KR102623461B1 - Pharmaceutical composition for prevention or treatment of allergic skin disease or skin pruritus - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for the prevention or treatment of allergic skin disease or skin itching, and more specifically, to a pharmaceutical composition for the prevention or treatment of allergic skin disease or skin itching containing ABT-751 as an active ingredient, ABT. It relates to a cosmetic composition for preventing or improving allergic skin disease or skin itching containing -751 as an active ingredient and a cosmetic composition for skin moisturizing containing ABT-751 as an active ingredient.
The composition for preventing, improving, or treating atopic dermatitis according to the present invention effectively increases the expression of filaggrin in skin cells and improves skin barrier function, thereby effectively improving the symptoms of allergic skin diseases including atopic dermatitis or skin itching. It was confirmed that it shows a therapeutic effect and has an excellent skin moisturizing effect.

Description

Pharmaceutical composition for prevention or treatment of allergic skin disease or skin pruritus}

The present invention relates to a pharmaceutical composition for the prevention or treatment of allergic skin disease or skin itching, and more specifically, to a pharmaceutical composition for the prevention or treatment of allergic skin disease or skin itching containing ABT-751 as an active ingredient, ABT. It relates to a cosmetic composition for preventing or improving allergic skin disease or skin itching containing -751 as an active ingredient and a cosmetic composition for skin moisturizing containing ABT-751 as an active ingredient.

Atopic dermatitis is a chronic, recurrent inflammatory skin disease that usually begins in infancy or childhood. It is a disease with a large number of patients, reaching 150 million people in 9 countries around the world, including the United States, the United Kingdom, and China.

In Korea, the number of patients who receive treatment for atopic dermatitis each year reaches 1 million, and the incidence is particularly high in infants and young children, with patients aged 0 to 4 accounting for nearly 30% of the total, and children under 9 years old accounting for half of the total. It reaches.

The reason for the increasing prevalence of atopic dermatitis at home and abroad is the high incidence rate, but also the lack of diagnostic methods and appropriate treatment that can accurately diagnose the etiology. Currently, the diagnosis of atopic dermatitis is mainly made through medical examination or visual examination, and steroids or immunosuppressants unrelated to the etiology are also used as treatments. In the case of immunosuppressants, the side effects are relatively not severe, but they cannot be prescribed for infants and children under 2 years of age, who have a particularly high incidence rate, so the treatment period for infants and children who desperately need early treatment at the onset of the disease is missed, resulting in the prolongation and deepening of the disease.

To date, antibody treatments are being developed by global pharmaceutical companies, but they can only be prescribed to seriously ill patients over the age of 12, and there are limitations in treating infants and young children. There is an urgent need to come up with a fundamental solution to this problem.

Republic of Korea Patent Publication No. 2020-0104238 (2020.09.03).

O'regan et al. Journal of Allergy and Clinical Immunology, 122(4):689-693 (2008).

The present researchers confirmed that colchicine not only improves the skin's inherent barrier function and moisture retention by controlling the expression level of filaggrin in the skin, but also exhibits disease prevention, treatment, and improvement effects in an atopic dermatitis model, and the present invention was completed.

In order to solve the above problems, the present invention provides a pharmaceutical composition for the prevention or treatment of allergic skin disease or skin itching containing ABT-751 as an active ingredient, allergic skin disease or skin itching containing ABT-751 as an active ingredient. An attempt was made to provide a cosmetic composition for preventing or improving skin itching and a cosmetic composition for skin moisturizing containing ABT-751 as an active ingredient.

The present invention provides a pharmaceutical composition for the prevention or treatment of allergic skin disease or skin itching containing a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient.

[Formula I]

In one example of the present invention, the allergic skin disease may be atopic dermatitis or contact dermatitis.

In one example of the present invention, the allergic skin disease may be atopic dermatitis.

In one example of the present invention, the pharmaceutical composition may be administered by dermal application.

