Composition comprising tubulin inhibitor for preventing or treating skin diseases

Abstract

The present invention relates to a pharmaceutical composition, quasi-drug composition, or cosmetic composition comprising a tubulin inhibitor, especially, ABT-751 or TN16, as an active ingredient, for preventing, alleviating, or treating skin diseases. In addition, the present invention relates to a cosmetic composition comprising a tubulin inhibitor as an active ingredient for skin moisturization, whitening, wrinkle alleviation, acne relief, skin barrier strengthening, or skin keratinocyte differentiation. The tubulin inhibitor of the present invention can be utilized as an active ingredient that exhibits activities, such as skin moisturization, whitening, wrinkle alleviation, acne relief, skin barrier strengthening, or skin keratinocyte differentiation, along with the prevention, treatment, or alleviation of skin diseases.

Description

Composition for preventing or treating skin diseases comprising a tubulin inhibitor

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

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

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

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

[0005]

[0006] The primary function of the skin is to act as a barrier, separating the internal environment from the external environment, protecting against external aggression and minimizing the loss of moisture and other essential body components. Skin tissue is structured in layers—the basal layer, the spinous and granular layer, and the cornified layer—and contains various cells, including skin cells and immune cells responsible for the body's unique immune function. Skin tissue maintains homeostasis through the balance and harmony of these cells, each performing its own role. Skin disease occurs when this homeostasis is disrupted.

[0007] Cytokines are known to be soluble molecules that play a crucial role in mediating communication between immune cells and between immune and non-immune cells. While cytokines generally contribute to host defense and immune homeostasis, abnormal cytokine production can lead to dysfunctional immune responses and immune-related diseases, including autoimmune, rheumatic, allergic, and inflammatory skin diseases. Cytokines have been reported to play a crucial role in the regulation of skin inflammation. These signals must be tightly regulated, and if dysregulated, they can lead to immune system dysfunction, excessive inflammation, and various immune-related pathologies. In this context, negative regulation of cytokine signaling may be an important target for the treatment of skin diseases.

[0008] Furthermore, tight junctions are known to play a crucial role in epidermal barrier function. Tight junctions are cell-cell junction structures that exist not only in monolayered and multilayered epithelia but also in endothelium. Tight junctions are composed of transmembrane proteins, such as the claudin family, tight junction-associated marvel proteins (e.g., occludin, junctional adhesion molecules), and tight junction plaque proteins (e.g., zonula occludens (ZO) proteins 1-3, MUPP-1, and cingulin), which are linked to the actin filament cytoskeleton. The primary function of tight junctions is to seal intercellular pathways to restrict the movement of molecules within the intercellular space, and they are also involved in cell differentiation, proliferation, cell polarity, and signaling processes.

[0009] Psoriasis, an inflammatory skin disease, is primarily caused by an abnormality in the immune system and is considered an autoimmune disease that can affect the entire body. Its main symptoms include rapid, higher-than-normal proliferation of skin cells, thickening the epidermis and the accumulation of white, scaly scales. This is followed by a red, millet-like rash, covered by layers of silvery, dandruff-like scales. The rash gradually grows, sometimes as large as the palm of a hand. (Skin Lesions) A red rash covered with silvery, flaky scales (plaque psoriasis) is the most common, and it primarily occurs on the scalp, elbows, knees, and lower back. (Itching and Pain) The affected area may feel itchy or tingly, and in severe cases, it can be painful. (Skin Cracking and Bleeding) As the lesion worsens, the skin cracks and bleeds. (Nail and joint damage) It can lead to psoriatic arthritis, which causes joint pain, swelling, and nail deformities.

[0010]

[0011] Accordingly, the present inventors confirmed that tubulin inhibitors, particularly ABT-751 or TN16, reduce inflammatory responses by inhibiting signal transduction of inflammatory cytokines (TNF-α, IL-6, IL-17, IL-23) and inhibit collagen degradation and photoaging by inhibiting the expression of matrix metalloproteinases (MMP-1, MMP-3, MMP-9) induced by ultraviolet (UV) rays. In addition, they confirmed that they effectively inhibit NF-κB signal transduction to regulate immune responses and inflammatory mechanisms and strengthen skin barrier function by increasing the expression of tight junction proteins (Claudin-1, ZO-1, Occludin). In addition, they confirmed that they exhibit a whitening effect by inhibiting tyrosinase activity and thus suppressing melanin production, 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]

[0014] To achieve the above purpose, the present invention provides a pharmaceutical composition for preventing or treating skin diseases, comprising a tubulin inhibitor, specifically, ABT-751 or TN16, as an active ingredient.

