Pharmaceutical compositions for keratosis and their uses

Abstract

This invention provides a novel pharmaceutical composition for keratosis, including Darier's disease and psoriasis. [Solution] The pharmaceutical composition for keratosis of the present invention contains 5-methyl-2-(1-piperazinyl)benzenesulfonic acid. The compound is its anhydride, its salt, its hydrate or solvate, or its hydrate or solvate, the hydrate being, for example, 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate. 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 hyperkeratosis, acantholysis, and abnormal keratinization of the epidermis, resulting in the appearance of keratotic papules (Non-Patent Literature 1). Pain and itching are frequently reported symptoms, and a foul odor is particularly noticeable in areas with excessive sweating and a tendency for secondary infections. Furthermore, because Darier's disease can become chronic and recur, it can lead to a decline in the patient's quality of life (QOL) and raise concerns about social disadvantages.

[0003] In particular, for Darier's disease, a type of keratosis, there is currently no fundamental cure recommended in guidelines. Treatment focuses on disease management and symptom management by avoiding triggers for exacerbations. Treatment methods involving the administration of retinoids, steroids, and vitamin D analogs have been proposed, but side effects have been reported, and no safe and consistently effective treatment has been found. Therefore, there remains a high unmet medical need for a safe and effective treatment. [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 [Overview of the project] [Problems that the invention aims to solve]

[0005] Therefore, the present invention aims to provide a novel pharmaceutical composition for keratosis, including Darier's disease and psoriasis, and a method for treating or preventing these conditions. [Means for solving the problem]

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

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

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

[0009] The present invention relates to the use of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid in the manufacture of pharmaceutical compositions for keratosis. [Effects of the Invention]

[0010] According to the pharmaceutical composition of the present invention, keratosis can be treated or prevented by including 5-methyl-2-(1-piperazinyl)benzenesulfonic acid. [Brief explanation of the drawing]

[0011] [Figure 1] Figure 1 is a graph showing the relationship between the administration of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate and epidermal thickness in in vitro epidermal tissue in Example 1. [Figure 2] Figure 2 is a graph showing the relationship between the administration of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate and biotin diffusion, which is an indicator of acantholysis, in Example 2. [Figure 3]Figure 3 is a graph showing the relationship between the administration of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate and the frequency of condensed nuclei, which is an index of abnormal keratinization, in Example 2. [Figure 4] Figure 4 is a graph showing the relationship between the administration of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate and the pathological evaluation of acantholysis in Example 2. [Figure 5A] Figure 5A is a graph showing the relationship between the 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] Figure 5B is a graph showing the relationship between the 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] Figure 6A is a graph showing the relationship between the 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] Figure 6B is a graph showing the relationship between the 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] Figure 7 is a graph showing the relationship between the administration of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate and the back skin thickness of hyperkeratotic mice in Example 3. [Figure 8] Figure 8 is a graph showing the relationship between the administration of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate and the average plasma concentration of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid in Example 5.

Modes for Carrying Out the Invention

[0012] The present invention can be exemplified by the following embodiments. [1] A pharmaceutical composition for keratosis containing 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 a keratosis in which the IL-22 signal 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, abnormal keratinization, and acantholysis in the keratosis.

[12] The pharmaceutical composition according to

[11] , wherein the hyperkeratosis is hyperkeratosis in which the IL-22 signal is involved.

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

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

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

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

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

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

[16] The pharmaceutical composition according to

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

[17] The pharmaceutical composition according to

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

[18] A pharmaceutical composition according to any one of items [1] to

[17] , wherein the number of doses per day is 1 to 3 times.

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

[17] , wherein the number of doses per day is two.

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

[19] , wherein the interval of administration is daily.

[0013]

[21] A method for treating or preventing keratosis, comprising the step of 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 its anhydride, its salt, its hydrate or solvate, or its hydrate or solvate.

[23] The method according to

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

[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 the claims

[21] to

[24] , wherein the method of administration 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 an anhydride of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid.

[27] The method according to any one of the items in

[21] to

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

[28] The method according to any one of the items in

[21] to

[27] , wherein the interval between doses is daily.

[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 its anhydride, its salt, its hydrate or solvate, or its hydrate or solvate.

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

[29] , wherein the 5-methyl-2-(1-piperazinyl)benzenesulfonic acid is its anhydrous, its salt, its hydrate, or its salt hydrate.

[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 pharmaceutical compositions for keratosis.

[34] The use according to

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

[35] The use according to

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

[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 acantholytics containing 5-methyl-2-(1-piperazinyl)benzenesulfonic acid.

[38] The pharmaceutical composition according to

[37] , wherein the 5-methyl-2-(1-piperazinyl)benzenesulfonic acid is its anhydrous, its salt, its hydrate or solvate, or its hydrate or solvate.

[39] The pharmaceutical composition according to

[37] , wherein the 5-methyl-2-(1-piperazinyl)benzenesulfonic acid is its anhydrous, its salt, its hydrate, or its salt hydrate.

[40] The pharmaceutical composition according to

[38] or

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

[0016] In this specification, "treatment of a disease" includes, for example, the meaning of curing a disease, achieving remission of a disease, alleviating a disease, or suppressing the progression of a disease, and also includes, for example, the meaning of treating symptoms caused by the disease (e.g., curing, achieving remission, alleviating, or suppressing symptoms). "Prevention of a disease" includes, for example, the meaning of preventing contracting a disease, preventing the onset of a disease, or preventing the recurrence of a disease, and also includes, for example, the meaning of preventing symptoms caused by the disease (e.g., preventing the onset of symptoms or preventing the recurrence of symptoms). In this specification, "treatment or prevention of a disease" can also be expressed as, for example, the suppression of a disease, and "treatment or prevention of symptoms caused by a disease" can also be expressed as, for example, the suppression of symptoms caused by a disease.

