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

Abstract

The present invention relates to a pharmaceutical composition for the prevention, improvement, or treatment of skin diseases, comprising a tubulin inhibitor, particularly ABT-751 or TN16, as an active ingredient; a quasi-drug composition or a cosmetic composition. Furthermore, the present invention relates to a cosmetic composition for skin moisturizing, whitening, wrinkle improvement, acne relief, skin barrier strengthening, or skin keratinocyte differentiation, comprising a tubulin inhibitor as an active ingredient. The tubulin inhibitor of the present invention can be used as an active ingredient that simultaneously exhibits activities such as skin moisturizing, whitening, wrinkle improvement, acne relief, skin barrier strengthening, or skin keratinocyte differentiation, along with the prevention, treatment, or improvement of skin diseases.

Description

Technical Field

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

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

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

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

Background Art

[0005] The main function of the skin is to play a defensive role in separating the internal environment from the external environment, protecting against attacks by external substances, and minimizing the loss of moisture and other essential body components to the external space. The skin tissue coexists with various cells such as immune cells that perform the human body's inherent immune function, together with skin cells having a layer structure composed of a basal layer, a spinous and granular layer, and a cornified layer. The skin tissue maintains skin homeostasis by such various cells fulfilling their respective roles while maintaining balance and harmony, and skin diseases correspond to cases where such skin homeostasis cannot be maintained.

[0006] Cytokines are known as 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 autoimmunity, rheumatism, allergies, and inflammatory skin diseases. Cytokines have been reported to play a vital role in regulating skin inflammation. Such signaling must be strictly regulated; failure to regulate it can lead to immune system dysfunction, excessive inflammation, and various immune-related pathological phenomena. From this perspective, negative regulation of cytokine signaling may be an important target for the treatment of skin diseases.

[0007] Furthermore, tight junctions are known to play an important role in epidermal barrier function. Tight junctions are cell-cell junction structures present not only in monolayer and multilayer epithelium but also in endothelium. Tight junctions consist of transmembrane proteins of the claudin series, tight junction-associated marble proteins (egoccludin, JAMs (junctional adhesion molecules)), and tight junction plaque proteins (egzonula occludens (ZO) proteins 1-3, MUPP-1, and cingulin). These are linked to the actin filament cytoskeleton. The main 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 signal transduction processes.

[0008] Psoriasis, a type of inflammatory skin disease, is known to be primarily caused by an abnormality in the immune system and is considered a type of autoimmune disease that can affect the entire body. The main symptoms are characterized by skin cells proliferating faster than normal, causing the epidermis to thicken and scaly white keratin to accumulate. Rice-grain-sized red rashes appear on the skin, and layers of silvery-white, dandruff-like keratin accumulate on the affected areas. The size of the rashes increases, and in severe cases, they can grow to the size of a palm. (Skin lesions) Red rashes covered with silvery-white keratin (plaque psoriasis) are the most common and mainly occur on the scalp, elbows, knees, and waist. (Itching and pain) The affected areas may be itchy or tingly, and in severe cases, painful. (Skin cracking and bleeding) As the disease progresses, the skin cracks and bleeding occurs. (Damage to nails and joints) This can progress to psoriatic arthritis, resulting in joint pain, swelling, and nail deformities. [Overview of the project] [Problems that the invention aims to solve]

[0009] Therefore, the inventors confirmed that tubulin inhibitors, particularly ABT-751 or TN16, suppress inflammatory cytokine signaling (TNF-α, IL-6, IL-17, IL-23), thereby reducing inflammatory responses, and inhibit the expression of ultraviolet (UV)-induced Matrix Metalloproteinases (MMP-1, MMP-3, MMP-9), thereby suppressing collagen degradation and photoaging. They also confirmed that these inhibitors effectively suppress NF-κB signaling, regulating immune responses and inflammatory mechanisms, and increasing the expression of Tight Junction proteins (Claudin-1, ZO-1, Occludin), thereby strengthening skin barrier function. Furthermore, they confirmed that these inhibitors exert a whitening effect by inhibiting tyrosinase activity and suppressing melanin production, thus completing the present invention.

[0010] Therefore, an object of the present invention is to provide pharmaceutical compositions and quasi-drug compositions for the prevention or treatment of skin diseases. Another object of the present invention is to provide cosmetic compositions for the prevention or improvement of skin diseases and cosmetic compositions for moisturizing the skin, whitening, wrinkle improvement, acne relief, strengthening the skin barrier, or differentiation of skin keratinocytes. [Means for solving the problem]

[0011] To achieve the above objective, the present invention provides a pharmaceutical composition for the prevention or treatment of skin diseases comprising a tubulin inhibitor, specifically ABT-751 or TN16, as an active ingredient.

[0012] Furthermore, the present invention provides a quasi-drug composition for the prevention or improvement of skin diseases, and a cosmetic composition for the prevention or improvement of skin diseases, both containing the tubulin inhibitor as an active ingredient.

