In addition, 2 mM CaCl2, 1 μM colchicine, 1 μM tapinarof, 1 μM fingolimod, and 1 μM tofacitinib were treated alone or in combination with ABT-751 or TN16. After 24 hours of drug treatment, the inhibitory effect of each compound on the Nfkb signaling pathway was measured using a kit (Promega) that measures the level of luciferase emission, and the results are shown in Fig. 3.

[0139] As shown in Fig. 3, it was confirmed that treatment with ABT-751 or TN-16 at a concentration of 1.0 μM inhibited the NF-kB signaling pathway. In addition, in the group 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, it was confirmed that the NF-kB signaling pathway inhibition effect was significantly increased compared to the group treated with ABT-751 or TN16 alone.

[0140]

[0141] [Example 4] Confirmation of skin cell differentiation potential according to combined treatment including tubulin inhibitor.

[0142] To determine the effect of combination therapy containing a tubulin inhibitor on human epidermal skin cells NHEK, cells were observed after drug treatment.

[0143] A group treated with 1.0 μM ABT-751 or TN16, a group treated with 2 mM CaCl2, 1 μM colchicine, 1 μM tapinarof, 1 μM fingolimod, and 1 μM tofacitinib alone, and a group treated with CaCl2, colchicine, tapinarof, fingolimod, and tofacitinib together with ABT-751 or TN16 were generated. The degree of differentiation was measured 5 days after drug treatment.

[0144] In the calcium and colchicine-treated groups, skin cells were stained with a Keratin-14 antibody and photographed using a fluorescence microscope. Keratin-10 protein is a well-known marker of skin cell differentiation. The results of the fluorescence microscopy are shown in Figure 4a.

[0145] Additionally, skin cells from the groups treated with tapinarop, fingolimod, or tofacitinib were photographed using a phase contrast microscope, and the results are shown in Fig. 4b.

[0146] As shown in Fig. 4a, it was confirmed that cell differentiation increased in the experimental group treated with ABT-751 or TN16 compared to the negative control group.

[0147] In addition, as shown in Figures 4a and 4b, it was confirmed that cell differentiation significantly increased in the experimental group treated with calcium, colchicine, tapinarop, finglimod, tofacitinib, and ABT-751 or TN16 in combination compared to the experimental group treated with ABT-751 or TN16 alone.

[0148]

[0149] [Example 5] Confirmation of whitening function using tyrosinase activity measurement

[0150] To determine whether the combination therapy including the tubulin inhibitor of the present invention exhibits a whitening function, tyrosinase activity was measured. Tyrosinase is an enzyme that produces melanin by oxidizing tyrosine in the presence of oxygen, and inhibition of tyrosinase activity inhibits melanin production. Tyrosine oxidation by tyrosinase produces an orange color. However, when tyrosinase activity is inhibited, the oxidation reaction of tyrosine through tyrosinase is reduced, so the color gradually becomes lighter and transparent depending on the degree of tyrosinase inhibition. The degree of tyrosinase inhibition was evaluated by measuring the decrease in absorbance value at 490 nm. In the tyrosinase activity inhibition experiment, ABT-751 or TN16 was treated at a concentration of 1.0 μM to 2 mM of L-tyrosine as a substrate.

[0151] Additionally, 2 mM CaCl2, 1 μM colchicine, 1 μM tapinarof, 1 μM fingolimod, or 1 μM tofacitinib were treated alone or in combination with ABT-751 or TN16. Kojic acid, known to inhibit tyrosinase activity, was used as a control group.

[0152] After measuring the absorbance at 490 nm, tyrosinase enzyme was added again and the reaction was performed at 37°C. After measuring the absorbance at 490 nm again, the absorbance value was calculated to determine the degree to which each compound inhibited tyrosinase activity. The results are shown in Figure 5.

[0153] As shown in Fig. 5, the degree of inhibition of tyrosinase activity was significantly increased in the experimental group treated with ABT-751 or TN16 in combination with one of calcium, colchicine, tapinarop, finglimod, and tofacitinib, compared to the degree of inhibition of tyrosinase activity in the group treated with ABT-751 or TN16 alone. This confirms that the combination treatment including a tubulin inhibitor can perform a whitening function by suppressing melanin production.

Claims

1. Tubulin inhibitor; and At least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof; A pharmaceutical composition for preventing or treating skin diseases, comprising as an effective ingredient.

2. A 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. [Chemical Formula I] [Chemical Formula II] 3. A composition according to claim 1, wherein the skin disease is at least one selected from the group consisting of atopic dermatitis, skin thermal damage, pruritus, acne, psoriasis, allergic dermatitis, contact dermatitis, exfoliative dermatitis, seborrheic dermatitis, seborrheic dermatitis, lichen planus, rosacea, pigmentation disorders, hypermelanosis, erythema, wounds, ulcers, bedsores, lupus, skin wrinkle-related diseases, and skin diseases caused by photodamage.

4. A composition according to claim 3, wherein the skin wrinkle-related disease is at least one selected from the group consisting of elasticity fibrosis, thinning of the skin, skin atrophy, reduction of collagen fibers and elastic fibers, loss of skin elasticity, dryness, wrinkle formation, and premature skin aging.

5. A composition according to claim 3, wherein the skin disease caused by photodamage is at least one selected from the group consisting of lentigines, freckles, hypopigmentation, hyperpigmentation, photodamage caused by acute or chronic UV radiation, and photosensitization.

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

7. A composition according to claim 1, wherein the skin disease is at least one selected from the group consisting of panniculitis, calluses, vitiligo, urticaria, folliculitis, sebaceous keratosis pilaris, eczema, styes, freckles, blemishes, rashes, athlete's foot, spots, stretch marks, freckles, prickly heat, dry skin, chilblains, suppuration, keratoses, dermatitis, and psoriatic arthritis. 8.Tubulin inhibitors; and At least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof; A pharmaceutical composition for preventing or improving skin diseases, containing as an effective ingredient. 9.Tubulin inhibitors; and At least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, or a pharmaceutically acceptable salt thereof; A cosmetic composition for preventing or improving skin diseases, comprising as an effective ingredient. 10.Tubulin inhibitors; and At least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof; A cosmetic composition for moisturizing the skin, whitening, improving wrinkles, alleviating acne, strengthening the skin barrier, or differentiating skin keratinocytes, containing as an effective ingredient. 11.Tubulin inhibitor; and At least one compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, tofacitinib, and pharmaceutically acceptable salts thereof; A method for preventing or treating a skin disease, comprising the step of administering to a subject in need thereof;

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