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

[0018] In this specification, treatment refers to, for example, a procedure performed on a subject diagnosed by a physician as having a disease or symptoms of a disease. In this specification, prevention refers to, for example, a procedure performed on a subject who does not have a disease or symptoms of a disease, and is intended to prevent the onset of the disease or symptoms of a disease. The subject may also be referred to as a test subject or subject, and may be a human (patient) or a non-human animal (sick animal). In this specification, the term "patient" may also include, for example, a non-human animal that is sick, and can be read as "sick animal". Furthermore, if the subject does not have the disease or symptoms of the disease in question, it may also be referred to as a healthy person (healthy human or healthy non-human animal) with respect to the disease or symptoms of the subject.

[0019] The present invention will be described below with specific examples, but the present invention is not limited to these examples. Unless otherwise specified, each embodiment described below can be used interchangeably with one another.

[0020] [Medicinal 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 above compound, but other components and conditions 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 may be used, for example, for therapeutic purposes, for preventive purposes, or for both therapeutic and preventive purposes. Hereinafter, the term "treatment / prevention" can be interpreted as meaning treatment, prevention, or treatment and prevention. The pharmaceutical composition for keratosis of the present invention allows for safe treatment / prevention with fewer side effects, for example.

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

[0023] The type of MPBS salt is not particularly limited and includes, for example, alkali metal salts, alkaline earth metal salts, amphoteric element salts, amine salts, inorganic acid salts, or organic acid salts. Examples of alkali metal salts include sodium salts or potassium salts, examples of alkaline earth metal salts include magnesium salts or calcium salts, and examples of amphoteric element salts include aluminum salts. Examples of amine salts 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 salts. Examples of the inorganic salt include hydrochloride, hydrobromide, sulfate, or phosphate, and examples of the organic salt include fumarate, succinate, oxalate, or lactate.

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

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

[0026] The aforementioned MPBS compounds are known compounds. These MPBS compounds can be synthesized, for example, by the methods described in Japanese Patent Publication No. 3-7263, Japanese Patent Publication No. 9-221479, European Patent Application Publication No. 390654, European Patent Application Publication No. 779283, U.S. Patent Publication No. 5053409, and U.S. Patent Publication No. 5990113, and are compounds readily available to those skilled in the art.

[0027] The aforementioned MPBS compounds may be substituted with compounds represented by the following general formula (I) or their salts, or their hydrates or solvates, as described in International Publication No. 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 an aryl group; R2 is a hydrogen atom, a C1-C6 alkyl group, or a C7-C 12The aralkyl group is an aralkyl group, and the 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, where n represents an integer from 1 to 4.

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

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

[0031] Examples of C1-C6 alkyl groups defined in R2 include the alkyl groups defined in R1 above. C7-C 12Examples of aralkyl groups include benzyl, phenethyl, and naphthylmethyl groups. These aralkyl groups may have one or more substituents selected from the group consisting of C1-C6 alkoxy groups such as cyano, nitro, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, tert-butoxy, pentyloxy, isopentyloxy, tert-pentyloxy, and hexyloxy groups; halogen atoms as defined by R1 above; alkyl groups as defined by R1 above; and amino groups.

[0032] Preferred examples of R2 include, for example, a hydrogen atom, a C1-C3 alkyl group, and a C7-C 12 Examples include aralkyl groups, and the aralkyl group may have one or more substituents selected from C1-C3 alkyl groups, C1-C3 alkoxy groups, and halogen atoms. More preferred examples include hydrogen atoms, C7-C 12 An aralkyl group is an example, and the aralkyl group may have one or more substituents selected from C1-C3 alkoxy groups, and is particularly preferably a hydrogen atom. Furthermore, in the above general formula (I), n is preferably 2.

[0033] In the keratosis pharmaceutical composition of the present invention, the MPBS may, for example, be in an ionized form. If the keratosis pharmaceutical composition of the present invention is, for example, a liquid as described later, and contains an aqueous solvent, a non-aqueous solvent, or a mixture thereof, the MPBS may be ionized regardless of whether it is an anhydrous, 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 proton solvent such as water. Specifically, the molecular species may take the form of a monovalent cation of the following formula (IIIa), a twin ion (twitter) having a positive and negative charge of the following formula (IIIb), or a monovalent anion of the following formula (IIIc). The monovalent cation molecular species can, for example, form a salt with an acid (anion), and the monovalent anion can, for example, form a salt with 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 as described above, for example. The pharmaceutical composition for keratosis of the present invention can be rephrased as, for example, a pharmaceutical composition used for the symptoms of keratosis, and more 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 described as, for example, the suppression of keratosis.

[0036] The keratosis targeted by the present invention is a disease characterized by hyperkeratosis. Hyperkeratosis refers to the proliferation of keratinocytes in the epidermis of the skin. The keratosis is a disease caused, for example, by an abnormality in the differentiation process of keratinocytes. The present invention is preferably applied to keratosis in which IL-22 signaling is involved. IL-22 signaling refers to a signal produced, for example, by IL-22 acting on the IL-22 receptor.

[0037] The keratosis covered by the present invention is not particularly limited and includes, for example, Darier's disease, psoriasis, and actinic keratosis.

[0038] Examples of psoriasis include psoriasis vulgaris, 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 symptoms to be suppressed are, for example, at least one selected from the group consisting of hyperkeratosis, abnormal keratosis, and acantholysis, and preferably includes hyperkeratosis. That is, the pharmaceutical composition for keratosis of the present invention may, for example, suppress hyperkeratosis, suppress abnormal keratosis, suppress acantholysis, suppress one symptom, suppress two or more symptoms, or suppress all of them. According to the pharmaceutical composition for keratosis of the present invention, for example, hyperkeratosis can be suppressed, hyperkeratosis and abnormal keratosis can be suppressed, hyperkeratosis and acantholysis can be suppressed, or hyperkeratosis, abnormal keratosis and acantholysis can be suppressed.