[0015] In addition, the present invention provides a pharmaceutical composition for preventing or improving skin diseases, and a cosmetic composition for preventing or improving skin diseases, comprising the tubulin inhibitor as an active ingredient.

[0016] In addition, the present invention provides a cosmetic composition for skin moisturizing, whitening, wrinkle improvement, acne relief, skin barrier strengthening, or skin keratinocyte differentiation, which contains the tubulin inhibitor as an active ingredient.

[0017]

[0018] The tubulin inhibitor ABT-751 or TN16 of the present invention reduces inflammatory responses by inhibiting signal transduction of inflammatory cytokines (TNF-α, IL-6, IL-17, IL-23), and reduces collagen degradation and photoaging by inhibiting the expression of matrix metalloproteinases (MMP-1, MMP-3, MMP-9) induced by ultraviolet (UV) rays. In addition, it effectively inhibits NF-κB signal transduction to regulate immune responses and inflammatory mechanisms, and strengthens the skin barrier function by increasing the expression of tight junction proteins (Claudin-1, ZO-1, Occludin). In addition, it exhibits a whitening effect by inhibiting tyrosinase activity and thus suppressing melanin production.

[0019] Accordingly, the tubulin inhibitor of the present invention can be used as an effective ingredient that simultaneously exhibits activities such as preventing, treating, or improving skin diseases, as well as moisturizing the skin, whitening, improving wrinkles, alleviating acne, strengthening the skin barrier, or differentiating skin keratinocytes.

[0020]

[0021] Figure 1 shows the effect of ABT-751 or TN-16 on inhibiting inflammatory cytokine (TNF-α, IL-6, IL-17, IL-23) signal transduction. The degree of signal transduction inhibition of each cytokine was measured at OD630mm, indicating that inflammatory cytokine signal transduction was significantly reduced when treated with ABT-751 or TN-16 compared to the control group.

[0022] Figure 2 shows the inhibitory effects of ABT-751 or TN-16 on UV irradiation-induced MMP-1, MMP-3, and MMP-9 expression. RT-qPCR results indicate that MMP protein expression levels in the ABT-751 or TN-16 treatment groups were reduced to less than 50% compared to the control group.

[0023] Figure 3 shows the inhibitory effect of ABT-751 or TN-16 on NF-κB signaling. As measured after drug treatment in cells in which a luciferase luminescent gene has been inserted into the NF-κB promoter, ABT-751 or TN-16 exhibits an NF-κB signaling inhibitory effect similar to that of the control group and TPCA1, a known NF-κB inhibitor.

[0024] Figure 4 shows the effect of ABT-751 or TN-16 on increasing the differentiation of human epidermal keratinocytes (NHEK). Observation of cells treated with the drugs for 5 days using a phase-contrast microscope revealed a significant increase in the differentiation of skin cells in the ABT-751 or TN-16 treatment group compared to the control group.

[0025] Figures 5a and 5b show the effects of ABT-751 or TN-16 on increasing the expression of tight junction proteins (Claudin-1, ZO-1, Occludin). RT-qPCR and Western Blot analysis results show that the expression levels of Claudin-1, ZO-1, and Occludin increased in the ABT-751 or TN-16 treatment group compared to the control group.

[0026] Figure 6 shows the whitening effect of ABT-751 or TN-16, which reduces melanin production by inhibiting tyrosinase activity. The results of the tyrosinase activity inhibition experiment show that ABT-751 or TN-16 exhibits the efficacy of inhibiting melanin production.

[0027]

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

[0029]

[0030] The present invention provides a pharmaceutical composition for preventing or treating skin diseases, which comprises a compound represented by the following chemical formula I or chemical formula II or a pharmaceutically acceptable salt thereof as an active ingredient.