[0013] Furthermore, the present invention provides a cosmetic composition containing the tubulin inhibitor as an active ingredient for moisturizing the skin, whitening, improving wrinkles, alleviating acne, strengthening the skin barrier, or differentiating skin keratinocytes. [Effects of the Invention]

[0014] The tubulin inhibitors ABT-751 or TN16 of the present invention suppress inflammatory cytokine signaling (TNF-α, IL-6, IL-17, IL-23) to reduce inflammatory responses, suppress ultraviolet (UV)-induced Matrix Metalloproteinase (MMP-1, MMP-3, MMP-9) expression, and reduce collagen degradation and photoaging. Furthermore, they effectively suppress NF-κB signaling to regulate immune responses and inflammatory mechanisms, and increase the expression of Tight Junction proteins (Claudin-1, ZO-1, Occludin) to enhance skin barrier function. They also exert a whitening effect by inhibiting tyrosinase activity and suppressing melanin production.

[0015] Therefore, the tubulin inhibitor of the present invention can be used as an active 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. [Brief explanation of the drawing]

[0016] [Figure 1] This figure shows the effect of ABT-751 or TN-16 on suppressing inflammatory cytokine (TNF-α, IL-6, IL-17, IL-23) signaling. The degree of signaling suppression for each cytokine was measured at OD630mm, and the results show that treatment with ABT-751 or TN-16 significantly reduced inflammatory cytokine signaling compared to the control group. [Figure 2] This figure shows the effect of ABT-751 or TN-16 on suppressing the expression of MMP-1, MMP-3, and MMP-9 induced by UV irradiation. Measurements using RT-qPCR show that MMP protein expression levels decreased to less than 50% in the ABT-751 or TN-16 treated group compared to the control group. [Figure 3] This figure shows the effect of ABT-751 or TN-16 on suppressing NF-κB signaling. Measurements taken after drug treatment in cells with a luciferase luminescence gene inserted into the NF-κB promoter showed that ABT-751 or TN-16 exhibited NF-κB signaling inhibitory effects similar to those of the control group and TPCA1, a known NF-κB inhibitor. [Figure 4] This figure illustrates the effect of ABT-751 or TN-16 on increasing the differentiation of human epidermal cells (NHEKs). Phase-contrast microscopy observation of cells treated with the drug for 5 days shows a significant increase in skin cell differentiation in the ABT-751 or TN-16 treated group compared to the control group. [Figure 5a]Figures 5a and 5b are diagrams regarding the effect of ABT-751 or TN-16 on increasing the expression of Tight Junction proteins (Claudin-1, ZO-1, Occludin). As a result of RT-qPCR and Western Blot analyses, it is shown that the expression levels of Claudin-1, ZO-1, and Occludin increase in the ABT-751 or TN-16 treatment groups compared to the control group. [Figure 5b] Figures 5a and 5b are diagrams regarding the effect of ABT-751 or TN-16 on increasing the expression of Tight Junction proteins (Claudin-1, ZO-1, Occludin). As a result of RT-qPCR and Western Blot analyses, it is shown that the expression levels of Claudin-1, ZO-1, and Occludin increase in the ABT-751 or TN-16 treatment groups compared to the control group. [Figure 6] These are diagrams regarding the whitening effect of ABT-751 or TN-16 on suppressing Tyrosinase activity and reducing melanin production. As a result of the Tyrosinase activity inhibition experiment, it is shown that ABT-751 or TN-16 has the efficacy of suppressing melanin production.

Modes for Carrying Out the Invention

[0017] Hereinafter, the present invention will be described in detail.

[0018] The present invention provides a pharmaceutical composition for preventing or treating skin diseases, comprising as an active ingredient a compound represented by the following Chemical Formula I or Chemical Formula II or a pharmaceutically acceptable salt thereof.

[0019] Furthermore, the present invention provides a method for preventing or treating skin diseases, comprising the step of administering to an individual in need thereof a tubulin inhibitor which is a therapeutically effective amount of a compound represented by the following Chemical Formula I or Chemical Formula II or a pharmaceutically acceptable salt thereof.

Chemical Formula

[0020] A tubulin inhibitor refers to a group of drugs that regulate or inhibit the function of tubulin protein, which is the main component of microtubules. Microtubules are part of the cytoskeleton that plays an important role within cells and are essential structures in processes such as cell division, cell migration, and intracellular transport. Different from such previous functions, the tubulin inhibitor in the present invention suppresses the signal transduction of inflammatory cytokines (TNF-α, IL-6, IL-17, IL-23) in epidermal cells to reduce the inflammatory response, suppresses the expression of Matrix Metalloproteinases (MMP-1, MMP-3, MMP-9) by ultraviolet (UV) rays, and reduces collagen degradation and photoaging. In addition, it effectively suppresses NF-κB signal transduction to regulate the immune response and inflammatory mechanism, increases the expression of Tight Junction proteins (Claudin-1, ZO-1, Occludin) to strengthen the skin barrier function. Moreover, it inhibits Tyrosinase activity to suppress melanin production, thereby exerting a whitening effect.

[0021] 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.

[0022] The Compound I is a compound with CAS Registry Number 141430-65-1 and is ABT-751 (N-[2-[(4-Hydroxyphenyl)amino]pyridin-3-yl]-4-methoxybenzenesulfonamide).