[0040] Hyperkeratosis is a proliferation of keratinocytes resulting from abnormal keratinization in the stratum corneum, stratum granulosum, and stratum spinosum of the skin, and can be observed as thickening of the stratum corneum. Examples of hyperkeratosis include hyperkeratosis involving IL-22 signaling. Abnormal keratinization refers to the individual keratinization of keratinocytes in the stratum spinosum of the skin, resulting in abnormal keratinocytes, and in the case of Darier's disease, it can be observed as rounded bodies. Acantholysis is a condition in which keratinocytes lose cell adhesion in the stratum spinosum of the skin, and can be observed as intercalation of the epidermis or blister formation, for example. Hyperkeratosis, abnormal keratinization, and acantholysis can be observed, for example, by taking a sample of the patient's diseased skin and following methods known to those skilled in the art.

[0041] In cases of keratosis caused by Darier's disease, pathological symptoms include, for example, hyperkeratosis in the stratum corneum, stratum granulosum, and stratum spinosum, abnormal keratinization in the stratum spinosum, and acantholysis in the stratum spinosum. The pharmaceutical composition for keratosis of the present invention can suppress these pathological symptoms simultaneously. In cases of keratosis caused by psoriasis, pathological symptoms include hyperkeratosis in the stratum corneum, stratum granulosum, and stratum spinosum. The pharmaceutical composition for keratosis of the present invention can suppress hyperkeratosis in these layers. In cases of actinic keratosis, pathological symptoms include, for example, atypia in the basal layer and / or stratum spinosum, and hyperkeratosis and abnormal keratinization in the epidermis. The pharmaceutical composition for keratosis of the present invention can suppress these symptoms in the epidermis.

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

[0043] The pharmaceutical composition for keratosis of the present invention can, for example, suppress or improve the clinical symptoms of the body caused by keratosis, and more specifically, it can suppress or improve at least one selected from the group consisting of lesional skin area, itching, pain, and odor in keratosis. According to the pharmaceutical composition for keratosis of the present invention, for example, by suppressing or improving the aforementioned pathological symptoms, the clinical symptoms of the body exemplified herein can be suppressed or improved.

[0044] The lesional skin area is, for example, the area where keratotic papules are observed. The pharmaceutical composition for keratosis of the present invention can suppress or improve the lesional skin area 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] Pruritus can be evaluated, for example, by using a numerical rating scale for pruritus, where the pruritus felt by the subject is assessed. In a numerical rating scale, for example, 0 indicates no pruritus, and 10 indicates the worst possible pruritus the subject can imagine, and the subject uses this scale to evaluate pruritus. The pharmaceutical composition for keratosis of the present invention can reduce the post-administration value of the numerical rating scale for pruritus 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 pre-administration value.

[0046] Pain can be evaluated, for example, by using a numerical rating scale for pain, where the pain felt by the subject is assessed. In a numerical rating scale, for example, 0 indicates no skin pain, and 10 indicates the worst skin pain the subject can imagine, and the subject uses this scale to evaluate skin pain. The pharmaceutical composition for keratosis of the present invention can reduce the post-administration value of the numerical rating scale for pain 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 pre-administration value.

[0047] Odor can be evaluated, for example, by using a numerical rating scale for odor, based on the odor perceived by the subject. In a numerical rating scale, for example, 0 indicates no odor, and 10 indicates the worst odor the subject can imagine, and the subject uses this scale to evaluate the odor. The pharmaceutical composition for keratosis of the present invention can reduce the value of 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 PGIS (Patient Global Impression of Severity), PGIC (Patient Global Impression of Change), CGIS (Clinician Global Impression of Severity), CGIC (Clinician Global Impression of Change), DLQI (Dermatology Life Quality Index), and Skindex-29 in the keratosis.

[0049] (1) The PGIS is a single questionnaire that evaluates the subject's overall impression of severity using a score, and the keratosis pharmaceutical composition of the present invention can improve the PGIS and improve the score. (2) PGIC is a single questionnaire that evaluates the degree of overall improvement in a subject's health status using a score, and the keratosis pharmaceutical composition of the present invention can improve PGIC and improve the score. (3) The CGIS is a single questionnaire in which a physician evaluates the overall impression of the severity of a subject using a score, and the keratosis pharmaceutical composition of the present invention can improve the CGIS and improve the score. (4) CGIC is a single questionnaire used by physicians to score the degree of overall improvement in a subject's health condition, and the keratosis pharmaceutical composition of the present invention can improve CGIC and improve the score. (5) The DLQI is a questionnaire in which subjects evaluate each question regarding their quality of life using a score. The pharmaceutical composition for keratosis of the present invention can improve the DLQI and thus improve the score. (6) Skindex-29 is a questionnaire in which subjects evaluate each question regarding their quality of life with a score. The pharmaceutical composition for keratosis of the present invention can improve Skindex-29 and improve the score.

[0050] The method of administering the pharmaceutical composition for keratosis of the present invention is not particularly limited and may be administered orally or parenterally. Parenteral administration may include, for example, transdermal, subcutaneous, intravenous, intra-arterial, intraperitoneal, intranasal, and intestinal administration.

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

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

[0053] The pharmaceutical composition for keratosis of the present invention may, for example, contain only the active ingredient, or may 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, for example, depending on the dosage form. Examples of additives include carriers, excipients, stabilizers, lubricants, sweeteners, preservatives, suspending agents, dispersants, thickeners, pH adjusters, antifoaming agents, and fragrances. Examples of carriers include liquids, solids, gels, and creams.

[0054] The target recipients of the pharmaceutical composition for keratosis of the present invention are not particularly limited, and include, for example, humans and non-human animals, with humans being preferred. Non-human animals include, for example, non-human mammals such as mice, rats, rabbits, and horses.