[0031] The present invention also provides a method for preventing or treating a skin disease, comprising administering to a subject in need thereof a therapeutically effective amount of a tubulin inhibitor, which is a compound represented by the following chemical formula I or chemical formula II or a pharmaceutically acceptable salt thereof.

[0032] [Chemical Formula I]

[0033]

[0034] [Chemical Formula II]

[0035]

[0036] 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.

[0037] In the present invention, unlike the previous functions, the tubulin inhibitor reduces the inflammatory response by suppressing the signal transduction of inflammatory cytokines (TNF-α, IL-6, IL-17, IL-23) in epidermal cells, and reduces collagen degradation and photoaging by suppressing the expression of matrix metalloproteinases (MMP-1, MMP-3, MMP-9) induced by ultraviolet (UV) rays. In addition, it effectively suppresses NF-κB signal transduction to regulate immune response and inflammatory mechanisms, and strengthens the skin barrier function by increasing the expression of tight junction proteins (Claudin-1, ZO-1, Occludin). In addition, it exhibits a whitening effect by inhibiting tyrosinase activity to suppress melanin production.

[0038] In the present invention, the tubulin inhibitor is ABT-751 represented by the following chemical formula I or TN-16 represented by the following chemical formula II.

[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]

[0043] 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).

[0044] 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.

[0045]

[0046] The tubulin inhibitor of the present invention is a concept that includes all pharmaceutically acceptable salts, isomers, derivatives, and analogs that exhibit the same or similar effects.

[0047] In the invention, pharmaceutically acceptable salts refer to salts commonly used in the pharmaceutical industry, and examples thereof include inorganic ion salts manufactured with calcium, potassium, sodium and magnesium, inorganic acid salts manufactured with hydrochloric acid, nitric acid, phosphoric acid, hydrobromic acid, iodic acid, perchloric acid and sulfuric acid, organic acid salts manufactured with acetic acid, trifluoroacetic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid and hydroiodic acid, sulfonic acid salts manufactured with methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid and naphthalenesulfonic acid, glycine, arginine, Examples thereof include, but are not limited to, amino acid salts manufactured from lysine, etc., and amine salts manufactured from trimethylamine, triethylamine, ammonia, pyridine, picoline, etc.

[0048] 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.

[0049] In the present invention, the tubulin inhibitor can exhibit activity that increases the regulation of cell tight junctions.

[0050]

[0051] 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.

[0052] In the present invention, the skin disease may be a skin disease or skin damage caused by a decrease in the expression of claudin 1, ZO-1 (Zonula Occludens-1), or Occludin protein.

[0053] In the present invention, the skin disease or skin damage may be at least one selected from the group consisting of, for example, psoriasis, skin thermal damage, acne, allergic dermatitis, contact dermatitis, exfoliative dermatitis, seborrheic dermatitis, seborrheic dermatitis, lichen planus, rosacea, pigmentation disorder, hypermelanosis, erythema, wound, ulcer, bedsore, lupus, skin wrinkle-related disease, and skin disease caused by photodamage, without limitation in type.

[0054] 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.

[0055] Additionally, the skin diseases caused by the photodamage may be lentigines, freckles, hypopigmentation, hyperpigmentation, photodamage due to acute or chronic UV radiation, or photosensitization.

[0056] Additionally, the skin disease of the present invention may be squamous cell carcinoma, basal cell carcinoma, benign epithelial tumor, and radiation dermatitis.

[0057] 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.

[0058] The skin disease of the present invention may be, for example, skin damage including psoriasis.

[0059] Histological characteristics of psoriatic lesions include a thickened epidermis due to abnormal keratinocyte proliferation and differentiation, as well as dermal infiltration and co-localization of T lymphocytes and dendritic cells. The pathogenesis of psoriasis is primarily defined as an immune response of Th1 and Th17 helper T cells, based on genetic and protein analysis. Accordingly, an increase in inflammatory cytokines directly related to skin disease, such as tumor necrosis factor (TNF-α) and interleukins IL-1α and IL-6, as well as IL-17α and IL-23, which are involved in the Th17 cell immune response, is observed. This increase in the inflammatory cytokines TNF-α, IL-1α, IL-6, IL-17α, and IL-23 can be defined as the inflammatory signaling profile of psoriasis, and therefore, drugs that can reduce these cytokines can be used as therapeutics for psoriasis.

