Pharmaceutical composition for keratosis and use thereof

Abstract

A new pharmaceutical composition for treating keratosis including Darier's disease and psoriasis is provided. The pharmaceutical composition for treating keratosis of the present invention contains 5-methyl-2-(1-piperazinyl)benzenesulfonic acid. The compound is an anhydride, a salt, a hydrate, or a solvate thereof, or a hydrate or solvate of a salt thereof, and the hydrate is, for example, 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate. The keratosis is, for example, Darier's disease or psoriasis.

Description

[Technical Field]

[0001] The present invention relates to a pharmaceutical composition for keratosis and a method for treating or preventing keratosis. [Background technology]

[0002] Darier's disease is a type of keratosis, a skin disorder characterized by the appearance of small keratotic papules due to hyperkeratosis, acantholysis, and abnormal keratinization in the epidermis (Non-Patent Document 1). Frequent symptoms include pain and itching, and a foul odor is often observed, particularly in flexed areas where sweating is frequent and secondary infections are more likely to occur. Furthermore, Darier's disease can become chronic and recur, leading to a decline in the quality of life (hereinafter also referred to as QOL) of patients and a potential social handicap.

[0003] In particular, there is currently no fundamental treatment for Darier's disease, a type of keratosis, recommended by guidelines, and treatment focuses on disease management and symptom management by avoiding triggers that exacerbate the condition. Treatment methods that involve administering retinoids, steroids, vitamin D analogs, etc. have been proposed, but side effects have been reported, and none have been safe or consistently effective. Therefore, there remains a high unmet medical need for safe and effective treatments. [Prior art documents] [Non-patent literature]

[0004] [Non-Patent Document 1] Susan M. Cooper and Susan M. Burge, Darier's Disease Epidemiology, Pathophysiology, and Management, Am J Clin Dermatol 2003; 4 (2): 97-105 Summary of the Invention [Problem to be solved by the invention]

[0005] Therefore, an object of the present invention is to provide a new pharmaceutical composition and a method for treating or preventing keratosis including Darier's disease and psoriasis. [Means for solving the problem]

[0006] The pharmaceutical composition for keratosis of the present invention contains 5-methyl-2-(1-piperazinyl)benzenesulfonic acid (also referred to as 5-methyl-2-(piperazin-1-yl)benzenesulfonic acid).

[0007] The method of the present invention for treating or preventing keratosis comprises the step of administering 5-methyl-2-(1-piperazinyl)benzenesulfonic acid to a subject.

[0008] The present invention is 5-methyl-2-(1-piperazinyl)benzenesulfonic acid for use in the treatment or prevention of keratosis.

[0009] The present invention is the use of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid in the manufacture of a pharmaceutical composition for treating keratosis. [Effects of the Invention]

[0010] According to the pharmaceutical composition of the present invention, by containing 5-methyl-2-(1-piperazinyl)benzenesulfonic acid, it is possible to treat or prevent keratosis. [Brief explanation of the drawings]

[0011] [Figure 1] FIG. 1 is a graph showing the relationship between administration of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate and epidermal thickness in in vitro epidermal tissue in Example 1. [Figure 2] FIG. 2 is a graph showing the relationship between administration of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate and biotin diffusion, which is an index of acantholysis, in Example 2. [Figure 3]FIG. 3 is a graph showing the relationship between the administration of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate and the frequency of pyknotic nuclei, which is an index of abnormal keratinization, in Example 2. [Figure 4] FIG. 4 is a graph showing the relationship between administration of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate and pathological evaluation of acantholysis in Example 2. [Figure 5A] FIG. 5A is a graph showing the relationship between administration of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate and the expression of keratinocyte differentiation marker molecules associated with abnormal keratinization in Example 2. [Figure 5B] FIG. 5B is a graph showing the relationship between administration of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate and the expression of keratinocyte differentiation marker molecules associated with abnormal keratinization in Example 2. [Figure 6A] FIG. 6A is a graph showing the relationship between administration of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate and the expression of cell adhesion marker molecules associated with acantholysis in Example 2. [Figure 6B] FIG. 6B is a graph showing the relationship between administration of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate and the expression of cell adhesion marker molecules associated with acantholysis in Example 2. [Figure 7] FIG. 7 is a graph showing the relationship between administration of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate and the dorsal skin thickness of hyperkeratotic mice in Example 3. [Figure 8] FIG. 8 is a graph showing the relationship between administration of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate and the mean plasma concentration of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid in Example 5. DETAILED DESCRIPTION OF THE INVENTION

[0012] The present invention can be exemplified by the following embodiments. [1] A pharmaceutical composition for treating keratosis, comprising 5-methyl-2-(1-piperazinyl)benzenesulfonic acid. [2] The pharmaceutical composition according to [1], wherein the 5-methyl-2-(1-piperazinyl)benzenesulfonic acid is an anhydride thereof, a salt thereof, a hydrate or solvate thereof, or a hydrate or solvate of a salt thereof. [3] The pharmaceutical composition according to [1], wherein the 5-methyl-2-(1-piperazinyl)benzenesulfonic acid is an anhydride thereof, a salt thereof, a hydrate thereof, or a hydrate of a salt thereof. [4] The pharmaceutical composition according to [2] or [3], wherein the hydrate is 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate. [5] The pharmaceutical composition according to any one of [1] to [4], wherein the keratosis is at least one selected from the group consisting of Darier's disease, psoriasis, and actinic keratosis. [6] The pharmaceutical composition according to any one of [1] to [4], wherein the keratosis is Darier's disease or psoriasis. [7] The pharmaceutical composition according to any one of [1] to [4], wherein the keratosis is Darier's disease. [8] The pharmaceutical composition according to any one of [1] to [4], wherein the keratosis is psoriasis. [9] The pharmaceutical composition according to [8], wherein the psoriasis is psoriasis vulgaris.

[10] The pharmaceutical composition according to any one of [1] to [9], wherein the keratosis is keratosis in which IL-22 signaling is involved.

[11] The pharmaceutical composition according to any one of [1] to

[10] , which suppresses at least one selected from the group consisting of hyperkeratosis, dyskeratosis, and acantholysis in the keratosis.

[12] The pharmaceutical composition according to

[11] , wherein the hyperkeratosis is hyperkeratosis involving IL-22 signaling.

[13] The pharmaceutical composition according to any one of [1] to

[12] , which improves the IGA score in the keratosis.

[14] The pharmaceutical composition according to any one of [1] to

[13] , which suppresses or improves at least one symptom selected from the group consisting of lesion skin area, itching, pain, and odor in the keratosis.

[15] The pharmaceutical composition according to any one of [1] to

[14] , wherein the dosage of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid is 1 to 1000 mg / day, and the dosage is calculated as the amount of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid anhydride.

[16] The pharmaceutical composition according to

[15] , wherein the dosage is 50 to 600 mg / day.

[17] The pharmaceutical composition according to

[15] , wherein the dosage is 100 to 400 mg / day.

[18] The pharmaceutical composition according to any one of [1] to

[17] , which is administered 1 to 3 times per day.

[19] The pharmaceutical composition according to any one of [1] to

[17] , which is administered twice a day.

[20] The pharmaceutical composition according to any one of [1] to

[19] , wherein the administration interval is daily.

[0013]

[21] A method for treating or preventing keratosis, comprising administering 5-methyl-2-(1-piperazinyl)benzenesulfonic acid to a subject.

[22] The method according to

[21] , wherein the 5-methyl-2-(1-piperazinyl)benzenesulfonic acid is an anhydride thereof, a salt thereof, a hydrate or solvate thereof, or a hydrate or solvate of a salt thereof.

[23] The method according to

[21] , wherein the 5-methyl-2-(1-piperazinyl)benzenesulfonic acid is an anhydride thereof, a salt thereof, a hydrate thereof, or a hydrate of a salt thereof.

[24] The method according to

[22] or

[23] , wherein the hydrate is 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate.

[25] The method according to any one of

[21] to

[24] , wherein the administration method is oral administration.

[26] The method according to any one of

[21] to

[25] , wherein the daily dose is 1 to 1000 mg / day, and the dose is calculated as the amount of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid anhydride.

[27] The method according to any one of

[21] to

[26] , wherein the number of administrations per day is 1 to 3 times.

[28] The method according to any one of

[21] to

[27] , wherein the administration interval is daily administration.

[0014]

[29] 5-Methyl-2-(1-piperazinyl)benzenesulfonic acid for use in the treatment or prevention of keratosis.

[30] The 5-methyl-2-(1-piperazinyl)benzenesulfonic acid according to

[29] , wherein the 5-methyl-2-(1-piperazinyl)benzenesulfonic acid is an anhydride thereof, a salt thereof, a hydrate or solvate thereof, or a hydrate or solvate of a salt thereof.

[31] The 5-methyl-2-(1-piperazinyl)benzenesulfonic acid according to

[29] , wherein the 5-methyl-2-(1-piperazinyl)benzenesulfonic acid is an anhydride thereof, a salt thereof, a hydrate thereof, or a hydrate of a salt thereof.

[32] The 5-methyl-2-(1-piperazinyl)benzenesulfonic acid according to

[30] or

[31] , wherein the hydrate is 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate.

[33] Use of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid in the manufacture of a pharmaceutical composition for keratosis.

[34] The use according to

[33] , wherein the 5-methyl-2-(1-piperazinyl)benzenesulfonic acid is an anhydride thereof, a salt thereof, a hydrate or solvate thereof, or a hydrate or solvate of a salt thereof.

[35] The use according to

[33] , wherein the 5-methyl-2-(1-piperazinyl)benzenesulfonic acid is an anhydride thereof, a salt thereof, a hydrate thereof, or a hydrate of a salt thereof.