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

[0056] When administering the pharmaceutical composition for keratosis of the present invention orally, the following conditions are exemplified. The following examples can be appropriately adjusted according to the patient's information, etc., for infants, toddlers, children, adults, or the elderly. The dosage of the MPBS can be expressed, for example, as the dosage of the MPBS converted to the MPBS anhydrous. The dosages exemplified below are the dosages of the MPBS converted to the MPBS anhydrous, and as a specific example, the dosage of the MPBS monohydrate converted to the MPBS anhydrous.

[0057] • Daily dose of the aforementioned MPBS (calculated as the amount of anhydrous MPBS) Lower limit: For example, 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: For example, 300mg, 400mg, 500mg, 600mg, 800mg, 1000mg, or 1200mg Range: For example, 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 dosages: 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 doses per day: For example, 1 to 3 times, preferably 1 or 2 times, more preferably 2 times. • Frequency of administration (interval): For example, daily

[0058] When administering the keratosis-fighting pharmaceutical composition of the present invention orally, the timing of administration can be freely set, for example, before meals, during meals, immediately after meals, after meals, between meals, upon waking, or before going to bed, and administration between meals is preferable. Inter-meal administration of the keratosis-fighting pharmaceutical composition of the present invention means, for example, that the time interval between meal intake and administration is at least 1 hour, preferably at least 2 hours, after meal intake, and / or, for example, that the time interval between administration and the intake of the next meal is at least 1 hour, 2 hours, 3 hours, 4 hours, or 4.5 hours, preferably 1 hour, after the previous meal. That is, the keratosis-fighting pharmaceutical composition of the present invention is administered, for example, between 1 or 2 hours after meal intake and 1 hour, 2 hours, 3 hours, 4 hours, or 4.5 hours before the intake 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, as described above, is a disease that causes hyperkeratosis. For this reason, the present invention can also be described as, for example, a pharmaceutical composition for hyperkeratosis that treats or prevents hyperkeratosis. Furthermore, the present invention can also be described as, for example, a pharmaceutical composition for hyperkeratosis and acantholysis that simultaneously treats or prevents hyperkeratosis and acantholysis. Furthermore, the present invention can also be described as a pharmaceutical composition for hyperkeratosis, acantholysis, and abnormal keratosis that simultaneously treats or prevents hyperkeratosis, acantholysis, and abnormal keratosis.

[0060] [Methods of treating or preventing keratosis] The present invention relates to a method for treating or preventing keratosis, comprising the step of administering 5-methyl-2-(1-piperazinyl)benzenesulfonic acid to a subject. As stated above, 5-methyl-2-(1-piperazinyl)benzenesulfonic acid (MPBS) may be any of the MPBS derivatives, i.e., MPBS anhydride, MPBS salt, MPBS hydrate, and / or MPBS solvate. Unless otherwise specified, the present invention relates to a method for treating or preventing keratosis, which is hereinafter referred to as the "method for treating / preventing keratosis of the present invention." The method for treating / preventing keratosis of the present invention is characterized by the administration of the MPBS derivatives, and other conditions and steps are not particularly limited.

[0061] In the treatment and prevention method 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 MPBS is, for example, the administration of the pharmaceutical composition for keratosis of the present invention. In the present invention, the MPBS, its composition, and the method of administration can be described by reference to the description of the pharmaceutical composition for keratosis of the present invention.

[0063] [Use of MPBS-type products] The present invention relates to 5-methyl-2-(1-piperazinyl)benzenesulfonic acid for use in the treatment or prevention of keratosis. As stated above, 5-methyl-2-(1-piperazinyl)benzenesulfonic acid (MPBS) may be any of the MPBS derivatives, i.e., MPBS anhydride, MPBS salt, MPBS hydrate, and / or MPBS solvate. In the present invention, the MPBS derivatives, their compositions, and methods of use thereof can be described by reference to the description in the above-mentioned pharmaceutical composition for keratosis of the present invention.

[0064] Furthermore, the present invention 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 MPBS compounds, i.e., the MPBS anhydride, the MPBS salt, the MPBS hydrate, and / or the MPBS solvate. In the present invention, the MPBS compounds, their compositions, and methods of use thereof can be described by reference to the description of the pharmaceutical composition for keratosis of the present invention.

[0065] [Medicinal compositions for agonisthraxation and their uses] The pharmaceutical composition for acantholysis of the present invention is characterized by containing 5-methyl-2-(1-piperazinyl)benzenesulfonic acid. As described above, the form of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid (MPBS) is not limited and may be any of the MPBS compounds, i.e., the MPBS anhydride, 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 MPBS compounds or two or more of them. The MPBS compounds can be used for the treatment or prevention of acantholysis. The pharmaceutical composition for acantholysis of the present invention is characterized by containing the MPBS compounds, but other components and conditions are not particularly limited. It is preferable that the pharmaceutical composition for acantholysis of the present invention contains the MPBS compounds 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 acantholytic pharmaceutical composition of the present invention can treat or prevent acantholysis. The acantholytic pharmaceutical composition of the present invention may be used, for example, for therapeutic purposes, for preventive purposes, or for both therapeutic and preventive purposes. Hereinafter, the term "treatment / prevention" can be interpreted as meaning treatment, prevention, or treatment and prevention. The acantholytic pharmaceutical composition of the present invention allows for safe treatment / prevention with fewer side effects, for example. Diseases that cause acantholysis include, for example, Darier's disease, actinic keratosis, Glover's disease, acantholytic vesicles, transient acantholytic dermatosis, pemphigus, keratoacanthoma, squamous cell carcinoma, varicella, herpes zoster, and herpes simplex.

[0067] In the acantholytic pharmaceutical composition of the present invention, the MPBS class, its composition, and method of use are not particularly limited, and for example, the description in the above-mentioned pharmaceutical composition for keratosis of the present invention can be referenced. When referencing, "keratosis" and "symptoms of keratosis" can be read as "acantholytic."