[0060]

[0061] In addition, the present invention may further include a compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, and tofacitinib, in addition to the above tubulin inhibitor.

[0062] A compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, and tofacitinib may be used in combination or complex with a tubulin inhibitor, and administration may be simultaneous or sequential.

[0063] The above calcium, colchicine, tapinarof, fingolimod, or tofacitinib is a concept that includes all of its salts, isomers, or derivatives, and analogues that exhibit the same or similar effects.

[0064]

[0065] In the present invention, the tubulin inhibitor can be provided as an active ingredient in a pharmaceutical composition for preventing or treating skin diseases.

[0066] The term “prevention” as used herein may include inhibiting the occurrence of a disease.

[0067] The term “treatment” as used herein includes inhibition, alleviation, or elimination of the development of a disease.

[0068] The term "included as an active ingredient" in this specification means that the tubulin inhibitor of this specification is added to an extent that it can exhibit the effect mentioned in this specification, and includes the formulation in various forms by adding various components as auxiliary components for drug delivery and stabilization, etc.

[0069] The pharmaceutical composition may contain the tubulin inhibitor in a pharmaceutically effective amount.

[0070] As used herein, the term “pharmaceutically effective amount” means an amount sufficient to achieve the efficacy or activity of an active ingredient.

[0071] 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.

[0072] 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.

[0073] 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.

[0074]

[0075] The subject of administration of the pharmaceutical composition provided herein may be a mammal including a human, a dog, a cat, a horse, a cow, a pig, a goat, a rabbit, a mouse, a rat, etc., or a cell, tissue, or culture thereof isolated therefrom. In one example, the subject may be a subject (a mammal such as a human) in need of prevention, improvement, and / or treatment of a skin disease as described above, or a subject having an inflammatory skin disease, or a cell, tissue, or culture thereof isolated therefrom.

[0076] According to an example, a pharmaceutical composition comprises the tubulin inhibitor in an amount of 1 to 80 wt%, 5 to 80 wt%, 5 to 75 wt%, 5 to 70 wt%, 5 to 65 wt%, 50 to 70 wt%, 55 to 65 wt%, 60 to 65 wt%, 10 to 60 wt%, 15 to 60 wt%, 20 to 60 wt%, 1 to 50 wt%, 5 to 50 wt%, 10 to 50 wt%, 15 to 50 wt%, 20 to 50 wt%, 1 to 40 wt%, 5 to 40 wt%, 10 to 40 wt%, 15 to 40 wt%, 20 to 40 wt%, 1 to 30 wt%, 5 to 30 wt%, 10 to It may comprise 30 wt%, 15 to 30 wt%, 20 to 30 wt%, 1 to 25 wt%, 5 to 25 wt%, 10 to 25 wt%, 15 to 25 wt%, 20 to 25 wt%, or 23 to 25 wt%.

[0077]

[0078] Alternatively, the composition may be a composition for external use in the skin.

[0079] 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.

[0080] 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.

[0081]

[0082] In addition, the present invention provides a pharmaceutical composition for preventing or improving skin diseases, which comprises a tubulin inhibitor, which is a compound represented by Chemical Formula I or Chemical Formula II or a pharmaceutically acceptable salt thereof, as an active ingredient.

[0083] In the present invention, a quasi-drug refers to any product other than a drug related to the treatment or prevention of a disease.

[0084] 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.

[0085] 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.

[0086] When the composition of the present invention is included in an over-the-counter drug for skin moisturizing purposes, the composition may be used as is or in combination with other over-the-counter drug ingredients, and may be used appropriately according to conventional methods. The mixing amount of the active ingredients may be appropriately determined depending on the intended use, and the over-the-counter drug composition according to the present invention may contain the tubulin inhibitor in an amount of 0.01 to 20 wt% based on the total weight of the composition.

[0087]

[0088] In addition, the present invention provides a cosmetic composition for preventing or improving skin diseases, which comprises a tubulin inhibitor, which is a compound represented by Chemical Formula I or Chemical Formula II or a pharmaceutically acceptable salt thereof, as an active ingredient.

[0089] 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.