[0023] ABT-751 is a compound that targets microtubules, and it has been reported that this drug shows the activity of suppressing the cell division of specific cancer cells and stopping the growth of cancer cells.

[0024] Furthermore, compound II is compound 33016-12-5, and is TN-16(3-[1-(phenylamino)ethylidene]-5-(phenylmethyl)-2,4-pyrrolidinedione).

[0025] TN-16 is a novel microtubule inhibitor with antitumor activity, synthesized by modifying tenuazonic acid (3-acetyl-5-sec-butyltetramic acid), a natural antibiotic isolated and characterized from Alternaria tenuis cultures. While TN-16 was found to possess potent anticancer properties in 1967, subsequent research has not uncovered any more potent drugs. Furthermore, in 1983, the mechanism of action of TN-16 was clarified, revealing that it inhibits microtubule formation and acts by binding to sensitive sites.

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

[0027] In this invention, pharmaceutically acceptable salts mean salts commonly used in the pharmaceutical industry, and examples include inorganic ion salts produced from calcium, potassium, sodium, and magnesium; inorganic salts produced from hydrochloric acid, nitric acid, phosphoric acid, bromate, iodate, perchloric acid, and sulfuric acid; acetic acid, trifluoroacetic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid, galactronic acid, glutamic acid, and glutamic acid. Examples include, but are not limited to, organic salts produced from taric acid, glucuronic acid, aspartic acid, ascorbic acid, carboxylic acids, vanillic acid, hydroiodic acid, etc.; sulfonates produced from methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, and naphthalenesulfonic acid, etc.; amino acid salts produced from glycine, arginine, lysine, etc.; and amine salts produced from trimethylamine, triethylamine, ammonia, pyridine, picoline, etc.

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

[0029] The tubulin inhibitor in this invention can exhibit activity that increases and regulates cell tight junctions.

[0030] In this invention, skin diseases are a group of diseases resulting from structural and functional abnormalities of the skin, and are a comprehensive concept encompassing abnormal conditions originating from the skin that can be triggered by various causes such as inflammation, infection, allergy, and immune abnormalities.

[0031] The skin disease in this invention may be a skin disease or skin injury caused by a decrease in the expression of claudin 1, ZO-1 (Zonula Occludens-1), or occludin protein.

[0032] The skin disease or skin injury in the present invention may be one or more selected from the group consisting of, for example, psoriasis, thermal skin injury, acne, allergic dermatitis, contact dermatitis, exfoliative dermatitis, seborrheic dermatitis, seborrheic scalp, lichen planus, rosacea, pigment disorders, melanosis, erythema, wounds, ulcers, pressure ulcers, lupus, wrinkle-related skin diseases, and skin diseases caused by photodamage, and the type is not limited.

[0033] The aforementioned wrinkle-related skin conditions may include elastic fibrosis, thinning of the skin, skin atrophy, reduction of collagen and elastic fibers, loss of skin elasticity, dryness, wrinkle formation, or premature skin aging.

[0034] Furthermore, the skin conditions caused by photodamage may include lentigines, freckles, hypopigmentation, hyperpigmentation, photodamage due to acute or chronic UV radiation, or photosensitization.

[0035] Furthermore, the skin diseases of the present invention may include squamous cell carcinoma, basal cell carcinoma, benign epithelial tumors, and radiation dermatitis.

[0036] Furthermore, the skin disease of the present invention may be panniculitis, calluses, vitiligo, urticaria, folliculitis, sebaceous keratosis pilaris, eczema, corns, melasma, rash, athlete's foot, spots, stretch marks, freckles, heat rash, dry skin, chilblains, suppuration, pox, scalp inflammation, or psoriatic arthritis.

[0037] The skin disease of the present invention may be, for example, a skin injury including psoriasis.

[0038] The histological characteristics of psoriatic lesions show not only abnormal keratinocyte proliferation and differentiation, but also thickened epidermis due to skin infiltration and co-localization of T lymphocytes and dendritic cells. The pathogenesis of psoriasis depends primarily on genetic and protein analysis results and is defined as the immune response of helper T cells, specifically Th1 and Th17 cells. This leads to an increase in inflammatory cytokines directly associated with skin disease, such as tumor necrosis factor (TNF-α), interleukins (IL-1α and IL-6), as well as increases in IL-17α and IL-23, which are involved in the immune response of Th17 cells. These increases in inflammatory cytokines, including TNF-α, IL-1α, IL-6, IL-17α, and IL-23, can be defined as the inflammatory signaling profile of psoriasis; therefore, drugs that can reduce these cytokines can be used as therapeutic agents for psoriasis.

[0039] Furthermore, the present invention may further include, in addition to the tubulin inhibitor, a compound selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, and tofacitinib.

[0040] The compounds selected from the group consisting of calcium, colchicine, tapinarof, fingolimod, and tofacitinib may be used in combination with or in combination with tubulin inhibitors, and may be administered simultaneously or sequentially.

[0041] The terms calcium, colchicine, tapinarof, fingolimod, or tofacitinib are concepts that include all salts, isomorphs, or derivatives thereof, as well as analogs that exhibit the same or similar effects.