[0068] The present invention provides a method for treating or preventing acantholysis, comprising the step of administering 5-methyl-2-(1-piperazinyl)benzenesulfonic acid to a subject. As described above, 5-methyl-2-(1-piperazinyl)benzenesulfonic acid (MPBS) is not limited in form and may be any of the MPBS derivatives, i.e., the MPBS anhydride, the MPBS salt, the MPBS hydrate, and / or the MPBS solvate. Unless otherwise specified, the present invention provides a method for treating or preventing acantholysis, hereinafter referred to as the "method for treating / preventing acantholysis of the present invention." The method for treating / preventing acantholysis of the present invention is characterized by the administration of the MPBS derivatives, and other conditions and steps are not particularly limited. The subjects targeted by the present invention are, for example, patients who have developed or are likely to 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, their compositions, and methods of administration thereof are not particularly limited, and the description of the pharmaceutical composition for keratosis of the present invention can be incorporated by reference. When incorporating such descriptions, "keratosis" and "symptoms of keratosis" can be replaced with "acantholysis."

[0070] The present invention relates to 5-methyl-2-(1-piperazinyl)benzenesulfonic acid for use in the treatment or prevention of acantholysis. As stated above, 5-methyl-2-(1-piperazinyl)benzenesulfonic acid (MPBS) may be any of the MPBS derivatives, i.e., MPBS anhydride, MPBS salt, MPBS hydrate, and / or MPBS solvate, without limiting its form. In the present invention, the MPBS derivatives, their compositions, and methods of use can be described by reference to the description in the above-mentioned pharmaceutical composition for keratosis of the present invention. When referenced, "keratosis" and "symptoms of keratosis" can be read as "acantholysis."

[0071] Furthermore, the present invention relates to the use of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid in the manufacture of a pharmaceutical composition for acantholysis. As described above, 5-methyl-2-(1-piperazinyl)benzenesulfonic acid (MPBS) is not limited in form and may be any of the MPBS compounds, i.e., the MPBS anhydride, the MPBS salt, the MPBS hydrate, and / or the MPBS solvate. In the present invention, the MPBS compounds, their compositions, and methods of use can be described by reference to the description in the pharmaceutical composition for keratosis of the present invention. When referencing, "keratosis" and "symptoms of keratosis" can be read as "cantholysis." [Examples]

[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] Suppression of epidermal thickening in an in vitro hyperkeratosis model The inhibitory effect of compound A on epidermal thickening in an epidermal hyperkeratosis model was evaluated.

[0074] When IL-22 is brought into contact with epidermal tissue, thickening is induced in the stratum spinosum, leading to 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 study design> An IL-22-induced human keratinocyte epidermal hyperkeratosis model was constructed using the following method. Human primary keratinocytes (foreskin, Lonza, 00192906) pooled from three lots of Caucasian neonatal donors were used as cells. Epilife culture medium (Thermo Fisher Scientific, M-EPI-500-A) was used to culture the human primary keratinocytes. Three-dimensional culture of the human primary keratinocytes was initiated using a gas-liquid interface culture insert and a 24-well plate (Day 0), and in vitro epidermal tissue was prepared after 14 days of culture (Day 0 to Day 14). The culture environment was maintained at 37°C and 5% CO2. Fourteen days after the start of the three-dimensional culture (Day 0) (Day 14), an aqueous solution of compound A (solvent: water) and an IL-22 solution (solvent: phosphate-buffered saline, hereafter sometimes referred to as PBS) were added to the culture medium of the in vitro epidermal tissue, and the tissue was cultured in three dimensions for a further three days (Day 14 to Day 17). In the culture medium, the final concentration of compound A 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 basically changed 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 aqueous solution of compound A and the IL-22 solution were added at the time of change. Seventeen days after the start of the three-dimensional culture (Day 17), the in vitro epidermal tissue was sampled and used for evaluation. In this specification, "final concentration" for each test substance solution and reagent means the concentration in the culture medium at the final stage after all test substance solutions and reagents have been added.

[0076] Each model group of the in vitro epidermal tissue was configured 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 with a 4% formaldehyde solution, dehydrated, and embedded in paraffin. From the paraffin-embedded tissue, sections 6 μm thick were prepared using a slicer (Leica microtome RM2245), and these were fixed to glass slides to obtain epidermal tissue slides.

[0079] For pathological evaluation, the epidermal tissue slides were stained with hematoxin-eosin (HE staining). For pathological analysis, three images were captured for each stained sample using an optical microscope (Nikon Eclipse (Ni-E)) connected to a digital camera (DS-Ri2). Image analysis using NIS-Elements AR software (Nikon) was performed to measure the epidermal thickness of each of the three images for each sample, and the average value was used as the representative value for that sample. Note that since hyperkeratosis caused by IL-22 is a factor in thickening of the stratum spinosum, and although hyperkeratosis of the stratum corneum occurs in 3D culture, it peels off each time, making quantitative evaluation difficult, so the stratum corneum was excluded from the measurement of epidermal thickness. The number of samples was 3 to 9 epidermal tissue slides per experiment (1 group). The results of the four example groups 1-1, 1-2, 1-3, and 1-4 in Table 1 and the control group 1-C were then combined and analyzed to evaluate the effect of compound A. Using the mean value of the normal group 1-N (without IL-22 supplementation) as 100%, the relative value (%), as well as their mean and standard deviation, were calculated for each group's measurements.

[0080] <Statistical analysis> Statistical analysis was performed using MATLAB®, with a significance level of 5% (two-sided). The pharmacological effects of compound A were assessed by double comparison between control group 1-C (IL-22 added / compound A not added) and each example group (IL-22 added / compound A added), using the Aspin-Welch t-test. The p-value was corrected for multiple comparisons using Bonferroni's method.