In one example of the present invention, the atopic dermatitis may be due to decreased expression of filaggrin.

In one example of the present invention, when the decrease in filaggrin expression is 5.0% or more compared to normal, the pharmaceutical composition may be administered to the patient.

In one example of the present invention, the pharmaceutical composition may be any one formulation selected from the group consisting of ointments, creams, lotions, gels, external solutions, pasta formulations, linear formulations, and aerol formulations.

In addition, the present invention provides a cosmetic composition for preventing or improving allergic skin disease or skin itching containing a compound represented by the following formula (I) as an active ingredient.

In one example of the present invention, the cosmetic composition includes a solution, external ointment, cream, foam, nourishing lotion, softening lotion, pack, softening water, emulsion, makeup base, essence, soap, liquid cleanser, bath salt, sunscreen cream, Any one formulation selected from the group consisting of sun oil, suspension, emulsion, paste, gel, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, patch and spray. You can.

Additionally, the present invention provides a cosmetic composition for moisturizing skin containing a compound represented by the following formula (I) as an active ingredient.

[Formula I]

Additionally, the present invention provides a quasi-drug composition for skin moisturizing containing a compound represented by the following formula (I) as an active ingredient.

[Formula I]

The composition for preventing, improving, or treating atopic dermatitis according to the present invention effectively increases the expression of filaggrin in skin cells and improves skin barrier function, thereby effectively improving the symptoms of allergic skin diseases including atopic dermatitis or skin itching. It was confirmed that it shows a therapeutic effect and has an excellent skin moisturizing effect.

Figure 1 relates to the increase in filaggrin expression by ABT-751 treatment,
Figure 2 relates to the increase in cell differentiation by ABT-751 treatment,
Figure 3 relates to the increase in filaggrin expression in the skin of HR-1 mice by ABT-751 treatment;
Figure 4 relates to the reduction of skin moisture loss and increase of skin barrier index in HR-1 mice by application of 4% formulation of ABT-751.

The present invention will be described in detail below with reference to the attached tables and drawings.

When drawings are described, they are provided as examples so that the idea of the present invention can be sufficiently conveyed to those skilled in the art. Accordingly, the present invention is not limited to the presented drawings and may be embodied in other forms, and the drawings may be exaggerated to clarify the spirit of the present invention.

At this time, if there is no other definition in the technical and scientific terms used, they have the meaning commonly understood by those skilled in the art to which this invention pertains, and the gist of the present invention is summarized in the following description and accompanying drawings. Descriptions of known functions and configurations that may be unnecessarily obscure are omitted.

Additionally, as used herein, the singular forms “a,” “an,” and “the” are intended to also include the plural forms, unless the context clearly dictates otherwise.

In addition, units used without special mention in the specification of the present invention are based on weight, and as an example, units of % or ratio mean weight % or weight ratio.

In addition, in the specification of the present invention, the expression “comprises” is an open description that has the same meaning as expressions such as “comprises,” “contains,” “has,” or “features,” and is additionally listed. It does not exclude elements, materials or processes that are not present. Also, “substantially… The expression “consists of” means that other elements, materials or processes not listed together with the specified elements, materials or processes do not have an unacceptably significant effect on at least one basic and novel technical idea of the invention. It means something that can exist in quantity. Additionally, the expression “consists of” means that only the elements, materials, or processes described are present.

Terms used in the specification of the present invention, “ingredient”, “composition”, “composition of compounds”, “compound”, “drug”, “pharmaceutically active agent”, “active agent”, “treatment”, “therapy”, “treatment” or "drug" refers to a compound or composition of compound(s) or substances that induces a desired pharmaceutical and/or physiological effect by local and/or systemic action when administered to a subject (human or animal). Used interchangeably.