[0042] In the present invention, the tubulin inhibitor can be provided as an active ingredient in a pharmaceutical composition for the prevention or treatment of skin diseases.

[0043] The term "prevention" as used herein may include suppressing the occurrence of disease.

[0044] In this specification, the term “treatment” includes the suppression, mitigation, or elimination of the progression of a disease.

[0045] In this specification, the term "contains as an active ingredient" means that the tubulin inhibitor specified herein is added to an extent that it can exert the effects described herein, and includes formulation into various forms by adding various components as adjuncts for purposes such as drug delivery and stabilization.

[0046] The pharmaceutical composition may contain the tubulin inhibitor in a therapeutically effective amount.

[0047] In this specification, the term "therapeutic dose" means an amount sufficient to achieve the efficacy or activity of the active ingredient.

[0048] The pharmaceutical composition may further comprise a pharmaceutically acceptable diluent or carrier. The diluent may be lactose, corn starch, soybean oil, amorphous cellulose, mannitol, or a combination thereof. The carrier may be an excipient, disintegrant, binder, lubricant, or a combination thereof. The excipient may be microcrystalline cellulose, lactose, low-substituted hydroxycellulose, or a combination thereof. The disintegrant may be carboxymethylcellulose calcium, sodium starch glycolate, anhydrous monohydrogen phosphate, or a combination thereof. The binder may be polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, hydroxypropylcellulose, or a combination thereof. The lubricant may be magnesium stearate, silicon dioxide, talc, or a combination thereof.

[0049] The aforementioned pharmaceutical composition may be formulated into oral or parenteral dosage forms. Oral dosage forms may include granules, powders, liquids, tablets, capsules, dried syrups, or combinations thereof. Parenteral dosage forms may include injections, ointments, aerosols, sprays, patches, etc., and the dosage form is not limited.

[0050] For example, the dosage of a pharmaceutical composition for a mammal throughout the day is approximately 0.0001-10000 mg / kg, 0.0001-5000 mg / kg, 0.0001-1000 mg / kg, 0.0001-900 mg / kg, 0.0001-800 mg / kg, 0.0001-700 mg / kg, 0.0001-600 mg / kg, 0.0001-500 mg / kg, 0.0001-400 mg / kg, 0.0001-300 mg / kg, 0 .0001~200mg / kg, 1~1000mg / kg, 1~900mg / kg, 1~800mg / kg, 1~700mg / kg, 1~600mg / kg, 1~500mg / kg, 1~450mg / kg, 1~400 mg / kg, 1~350mg / kg, 1~300mg / kg, 1~250mg / kg, 10~1000mg / kg, 10~900mg / kg, 10~800mg / kg, 10~700mg / kg, 10~600mg / k g, 10~500mg / kg, 10~450mg / kg, 10~400mg / kg, 10~350mg / kg, 10~300mg / kg, 10~250mg / kg, 100~1000mg / kg, 100~900mg / kg, 100~800mg / kg, 100~700mg / kg, 100~600mg / kg, 100~500mg / kg, 100~450mg / kg, 100~400mg / kg, 100~350mg / kg, 100~ The drug can be administered in doses of 300 mg / kg, 100-250 mg / kg, 200-1000 mg / kg, 200-900 mg / kg, 200-800 mg / kg, 200-700 mg / kg, 200-600 mg / kg, 200-500 mg / kg, 200-450 mg / kg, 200-400 mg / kg, 200-350 mg / kg, 200-300 mg / kg, or 200-250 mg / kg, but is not limited thereto. The frequency of administration of the pharmaceutical composition of this application is not particularly limited, but can be administered once a day or in divided doses several times a day. The aforementioned dosages do not limit the scope of this application in any respect.

[0051] The subjects of the pharmaceutical compositions provided herein may be mammals, including humans, dogs, cats, horses, cattle, pigs, goats, rabbits, mice, rats, etc., or cells, tissues, or cultures thereof isolated therefrom. For example, the subjects may be individuals (such as humans) that require prevention, improvement, and / or treatment of skin diseases as described above, or that have inflammatory skin diseases, or cells, tissues, or cultures thereof isolated therefrom.

[0052] A pharmaceutical composition, as an example, contains the tubulin inhibitor in amounts of 1-80% by weight, 5-80% by weight, 5-75% by weight, 5-70% by weight, 5-65% by weight, 50-70% by weight, 55-65% by weight, 60-65% by weight, 10-60% by weight, 15-60% by weight, 20-60% by weight, 1-50% by weight, 5-50% by weight, 10-50% by weight, 15-50% by weight, and 20% by weight. It may also contain ~50% by weight, 1~40% by weight, 5~40% by weight, 10~40% by weight, 15~40% by weight, 20~40% by weight, 1~30% by weight, 5~30% by weight, 10~30% by weight, 15~30% by weight, 20~30% by weight, 1~25% by weight, 5~25% by weight, 10~25% by weight, 15~25% by weight, 20~25% by weight, or 23~25% by weight.

[0053] Alternatively, the composition may be a topical skin preparation composition.

[0054] The aforementioned topical skin 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.