[36] The use according to

[34] or

[35] , wherein the hydrate is 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate.

[0015]

[37] A pharmaceutical composition for acantholysis comprising 5-methyl-2-(1-piperazinyl)benzenesulfonic acid.

[38] The pharmaceutical composition according to

[37] , wherein the 5-methyl-2-(1-piperazinyl)benzenesulfonic acid is an anhydride thereof, a salt thereof, a hydrate or solvate thereof, or a hydrate or solvate of a salt thereof.

[39] The pharmaceutical composition according to

[37] , wherein the 5-methyl-2-(1-piperazinyl)benzenesulfonic acid is an anhydride thereof, a salt thereof, a hydrate thereof, or a hydrate of a salt thereof.

[40] The pharmaceutical composition according to

[38] or

[39] , wherein the hydrate is 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate.

[0016] As used herein, "treatment of a disease" includes, for example, curing a disease, remission of a disease, alleviating a disease, or suppressing the progression of a disease, and "treatment of a disease" also includes, for example, treating symptoms caused by the disease (e.g., curing, remission, alleviating, or suppressing symptoms). "Prevention of a disease" includes, for example, preventing contraction of a disease, preventing the onset of a disease, and preventing the recurrence of a disease, and "prevention of a disease" also includes, for example, preventing symptoms caused by the disease (e.g., preventing the onset of symptoms or preventing the recurrence of symptoms). As used herein, "treatment or prevention of a disease" can also be referred to as, for example, suppressing a disease, and "treatment or prevention of symptoms caused by a disease" can also be referred to as, for example, suppressing symptoms caused by a disease.

[0017] The "epidermis" of the skin is usually composed of the stratum corneum, stratum granulosum, stratum spinosum, and stratum basale.

[0018] As used herein, treatment refers to, for example, treatment for a subject diagnosed by a physician as having a disease or a symptom of a disease. Furthermore, as used herein, prevention refers to, for example, treatment for a subject who does not have a disease or a symptom of a disease, and refers to treatment aimed at preventing the onset of a disease or a symptom of a disease. The subject may also be referred to as, for example, a test subject or a subject, and may be a human (patient) or a non-human animal (animal patient). As used herein, the term "patient" may also include, for example, the meaning of a non-human animal patient, and can be interpreted as "animal patient." Furthermore, when the subject does not have the disease or its symptoms, they are also referred to as a healthy subject (a healthy person or a healthy non-human animal) with respect to the disease or symptom of interest.

[0019] The present invention will be described below with reference to specific examples, but the present invention is not limited to these examples. The embodiments exemplified below can be mutually incorporated unless otherwise specified.

[0020] [Pharmaceutical composition for keratosis] The pharmaceutical composition for keratosis of the present invention is characterized by containing 5-methyl-2-(1-piperazinyl)benzenesulfonic acid. The pharmaceutical composition for keratosis of the present invention is characterized by containing the compound, and other components, conditions, etc. are not particularly limited.

[0021] As described above, the pharmaceutical composition for keratosis of the present invention can treat or prevent keratosis. The pharmaceutical composition for keratosis of the present invention can be used, for example, for the purpose of treatment, for the purpose of prevention, or for both the purpose of treatment and prevention. Hereinafter, in this specification, the term "treatment / prevention" can be interpreted as meaning either treatment, prevention, or both treatment and prevention. The pharmaceutical composition for keratosis of the present invention can, for example, provide safe treatment / prevention with few side effects.

[0022] Hereinafter, 5-methyl-2-(1-piperazinyl)benzenesulfonic acid will be referred to as MPBS. The form of MPBS is not limited, and may include anhydrous MPBS (also referred to as MPBS anhydrous), a salt of MPBS (also referred to as MPBS salt), a hydrate of MPBS or a hydrate of the MPBS salt (also referred to as MPBS hydrate), and a solvate of MPBS or a solvate of the MPBS salt (also referred to as MPBS solvate). These are collectively referred to as the MPBSs of the present invention. The MPBSs contained in the pharmaceutical composition of the present invention may be, for example, the MPBS anhydrous, the MPBS salt, the MPBS hydrate, or the MPBS solvate, and may contain only one or more of these. The pharmaceutical composition of the present invention preferably contains the MPBSs as an active ingredient.

[0023] The type of MPBS salt is not particularly limited and may be, for example, an alkali metal salt, an alkaline earth metal salt, an amphoteric element salt, an amine salt, an inorganic acid salt, or an organic acid salt. Examples of the alkali metal salt include sodium salt or potassium salt, examples of the alkaline earth metal salt include magnesium salt or calcium salt, and examples of the amphoteric element salt include aluminum salt. Examples of the amine salt include lower alkylamine salts such as triethylamine salt; hydroxy lower alkylamine salts such as 2-hydroxyethylamine salt, bis-(2-hydroxyethyl)amine salt, tris(hydroxymethyl)aminomethane salt, or N-methyl-D-glucamine salt; cycloalkylamine salts such as dicyclohexylamine salt; benzylamine salts such as N,N-dibenzylethylenediamine salt; or dibenzylamine salt. Examples of the inorganic acid salts include hydrochloride, hydrobromide, sulfate, and phosphate, and examples of the organic acid salts include fumarate, succinate, oxalate, and lactate.

[0024] The type of MPBS hydrate is not particularly limited and may be, for example, a monohydrate. The type of MPBS solvate is also not particularly limited. The solvent capable of forming the MPBS solvate is not particularly limited and may be, for example, a non-aqueous solvent, specific examples of which include alcohols such as methanol, ethanol, and isopropyl alcohol, acetone, ethyl acetate, and methylene chloride.

[0025] In the pharmaceutical composition for keratosis of the present invention, the MPBS is preferably the MPBS anhydrate or the MPBS hydrate, more preferably the MPBS hydrate, and specifically, 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate (also referred to as MPBS monohydrate).

[0026] The MPBSs are known compounds and can be synthesized by the methods described in, for example, JP-A-3-7263, JP-A-9-221479, EP-A-390654, EP-A-779283, U.S. Pat. No. 5,053,409, and U.S. Pat. No. 5,990,113, and are readily available to those skilled in the art.

[0027] The MPBSs can be substituted with a compound represented by the following general formula (I), or a salt thereof, or a hydrate or solvate thereof, which is described in International Publication WO03 / 011296.

[0028] [ka] In the formula, R1 is a hydrogen atom, a C1-C6 alkyl group, a C3-C7 cycloalkyl group, a C1-C4 halogenated alkyl group, a halogen atom, or a C6-C 12 R2 represents a hydrogen atom, a C1-C6 alkyl group, or a C7-C 12The aralkyl group may have one or more substituents selected from the group consisting of a cyano group, a nitro group, a C1-C6 alkoxy group, a halogen atom, a C1-C6 alkyl group, and an amino group, and n represents an integer of 1 to 4.

[0029] In the general formula (I), examples of the C1-C6 alkyl group defined by R1 include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, tert-pentyl, hexyl, and isohexyl groups. Examples of the C3-C7 cycloalkyl group include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl groups. Examples of the C1-C4 halogenated alkyl group include trifluoromethyl, trifluoroethyl, and pentafluoroethyl groups. Examples of the halogen atom include fluorine, chlorine, and bromine atoms. C6-C 12 Examples of the aryl group include a phenyl group and a naphthyl group.

[0030] Preferred examples of R1 include a hydrogen atom, a C1-C6 alkyl group, a C5-C6 cycloalkyl group, a trifluoromethyl group, a halogen atom, and a phenyl group, more preferred examples include a C1-C3 alkyl group, a cyclohexyl group, a trifluoromethyl group, a chlorine atom, a bromine atom, and a phenyl group, and even more preferred examples include a methyl group or a propyl group, with a methyl group being particularly preferred.

[0031] Examples of the C1-C6 alkyl group defined by R2 include the alkyl groups defined above for R1. 12Examples of the aralkyl group include a benzyl group, a phenethyl group, and a naphthylmethyl group. This aralkyl group may have one or more substituents selected from the group consisting of a C1-C6 alkoxy group such as a cyano group, a nitro group, a methoxy group, an ethoxy group, a propoxy group, an isopropoxy group, a butoxy group, an isobutoxy group, a tert-butoxy group, a pentyloxy group, an isopentyloxy group, a tert-pentyloxy group, or a hexyloxy group; a halogen atom as defined above for R1; an alkyl group as defined above for R1; and an amino group.

[0032] Preferred examples of R2 include a hydrogen atom, a C1-C3 alkyl group, and a C7-C 12 The aralkyl group may have one or more substituents selected from a C1-C3 alkyl group, a C1-C3 alkoxy group, and a halogen atom, and more preferred examples thereof include a hydrogen atom, a C7-C 12 The aralkyl group may have one or more substituents selected from C1-C3 alkoxy groups, and is preferably a hydrogen atom. In addition, in the above general formula (I), n is preferably 2.

[0033] In the pharmaceutical composition for keratosis of the present invention, the MPBSs may be, for example, in an ionized form. When the pharmaceutical composition for keratosis of the present invention is, for example, a liquid and contains an aqueous solvent, a non-aqueous solvent, or a mixture thereof, as described below, the MPBSs may be ionized regardless of the type of anhydride, salt, hydrate, or solvate. The molecular form of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid (MPBS) is, for example, represented by the following formula (II), and can become an ionized molecular species in a protic solvent such as water. Specific examples of the molecular species include a monovalent cation represented by the following formula (IIIa), a zwitterion (zwitterion) having a positive charge and a negative charge represented by the following formula (IIIb), and a monovalent anion represented by the following formula (IIIc). The monovalent cation molecular species can form a salt with, for example, an acid (anion), and the monovalent anion can form a salt with, for example, a base (cation).