As used herein, the terms “treatment” or “therapy” (as well as different forms thereof) include prophylactic (eg, prophylactic treatment), curative, or palliative treatment. As used herein, the term “treating” includes alleviating or reducing at least one detrimental or negative effect or symptom of a condition, disease or disorder. The terms “prevention,” “improvement,” and “treatment” of the present invention should be interpreted in the broadest sense, and “prevention” refers to patients who may be exposed to or susceptible to a disease but have not yet experienced or revealed symptoms of the disease. This means preventing one or more of the clinical symptoms of the disease from progressing. “Treatment” means any action that arrests or reduces the development of a disease or one or more clinical symptoms thereof.

In the present invention, “sample” or “sample” refers to an object for analysis and is used with the same meaning throughout the specification.

The present invention provides a pharmaceutical composition for the prevention or treatment of allergic skin disease or skin itching containing a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient.

[Formula I]

The compound has CAS registry number 141430-65-1 and is ABT-751 ( N -[2-[(4-Hydroxyphenyl)amino]pyridin-3-yl]-4-methoxybenzenesulfonamide).

In the present invention, pharmaceutically acceptable salts refer to salts commonly used in the pharmaceutical industry, such as inorganic ionic salts made of calcium, potassium, sodium and magnesium, hydrochloric acid, nitric acid, phosphoric acid, bromic acid, and iodine. Inorganic acid salts made from acids, 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, brown acid. Organic acid salts made from lacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, hydroiodic acid, etc., methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, and p-toluene. Examples include sulfonates made from sulfonic acid and naphthalenesulfonic acid, amino acid salts made from glycine, arginine, lysine, etc., and amine salts made from trimethylamine, triethylamine, ammonia, pyridine, picoline, etc. , but is not limited to this.

According to the results of an embodiment of the present invention, the pharmaceutical composition not only improves the skin barrier function by increasing the expression level of filaggrin, which is highly related to the onset of allergic skin diseases, and enhances the skin moisturizing effect, but also improves allergic skin disease. It shows excellent effects in the prevention and treatment of allergic skin diseases, skin itching, or both through effects such as controlling the inflammatory response of skin diseases and reducing skin itchiness.

Allergic skin disease refers to pathological symptoms caused by allergic reactions mediated by mast cell activation, such as mast cell degranulation. Representative examples of allergic skin diseases include atopic dermatitis and contact dermatitis. dermatitis), etc.

Skin pruritus is a disease that includes itching caused by a decrease in antibacterial activity and deterioration of barrier function due to a decrease in the lipid content of the stratum corneum of the skin, or itching caused by external stimuli such as temperature changes, chemical substances, electrical stimulation, etc.

In the present invention, the term “anti-allergy” is used to include improvement (reduction of symptoms), treatment, and prevention (inhibition or delay of onset) of allergic skin diseases.

The allergic skin disease may be, for example, atopic dermatitis. Atopic dermatitis shows symptoms such as dry eczematous skin, papules, etc., and lesion samples from atopic patients show epithelial hyperplasia, epidermal hyperplasia, and lymphocytes and mast cells. Accumulation, etc. is confirmed. Patients with atopic dermatitis may suffer from severe skin itching, which causes inflammation of skin lesions and further worsens clinical symptoms.

The skin is anatomically located on the outermost side of the body, protecting the body from the external environment by blocking direct entry into the body of pathogenic microorganisms, viruses, and chemicals in the air, and performs an important barrier function to prevent excessive leakage of body moisture. I'm doing it. Skin tissue has a layered structure consisting of a basal layer, a spinous and granular layer, and a cornified layer, and special expression markers are well known for each layer structure. Among them, Keratin 1 (K1) and Keratin 10 (K10) are expressed in the differentiation layer, and Filaggrin is expressed in the uppermost epithelium, including the stratum corneum, and is essential for the formation of the skin's inherent barrier function as described above. It is one of the most essential proteins.