[0081] These results are shown in Figure 1. Figure 1 is a graph showing the epidermal thickness in in vitro epidermal tissue. In Figure 1, the vertical axis represents the thickness of reconstructed human epidermis, and is a relative value (%) with the normal group 1-N without IL-22 added 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 without IL-22. In contrast, in the example group to which compound A was added along with IL-22, the epidermal thickness decreased in a concentration-dependent manner compared to the control group 1-C. From these results, it was found that compound A can suppress IL-22-induced hyperkeratosis in the epidermis in a concentration-dependent manner at concentrations of at least 1 μmol / L or higher. Furthermore, the hyperkeratosis-suppressing effect of compound A was statistically significant at concentrations of 10 μmol / L or higher.

[0082] [Example 2] Suppression of acantholysis and abnormal keratinization in an in vitro epidermal acantholysis model The inhibitory effects of compound A on acantholysis and abnormal keratinization in an epidermal acantholysis model were evaluated. In this example, acantholysis was evaluated using intercellular adhesion as an indicator and pathological evaluation, while abnormal keratinization was evaluated using the frequency of concentrated nuclei as an indicator and pathological evaluation.

[0083] <Test cells and study design> A human keratinocyte epidermal acantholysis model was constructed using the following method. Human primary keratinocytes (foreskin, Lonza, 00192906) pooled from three lots of Caucasian neonatal donors were used as the cells. EpiLife® culture medium (Thermo Fisher Scientific, M-EPI-500-A) was used to culture the aforementioned human primary keratinocytes. Thasigargin was used to induce acantholysis and abnormal keratinization.

[0084] The aforementioned primary human keratinocytes were cultured in three dimensions using a gas-liquid interface culture insert and a 24-well plate (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. Fourteen days after the start of the three-dimensional culture (Day 0) (Day 14), an aqueous solution of compound A (solvent: water) and a thapsigargin solution (solvent: DMSO) were added to the culture medium of the in vitro epidermal tissue, and the tissue was cultured in three dimensions for a further 3 days (Day 14-Day 17). In the culture medium, the final concentration of compound A was 0, 1, 3, 10, or 30 μmol / L, and the final concentration of thapsigargin was 0 or 30 nmol / L. In the three-dimensional culture, the culture medium was basically changed daily from Day 0 to Day 17 (except on weekends, Saturday and Sunday), and from Day 14 to Day 17, the culture medium was changed daily, and the aqueous solution of compound A and the thapsigargin solution were also added during the 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 determined 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 cell adhesion strength based on biotin diffusion, acantholysis score evaluation based on pathological observation, and immunofluorescence staining of cell adhesion marker proteins. Abnormal keratinization was evaluated by the frequency of concentrated nuclei based on pathological observation and immunofluorescence staining of keratinocyte differentiation marker proteins. Specifically, the in vitro epidermal tissue sampled from the same well of the 24-well plate was used to prepare slides, which were then evaluated.

[0088] (1) Biotin diffusion The sampled in vitro epidermal tissue was rinsed twice with 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 impermeability marker molecule. In this example, in particular, to confirm cell adhesion in the spinous layer of the epidermal tissue, the biotin marker was added to the basal layer side 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 PBS / CaCl2 (1 mmol / L) / Glycine (100 mmol / L) solution (hereinafter the same). Here, sample numbers were coded to blind the subsequent steps. The in vitro epidermal tissue was then fixed with a 4% formaldehyde solution, dehydrated, and embedded in paraffin. From the paraffin-embedded tissue, 6 μm thick sections were prepared using a slicer (Leica microtome RM2245), and these were fixed onto glass slides to form epidermal tissue slides.

[0089] When acantholysis occurs, the amount of biotin in the epidermal layer relatively increases due to a decrease in cell adhesion. Therefore, the intercellular adhesion of the epidermis in the in vitro epidermal tissue was evaluated by detecting the biotin marker using the prepared epidermal tissue slides. Specifically, the epidermal tissue slides were first deparaffinized, rehydrated, and then left to stand in the presence of fluorescently labeled streptavidin (Thermo Fisher Scientific, S32354) to detect the biotin (the molecular marker) that had diffused into the epidermal layer of the in vitro epidermal tissue.

[0090] Specifically, under dark conditions at 4°C, three images were captured for each sample (the epidermal tissue slide) using an optical microscope (Nikon Eclipse (Ni-E)) connected to a digital camera (DS-Ri2). Image analysis was performed using NIS-Elements AR software (Nikon), quantifying the biotin fluorescence intensity 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 The 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 by the ROI surface area A. For each sample, the average value was calculated from the biotin fluorescence intensity of each of the three images, and this average value was used as the representative value for the three images. The number of samples was nine epidermal tissue slides per experiment (one in vitro epidermal tissue). The measured values ​​of the normal group 2-N without thapsigargin were set as 100%, and the relative values ​​(%), their average values, and standard deviations were calculated for the measured values ​​of each group.

[0091] (2) Pathological evaluation The sampled in vitro epidermal tissue was rinsed twice with the PBS / CaCl2 solution, fixed with a 4% formaldehyde solution, dehydrated, and then embedded in paraffin. From the paraffin-embedded tissue, 6 μm thick sections were prepared using a slicer (Leica microtome RM2245), and these were fixed to glass slides to form epidermal tissue slides.

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

[0093] The occurrence of abnormal keratinization relatively increases the frequency of concentrated nuclei. Therefore, for the pathological evaluation of abnormal keratinization, the aforementioned pathological evaluation samples were used, and the number of concentrated nuclei in the field of view was counted. The number of concentrated nuclei was counted in 3 fields (3 images) for each sample, and the average value was used as the representative value for that sample. The number of samples was 9 epidermal tissue slides per experiment (1 group). In addition, for the pathological evaluation of acantholysis, the aforementioned pathological evaluation samples were used, and observations were made to qualitatively record the presence or absence of findings of acantholysis. The qualitative assessment of findings of acantholysis 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 mean and standard deviation of the score values ​​for each group were calculated. 0: No cracks 1: There is a crack in one place. 2: There are two fissures. 3: There are three or more fissures.

[0094] (3) Immunofluorescence staining Immunofluorescence staining was performed on the aforementioned epidermal tissue slides using antibodies corresponding to each target marker.