[0034] [ka] [ka]

[0035] The pharmaceutical composition for keratosis of the present invention can be used, for example, for the treatment or prevention of keratosis, and the treatment and prevention are, for example, as described above. The pharmaceutical composition for keratosis of the present invention can be read, for example, as a pharmaceutical composition used for the symptoms of keratosis, and specifically, as a pharmaceutical composition used for the treatment or prevention of the symptoms of keratosis. Furthermore, the treatment or prevention of keratosis can also be said, for example, to be the suppression of keratosis.

[0036] The keratosis that is the subject of the present invention is a disease that causes hyperkeratosis. Hyperkeratosis refers to hyperkeratosis in the epidermis of the skin. The keratosis is, for example, a disease that occurs due to an abnormality in the differentiation process of keratinocytes. The present invention is preferably applied to, for example, keratosis in which IL-22 signaling is involved. The IL-22 signaling refers to, for example, a signal that occurs when IL-22 acts on the IL-22 receptor.

[0037] The keratosis that is the subject of the present invention is not particularly limited, and examples thereof include Darier's disease, psoriasis, and actinic keratosis.

[0038] Examples of psoriasis include plaque psoriasis, psoriatic arthritis, guttate psoriasis, erythrodermic psoriasis, pustular psoriasis, and palmoplantar pustulosis.

[0039] The pharmaceutical composition for keratosis of the present invention can, for example, suppress the pathological symptoms of keratosis. The pathological symptom to be suppressed is, for example, at least one selected from the group consisting of hyperkeratosis, dyskeratosis, and acantholysis, and preferably includes hyperkeratosis. That is, the pharmaceutical composition for keratosis of the present invention may, for example, suppress hyperkeratosis, dyskeratosis, or acantholysis, or may suppress any one symptom, two or more symptoms, or all symptoms. The pharmaceutical composition for keratosis of the present invention can, for example, suppress hyperkeratosis, hyperkeratosis and dyskeratosis, hyperkeratosis and acantholysis, or hyperkeratosis, dyskeratosis, and acantholysis.

[0040] Hyperkeratosis is hyperkeratosis caused by abnormal keratinization in the stratum corneum, granular layer, and spinous layer of the skin, and can be observed as thickening of the stratum corneum. Examples of hyperkeratosis include hyperkeratosis involving IL-22 signaling. Dyskeratinization refers to the individual keratinization of keratinocytes in the spinous layer of the skin, which can be observed as round bodies in Darier's disease. Acantholysis is a state in which keratinocytes lose cell adhesion in the spinous layer of the skin, and can be observed as intraepidermal cracks or blister formation. Hyperkeratosis, dyskeratinization, and acantholysis can be observed, for example, by collecting lesional skin from a patient and using methods known to those skilled in the art.

[0041] When the keratosis is Darier's disease, pathological symptoms observed include, for example, hyperkeratosis in the stratum corneum, granular layer, and spinous layer, dyskeratosis in the spinous layer, and acantholysis in the spinous layer. The pharmaceutical composition for keratosis of the present invention can, for example, simultaneously suppress these pathological symptoms. Furthermore, when the keratosis is psoriasis, pathological symptoms observed include hyperkeratosis in the stratum corneum, granular layer, and spinous layer. The pharmaceutical composition for keratosis of the present invention can, for example, suppress hyperkeratosis in these layers. Furthermore, when the keratosis is actinic keratosis, pathological symptoms observed include, for example, atypia in the basal layer and / or spinous layer, and hyperkeratosis and dyskeratosis in the epidermis. The pharmaceutical composition for keratosis of the present invention can, for example, suppress these symptoms in the epidermis.

[0042] The pharmaceutical composition for keratosis of the present invention can improve, for example, the IGA (Investigator's Global Assessment) score of a patient. Here, the IGA score is not particularly limited. IGA scores are generally classified into multiple stages, and a higher score indicates more severe symptoms.

[0043] The pharmaceutical composition for keratosis of the present invention can, for example, suppress or improve clinical physical symptoms caused by the keratosis, and specifically, for example, can suppress or improve at least one selected from the group consisting of lesion skin area, pruritus (itching), pain (ache), and odor in the keratosis. The pharmaceutical composition for keratosis of the present invention can suppress or improve, for example, the aforementioned pathological symptoms, thereby suppressing or improving the clinical physical symptoms exemplified herein.

[0044] The area of ​​lesioned skin is, for example, the area where keratotic papules are observed. The pharmaceutical composition for keratosis of the present invention can suppress or improve the area of ​​lesioned skin to, for example, 90% or less, 80% or less, 70% or less, 60% or less, 50% or less, 40% or less, 30% or less, 20% or less, or 10% or less compared to before administration.

[0045] Itching can be evaluated, for example, by measuring the itching felt by a subject using a numerical rating scale for itching. For example, 0 indicates no itching and 10 indicates the worst itching imaginable to the subject, and the subject evaluates the itching using this numerical rating scale. The pharmaceutical composition for keratosis of the present invention can reduce the value on the numerical rating scale for itching after administration by, for example, 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, or 9 or more, compared to the value before administration.

[0046] Pain can be evaluated, for example, by using a numerical rating scale for pain, where the pain felt by the subject is evaluated using a numerical rating scale, where 0 indicates no skin pain and 10 indicates the worst skin pain the subject can imagine. The pharmaceutical composition for keratosis of the present invention can reduce the value on the numerical rating scale for pain after administration by, for example, 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, or 9 or more, compared to the value before administration.

[0047] The odor can be evaluated, for example, by using a numerical rating scale for odor, where the odor perceived by the subject is evaluated by the subject, with 0 indicating no odor and 10 indicating the worst odor the subject can imagine. The pharmaceutical composition for keratosis of the present invention can reduce the value on the numerical rating scale for odor after administration by, for example, 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, or 9 or more, compared to the value before administration.

[0048] The pharmaceutical composition for keratosis of the present invention improves, for example, at least one selected from the group consisting of Patient Global Impression of Severity (PGIS), Patient Global Impression of Change (PGIC), Clinician Global Impression of Severity (CGIS), Clinician Global Impression of Change (CGIC), Dermatology Life Quality Index (DLQI), and Skindex-29 in the case of keratosis.

[0049] (1) The PGIS is a single questionnaire that evaluates the subject's overall impression of severity using a score, and the pharmaceutical composition for keratosis of the present invention can improve the PGIS and thereby improve the score. (2) The PGIC is a single questionnaire that evaluates the degree of improvement in overall health status by a subject using a score, and the pharmaceutical composition for keratosis of the present invention can improve the PGIC and thereby improve the score. (3) The CGIS is a single questionnaire in which a doctor evaluates a subject's overall impression of the severity of their condition using a score, and the pharmaceutical composition for keratosis of the present invention can improve the CGIS and thereby improve the score. (4) The CGIC is a single questionnaire in which a doctor evaluates the degree of improvement in the overall health condition of a subject using a score, and the pharmaceutical composition for keratosis of the present invention can improve the CGIC and improve the score. (5) DLQI is a questionnaire in which subjects answer questions about QOL with a score. The pharmaceutical composition for keratosis of the present invention can improve the DLQI and improve the score. (6) Skindex-29 is a questionnaire in which subjects answer questions about their quality of life (QOL) and are evaluated by a score. The pharmaceutical composition for keratosis of the present invention can improve Skindex-29 and thereby improve the score.

[0050] The method of administration of the pharmaceutical composition for keratosis of the present invention is not particularly limited, and may be oral or parenteral administration. Parenteral administration includes, for example, transdermal, subcutaneous, intravenous, intraarterial, intraperitoneal, intranasal, and intraintestinal administration.

[0051] The dosage form of the pharmaceutical composition for keratosis of the present invention is not particularly limited and can be appropriately determined depending on, for example, the method of administration. Examples of dosage forms include liquid, gel, cream, and solid. Examples of oral dosage forms include granules, fine granules, powders, tablets, capsules (e.g., hard capsules and soft capsules), syrups, emulsions, suspensions, liquids, and jellies. Examples of parenteral dosage forms include injections, suppositories, and transdermal agents.

[0052] The pharmaceutical composition for keratosis of the present invention is not particularly limited in other components as long as it contains the MPBSs. The pharmaceutical composition for keratosis of the present invention may contain, for example, only the MPBSs as an active ingredient, or may contain, in addition to the MPBSs, other active ingredients for keratosis.

[0053] The pharmaceutical composition for keratosis of the present invention may contain, for example, only the active ingredient, or may further contain additives in addition to the active ingredient. The additives are preferably, for example, pharmaceutically acceptable substances. The type of additive is not particularly limited and can be appropriately selected depending on, for example, the dosage form. Examples of the additives include carriers, excipients, stabilizers, lubricants, sweeteners, preservatives, suspending agents, dispersants, thickeners, pH adjusters, antifoaming agents, and flavorings. Examples of the carriers include liquids, solids, gels, and creams.

[0054] The subject to which the pharmaceutical composition for keratosis of the present invention is administered is not particularly limited and may be, for example, a human or a non-human animal, preferably a human. Non-human animals include non-human mammals such as mice, rats, rabbits, and horses.

[0055] The conditions for administering the pharmaceutical composition for keratosis of the present invention are not particularly limited and can be appropriately determined depending on, for example, the administration method and patient information, such as age, sex, weight, the presence or absence of keratosis and its symptoms, the severity of keratosis and its symptoms, and medical history.