Recently, a correlation between atopic dermatitis and filaggrin gene abnormalities has been discovered in patients with atopic dermatitis, and in particular, in Europe, mutations in the filaggrin gene have been detected in more than half of all atopic dermatitis patients. In Japan, it was discovered that mutations in the filaggrin gene exist in more than 25% of atopic patients, and in many Asian countries, including China and Korea, mutations in the filaggrin gene were detected in patients with dermatitis including atopy. It has been reported that abnormalities in the filaggrin gene cause decreased expression of filaggrin protein in the skin and loss of skin barrier function, and facilitate the penetration of antigens such as allergic irritants, leading to the development of dermatitis. In fact, among atopic dermatitis patients with filaggrin abnormalities, it is very easy to progress to allergic diseases such as asthma and rhinitis, and there is a need for the development of diagnostic methods and treatments that can determine the presence or absence of filaggrin gene abnormalities.

The atopic dermatitis may be due to decreased expression of filaggrin, and when the decreased expression of filaggrin is 5.0% or more compared to normal, the pharmaceutical composition may be administered to the patient.

In a specific embodiment of the present invention, the compound represented by the formula (I) enhances the expression of filaggrin and improves the moisturizing effect of the skin, showing excellent improvement effects in terms of the skin barrier index and skin composite index. In addition, it not only significantly suppresses the inflammatory response in allergic skin disease models, but also suppresses epidermal hyperplasia and epidermal proliferation, regulates inflammatory cytokines, and reduces skin itchiness, thereby reducing allergic skin disease and/or itching. It shows excellent prevention, treatment and improvement effects. Here, inflammatory cytokine refers to a cytokine that induces an inflammatory response occurring in the body, and is used in the conventional sense in the technical field to which the present invention pertains. For example, IL-2, IL-4, and IL-13 may act as inflammatory cytokines in causing allergic skin diseases.

For example, if the expression of filaggrin is reduced by more than 5.0%, preferably more than 5.3%, and more preferably more than 5.6% compared to normal people, it can be determined that the patient is suffering from atopic dermatitis, and accordingly, the patient The pharmaceutical composition according to the present invention can be administered for the treatment of atopic dermatitis.

In addition to containing the compound represented by Formula (I) or a pharmaceutically acceptable salt thereof, the pharmaceutical composition may further include appropriate carriers, excipients, and diluents commonly used in pharmaceutical compositions.

Carriers, excipients and diluents that may be included in the composition include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, These include methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxy benzoate, propylhydroxy benzoate, talc, magnesium stearate and mineral oil. When formulating the composition, it can be prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants.

The pharmaceutical composition can be formulated and used in various forms according to conventional methods. When formulated as a transdermal administration agent, suitable formulations include, for example, ointments, creams, lotions, gels, external solutions, paste preparations, and liniments. Agents and air agents are preferred, but are not limited thereto.

In addition to the above carrier, the pharmaceutical composition according to the present invention may further include preservatives, stabilizers, wetting agents, emulsifiers, solubilizers, sweeteners, colorants, osmotic pressure regulators, antioxidants, etc.

Regarding the formulation of pharmaceutical compositions, it is known in the art, and specifically, references can be made to the literature [Remington's Pharmaceutical Sciences (19th ed., 1995)]. The above documents are considered part of this specification.

The pharmaceutical composition according to the present invention can be administered in a pharmaceutically effective amount.

In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat the disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is determined by the type, severity, and activity of the patient's disease. , can be determined based on factors including sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, drugs used simultaneously, and other factors well known in the field of medicine. The pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiple times. Taking all of the above factors into consideration, it is important to administer an amount that can achieve the maximum effect with the minimum amount without side effects, and this can be easily determined by a person skilled in the art.

The pharmaceutical composition of the present invention can be administered to an individual through various routes. The method of administration can be, for example, administration in various oral and parenteral formulations, and when formulated, it can be prepared using the above-mentioned excipients that are commonly used. Preferably, in the present invention, it is administered as a parenteral agent and is applied to the skin. It can be used as a coating agent for The administration method of the pharmaceutical composition of the present invention is determined depending on the type of drug that is the active ingredient, along with various related factors such as the disease to be treated, the route of administration, the patient's age, gender and weight, and the severity of the disease.