[0095] (i) As in (2) above, sections were fixed to slides to prepare epidermal tissue slides. The epidermal tissue slides were deparaffinized and rehydrated, and then the antigens were activated by heating with 0.01 mol / L sodium citrate buffer (pH 6.0). After rinsing the antigen-activated epidermal tissue slides with PBS, to avoid nonspecific labeling, they were saturated with PBS / 0.1% Tween 20 solution containing 5% normal goat serum (NGS) for 1 hour in a constant temperature and humidity bath. These epidermal tissue slides were left to stand overnight in a constant temperature and humidity bath at 4°C with primary antibodies (anti-filaggrin (FLG) antibody and anti-cytokeratin 10 (K-10) antibody). After continuous washing of the epidermal tissue slides with PBS / 0.1% Tween 20 solution and PBS, the epidermal tissue slides were left to stand for 1 hour in a constant temperature and humidity bath at room temperature with anti-mouse secondary antibody and anti-rabbit secondary antibody conjugated with Alexa Fluor dye. Finally, the epidermal tissue slides were rinsed with PBS. The primary and secondary antibodies used for co-immunostaining of FLG / K-10, a keratinocyte differentiation-related protein, are as shown in the table below.

[0096] [Table 3]

[0097] (ii) The epidermal tissue slides were deparaffinized and rehydrated in the same manner as in (i) above, and the antigens were activated by heating with pepsin solution (Sigma, R2283) at 56°C for 15 minutes. After rinsing the antigen-retrieved epidermal tissue slides with PBS, they were saturated with PBS / 0.1% Tween20 solution containing 2% bovine serum albumin (BSA) for 1 hour in a constant temperature and humidity bath to avoid nonspecific labeling. These epidermal tissue slides were left to stand overnight in a constant temperature and humidity bath at 4°C with primary antibodies (anti-desmoglein 1 (DSG1) antibody and anti-claudin 1 (CLDN1) antibody). After sequential washing of the epidermal tissue slides with PBS / 0.1% Tween20 solution and PBS, the epidermal tissue slides were left to stand for 1 hour in a constant temperature and humidity bath at room temperature with anti-goat secondary antibodies and anti-rabbit secondary antibodies conjugated with Alexa Fluor dye. Finally, the epidermal tissue slides were rinsed with PBS. The primary and secondary antibodies used for co-immunostaining of the cell adhesion-related protein DSG1 / CLDN1 are as shown in the table below.

[0098] [Table 4]

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

[0100] Specifically, three images of each sample (the epidermal tissue slide) were taken using an optical microscope (Nikon Eclipse (Ni-E)) 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) to quantify the fluorescence staining area in three images per sample. For each image, FLG was manually demarcated with the epidermal tissue including the stratum corneum as the region of interest (ROI), and K-10, DSG1, and CLDN1 were manually demarcated with the epidermal tissue excluding the stratum corneum as the region of interest (ROI), and their surface area A was measured (unit: μm 2 ). Next, the area B of the stained region within the demarcated region of interest (ROI) was measured (unit: μm 2 ). The immunostaining area was quantified as the numerical value (B / A) obtained by dividing the stained area B by the ROI surface area A, that is, the relative value (%) of the stained area, and the mean value and standard deviation of each group were calculated. For each sample, the mean value was obtained 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 significance level of 2.5% for Williams multiple comparison tests (one-sided) and 5% for other testing methods (two-sided). The success of the formation of the epidermal acantholysis model by thapsigargin addition was determined by comparing two groups: normal group 2-N (no thapsigargin / no compound A) and control group 2-C (thapsigargin added / no compound A). Student's t-test was used for the statistical analysis. For in vitro epidermal tissue where no significant change was observed, it was determined that the formation of the epidermal acantholysis model had failed, and no further analysis was performed. The pharmacological effect of compound A was determined by comparing control group 2-C (thapsigargin added / no compound A) with each example group (thapsigargin added / compound A added). Williams multiple comparison tests were used for the statistical analysis. For the evaluation of immunofluorescence staining, only example group 2-3 (compound A, 10 μmol / L) was used, so Student's t-test was used as the statistical assay.

[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 representing the relative value (%) of biotin. Figure 3 is a graph showing the results of enriched nuclei frequency, an indicator of abnormal keratinization, with the vertical axis representing the number of enriched nuclei in the image. Figure 4 is a graph showing the results of pathological evaluation of acantholysis, with the vertical axis representing the acantholysis score. Figures 5A and 5B are graphs showing the results of keratinocyte differentiation markers associated with abnormal keratinization, with Figure 5A showing the results for FLG and Figure 5B showing the results for K-10, where the vertical axis represents the relative value (%) of 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, where the vertical axis represents the relative value (%) of stained area.

[0103] First, let's discuss acantholysis. As shown in Figure 2, the amount of biotin in the epidermal layer, an indicator of acantholysis, increased significantly in the control group 2-C, which was treated with thapsigargin, compared to the normal group 2-N, which was not treated with thapsigargin. This indicates that the addition of thapsigargin caused acantholysis, and the decrease in intercellular adhesion led to an increase in the amount of biotin in the epidermal layer. In contrast, in the example group, which was treated with compound A along with thapsigargin, the amount of biotin in the epidermal layer decreased in a concentration-dependent manner at least above 1 μmol / L compared to the control group 2-C. Furthermore, as shown in Figure 4, similar results were obtained in the pathological evaluation. That is, compared to the normal group 2-N, the acantholysis score in the control group 2-C increased significantly with the addition of thapsigargin, but in the example group, the addition of compound A suppressed the increase in the acantholysis score in a concentration-dependent manner at least above 1 μmol / L. Furthermore, as shown in Figures 6A and 6B, similar results were obtained in immunofluorescence staining of cell adhesion-related proteins DSG1 and CLDN1. Compared to the normal group 2-N, the immunofluorescence staining area in the control group 2-C decreased with the addition of thapsigargin. This indicates that the addition of thapsigargin weakened or disrupted desmosomes and tight junctions, which are one of the modes of cell adhesion. In contrast, in the group of examples in which compound A was added along with thapsigargin, the immunofluorescence staining area increased compared to the control group 2-C. This indicates that compound A was able to normalize the abnormal cell adhesion induced by thapsigargin.