[0056] When the pharmaceutical composition for keratosis of the present invention is orally administered, the following conditions can be exemplified. The following examples can be adjusted appropriately for infants, young children, children, adults, or the elderly depending on the patient's information, etc. The dosage of the MPBSs can be expressed, for example, as a dosage converted into the MPBS anhydrous form. The dosages exemplified below are dosages of the MPBSs converted into the MPBS anhydrous form, and specifically, for the MPBS monohydrate, are dosages converted into the MPBS anhydrous form.

[0057] Daily dosage of the MPBSs (equivalent to the anhydrous MPBS) Lower limit: e.g., 1 mg, 10 mg, 20 mg, 25 mg, 30 mg, 50 mg, 60 mg, 100 mg, 150 mg, 200 mg, or 300 mg Upper limit: e.g., 300mg, 400mg, 500mg, 600mg, 800mg, 1000mg, or 1200mg Range: e.g., 1-1000mg, 10-1000mg, 20-1000mg, 25-1000mg, 30-1000mg, 50-1000mg, 60-1000mg, 100-1000mg, 1-600mg, 10-600mg, 25-600mg, 30-600mg, 50-600mg, 100-600mg, 10-500mg, 20-500mg, 25-500mg , 30~500mg, 50~500mg, 60~500mg, 100~500mg, 200~500mg, 10~400mg, 20~400mg, 25~400mg, 30~400m g, 50~400mg, 60~400mg, 100~400mg, 200~400mg, 10~300mg, 20~300mg, 25~300mg, 30~300mg, 50~300m g, 60~300mg, 100~300mg, 200~300mg, 10~200mg, 20~200mg, 25~200mg, 30~200mg, 50~200mg, 60~200 mg, 100~200mg, 10~100mg, 20~100mg, 25~100mg, 30~100mg, 50~100mg, 60~100mg, 300~400mg, 300~5 00mg, 300-600mg, 300-800mg, 300-1000mg, 300-1200mg, 400-600mg, 400-800mg, 400-1000mg, 400-1200mg, 500-600mg, 500-800mg, 500-1000mg, 500-1200mg, 600-800mg, 600-1000mg, or 600-1200mg Examples of specific dosage amounts: for example, 1 mg, 10 mg, 20 mg, 25 mg, 30 mg, 40 mg, 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 700 mg, 800 mg, 900 mg, 1000 mg, or 1200 mg. Number of administrations per day: for example, 1 to 3 times, preferably 1 or 2 times, more preferably 2 times Frequency of administration (interval): e.g., daily

[0058] When the pharmaceutical composition for keratosis of the present invention is orally administered, the timing of administration can be freely set, for example, before meals, during meals, immediately after meals, after meals, between meals, upon waking up, or before bedtime, and administration between meals is preferred. Administration between meals of the pharmaceutical composition for keratosis of the present invention means, for example, administration at least 1 hour, preferably at least 2 hours, after ingestion at mealtime, with respect to the time interval between ingestion and administration, and / or, for example, administration at least 1 hour, 2 hours, 3 hours, 4 hours, or 4.5 hours, preferably 1 hour, after administration, with respect to the time interval between administration and ingestion of the next meal. That is, the pharmaceutical composition for keratosis of the present invention is administered, for example, between 1 or 2 hours after ingestion of a meal and 1 hour, 2 hours, 3 hours, 4 hours, or 4.5 hours before ingestion of the next meal.

[0059] As described above, the pharmaceutical composition for keratosis of the present invention can be used to treat or prevent the symptoms of keratosis, which is a disease that causes hyperkeratosis as described above. Therefore, the present invention can also be referred to as, for example, a pharmaceutical composition for hyperkeratosis that treats or prevents hyperkeratosis. The present invention can also be referred to as, for example, a pharmaceutical composition for hyperkeratosis / acantholysis that simultaneously treats or prevents hyperkeratosis and acantholysis. The present invention can also be referred to as, for example, a pharmaceutical composition for hyperkeratosis / acantholysis / dyskeratosis that simultaneously treats or prevents hyperkeratosis, acantholysis, and dyskeratosis.

[0060] [Method for treating or preventing keratosis] The method for treating or preventing keratosis of the present invention comprises the step of administering 5-methyl-2-(1-piperazinyl)benzenesulfonic acid to a subject. As described above, the form of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid (MPBS) is not limited, and may be any of the MPBSs, i.e., the MPBS anhydrate, the MPBS salt, the MPBS hydrate, and / or the MPBS solvate. Unless otherwise specified, the method for treating or preventing keratosis of the present invention will hereinafter be referred to as the method for treating or preventing keratosis of the present invention. The method for treating or preventing keratosis of the present invention is characterized by administering the MPBSs, and other conditions and steps are not particularly limited.

[0061] In the treatment and prevention methods of the present invention, the subject is a patient, and the patient may be, for example, a patient who has developed keratosis or a patient who has not developed keratosis.

[0062] In the treatment and prevention method of the present invention, the administration of the MPBSs is, for example, the administration of the pharmaceutical composition for keratosis of the present invention. In the present invention, the MPBSs, their compositions, administration methods, etc. can be referenced from the descriptions of the pharmaceutical composition for keratosis of the present invention.

[0063] [Use of MPBSs] The present invention relates to 5-methyl-2-(1-piperazinyl)benzenesulfonic acid for use in the treatment or prevention of keratosis. As described above, the form of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid (MPBS) is not limited, and it may be any of the MPBSs, i.e., the MPBS anhydrate, the MPBS salt, the MPBS hydrate, and / or the MPBS solvate. In the present invention, the descriptions of the pharmaceutical composition for keratosis of the present invention can be used to refer to the MPBSs, compositions thereof, and methods of use thereof.

[0064] The present invention also relates to the use of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid in the manufacture of a pharmaceutical composition for keratosis. As described above, the form of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid (MPBS) is not limited, and may be any of the MPBSs, i.e., the MPBS anhydrate, the MPBS salt, the MPBS hydrate, and / or the MPBS solvate. In the present invention, the descriptions of the pharmaceutical composition for keratosis of the present invention can be used to refer to the MPBSs, compositions thereof, and methods of use thereof.

[0065] [Pharmaceutical composition for acantholysis and its use] The pharmaceutical composition for acantholysis of the present invention is characterized by containing 5-methyl-2-(1-piperazinyl)benzenesulfonic acid. As mentioned above, the form of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid (MPBS) is not limited and may be any of the MPBSs, i.e., the MPBS anhydrate, the MPBS salt, the MPBS hydrate, and / or the MPBS solvate. The pharmaceutical composition for acantholysis of the present invention may contain, for example, only one of the MPBSs or two or more of the MPBSs. The MPBSs can be used for the treatment or prevention of acantholysis. The pharmaceutical composition for acantholysis of the present invention is characterized by containing the MPBSs; other components and conditions are not particularly limited. The pharmaceutical composition for acantholysis of the present invention preferably contains the MPBSs as an active ingredient. When the pharmaceutical composition for acantholysis of the present invention is used for the treatment or prevention of acantholysis, the type of disease that causes acantholysis as a pathological symptom is not particularly limited.

[0066] As described above, the pharmaceutical composition for acantholysis of the present invention can treat or prevent acantholysis. The pharmaceutical composition for acantholysis of the present invention can be used, for example, for treatment, prevention, or both treatment and prevention. Hereinafter, the term "treatment / prevention" can be interpreted as meaning either treatment, prevention, or both treatment and prevention. The pharmaceutical composition for acantholysis of the present invention enables safe treatment / prevention with few side effects. Diseases that can cause acantholysis include, for example, Darier's disease, actinic keratosis, Grover's disease, acantholytic bullae, transient acantholytic dermatosis, pemphigus, keratoacanthoma, squamous cell carcinoma, chickenpox, shingles, and herpes simplex.

[0067] In the pharmaceutical composition for acantholysis of the present invention, the MPBSs, their composition, and method of use are not particularly limited, and for example, the description of the pharmaceutical composition for keratosis of the present invention described above can be used. When using this description, "keratosis" and "symptoms of keratosis" can be read as "acantholysis."

[0068] The method for treating or preventing acantholysis of the present invention includes administering 5-methyl-2-(1-piperazinyl)benzenesulfonic acid to a subject. As described above, the form of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid (MPBS) is not limited, and may be any of the MPBSs, i.e., the MPBS anhydrate, the MPBS salt, the MPBS hydrate, and / or the MPBS solvate. The method for treating or preventing acantholysis of the present invention will hereinafter be referred to as the method for treating or preventing acantholysis of the present invention, unless otherwise specified. The method for treating or preventing acantholysis of the present invention is characterized by administering the MPBSs; other conditions and steps are not particularly limited. The subject of the present invention may be, for example, a patient who has or may develop acantholysis as a pathological symptom, and the type of disease that causes acantholysis as a pathological symptom is not particularly limited.

[0069] In the present invention, the MPBSs, compositions thereof, and administration methods thereof are not particularly limited, and the descriptions of the pharmaceutical composition for keratosis of the present invention can be used. When using the descriptions, "keratosis" and "symptoms of keratosis" can be read as "acantholysis."

[0070] The present invention relates to 5-methyl-2-(1-piperazinyl)benzenesulfonic acid for use in the treatment or prevention of acantholysis. As described above, 5-methyl-2-(1-piperazinyl)benzenesulfonic acid (MPBS) is not limited in form and may be any of the MPBSs, i.e., the MPBS anhydrate, the MPBS salt, the MPBS hydrate, and / or the MPBS solvate. In the present invention, the MPBSs, their compositions, and methods of use can be incorporated by reference to the description of the pharmaceutical composition for keratosis of the present invention. When incorporated by reference, "keratosis" and "symptoms of keratosis" can be read as "acantholysis."