In addition, the present invention provides a cosmetic composition for preventing or improving allergic skin disease or skin itching containing a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient.

[Formula I]

The cosmetic composition includes solution, external ointment, cream, foam, nourishing lotion, softening lotion, pack, softening water, emulsion, makeup base, essence, soap, liquid cleanser, bath agent, sunscreen cream, sun oil, suspension, emulsion, It may be any one formulation selected from the group consisting of paste, gel, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, patch, and spray, but is not limited thereto.

In addition, the cosmetic composition of the present invention may additionally include one or more cosmetically acceptable carriers that are blended with general skin cosmetics, and common ingredients include, for example, oil, water, surfactant, moisturizer, lower alcohol, Thickeners, chelating agents, pigments, preservatives, fragrances, etc. may be appropriately mixed, but are not limited thereto.

Cosmetically acceptable carriers included in the cosmetic composition of the present invention vary depending on the formulation.

When the formulation of the present invention is an ointment, paste, cream or gel, the carrier ingredients include animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide or Mixtures of these can be used.

When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silcate, polyamide powder, or mixtures thereof may be used as the carrier ingredient, and especially in the case of a spray, chloro May contain propellants such as fluorohydrocarbons, propane/butane or dimethyl ether.

When the formulation of the invention is a solution or emulsion, a solvent, solubilizing agent or emulsifying agent is used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 , 3-butyl glycol oil can be used, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol aliphatic esters, polyethylene glycol or fatty acid esters of sorbitan. .

When the formulation of the present invention is a suspension, the carrier ingredients include water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester, and miso. Crystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracant may be used.

When the formulation of the present invention is soap, alkali metal salts of fatty acids, hemiester salts of fatty acids, fatty acid protein hydrolyzates, isethionates, lanolin derivatives, fatty alcohols, vegetable oils, glycerol, sugars, etc. may be used as carrier ingredients. You can.

When the formulation of the present invention is a surfactant-containing cleansing agent, the carrier ingredients include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosite, and fatty acid amide. Ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester can be used.

The cosmetic composition according to the present invention may contain 0.01 to 20% by weight of colchicine based on the total weight of the composition.

Additionally, the present invention provides a cosmetic composition for moisturizing skin containing a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient.

[Formula I]

The skin moisturizing may mean increasing the moisture in the skin and maintaining a moist state, and the cosmetic composition for skin moisturizing according to the present invention has an excellent skin moisturizing effect of suppressing or reducing the loss of skin moisture, and moisturizing. It has an excellent effect on skin moisturization by controlling the expression of related factors such as filaggrin, involucrin, and loricrin.

Additionally, the present invention provides a quasi-drug composition for skin moisturizing containing a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient.

[Formula I]

The above-mentioned quasi-drug composition is a quasi-drug composition for the purpose of moisturizing the skin.

The above-mentioned quasi-drugs refer to products that have a milder effect than pharmaceuticals among products used for the purpose of diagnosing, treating, improving, alleviating, treating, or preventing diseases in humans or animals. For example, according to the Pharmaceutical Affairs Act, quasi-drugs are used as medicines. Excluding articles used, textile and rubber products used for the treatment or prevention of diseases in humans and animals; items that have a minor or no direct effect on the human body; items that are not instruments or machines and similar items; sterilization and other products used to prevent infectious diseases; This may include pesticides, etc., but is not limited thereto.

The type or formulation of the quasi-drug composition of the present invention is not particularly limited, but preferably may be a disinfectant cleaner, shower foam, gargle, wet tissue, detergent soap, hand wash, humidifier filler, mask, ointment, or filter filler.

When the composition of the present invention is included in a quasi-drug for skin moisturizing purposes, the composition can be used as is or together with other quasi-drug ingredients, and can be used appropriately according to conventional methods. The mixing amount of the active ingredient can be appropriately determined depending on the purpose of use, and the quasi-drug composition according to the present invention may contain the compound represented by the above formula (I) in an amount of 0.01 to 20% by weight based on the total weight of the composition.