[0055] In the present invention, the topical skin preparation can be appropriately formulated as needed with ingredients commonly used in topical skin preparations such as cosmetics and pharmaceuticals, such as aqueous components, oily components, powder components, alcohols, humectants, thickeners, UV absorbers, whitening agents, preservatives, antioxidants, surfactants, fragrances, colorants, various skin nutrients, metal ion chelating agents, sugars, or combinations thereof.

[0056] Furthermore, the present invention provides a quasi-drug composition for the prevention or improvement of skin diseases, comprising 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.

[0057] In this invention, "quasi-drug" refers to all products that are not pharmaceuticals related to the treatment or prevention of disease.

[0058] In this invention, "quasi-drug" refers to all products that are not pharmaceuticals but are related to the treatment or prevention of disease. The term "quasi-drug" means articles used for the purpose of diagnosing, treating, improving, alleviating, managing, or preventing diseases in humans or animals, and which have a milder effect than pharmaceuticals. For example, according to the Pharmaceuticals and Medical Devices Act, a quasi-drug is an article that is not used for pharmaceutical purposes, and may include, but is not limited to, textile and rubber products used for the treatment or prevention of diseases in humans and animals, items that have a mild or no direct effect on the human body and are not instruments or machines, similar items, and disinfectants and insecticides for preventing infectious diseases.

[0059] The type and dosage form of the quasi-drug composition of the present invention are not particularly limited, but preferably include disinfectant cleansers, shower foams, mouthwashes, wet wipes, detergents, soaps, hand washes, humidifier fillers, masks, ointments, or filter fillers.

[0060] When the composition of the present invention is included in a quasi-drug for moisturizing the skin, the composition can be used as is or in combination with other quasi-drug ingredients, and can be used appropriately according to conventional methods. The amount of active ingredients mixed can be appropriately determined according to the purpose of use, and the quasi-drug composition according to the present invention may contain the tubulin inhibitor in an amount of 0.01 to 20% by weight relative to the total weight of the composition.

[0061] Furthermore, the present invention provides a cosmetic composition for the prevention or improvement of skin diseases, comprising 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. In this specification, the term “improvement” may mean any action that alleviates or treats a condition, such as any action that reduces the severity of the symptoms.

[0062] The tubulin inhibitor may be present in amounts of 0.0001 to 80% by weight, 0.01 to 70% by weight, 0.01 to 60% by weight, 0.01 to 50% by weight, 0.01 to 40% by weight, 0.01 to 30% by weight, 0.1 to 70% by weight, 0.1 to 60% by weight, 0.1 to 50% by weight, 0.1 to 40% by weight, 0.1 to 30% by weight, 0.5 to 70% by weight, 0.5 to 60% by weight, 0.5 to 50% by weight, 0.5 to 40% by weight, 0.5 to 30% by weight, 1 to 70% by weight, 1 to 60% by weight, 1 to 50% by weight, 1 to 40% by weight, or 1 to 30%, for example, 30% by weight, relative to the total weight of the cosmetic composition, but is not limited thereto.

[0063] The cosmetic composition is not particularly limited to a specific dosage form, and the dosage form can be appropriately selected according to the purpose. The cosmetic composition may, for example, have a solubilizing dosage form, an emulsifying dosage form, or a dispersing dosage form. The cosmetic composition may, but is not limited to, a softening lotion, a nourishing lotion, a massage cream, a nourishing cream, an essence, a pack, a gel, an ampoule, or a skin-applied cosmetic dosage form.

[0064] In addition to the active ingredients disclosed herein, the cosmetic composition may further contain ingredients commonly used in cosmetic compositions, such as common auxiliary agents and carriers including antioxidants, stabilizers, solubilizers, surfactants, dispersants, emulsifiers, preservatives, vitamins, pigments, and fragrances.

[0065] Furthermore, the present invention provides a cosmetic composition for moisturizing the skin, whitening, improving wrinkles, alleviating acne, strengthening the skin barrier, or differentiating skin keratinocytes, comprising a tubulin inhibitor, which is a compound represented by chemical formula I or chemical formula II, or a pharmaceutically acceptable salt thereof, as an active ingredient.

[0066] In this specification, the term "skin moisturizing" may mean any action that retains moisture in the skin or prevents moisture loss.

[0067] In this specification, the term "whitening" may mean improving the brightness of skin tone or reducing hyperpigmentation by suppressing melanin production.

[0068] In this specification, the term “wrinkle improvement” may mean an improvement in skin elasticity, a reduction in the depth of wrinkles, or an improvement in the smoothness of the skin surface.

[0069] In this specification, the term “acne relief” may mean suppressing acne inflammation, redness, or excessive sebum production, or promoting the healing of acne lesions.

[0070] In this specification, the term "skin barrier enhancement" may mean all actions that improve the function of the skin barrier, which is located on the outermost layer of the skin and prevents the loss of moisture and nutrients.

[0071] In this specification, the term “keratinocyte differentiation” may mean the process by which keratinocytes mature and form the stratum corneum, or thereby enhance the function of the skin barrier.

[0072] Another aspect of the present invention provides a method for preventing, improving, or treating a skin disease, comprising the step of administering an effective amount of the tubulin inhibitor to an individual in need.

[0073] The term "effective quantity" refers to a quantity that is effective enough to produce the effects mentioned above.