[0071] The present invention also relates to the use of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid in the preparation of a pharmaceutical composition for acantholysis. As described above, the form of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid (MPBS) is not limited, and may be any of the MPBSs, i.e., the MPBS anhydrate, the MPBS salt, the MPBS hydrate, and / or the MPBS solvate. In the present invention, the MPBSs, their compositions, and methods of use can be referenced from the description of the pharmaceutical composition for keratosis of the present invention. When referenced, "keratosis" and "symptoms of keratosis" can be read as "acantholysis." [Example]

[0072] In the following examples, 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate (hereinafter, sometimes referred to as Compound A in the examples) was used as the MPBS.

[0073] [Example 1] Inhibition of epidermal thickening in an in vitro hyperkeratosis model The compound A was evaluated for its inhibitory effect on acanthosis in an epidermal hyperkeratosis model.

[0074] Contact of epidermal tissue with IL-22 induces hyperplasia in the spinous layer, resulting in hyperkeratosis. Therefore, in this example, Compound A was added to epidermal tissue constructed in vitro in parallel with the addition of IL-22, and the inhibitory effect of Compound A on hyperkeratosis was confirmed.

[0075] <Test cells and test design> An IL-22-induced human keratinocyte epidermal hyperkeratinization model was prepared using the following method. Human primary keratinocytes (foreskin, Lonza, 00192906) were pooled from three lots of Caucasian newborn donors. Epilife culture medium (Thermo Fisher Scientific, M-EPI-500-A) was used to culture the primary keratinocytes. Three-dimensional culture of the primary keratinocytes was initiated (Day 0) using air-liquid interface culture inserts and 24-well plates, and in vitro epidermal tissue was generated by culturing for 14 days (Day 0–Day 14). The culture environment was maintained at 37°C and 5% CO2. On Day 14 after the initiation of the three-dimensional culture (Day 0), the Compound A aqueous solution (solvent: water) and IL-22 solution (solvent: phosphate-buffered saline, hereinafter sometimes referred to as PBS) were added to the in vitro epidermal tissue culture medium, and the three-dimensional culture was continued for another three days (Days 14 to 17). The final concentration of Compound A in the culture medium was 0, 1, 3, 10, or 30 μmol / L, and the final concentration of IL-22 was 0 or 20 ng / mL. During the three-dimensional culture, the culture medium was changed essentially daily from Day 0 to Day 13 (except on weekends), and from Day 14 to Day 17, the culture medium was changed daily, and the Compound A aqueous solution and IL-22 solution were added at each change. On Day 17 after the initiation of the three-dimensional culture (Day 17), the in vitro epidermal tissue was sampled and used for evaluation. In this specification, the "final concentration" of each test substance solution, reagent, etc. means the concentration in the culture medium at the final stage after all test substance solutions, reagents, etc. have been added.

[0076] Each of the in vitro epidermal tissue model groups was prepared by combining the concentrations of Compound A and IL-22 in the culture medium as shown in the table below.

[0077] [Table 1]

[0078] <Evaluation method> The sampled in vitro epidermal tissue was fixed in a 4% formaldehyde solution, dehydrated, and embedded in paraffin. 6 μm-thick sections were prepared from the paraffin-embedded tissue using a slicer (Leica microtome RM2245), and fixed on glass slides to prepare epidermal tissue slides.

[0079] For pathological evaluation, the epidermal tissue slides were stained with hematoxylin-eosin (HE staining). For pathological analysis, three images were captured for each stained sample using a Nikon Eclipse (Ni-E) optical microscope connected to a digital camera (DS-Ri2). Image analysis was performed using NIS-Elements AR software (Nikon). Epidermal thickness was measured for each of the three images per sample, and the average was used as the representative value for that sample. Because IL-22-induced hyperkeratosis is a cause of acanthosis, and because hyperkeratosis of the stratum corneum occurs in 3D culture, the stratum corneum peels off each time, making quantitative evaluation difficult, epidermal thickness measurements were performed excluding the stratum corneum. Three to nine epidermal tissue slides were used per experiment (group). The effects of Compound A were evaluated by integrating and analyzing the results of the four experimental groups (Groups 1-1, 1-2, 1-3, and 1-4) and the control group (Group 1-C) in Table 1. The mean value of the normal group 1-N to which IL-22 was not added was set as 100%, and the relative value (%), as well as the mean value and standard deviation, were calculated for the measured values ​​of each group.

[0080] <Statistical analysis> Statistical analysis was performed using MATLAB (registered trademark), with a two-sided significance level of 5%. The pharmacological effect of Compound A was assessed by double comparison between control group 1-C (IL-22 added / no Compound A added) and each example group (IL-22 added / Compound A added), using the Aspin-Welch t-test. p values ​​were corrected for multiple comparisons using the Bonferroni method.

[0081] These results are shown in Figure 1. Figure 1 is a graph showing epidermal thickness in in vitro epidermal tissue. In Figure 1, the vertical axis represents the thickness of the reconstituted human epidermis, expressed relative to the normal group 1-N to which IL-22 was not added (%), which is set to 100%. As shown in Figure 1, the control group 1-C to which IL-22 was added showed a significant increase in epidermal thickness compared to the normal group 1-N to which IL-22 was not added. In contrast, the experimental group to which Compound A was added in addition to IL-22 showed a concentration-dependent decrease in epidermal thickness compared to Control group 1-C. These results demonstrate that Compound A can inhibit IL-22-induced epidermal hyperkeratosis in a concentration-dependent manner at concentrations of at least 1 μmol / L or higher. Furthermore, the hyperkeratosis-inhibitory effect of Compound A was statistically significant at concentrations of 10 μmol / L or higher.

[0082] [Example 2] Inhibition of acantholysis and dyskeratosis in an in vitro epidermal acantholysis model The inhibitory effects of Compound A on acantholysis and dyskeratosis in an epidermal acantholysis model were evaluated. In this example, acantholysis was evaluated using intercellular adhesion as an index and pathological evaluation, and dyskeratosis was evaluated using the frequency of pyknotic nuclei as an index and pathological evaluation.

[0083] <Test cells and test design> A human keratinocyte epidermal acantholysis model was established as follows. Human primary keratinocytes (foreskin, Lonza, 00192906) were pooled from three lots of Caucasian newborn donors. The human primary keratinocytes were cultured in EpiLife™ medium (Thermo Fisher Scientific, M-EPI-500-A). Thapsigargin was used to induce acantholysis and dyskeratinization.

[0084] The human primary keratinocytes were cultured in three dimensions using air-liquid interface culture inserts and 24-well plates (Day 0), and in vitro epidermal tissue was prepared by culturing for 14 days. The culture environment was maintained at 37°C and 5% CO2. 14 days after the initiation of the three-dimensional culture (Day 0) (Day 14), the compound A aqueous solution (solvent: water) and thapsigargin solution (solvent: DMSO) were added to the in vitro epidermal tissue culture medium, and the three-dimensional culture was continued for an additional 3 days (Days 14 to 17). The final concentration of compound A in the culture medium was 0, 1, 3, 10, or 30 μmol / L, and the final concentration of thapsigargin was 0 or 30 nmol / L. During the three-dimensional culture, the culture medium was basically changed every day (except on weekends) from Day 0 to Day 17, and was changed every day from Day 14 to Day 17. The Compound A aqueous solution and the thapsigargin solution were also added at each change. Seventeen days after the start of the three-dimensional culture (Day 17), the in vitro epidermal tissue was sampled and used for evaluation.

[0085] Each of the in vitro epidermal tissue model groups was prepared by combining the concentrations of Compound A and thapsigargin in the culture medium as shown in the table below.

[0086] [Table 2]

[0087] <Evaluation method> As shown below, acantholysis was evaluated by measuring cell adhesion force based on biotin diffusion, scoring the acantholysis score based on pathological observation, and immunofluorescent staining of cell adhesion marker proteins. Dyskeratinization was evaluated by measuring the frequency of pyknotic nuclei based on pathological observation and immunofluorescent staining of keratinocyte differentiation marker proteins. Specifically, slides were prepared using the in vitro epidermal tissues sampled from the same well of the 24-well plate, and evaluation was performed.

[0088] (1) Biotin diffusion The sampled in vitro epidermal tissue was rinsed twice with a PBS / CaCl2 (1 mmol / L) solution (hereinafter the same), and then a biotin marker (EZ-link™ Sulfo-NHS-LC-Biotin, Thermo Fisher Scientific) was added to the epidermal tissue as a cell membrane-impermeable marker molecule. In this example, the biotin marker was added to the basal layer side of the epidermal tissue to specifically confirm cell adhesion in the spinous layer of the epidermal tissue. The biotin marker was suspended in PBS to a concentration of 2 mg / mL, and this suspension was added to the basal layer side. The in vitro epidermal tissue was then left to stand at 37°C for 30 minutes and then rinsed twice with a PBS / CaCl2 (1 mmol / L) / Glycine (100 mmol / L) solution (hereinafter the same). Sample numbers were coded to ensure blinding of subsequent procedures. The in vitro epidermal tissue was then fixed in a 4% formaldehyde solution, dehydrated, and embedded in paraffin. 6-μm-thick sections were prepared from the paraffin-embedded tissue using a slicer (Leica microtome RM2245), and fixed on glass slides to prepare epidermal tissue slides.

[0089] During acantholysis, the amount of biotin in the epidermal layer increases relative to the amount of biotin in the epidermal tissue due to a decrease in cell adhesion. Therefore, the epidermal tissue slides were prepared and the biotin marker was detected to evaluate the intercellular adhesion of the epidermis in the in vitro epidermal tissue. Specifically, the epidermal tissue slides were first deparaffinized, rehydrated, and then placed in the presence of fluorescently labeled streptavidin (Thermo Fisher Scientific, S32354) to detect biotin (the molecular marker) diffused into the epidermal layer of the in vitro epidermal tissue.