The present invention also provides a method of treating allergic skin disease or skin itching, comprising administering to a patient a pharmaceutical composition containing a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof.

[Formula I]

The treatment method includes, for example, a) measuring the level of filaggrin expression in the patient; and b) when the filaggrin expression level decreases by more than 5.0% compared to the normal level, determining that administration of the pharmaceutical composition to the patient is necessary. The pharmaceutical composition may be administered to the patient, but is not limited to this. Depending on the amount of green reduced, the amount and administration cycle of the pharmaceutical composition administered to the patient may vary. For example, according to an embodiment of the present invention, at a concentration of 0.5 to 2.5 μM, once a day for 1 to 4 weeks. It can be administered, but is not limited to this.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as limited by these examples.

[Materials, reagents, strains and devices, etc.]

- Normal Human Epidermal Keratinocyte (NHEK) is a human fetal epidermal cell line that was purchased and used from ATCC.

These cells were obtained through primary culture and were used to confirm intracellular biochemical changes caused by ABT-751 under normal conditions, not disease conditions.

- Experimental animals: hos:HR-1 (hairless) mice have the advantage of being able to conduct skin-related research by applying drugs directly to the skin or inducing skin diseases with chemical or biological allergens because they do not have hair in skin disease research. there is.

In addition, at 8-10 weeks of age, the skin thickness is about 0.4 mm, which is similar to human skin, which has the advantage of being useful for studying the pathological mechanisms of skin diseases and confirming biochemical changes in skin tissue.

[Test Example 1]

1.1 RNA extraction and measurement of filaggrin expression level using RT-qPCR

Human epidermal cell line (NHEK) or mouse skin tissue to which ABT-751 or control sample was applied was lysed using TRI reagent (MRC), and intracellular RNA was extracted using Rneasy Mini Kit (Qiagen).

The extracted RNA was reverse transcribed using the ImProm-II TM Reverse Transcription Kit (Promega) to synthesize cDNA, and then the amount of filaggrin was measured using the Real-Time PCR Detection System (Bio-Rad, CFX96) using quantitative PCR (quantitative PCR). measured by qPCR).

1-2. Protein extraction and western blot

A portion of the collected tissue was homogenized using a tissue homogenizer (Daehan Science Co., Ltd.), then lysed using a cell lysis buffer (RIPA buffer, Invitrogen Co., Ltd.), centrifuged in a centrifuge (Labogene Co., Ltd.), and the upper layer The liquid was transferred to a new tube and used as a protein sample.

30 μg of protein sample was loaded onto a PAGE gel (Invitrogen) with a concentration gradient of 4% to 12%, and proteins were analyzed by performing SDS-PAGE using a running buffer (Invitrogen) and an electrophoresis machine (Bio-Rad). They were separated by molecular weight.

The separated protein was transferred to a PVDF thin film (Bio-Rad) and then blocked with 5% blocking solution (Skim milk; BD).

The thin film was reacted with a filaggrin antibody (Santa Cruz) or a GAPDH antibody (Abcam) for 16 hours, washed, and then reacted again with a secondary antibody for 1 hour and washed.

After reacting Bakak with ECL solution (Thermo Fisher), the reacted protein was detected using a luminescence reaction measurement device (Fusion Solo, Vilber).

[Test Example 2] Skin barrier function test

2 hours after application of ABT-751, the amount of skin moisture loss, skin barrier index, and overall index at the application site were measured and recorded for 3 weeks using a precision measuring device (Courage-khazaka electronic GmbH, MPA10).

[Example 1] Filaggrin expression level and cell differentiation in human epidermal cell lines by ABT-751 treatment

Human epidermal cell line (NHEK) in culture was treated with ABT-751, the level of filaggrin expression was measured, and changes in cell differentiation were confirmed.