[0074] The individual may be a mammal, such as a human, cattle, horse, pig, dog, sheep, goat, or cat. The individual may be one that requires preventive, ameliorative, or therapeutic effects for a skin disease.

[0075] In this specification, the term “administer” is interchangeable with “introduce” and “implant,” and may mean the placement of a particular composition into an individual by a method or route that results in at least partial localization of the composition to a desired site. Administration may be by methods known in the art. The method of administration, including the route and number of doses, can be appropriately selected by a skilled technician. Administration may be directly administered to the individual by any means, such as intravenous, intramuscular, oral, transdermal, mucosal, intranasal, oral, intratracheal, or subcutaneous administration. The administration may be systemic or topical. The administration may include topical application to the skin.

[0076] In the present invention, administration may involve administering a composition according to one specific example at a dose of 0.1 mg to 1,000 mg per individual per day, for example, 0.1 mg to 500 mg, 0.1 mg to 100 mg, 0.1 mg to 50 mg, 0.1 mg to 25 mg, 1 mg to 1,000 mg, 1 mg to 500 mg, 1 mg to 100 mg, 1 mg to 50 mg, 1 mg to 25 mg, 5 mg to 1,000 mg, 5 mg to 500 mg, 5 mg to 100 mg, 5 mg to 50 mg, 5 mg to 25 mg, 10 mg to 1,000 mg, 10 mg to 500 mg, 10 mg to 100 mg, 10 mg to 50 mg, or 10 mg to 25 mg. However, the dosage can be prescribed in various ways depending on factors such as the formulation method, administration method, patient's age, weight, sex, condition, diet, administration time, administration route, excretion rate, and response sensitivity, and a person skilled in the art can appropriately adjust the dosage considering these factors. The number of administrations can be once a day or two or more times a day within the range of clinically acceptable side effects, and it can be administered to one or more sites, and the total number of administration days in a single treatment can be from 1 to 30 days, administered daily or at intervals of 2 to 5 days. If necessary, the same treatment can be repeated after an appropriate period. For non-human animals, the dosage can be the same as for humans per kg, or the dosage can be converted using, for example, the volume ratio of organs (such as the heart) of the target animal and humans (e.g., average value) and administered.

[0077] Another embodiment provides the use of the tubulin inhibitor for use in manufacturing compositions for the prevention, improvement, or treatment of skin diseases.

[0078] Another embodiment provides the use of the tubulin inhibitor for use in manufacturing agents for the prevention, improvement, or treatment of skin diseases.

[0079] Another embodiment provides a food composition for preventing or improving skin diseases, comprising the tubulin inhibitor of the present invention as an active ingredient.

[0080] A food composition according to one embodiment of the present invention may be in liquid or solid form, and may be in the form of tablets, capsules, soft capsules, pills, granules, beverages (drinks), diet bars, chocolates, caramel, or confectionery, and the form is not particularly limited. In addition to the active ingredients, the food composition of the present invention may optionally contain excipients, sugars, flavorings, colorings, oils and fats, proteins, etc.

[0081] Repetitive content has been omitted in consideration of the complexity of this specification, and terms not otherwise defined herein have the meanings commonly used in the art to which this invention pertains.

[0082] The present invention will be described in detail below with reference to examples. However, the following examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following examples.

[0083] [Example 1] Mechanism of suppression of inflammatory cytokines by tubulin inhibitors We purchased HEK293 cell lines (Promega) in which genes with chromogenic function were inserted into the respective signaling promoters of TNF-α, IL-2, IL-4 / 13, IL-6, IL-12, IL-17, and IL-23. Each of these cell lines allows us to measure the degree of signal transduction based on the degree to which each signal is induced and the resulting blue coloration.

[0084] Each HEK-Blue cell line was treated with either 1 μM ABT-751 or 1 μM TN-16, and then treated with a signaling-inducing substance appropriate for each cell line 6 hours later. The signaling levels of each induced inflammatory response were measured at OD630 mm, and the results are shown in Figure 1.

[0085] As can be seen in Figure 1, treatment with 1 μM ABT-751 or 1 μM TN-16 drugs resulted in decreased signaling of the inflammatory cytokine TNF-α and IL-6, IL-17, and IL-23.

[0086] This confirms the efficacy of ABT-751 or TN-16 drugs in reducing TNF-α and IL-6, IL-17, and IL-23 cytokines, which are skin inflammation signaling profiles, from the site of inflammation.

[0087] [Example 2] Confirmation of the collagen degradation inhibitory effect of a tubulin inhibitor. Photoaging refers to the skin aging phenomenon caused by exposure to ultraviolet rays from the sun. This is a form of aging distinct from biological aging (senescence or biological aging), which is caused by a gradual decline in biological functions, and generally acts as a major cause of wrinkles and dryness, elastic fibrosis, and hyperpigmentation.

[0088] The sun emits two types of ultraviolet radiation: UV-A and UV-B. UV-A has wavelengths of 320-400 nm, while UV-B has wavelengths of 290-320 nm. Collagen is a prime example of tissue damaged by this type of UV radiation. Collagen is the main component of connective tissue, primarily found in bones and skin, and is broadly classified into five types. Type 1 collagen is the most abundant type found in the connective tissue of skin, followed by Type 3.