[0090] Specifically, three images were captured for each sample (the epidermal tissue slide) using a Nikon Eclipse (Ni-E) optical microscope connected to a digital camera (DS-Ri2) in the dark at 4°C. Image analysis was performed using NIS-Elements AR software (Nikon), and biotin fluorescence intensity was quantified in the three images per sample. For each image, the epidermal tissue, excluding the stratum corneum, was manually demarcated as a region of interest (ROI), and its surface area, A, was measured (unit: μm 2 Next, the area B of the biotin-stained region within the delimited region of interest (ROI) was measured (unit: μm 2 ). Biotin fluorescence intensity was quantified as the staining intensity per unit surface area, calculated by multiplying the average staining intensity C of the biotin-stained region by the biotin-stained area B and dividing the result by the ROI surface area A. For each sample, the average value of the biotin fluorescence intensity for each of the three images was calculated, and this average value was used as the representative value for the three images. Nine epidermal tissue slides were used per experiment (one in vitro epidermal tissue). The measured values ​​for the normal group 2-N, to which thapsigargin had not been added, were set as 100%, and the relative values ​​(%), as well as the average values ​​and standard deviations, for the measured values ​​for each group were calculated.

[0091] (2) Pathological evaluation The in vitro epidermal tissue samples were rinsed twice with the PBS / CaCl solution, fixed in 4% formaldehyde, dehydrated, and embedded in paraffin. 6 μm-thick sections were prepared from the paraffin-embedded tissue using a slicer (Leica microtome RM2245) and fixed on glass slides to prepare epidermal tissue slides.

[0092] The epidermal tissue slides were stained with HE and used as samples for pathological evaluation. For pathological analysis, images of each stained sample were captured using a Nikon Eclipse (Ni-E) optical microscope connected to a digital camera (DS-Ri2). The captured images were analyzed to visually evaluate dyskeratosis and acantholysis in each sample.

[0093] The occurrence of dyskeratosis relatively increases the frequency of pyknotic nuclei. Therefore, for the pathological evaluation of dyskeratosis, the pathological evaluation samples were used, and the number of pyknotic nuclei in each field was counted. The pyknotic nuclei were counted in three fields (three images) for each sample, and the average value was used as the representative value for that sample. Nine epidermal tissue slides were used per experiment (one group). For the pathological evaluation of acantholysis, the pathological evaluation samples were used, and observations were made, and the presence or absence of acantholysis findings was qualitatively recorded. The qualitative evaluation of acantholysis findings was performed by scoring the number of fissures that appeared at the boundary between the basal layer and the spinous layer. The scoring criteria were as follows, and the average and standard deviation of the scores for each group were calculated. 0: No fissure 1: One fissure 2: Two fissures 3: Three or more fissures

[0094] (3) Immunofluorescence staining The epidermal tissue slides were subjected to immunofluorescence staining using antibodies corresponding to each marker of interest.

[0095] (i) As in (2), sections were fixed to slides to prepare epidermal tissue slides. The epidermal tissue slides were deparaffinized and rehydrated, and then heated with 0.01 mol / L sodium citrate buffer (pH 6.0) to activate antigens. The antigen-activated epidermal tissue slides were rinsed with PBS and then saturated with 5% normal goat serum (NGS) in a PBS / 0.1% Tween 20 solution in a thermo-humidifier for 1 hour to prevent nonspecific labeling. The epidermal tissue slides were incubated overnight at 4°C with primary antibodies (anti-filaggrin (FLG) antibody and anti-cytokeratin 10 (K-10) antibody). After sequential washing with PBS / 0.1% Tween 20 and PBS, the epidermal tissue slides were incubated with Alexa Fluor dye-conjugated anti-mouse and anti-rabbit secondary antibodies in a thermo-humidifier at room temperature for 1 hour. Finally, the epidermal tissue slide was rinsed with PBS. The primary and secondary antibodies used for co-immunostaining of keratinocyte differentiation-associated proteins FLG / K-10 were as shown in the table below.

[0096] [Table 3]

[0097] (ii) The epidermal tissue slides were deparaffinized and rehydrated as described in (i) above, and then heated with pepsin solution (Sigma, R2283) at 56°C for 15 minutes to activate the antigens. The antigen-activated epidermal tissue slides were then rinsed with PBS and saturated with PBS / 0.1% Tween 20 containing 2% bovine serum albumin (BSA) in a thermo-humidifier for 1 hour to prevent nonspecific labeling. The epidermal tissue slides were then incubated overnight at 4°C with primary antibodies (anti-desmoglein 1 (DSG1) antibody and anti-claudin 1 (CLDN1) antibody). After sequential washing with PBS / 0.1% Tween 20 and PBS, the epidermal tissue slides were incubated with Alexa Fluor dye-conjugated anti-goat and anti-rabbit secondary antibodies in a thermo-humidifier at room temperature for 1 hour. Finally, the epidermal tissue slides were rinsed with PBS. The primary and secondary antibodies used for co-immunostaining of DSG1 / CLDN1, which are cell adhesion-related proteins, are as shown in the table below.

[0098] [Table 4]

[0099] Keratinocyte nuclei can be detected using DAPI (4',6-diamidino-2-phenylindole), a fluorescent molecule that binds to the adenine and thymine bases of DNA. Therefore, after the secondary antibody treatment and final wash, the epidermal tissue slides (i) and (ii) were treated with ProLong™ Diamond Antifade Mountant (Thermo Fisher Scientific, P36962) containing DAPI, and immunofluorescent staining images were obtained.

[0100] Specifically, three images were taken for each sample (the epidermal tissue slide) using a Nikon Eclipse (Ni-E) optical microscope connected to a digital camera (DS-Ri2) in the dark at 4°C. Image analysis was performed using NIS-Elements Advanced Research Imaging software (Nikon), and the fluorescently stained area in the three images per sample was quantified. For each image, the epidermal tissue including the stratum corneum was manually defined as a region of interest (ROI) for FLG, and the epidermal tissue excluding the stratum corneum was manually defined as a region of interest (ROI) for K-10, DSG1, and CLDN1, and their surface areas, A, were measured (unit: μm 2 Next, the area B of the stained region within the delimited region of interest (ROI) was measured (unit: μm 2 The immunostained area was quantified as the relative value (%) of the stained area (B / A), i.e., the stained area (B / A) divided by the ROI surface area (A), and the mean value and standard deviation for each group were calculated. For each sample, the mean value was calculated from the biotin fluorescence intensity of each of the three images, and this mean value was used as the representative value of the three images. The number of samples was nine epidermal tissue slides per experiment (one in vitro epidermal tissue).

[0101] <Statistical analysis> Statistical analysis was performed using SAS, with a one-sided significance level of 2.5% for the Williams multiple comparison test and a two-sided significance level of 5% for other test methods. The success of the formation of the epidermal acantholysis model by adding thapsigargin was determined by comparing two groups: normal group 2-N (no thapsigargin added / no compound A added) and control group 2-C (thapsigargin added / no compound A added), using a Student's t-test. For in vitro epidermal tissues where no significant changes were observed, the formation of the epidermal acantholysis model was determined to have failed, and further analysis was not performed. The pharmacological effect of Compound A was also determined by comparing control group 2-C (thapsigargin added / no compound A added) with each example group (thapsigargin added / compound A added), using a Williams multiple comparison test. In the evaluation of immunofluorescence staining, only Example Group 2-3 (10 μmol / L of the above-mentioned Compound A) was used, and therefore the test method used was Student's t-test.

[0102] These results are shown in Figures 2, 3, 4, 5A, 5B, 6A, and 6B. Figure 2 is a graph showing the results of biotin diffusion, an indicator of acantholysis, with the vertical axis indicating the relative amount of biotin (%). Figure 3 is a graph showing the results of pyknotic nuclei frequency, an indicator of dyskeratinization, with the vertical axis indicating the number of pyknotic nuclei in the image. Figure 4 is a graph showing the results of pathological evaluation of acantholysis, with the vertical axis indicating the acantholysis score. Figures 5A and 5B are graphs showing the results of keratinocyte differentiation markers associated with dyskeratinization, with Figure 5A showing the results for FLG and Figure 5B showing the results for K-10, each with the vertical axis indicating the relative stained area (%). Figures 6A and 6B are graphs showing the results of cell adhesion markers associated with acantholysis, with Figure 6A showing the results for DSG1 and Figure 6B showing the results for CLDN1, each with the vertical axis indicating the relative stained area (%).

[0103] First, let us consider acantholysis. As shown in Figure 2, the amount of biotin in the epidermal layer, which is an indicator of acantholysis, was significantly increased in the control group 2-C to which thapsigargin was added, compared to the normal group 2-N to which thapsigargin was not added. This indicates that the addition of thapsigargin caused acantholysis, and the amount of biotin in the epidermal layer increased due to a decrease in intercellular adhesion. In contrast, in the experimental group to which thapsigargin and the compound A were added, the amount of biotin in the epidermal layer decreased in a concentration-dependent manner at least at 1 μmol / L or more compared to the control group 2-C. Furthermore, as shown in Figure 4, similar results were obtained in pathological evaluation. That is, compared to the normal group 2-N, the acantholysis score of the control group 2-C was significantly increased by the addition of thapsigargin, but in the experimental group, the addition of the compound A was able to suppress the increase in acantholysis score in a concentration-dependent manner at least at 1 μmol / L or more. Furthermore, as shown in Figures 6A and 6B, similar results were obtained in immunofluorescent staining of the cell adhesion-related proteins DSG1 and CLDN1, where the immunofluorescent staining area of ​​control group 2-C was reduced by the addition of thapsigargin compared to normal group 2-N. This indicates that the addition of thapsigargin weakened or disrupted desmosomes and tight junctions, which are modes of cell adhesion. In contrast, the immunofluorescent staining area increased in the experimental group to which compound A was added in addition to thapsigargin compared to control group 2-C. This indicates that compound A was able to normalize the cell adhesion abnormalities induced by thapsigargin.