[0090] The tubulin inhibitor is present in an amount of 0.0001 to 80 wt%, 0.01 to 70 wt%, 0.01 to 60 wt%, 0.01 to 50 wt%, 0.01 to 40 wt%, 0.01 to 30 wt%, 0.1 to 70 wt%, 0.1 to 60 wt%, 0.1 to 50 wt%, 0.1 to 40 wt%, 0.1 to 30 wt%, 0.5 to 70 wt%, 0.5 to 60 wt%, 0.5 to 50 wt%, 0.5 to 40 wt%, 0.5 to 30 wt%, 1 to 70 wt%, 1 to 60 wt%, 1 to 50 wt%, 1 to 40 wt%, or 1 to 30%, based on the total weight of the cosmetic composition. For example, it may be included at 30 wt%, but is not limited thereto.

[0091] 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.

[0092] 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.

[0093]

[0094] In addition, the present invention provides a cosmetic composition for skin moisturizing, whitening, wrinkle improvement, acne relief, skin barrier strengthening, or skin keratinocyte differentiation, which comprises a tubulin inhibitor, which is a compound represented by Chemical Formula I or Chemical Formula II or a pharmaceutically acceptable salt thereof, as an active ingredient.

[0095] As used herein, the term “skin moisturizing” may mean any action that maintains skin moisture or prevents moisture loss.

[0096] The term "whitening" as used herein may mean improving the brightness of skin tone or alleviating pigmentation by inhibiting melanin production.

[0097] 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.

[0098] The term "acne relief" as used herein may mean suppressing inflammation, redness, or excessive sebum production of acne, or promoting healing of acne lesions.

[0099] 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.

[0100] 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.

[0101]

[0102] Another aspect of the present invention provides a method for preventing, improving, or treating a skin disease comprising administering an effective amount of the tubulin inhibitor to a subject in need thereof.

[0103] The term "effective amount" means an amount effective enough to produce the effect mentioned above.

[0104] 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.

[0105] 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.

[0106] 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.

[0107] 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.

[0108] Another aspect provides the use of the tubulin inhibitor for preparing a composition for preventing, improving or treating a skin disease.

[0109] Another aspect provides the use of the tubulin inhibitor for the manufacture of a medicament for preventing, improving or treating a skin disease.

[0110] Another aspect provides a food composition for preventing or improving skin diseases, comprising the tubulin inhibitor of the present invention as an active ingredient.

[0111] 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.

[0112]

[0113] 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.

[0114]

[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] Mechanism of Inflammatory Cytokine Suppression by Tubulin Inhibitors

[0118] We purchased HEK293 cell lines (Promega) each with a gene that has a chromogenic function inserted into the signal transduction promoters of TNF-α, IL-2, IL-4 / 13, IL-6, IL-12, IL-17, and IL-23. The degree of signal transduction can be measured based on the degree of blue color development in each cell line when each signal transduction is induced.

[0119] Each HEK-Blue cell line was treated with 1 μM ABT-751 or 1 μM TN-16, and then treated with a signal transduction inducer appropriate for each cell line after 6 hours. The degree of signal transduction of each induced inflammatory response was measured at OD630 mm, and the results are shown in Figure 1.

[0120] As can be seen in Fig. 1, when treated with 1 μM ABT-751 or 1 μM TN-16 drugs, a decrease in signal transduction of inflammatory cytokines TNF-α, IL-6, IL-17, and IL-23 was confirmed.

[0121] This confirmed the efficacy of ABT-751 or TN-16 drugs in reducing the skin inflammation signal profile of TNF-α and IL-6, IL-17, and IL-23 cytokines from the inflamed area.

[0122]

[0123] [Example 2] Confirmation of the collagen degradation inhibition effect of tubulin inhibitors.

[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 was applied to human NHEK cells, respectively, and UV irradiation was performed 30 minutes later to induce photoaging. To compare and verify ABT-751 or TN-16, 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 by measuring the change in the expression level of Matrix Metalloproteinases (MMP) proteins described above. That is, the expression level of MMP proteins was measured in each of the control group 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, and the degree to which the expression level of MMP proteins was suppressed by applying ABT-751 or TN-16, respectively, was measured.