[0089] Matrix metalloproteinases (MMPs) are proteolytic enzymes that break down collagen and other components of connective tissue, and their expression increases when exposed to UV ultraviolet light. Therefore, when the skin is exposed to UV light, the expression of these Matrix Metalloproteinases (MMPs) increases, destroying collagen in the dermis and accelerating photoaging, such as wrinkles and hyperpigmentation.

[0090] Human NHEK cells were coated with either ABT-751 or TN-16, and photoaging was induced by UV irradiation 30 minutes later. To compare and validate ABT-751 and TN-16, a negative control group (untreated) and a positive control group (irradiated only with UV-A) were included in the experiment.

[0091] After the application and UV ultraviolet experiments were completed, RNA was extracted from skin cells. The extracted RNA was reverse transcribed into cDNA, and the amounts of MMP-1, MMP-3, and MMP-9 present in the skin tissue were measured using quantitative PCR (qPCR) with a Real-time PCR machine.

[0092] Specifically, the photoaging-inhibiting effect of ABT-751 or TN-16 was measured by the change in Matrix Metalloproteinase (MMP) protein expression levels described above. That is, the expression levels of MMP proteins were measured in a control group that was irradiated with ultraviolet light only, and in experimental groups that were irradiated with ultraviolet light after being coated with ABT-751 or TN-16, respectively. The degree to which MMP protein expression was suppressed by the application of ABT-751 or TN-16 was then measured.

[0093] The results are shown in Figure 2.

[0094] MMP-1 and MMP-9 are enzymes that promote the degradation of type 1 and type 3 collagen, which are mainly expressed in skin tissue, while MMP-3 is a regulatory enzyme that degrades type 1 collagen and adjusts changes in MMP-1 expression levels. These MMP proteins have a very important impact on photoaging.

[0095] As can be seen in Figure 2, when ABT-751 or TN-16 was applied to the skin, the expression levels of MMP-1, MMP-3, and MMP-9 were all reduced to approximately 50% or less compared to the control group irradiated only with ultraviolet light. Thus, it has been verified that the application of ABT-751 or TN-16 suppresses the expression of Matrix Metalloproteinases (MMP) proteins, which indicates that ABT-751 or TN-16 may be effective for wrinkle-related diseases caused by collagen reduction or skin diseases caused by photodamage.

[0096] [Example 3] Confirmation of the NF-κB inhibitory effect of a tubulin inhibitor. NF-κB, a DNA transcription factor, is present in the cell matrix and exists in a form bound to IκB, which suppresses NF-κB. NF-κB is involved in both congenital and acquired immune responses and is a representative pro-inflammatory cytokine exhibited in many inflammatory diseases.

[0097] The NF-κB mechanism involves a group of proteins (protein family) involved in regulating inflammatory responses, immune system modulation, apoptosis, cell proliferation, and epithelial differentiation. They regulate the expression of various genes and form the central axis of intracellular signaling pathways.

[0098] In this study, intracellular biochemical changes were observed in HEK293 cell lines by treatment with ABT-751 or TN-16 to assess inflammatory responses.

[0099] Human embryonic kidney cells (HEKs) were given a luciferase luminescence gene with an NF-κB promoter using the Fugene4K reagent (Promega). Six hours later, TNF-α was used to induce inflammation and NF-κB signaling. Eighteen hours later, the cells were treated with either ABT-751 or TN-16 at a concentration of 1.0 μM.

[0100] 24 hours after drug treatment, the inhibitory effect of each tubulin inhibitor on the NF-κB signaling mechanism was measured using a luciferase luminescence intensity measurement kit (Promega), and the results are shown in Figure 3.

[0101] As shown in Figure 3, it was confirmed that treatment with ABT-751 or TN-16 according to the present invention suppresses NF-κB signaling in a similar manner to TPCA1, a known NF-κB signaling inhibitor that is a control group substance.

[0102] [Example 4] Confirmation of the skin cell differentiation ability of a tubulin inhibitor Human epidermal cell lines (NHEK) were used, and after applying ABT-751 or TN-16, the degree of skin cell differentiation was observed and compared with a control group.

[0103] After treating skin cells with either 1 μM ABT-751 or TN-16, respectively, the degree of skin cell differentiation was observed daily using a phase-contrast microscope. Figure 4 shows the results of cells on day 5 of differentiation, captured with a phase-contrast microscope camera. Compared to the untreated control group, a significant increase in the degree of differentiation can be confirmed in skin cells treated with ABT-751 or TN-16.

[0104] [Example 5] Confirmation of the tight junction-related efficacy of a tubulin inhibitor. Tight junctions are crucial structures that form close 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 playing a specific role. Claudin-1 is a core membrane protein of tight junctions that regulates selective permeability, Occludin enhances structural stability, and ZO-1 acts as a scaffold protein, linking the cytoskeleton to other proteins.