[0104] From these results, it can be found that by adding compound A, the weakening of intercellular adhesive force induced by thapsigargin in in vitro epidermal tissue can be improved concentration-dependently, that is, acantholysis can be suppressed.In addition, as a specific example, it can be found that compound A suppresses the decrease of intercellular adhesive force at 10 μ mol / L, and suppresses the increase of the pathological evaluation score of acantholysis at 1 μ mol / L and above.

[0105] Next, we will discuss abnormal keratinization. As shown in Figure 3, the number of pyknotic nuclei, an indicator of abnormal keratinization, was significantly increased in the control group 2-C, which was treated with thapsigargin, compared with the normal group 2-N, which was not treated with thapsigargin. This indicates that the addition of thapsigargin caused abnormal keratinization and increased the number of pyknotic nuclei. In contrast, in the experimental group, to which thapsigargin and Compound A were added, the amount of biotin in the epidermal layer decreased in a concentration-dependent manner, at least at concentrations of 1 μmol / L or more of Compound A, compared with the control group 2-C. Furthermore, as shown in Figures 5A and 5B, immunofluorescent staining of the differentiation marker FLG, which is expressed in keratinocytes in the stratum granulosum and stratum corneum, and the differentiation marker K-10, which is expressed in keratinocytes in the stratum spinosum and granular layer, showed a statistically significant increase and decrease, respectively, in the immunofluorescent stained areas of the control group 2-C compared with the normal group 2-N, due to the addition of thapsigargin. This indicates that the addition of thapsigargin causes the differentiation state of keratinocytes forming the spinous layer to become non-uniform, making them more susceptible to abnormal keratinization. In contrast, in the Example group to which 10 μmol / L of Compound A was added in addition to thapsigargin, the immunofluorescent staining area for FLG and the immunofluorescent staining area for K-10 were statistically significantly decreased and increased, respectively, compared to Control Group 2-C. This indicates that Compound A was able to normalize the abnormality in keratinocyte differentiation induced by thapsigargin.

[0106] These results demonstrate that the addition of Compound A can improve the increase in the frequency of pyknotic nuclei induced by thapsigargin in in vitro epidermal tissue in a concentration-dependent manner, i.e., inhibits dyskeratosis. Specifically, Compound A was found to be statistically significant at concentrations of 3 μmol / L or higher in inhibiting the increase in pyknotic nuclei, an index of dyskeratosis.

[0107] Furthermore, these results indicate that Compound A can suppress both dyskeratosis and acantholysis. Therefore, it was found that, among keratosis, not only diseases that cause only one of dyskeratosis or acantholysis, but also diseases that cause both (e.g., Darier's disease), simply administering Compound A can simultaneously suppress both symptoms. Furthermore, in Example 1, a hyperkeratinization model was prepared using the same keratinocytes, and the suppression of hyperkeratinization by Compound A was also confirmed. Therefore, it can be said that Compound A exhibits an inhibitory effect on all three major pathological symptoms of Darier's disease, namely, hyperkeratinization, acantholysis, and dyskeratosis.

[0108] [Example 3] Inhibition of hyperkeratosis in an in vivo hyperkeratosis model The compound A was evaluated for its inhibitory effect on dorsal skin thickening in a hyperkeratosis model.

[0109] By applying imiquimod to mouse skin, hyperkeratosis accompanied by an inflammatory reaction is induced, and a psoriasis-like dermatitis model mouse with hyperkeratosis (thickening of the skin) can be obtained. Therefore, in this example, the compound A was administered to mice in parallel with the application of imiquimod, which induces hyperkeratosis, and the inhibitory effect of compound A on hyperkeratosis was confirmed.

[0110] <Test cells and test design> Seven-week-old male BALB / c mice (Charles River Japan) were used. The mice were observed for general condition over a six-day quarantine and acclimation period. After the acclimation period (at 8 weeks of age), the mice were weighed and divided into three groups using stratified randomization to ensure uniform mean body weights across each group. The dorsal hair of mice in one of the experimental groups was shaved. Three days later, under 2% inhalation anesthesia using isoflurane (Mylan Pharmaceuticals, Inc.), imiquimod 5% cream (Mochida Pharmaceutical Co., Ltd.) was applied to the dorsal skin of the mice once daily for a total of four days to induce psoriasiform dermatitis. Psoriasiform dermatitis manifests as hyperkeratosis. The application dose of imiquimod 5% cream per application was 62.5 mg (3.125 mg of imiquimod). After each application of imiquimod 5% cream, the application site was gently wiped with absorbent cotton soaked in lukewarm water approximately 4 hours after application. Concurrently, Compound A was orally administered at a dose of 30 mg (Compound A) / kg twice daily for 4 days (Day 1 to Day 5) on the same day as the application of imiquimod 5% cream. The twice-daily administration consisted of the first administration before the application of imiquimod 5% cream and the second administration at least 10 hours after the first administration of the same day. A 3 mg / mL solution of Compound A (solvent: water for injection) was used for oral administration of Compound A.

[0111] The remaining two groups were a normal group and a control group. Normal group 3-N was not challenged with imiquimod after shaving the back, nor was it administered with Compound A. Control group 3-C was challenged with imiquimod in the same manner as the example group, but was not administered with Compound A. Instead, 10 mL / kg of a 1% Tween 80 solution not containing Compound A was orally administered.

[0112] [Table 5]

[0113] <Evaluation method> The first application of imiquimod 5% cream was designated Day 1, and the dorsal skin thickness of the mice was measured before dissection on Day 5. Specifically, the mice were placed under 2% isoflurane inhalation anesthesia, and the dorsal skin thickness was measured using a digital caliper (Mitutoyo Corporation). Measurement values ​​were recorded to two decimal places and expressed in mm. The number of animals per group was 9 for the control group and the experimental group, and 4 for the normal group, and the effect of Compound A was evaluated. The mean and standard deviation of the dorsal skin thickness for each group were calculated.

[0114] <Statistical analysis> Statistical analysis was performed using SAS, and intergroup comparison was performed with a two-sided significance level of 5%. The pharmacological action of Compound A was evaluated by comparing the control group 3-C (imiquimod applied / not administered Compound A) with the example group (imiquimod applied / administered Compound A), and the test method used was Student's t-test.

[0115] These results are shown in Figure 7. Figure 7 is a graph showing the dorsal skin thickness on Day 5 in hyperkeratotic mice, with the vertical axis representing the thickness of the dorsal skin (mm). As shown in Figure 7, compared with the normal group 3-N in which psoriasis-like dermatitis was not induced, the control group 3-C in which psoriasis-like dermatitis was induced with imiquimod showed significant thickening of the dorsal skin, i.e., hyperkeratosis. In contrast, the example group 3-1, in which compound A was administered on the same day as imiquimod application, showed a statistically significant decrease in dorsal skin thickness compared with the control group 3-C. These results demonstrate that compound A can suppress imiquimod-induced skin thickening, i.e., hyperkeratosis.

[0116] [Example 4] Involvement of IL-22 in Darier's disease We performed gene expression analysis in formalin-fixed, paraffin-embedded (FFPE) human skin samples from patients with Darier's disease and psoriasis vulgaris. Specifically, we evaluated the involvement of IL-22 in Darier's disease, a type of keratosis known to be involved in psoriasis vulgaris, by comparing it with psoriasis vulgaris.

[0117] <Test cells and test design> Image acquisition and library preparation were performed on FFPE block samples (Aurus Biosciences, Inc.) with a diagnosis history, as described below. Patient-derived FFPE block samples were obtained from a chest skin biopsy specimen of a black female in her 70s with psoriasis vulgaris, and from a back skin biopsy specimen of a white female in her 40s with Darier's disease. 5 μm-thick sections were cut from the FFPE block samples using a slicer, and a section adhesion test was performed using Visium Test Slides (10X Genomics). After confirming minimal peeling or peeling of the sections, slides were prepared using a commercially available kit (Visium Spatial for FFPE Gene Expression Kit, Human Transcriptome, 10X Genomics). After HE staining, digital slide data was acquired using a scanner (Nanozoomer, Hamamatsu Photonics K.K.). Furthermore, a library was prepared from the slides using a commercially available kit (Visium Spatial for FFPE Gene Expression Starter Kit, Human Transcriptome, 10X Genomics), and the library was subjected to next-generation sequencing analysis.

[0118] <Data analysis and evaluation methods> Spatial transcriptome analysis of sequences was performed using Space Ranger (10X Genomics). Gene expression intensities were calculated by log2-transforming UMI counts normalized by the total UMI counts for the same barcode sequence. To estimate the contribution of IL-22 to pathogenesis, we compared the gene expression intensities of the IL-22 receptor IL22RA1 in the epidermis of patients with Darier's disease with those of psoriasis vulgaris, where IL-22 has been reported to significantly contribute to the progression of hyperkeratosis. As keratinization progresses, the number of spots classified as epidermal in Visium increases and layers are formed. Therefore, we classified the epidermis into upper and basal layers and calculated the expression intensity of IL22RA1 in each layer of the epidermis. We then calculated the ratio of IL22RA1 expression in the upper layer to the basal layer, where IL22RA1 expression is low.

[0119] These results are shown in Table 6. As shown by the IL22RA1 epidermal expression ratio in Table 6, Darier's disease patients showed strong IL22RA1 (IL-22 receptor) gene expression in the upper layers of the epidermis compared to the basal layer, similar to patients with psoriasis vulgaris, in which IL-22 is known to be involved. This indicates that IL-22 is involved in Darier's disease as well as psoriasis vulgaris.