[0130] The results are shown in Figure 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 confirmed 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. In this way, it was verified that the expression levels of Matrix Metalloproteinases (MMP) proteins were suppressed by the application of ABT-751 or TN-16, and this result indicates that ABT-751 or TN-16 may be effective for wrinkle-related diseases caused by collagen reduction or skin diseases caused by photodamage.

[0133]

[0134] [Example 3] Confirmation of the Nfkb inhibitory effect of a tubulin inhibitor.

[0135] 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.

[0136] 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.

[0137] In this study, we examined intracellular biochemical changes in HEK293 cells by treating them with ABT-751 or TN-16 to test the inflammatory response.

[0138] After inserting a luciferase gene with the NFkb promoter into human embryonic kidney cells (HEK) using Fugene4K reagent (Promega), inflammation was induced by treatment with TNF-α 6 hours later, and Nfkb signaling was induced. After 18 hours, treatment was performed with ABT-751 or TN-16 at a concentration of 1.0 μM.

[0139] After 24 hours of drug treatment, the Nfkb signaling pathway inhibition effect according to each tubulin inhibitor treatment was measured using a kit (Promega) that measures the degree of luciferase luminescence, and the results are shown in Figure 3.

[0140] As shown in FIG. 3, it was confirmed that treatment with ABT-751 or TN-16 according to the present invention inhibited signal transduction of NF-kB similarly to TPCA1, a control substance known as an NF-kB signal transduction inhibitor.

[0141]

[0142] [Example 4] Confirmation of the skin cell differentiation ability of a tubulin inhibitor

[0143] Using a human epidermal cell line (NHEK), the degree of skin cell differentiation was observed after applying ABT-751 or TN-16 and compared with the control group.

[0144] After treatment with 1 μM concentrations of ABT-751 or TN-16, skin cell differentiation was observed daily using a phase-contrast microscope. Figure 4 shows images of cells on day 5 of differentiation captured using a phase-contrast microscope camera. Compared to the untreated control group, skin cells treated with ABT-751 or TN16 showed a marked increase in differentiation.

[0145]

[0146] [Example 5] Confirmation of the efficacy of tubulin inhibitors on cell tight junctions.

[0147] Tight junctions (TJs) form intimate connections between epithelial and endothelial cells, regulating intercellular transport and maintaining barrier function. These structures are primarily composed of proteins such as Claudin-1, ZO-1, and Occludin, each with specific roles. Claudin-1, a core membrane protein of tight junctions, regulates selective permeability, Occludin enhances structural stability, and ZO-1, a scaffold protein, connects other proteins to the cytoskeleton.

[0148] The expression of tight junction proteins, Claudin-1, ZO-1, and Occludin, is essential for maintaining skin barrier function and strengthening defense against external stimuli. Reduced expression or dysfunction of these proteins can lead to the development or exacerbation of various skin diseases, such as atopic dermatitis, psoriasis, and dry skin. Therefore, therapeutic strategies that restore or regulate tight junction protein expression are attracting attention as important approaches for the prevention and treatment of skin diseases.

[0149] To confirm whether the tubulin inhibitor of the present invention is involved in tight junctions, the expression levels of Claudin-1, ZO-1, and Occludin were measured using RT-qPCR (reverse transcription real-time polymerase chain reaction).

[0150] Six hours after NHEK was treated with 1 μM concentration of ABT-751 or TN-16, intracellular RNA was extracted using the RNeasy mini kit (Qiagen). The extracted RNA was reverse transcribed into cDNA using the ImProm-II ™ Reverse Transcription kit (Promega), and the expression levels of Claudin-1, ZO-1, and Occludin in skin cells were measured by quantitative PCR (qPCR) using a Real-Time PCR Detection System (Bio-Rad, CFX96). The results are shown in Fig. 5a.

[0151] Additionally, intracellular proteins were extracted from NHEK cells treated with 1 μM concentration of ABT-751 or TN-16, respectively, after 24 hours. The extracted proteins were used to measure the expression levels of Claudin-1, ZO-1, and Occludin proteins present in skin cells using Western blot.

[0152] A portion of the collected tissue was homogenized using a tissue homogenizer (Daehan Science), lysed using a cell lysis buffer (RIPA buffer, Invitrogen), centrifuged using a centrifuge (Labogene), and the supernatant was transferred to a new tube and used as a protein sample.