[0105] The expression of tight junction proteins, Claudin-1, ZO-1, and Occludin, is essential for maintaining skin barrier function and enhancing protection against external stimuli. Decreased expression or abnormal function of these proteins can lead to the development or exacerbation of various skin diseases, such as atopic dermatitis, psoriasis, and xerosis. Therefore, therapeutic strategies that restore or regulate the expression of tight junction proteins are attracting attention as important approaches in the prevention and treatment of skin diseases.

[0106] 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).

[0107] Intracellular RNA was extracted from NHEK samples coated with 1 μM ABT-751 or TN-16 after 6 hours 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 quantitatively using PCR (qPCR) with the Real-Time PCR Detection System (Bio-Rad, CFX96). The results are shown in Figure 5a.

[0108] Furthermore, intracellular proteins were extracted from NHEK samples coated with 1 μM ABT-751 or TN-16, respectively, 24 hours later. The expression levels of Claudin-1, ZO-1, and Occludin proteins present in skin cells were measured using Western blotting on the extracted proteins.

[0109] A portion of the collected tissue was homogenized using a tissue homogenizer (DAIHAN Scientific), then dissolved using a cell lysis buffer solution (RIPA buffer, Invitrogen), centrifuged using a centrifuge (Labogene), and the supernatant was transferred to a new tube to be used as a protein sample.

[0110] A 30 μg protein sample was loaded onto a PAGE gel (Invitrogen) with a concentration gradient of 4% to 12%, and the proteins were separated by molecular weight using SDS-PAGE with running buffer (Invitrogen) and electrophoresis (Bio-Rad). The separated proteins were transferred to a PVDF thin film (Bio-Rad) and then blocked with a 5% blocking solution (Skim milk; BD).

[0111] The thin film was reacted with claudin-1, ZO-1, and occludin antibody (Santa Cruz) or Actin antibody (Abcam) for 16 hours, washed, and then reacted again with a secondary antibody for 1 hour, followed by washing.

[0112] After reacting the thin film with an ECL solution (Thermo Fisher), the reacted proteins were detected using a luminescence reaction analyzer (Fusion Solo, Vilber). The results are shown in Figure 5b.

[0113] The results in Figures 5a and 5b show that the expression levels of Claudin-1, ZO-1, and Occludin are increased in skin cells treated with ABT-751 or TN16 compared to the control group, indicating that ABT-751 or TN-16, which are tubulin inhibitors of the present invention, enhance the tight junctions of cells.

[0114] [Example 6] Confirmation of the whitening effect of a tubulin inhibitor Tyrosinase is an enzyme involved in the most important initial rate-determining step in the melanin biosynthesis pathway in the human body, and many skin-whitening ingredients work by inhibiting this enzyme. This test is a method to evaluate the degree to which the activity of the tyrosinase enzyme is inhibited in a test tube.

[0115] To confirm the whitening effect of ABT-751 or TN-16, the effect was measured in in vitro tyrosinase activity inhibition experiments during in vitro experiments. Kojic acid, which is known to inhibit tyrosinase activity, was used as a comparison group.

[0116] Through tyrosinase activity inhibition experiments, each of ABT-751 or TN-16 was treated with a 1.0 μM concentration in a 2 mM L-tyrosine matrix, and the absorbance was measured at 490 nm. Then, tyrosinase enzyme was added again and the reaction was carried out at 37°C. After measuring the absorbance at 490 nm, the absorbance value was calculated to confirm the degree to which each of ABT-751 or TN-16 inhibited tyrosinase activity. The results are shown in Figure 6.

[0117] As can be seen from Figure 6, the tubulin inhibitors of the present invention, ABT-751 or TN-16, exhibit melanin production inhibitory efficacy.

Claims

1. A pharmaceutical composition for the prevention or treatment of skin diseases, comprising as an active ingredient a tubulin inhibitor, which is a compound represented by the following chemical formula I or chemical formula II, or a pharmaceutically acceptable salt thereof. 【Chemistry 1】

2. The tubulin inhibitor regulates and increases cell tight junctions, the pharmaceutical composition for the prevention or treatment of a skin disease according to claim 1.

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

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

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

6. The composition according to claim 4, wherein the skin disease caused by photodamage is one or more selected from the group consisting of lentigines, freckles, hypopigmentation, hyperpigmentation, photodamage due to acute or chronic UV radiation, and photosensitization.

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

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

9. A quasi-drug composition for improving skin diseases, comprising as an active ingredient a tubulin inhibitor, which is a compound represented by the following chemical formula I or chemical formula II, or a pharmaceutically acceptable salt thereof. 【Chemistry 2】

10. A cosmetic composition for the prevention or improvement of skin diseases, comprising as an active ingredient a tubulin inhibitor which is a compound represented by the following chemical formula I or chemical formula II, or a pharmaceutically acceptable salt thereof. 【Transformation 3】

11. A cosmetic composition for moisturizing, whitening, wrinkle improvement, acne relief, strengthening the skin barrier, or differentiation of skin keratinocytes, comprising as an active ingredient a tubulin inhibitor which is a compound represented by the following chemical formula I or chemical formula II or a pharmaceutically acceptable salt thereof. 【Chemistry 4】

12. A method for preventing or treating a skin disease, comprising the step of administering 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, to an individual in need. 【Transformation 5】

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