[0120] [Table 6]

[0121] The above results confirmed the involvement of IL-22 in both psoriasis vulgaris and Darier's disease, which are major keratosis diseases. In Example 1, it was demonstrated that IL-22 induced hyperkeratosis and that Compound A inhibited hyperkeratosis. From this, it can be said that the inhibitory effect of Compound A on hyperkeratosis in Example 1 also exhibits an inhibitory effect on both psoriasis vulgaris and Darier's disease.

[0122] [Example 5] Evaluation of pharmacokinetics The pharmacokinetics of Compound A was evaluated using the compound A. In the present examples, the amount of Compound A used is expressed in terms of the amount of MPBS anhydrous (hereinafter, sometimes referred to as Compound B in the examples).

[0123] A phase 1, double-blind, placebo-controlled, dose-escalating oral study was conducted in healthy males and females. In this study, Compound A was orally administered once or twice daily to subjects. The single dose of Compound A was 50 mg or 100 mg, equivalent to the amount of Compound B. Twenty-four subjects (16 males and 8 females) were enrolled in the study, and 24 completed the study. Table 7 below shows the study groups and treatment details. The study groups were divided into three groups, A to C, as shown in Table 7. Each study group consisted of eight subjects, six of whom received Compound A and two of whom received a placebo. Compound A was administered in tablets containing 5 mg of Compound A, equivalent to the amount of Compound B (hereinafter referred to as Compound A 5 mg tablets). Test groups A and B were administered 10 tablets of the compound A 5 mg tablets or placebo tablets per dose, and test group C was administered 20 tablets of the compound A 5 mg tablets or placebo tablets per dose.

[0124] [Table 7]

[0125] Each subject received a total of 16 doses of Compound A 5 mg tablets or placebo tablets during the study period. These were divided into 16 doses: one in the morning on Day 1, one in the morning and one in the evening from Days 3 to 9, and one in the morning on Day 10. From Days 3 to 10, Compound A 5 mg tablets or placebo tablets were administered at 12-hour intervals. The morning dose was administered approximately 2 hours after breakfast, and the afternoon dose was administered approximately 2 hours after dinner. Breakfast, lunch, afternoon snack, and dinner were served approximately 2 hours before, 4 hours after, 7 hours after, and 10 hours after the morning dose, respectively. Meals were limited to no more than 2500 kcal for men and 2000 kcal for women. Water was prohibited for two hours after administration on Day 1 and Day 10, but was otherwise freely available at any time. The study drug (the Compound A 5 mg tablet or placebo tablet) was administered with 200 mL of water while standing. On Day 1 and Day 10, subjects were prohibited from lying supine until two hours after administration, except when following the study procedures or when instructed by a physician.

[0126] Blood samples were collected over time from test groups A and B (hereinafter referred to as the 50 mg group), in which 50 mg of compound A was administered as compound B per dose, and from test group C (hereinafter referred to as the 100 mg group), in which 100 mg of compound B was administered as compound B per dose, to measure the plasma concentration of compound B. The mean plasma concentrations from day 9 (Day 9) to day 10 (Day 10) are shown in Figure 8 and Table 8. Figure 8 is a graph showing the mean plasma concentration of compound B, with the vertical axis representing the plasma concentration of compound B (ng / mL) and the horizontal axis representing the time elapsed since administration on day 9 (Day 9), with the time immediately before administration on day 9 (Day 9) being designated as time 0. The results for the 50 mg group are the average of 12 cases from test groups A and B, excluding those administered placebo, and the results for the 100 mg group are the average of 6 cases from test group C, excluding those administered placebo.

[0127] [Table 8]

[0128] In the results of the thapsigargin-induced human keratinocyte epidermal acantholysis model in Example 2, the effective pharmacological concentration of Compound A was about 1 μmol / L (about 256 ng / mL) to about 10 μmol / L (about 2560 ng / mL). Therefore, from the concentration change (drug concentration change) of Compound B in human plasma after administration of Compound A shown in FIG. 8 and the effective pharmacological concentration in Example 2, the clinically effective dose of Compound A is, for example, about 50 mg or more, converted into the amount of Compound B, orally administered twice a day, and further 50 to 200 mg, orally administered twice a day.

[0129] [Example 6] In vivo clinical trial Compound A will be administered to Darier's disease patients to conduct a phase 2 clinical trial. Compound A 50 mg tablets containing 50 mg of compound A converted into the amount of compound B will be used for administration of compound A.

[0130] <Subject> Darier's disease patients aged 18 to 75 <Test Overview> Randomized double-blind clinical trial Placebo tablets administered: 24 cases 100mg group: 12 cases One 50 mg tablet of Compound A was orally administered twice a day (100 mg / day as the amount of Compound B). 200mg group: 24 patients Two 50 mg tablets of Compound A were orally administered twice a day (200 mg / day of Compound B). 400mg group: 24 cases Four 50 mg tablets of Compound A were orally administered twice a day (400 mg / day of Compound B).

[0131] <Test items> In clinical trials of Compound A in patients with Darier's disease, evaluations are conducted on the Investigator's Global Assessment (IGA) score, body surface area, scores for itching, pain, and odor, Patient Global Impression of Severity (PGIS) score, Patient Global Impression of Change (PGIC) score, Clinician Global Impression of Severity (CGIS) score, Clinician Global Impression of Change (CGIC) score, Dermatology Life Quality Index (DLQI) score, and Skindex-29 score. Compared to a placebo group, administration of Compound A is expected to improve at least one of the test items.

[0132] In the above-mentioned Examples 1 to 4, the inhibitory effect on acanthosis in epidermal hyperkeratosis and the inhibitory effect on acantholysis and dyskeratosis in epidermal acantholysis have been confirmed. Therefore, it can be said that various symptom scores for Darier's disease patients are improved by administering Compound A.

[0133] [Example 7] Evaluation of the effect of food on pharmacokinetics The compound A was administered in a fasted state or after a meal to evaluate the effect of food on the pharmacokinetics.

[0134] A phase 1, double-blind, placebo-controlled, four-period crossover study was conducted on healthy male subjects. In this study, subjects received a single dose of Compound A in each administration period (1 to 4) for evaluation. The single dose of Compound A was 25 mg, equivalent to the amount of Compound B. Nine subjects were enrolled in the study. Table 9 below shows the test groups and treatment details. The test groups were composed of three groups, Test Groups 1 to 3, as shown in Table 9. Each test group consisted of three subjects, and in each administration period, six subjects received Compound A and three subjects received a placebo. Compound A was administered using the 5 mg Compound A tablets described in Example 5.

[0135] [Table 9]

[0136] Each subject was orally administered a 5 mg tablet of Compound A or a placebo tablet once in the morning of Day 1 of the study for each dosing period. Food intake was prohibited until 4 hours after dosing, and water intake until 2 hours after dosing. The study drug (5 mg tablet of Compound A or placebo tablet) was administered with 240 mL of water while standing. For both dosing periods, dinner was provided the day before the study (Day 0), and on Day 1, lunch, an afternoon snack, and dinner were provided 4.5, 7.5, and 11 hours after dosing, respectively. However, only in the third dosing period, a high-fat meal was provided in the morning of Day 1. The high-fat meal was provided 20 minutes before dosing, consumed over 15 minutes, and finished 5 minutes before dosing.

[0137] Table 10 shows the pharmacokinetic parameters when 25 mg of compound A was administered after fasting (1st, 2nd, and 4th administration periods) or after a high-fat meal (3rd administration period). In Table 10, column A indicates the type of parameter, column B indicates the average for the 3rd administration period (25 mg after meals), and column C indicates the average for the 2nd administration period (25 mg fasting). In columns B and C, Tmax is the median, minimum, and maximum (in parentheses), and the rest are the arithmetic mean and standard deviation (in parentheses). In column D, Tmax is the least squares mean difference and 95% confidence interval (in parentheses), and the rest are the least squares mean ratio and 95% confidence interval (in parentheses).

[0138] [Table 10]

[0139] The area under the drug concentration curve (AUC) was measured from time 0 to the last quantifiable time point after the high-fat meal. 0-t ), the area under the drug concentration curve from time 0 to time infinity (AUC 0-∞ ), and maximum blood concentration (C max ) decreased by 23%, 27%, and 21%, respectively. Tmax ) was slightly faster with the high-fat diet than with fasting.

[0140] Although the present invention has been described above with reference to the embodiments and examples, the present invention is not limited to the above-described embodiments and examples. Various modifications that can be understood by those skilled in the art can be made to the configuration and details of the present invention within the scope of the present invention.

[0141] This application claims priority based on Japanese Patent Application No. 2024-104616, filed June 28, 2024, the disclosure of which is incorporated herein in its entirety. [Industrial Applicability]

[0142] According to the pharmaceutical composition of the present invention, by containing 5-methyl-2-(1-piperazinyl)benzenesulfonic acid, it is possible to treat or prevent keratosis.

Claims

1. A pharmaceutical composition for treating keratosis, comprising 5-methyl-2-(1-piperazinyl)benzenesulfonic acid.

2. 2. The pharmaceutical composition of claim 1, wherein the keratosis is at least one selected from the group consisting of Darier's disease, psoriasis, and actinic keratosis.

3. The pharmaceutical composition of claim 1, wherein the keratosis is Darier's disease.

4. The pharmaceutical composition according to claim 1 , which inhibits at least one selected from the group consisting of hyperkeratosis, dyskeratosis, and acantholysis in the keratosis.

5. The pharmaceutical composition according to any one of claims 1 to 3, wherein the dosage of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid is 1 to 1000 mg / day.

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