ABT-751 was treated with NHEK at concentrations of 0, 0.5, 1.0, and 2.5 μM, and filaggrin expression level and cell differentiation were observed 24 hours after treatment.

The results are shown in Figures 1 and 2.

From this, it was confirmed that the level of filaggrin expression increased depending on the ABT-751 treatment concentration (see Figure 1), and the degree of cell differentiation significantly increased in the ABT-751 treated group compared to the ABT-751 untreated group (control group). Confirmed (see Figure 2).

[Example 2] Filagrin expression level and cell differentiation in animal cells by ABT-751 treatment

The same amount of 0, 1, 2, and 4% formulations of ABT-751 was applied to the back skin of hairless hos:HR-1 mice (hereinafter referred to as HR-1 mice) once a day for 3 weeks.

After each application, the skin moisture loss and skin barrier index were measured by the method of the above test example.

After 2 weeks of application, HR-1 mice were euthanized in a carbon dioxide chamber, and back skin tissue was collected.

2-1 Measurement of filaggrin expression level

RNA and protein were extracted from the back skin tissue to which ABT-751 was applied by the method of the above test example, and the expression level of filaggrin was measured, and the results are shown in Figure 3.

From this, it was confirmed that the expression level of filaggrin in skin tissue increased in a concentration-dependent manner after application of the remaining formulations of ABT-751 compared to the control group (0% formulation ABT-751).

2-2. Measurement of skin moisture loss and skin barrier index

The results of measuring skin moisture loss and skin barrier index after application of the control group (0% formulation ABT-751) and 4% formulation ABT-751 are shown in Figure 4.

From this, it was confirmed that application of 4% formulation of ABT-751 reduced skin moisture loss upon application and increased skin barrier index in HR-1 mice.

In particular, it was confirmed that as the number of days of application passed, the decrease in skin moisture loss and the increase in skin barrier index increased.

As above, specific parts of the content of the present invention have been described in detail, and those skilled in the art will understand that these specific techniques are merely preferred embodiments and do not limit the scope of the present invention. It will be obvious. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims (11)

A pharmaceutical composition for the prevention or treatment of allergic skin disease or skin itching containing a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula I]
According to clause 1,
The allergic skin disease is atopic dermatitis or contact dermatitis, and a pharmaceutical composition for preventing or treating allergic skin disease or skin itching. According to clause 1,
The allergic skin disease is atopic dermatitis, and a pharmaceutical composition for preventing or treating allergic skin disease or skin itching. According to clause 1,
The pharmaceutical composition is a pharmaceutical composition for the prevention or treatment of allergic skin disease or skin itching, which is administered by skin application. According to clause 2,
A pharmaceutical composition for preventing or treating allergic skin disease or skin itching, wherein the atopic dermatitis is caused by reduced expression of filaggrin. According to clause 5,
A pharmaceutical composition for the prevention or treatment of allergic skin disease or skin itching, wherein the pharmaceutical composition is administered to a patient when the decrease in filaggrin expression is 5.0% or more compared to normal. According to clause 1,
The pharmaceutical composition is for the prevention or treatment of allergic skin disease or skin itching, which is any one formulation selected from the group consisting of ointments, creams, lotions, gels, external solutions, paste preparations, liniment preparations, and aerosol preparations. Pharmaceutical composition. A cosmetic composition for preventing or improving allergic skin disease or skin itching containing a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula I]
According to clause 8,
The cosmetic composition includes solution, external ointment, cream, foam, nourishing lotion, softening lotion, pack, softening water, emulsion, makeup base, essence, soap, liquid cleanser, bath agent, sunscreen cream, sun oil, suspension, emulsion, Allergic skin disease or itching skin, any formulation selected from the group consisting of paste, gel, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, patch and spray. Cosmetic composition for preventing or improving. A cosmetic composition for moisturizing skin containing a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula I]
A quasi-drug composition for skin moisturizing containing a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula I]