[0153] 30 μg of protein sample was loaded onto a 4% to 12% gradient PAGE gel (Invitrogen), and SDS-PAGE was performed using Running buffer (Invitrogen) and an electrophoresis machine (Bio-Rad) to separate proteins by molecular weight. The separated proteins were transferred to a PVDF membrane (Bio-Rad) and then blocked with 5% blocking solution (Skim milk; BD).

[0154] The thin film was reacted with claudin-1, ZO-1, occludin antibody (Santa Cruz) or Actin antibody (Abcam) for 16 hours, washed, and then reacted with secondary antibody for 1 hour and washed.

[0155] After reacting the thin film with ECL solution (Thermo Fisher), the reacted protein was detected using a luminescence reaction measuring device (Fusion Solo, Vilber). The results are shown in Fig. 5b.

[0156] The results of Figures 5a and 5b show that the expression levels of Claudin-1, ZO-1, and Occludin increased in skin cells treated with ABT-751 or TN16 compared to the control group, indicating that ABT-751 or TN-16, a tubulin inhibitor of the present invention, strengthens the tight junctions of cells.

[0157]

[0158] [Example 6] Confirmation of the whitening effect of a tubulin inhibitor

[0159] Tyrosinase is a key enzyme involved in the initial, rate-determining step of melanin biosynthesis in the human body. Many whitening ingredients have mechanisms of action that inhibit this enzyme. This test assesses the degree to which tyrosinase enzyme activity is inhibited in vitro.

[0160] The whitening effects of ABT-751 or TN-16 were measured through in vitro tyrosinase inhibition experiments. Kojic acid, known to inhibit tyrosinase activity, was used as a control group.

[0161] In a tyrosinase activity inhibition experiment, 2 mM of the substrate L-tyrosine was treated with 1.0 μM concentration of each ABT-751 or TN-16, and the absorbance was measured at 490 nm. Then, 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 confirm the degree to which each ABT-751 or TN-16 inhibited tyrosinase activity. The results are shown in Fig. 6.

[0162] As can be seen in Fig. 6, the tubulin inhibitor of the present invention, ABT-751 or TN-16, exhibits an inhibitory effect on melanin production.

Claims

1. A pharmaceutical composition for preventing or treating skin diseases, comprising a tubulin inhibitor, which is a compound represented by the following chemical formula I or chemical formula II or a pharmaceutically acceptable salt thereof, as an active ingredient. [Chemical Formula I] [Chemical Formula II] 2. A pharmaceutical composition for preventing or treating skin diseases, wherein the tubulin inhibitor according to claim 1 increases tight junctions between cells.

3. A pharmaceutical composition according to claim 1, wherein the skin disease is a skin disease caused by decreased expression of claudin 1, Zonula Occludens-1 (ZO-1), or Occludin protein.

4. A composition according to claim 1, wherein the skin disease is at least one selected from the group consisting of psoriasis, skin thermal damage, acne, 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.

5. A composition according to claim 4, 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.

6. A composition according to claim 4, 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.

7. 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.

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

9. A pharmaceutical composition for improving skin diseases, comprising a tubulin inhibitor, which is a compound represented by the following chemical formula I or chemical formula II or a pharmaceutically acceptable salt thereof, as an active ingredient. [Chemical Formula I] [Chemical Formula II] 10. A cosmetic composition for preventing or improving skin diseases, comprising a tubulin inhibitor represented by the following chemical formula I or chemical formula II or a pharmaceutically acceptable salt thereof as an active ingredient. [Chemical Formula I] [Chemical Formula II] 11. A cosmetic composition for moisturizing the skin, whitening, improving wrinkles, alleviating acne, strengthening the skin barrier, or differentiating skin keratinocytes, comprising a tubulin inhibitor represented by the following chemical formula I or chemical formula II or a pharmaceutically acceptable salt thereof as an active ingredient. [Chemical Formula I] [Chemical Formula II] 12. A method for preventing or treating a skin disease, comprising administering to a subject in need thereof a therapeutically effective amount of a tubulin inhibitor represented by the following chemical formula I or chemical formula II or a pharmaceutically acceptable salt thereof. [Chemical Formula I] [Chemical Formula II]

Images