[0104] These results indicate that the addition of compound A can improve the attenuation of intercellular adhesion induced by thapsigargin in in vitro epidermal tissue in a concentration-dependent manner, that is, it suppresses acantholysis. Specifically, compound A was found to be statistically significant in suppressing the decrease in intercellular adhesion at 10 μmol / L, and statistically significant in suppressing the increase in the pathological evaluation score of acantholysis from 1 μmol / L.

[0105] Next, we will discuss abnormal keratinization. As shown in Figure 3, the number of condensed nuclei, an indicator of abnormal keratinization, increased significantly in the control group 2-C, to which thapsigargin was added, compared with the normal group 2-N, to which thapsigargin was not added. This means that the addition of thapsigargin caused abnormal keratinization and increased the number of condensed nuclei. In contrast, in the group of examples to which compound A was added along with thapsigargin, the amount of biotin in the epidermal layer decreased in a concentration-dependent manner, at least at compound A concentrations of 1 μmol / L or higher, compared with the control group 2-C. Furthermore, as shown in Figures 5A and 5B, immunofluorescence staining of the differentiation marker FLG expressed in keratinocytes of the granular layer and stratum corneum, and immunofluorescence staining of the differentiation marker K-10 expressed in keratinocytes of the spinous layer and granular layer, showed that the immunofluorescence staining area in the control group 2-C increased and decreased statistically significantly with the addition of thapsigargin, compared with the normal group 2-N. In other words, the addition of thapsigargin resulted in heterogeneous differentiation of keratinocytes forming the stratum spinosum, creating a state prone to abnormal keratinization. In contrast, in the group of examples in which compound A was added at a concentration of 10 μmol / L along with thapsigargin, the immunofluorescence staining area of ​​FLG and K-10 were statistically significantly reduced and increased, respectively, compared to the control group 2-C. This indicates that compound A was able to normalize the abnormal keratinocyte differentiation induced by thapsigargin.

[0106] These results indicate that the addition of compound A can improve, in a concentration-dependent manner, the increased frequency of thapsigargin-induced enriched nuclei in in vitro epidermal tissue, meaning that abnormal keratinization is suppressed. Specifically, compound A was found to be statistically significant in suppressing the increase of enriched nuclei, an indicator of abnormal keratinization, from 3 μmol / L.

[0107] Furthermore, these results show that compound A can suppress both abnormal keratinization and acantholysis. Therefore, it was found that even in keratosis, not only in diseases that cause only one of either abnormal keratinization or acantholysis, but also in diseases that cause both (for example, Darier's disease), administering compound A alone can suppress both symptoms simultaneously. In addition, in Example 1, a hyperkeratosis model was created using the same keratinocytes, and the suppression of hyperkeratosis by compound A was also confirmed. Thus, it can be said that compound A shows an inhibitory effect on all three major pathological symptoms of Darier's disease, namely hyperkeratosis, acantholysis, and abnormal keratinization.

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

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

[0110] <Test cells and study design> Seven-week-old male BALB / c mice (Charles River, Japan) were used. The general condition of the mice was observed for a 6-day quarantine and acclimatization period. After the acclimatization period (at 8 weeks of age), the weight of the mice was measured, and they were divided into three groups using stratified randomization so that the mean weight was uniform across all groups. In one of the example groups, the hair on the backs of the mice was shaved, and three days later, under 2% inhalation anesthesia using isoflurane (Mylan Pharmaceuticals Co., Ltd.) as an anesthetic, imiquimod 5% cream (Mochida Pharmaceutical Co., Ltd.) was applied to the skin of the backs of the mice once a day for a total of 4 days to induce psoriasis-like dermatitis in the mice. Psoriasis-like dermatitis presents with hyperkeratosis as a pathological symptom. The amount of imiquimod 5% cream applied per application was 62.5 mg (3.125 mg as imiquimod). After applying imiquimod 5% cream, the application site was gently wiped with cotton wool soaked in lukewarm water approximately 4 hours after each application. Simultaneously, compound A was administered orally 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 two daily doses were administered with the first dose before the application of imiquimod 5% cream and the second dose at least 10 hours after the first dose of the day. For oral administration of compound A, a 3 mg / mL solution of compound A (solvent: sterile water for injection) was used.

[0111] The remaining two groups were designated as the normal group and the control group. In the normal group 3-N, the hair on the back was shaved, but no induction with imiquimod was performed, nor was compound A administered. In the control group 3-C, induction with imiquimod was performed in the same manner as in the example group, but compound A was not administered, and 10 mL / kg of a 1% Tween 80 solution without compound A was orally administered.

[0112] [Table 5]

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

[0114] <Statistical analysis> Statistical analysis was performed using SAS, with a significance level of 5% (two-sided) for group comparisons. The pharmacological effect of compound A was determined by comparing the control group 3-C (imiquimod application / no administration of compound A) with the example group (imiquimod application / administration of compound A), using 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 dorsal skin thickness (mm). As shown in Figure 7, compared to 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, in example group 3-1, in which compound A was administered on the same day as imiquimod application, the dorsal skin thickness was statistically significantly reduced compared to the control group 3-C. These results indicate that compound A can suppress imiquimod-induced skin thickening, i.e., hyperkeratosis.

[0116] [Example 4] Involvement of IL-22 in Darier's disease Gene expression analysis was performed on human formalin-fixed paraffin-embedded (FFPE) skin from patients with Darier's disease and psoriasis vulgaris. Specifically, the involvement of IL-22 in Darier's disease, another keratosis, was evaluated by comparing it with psoriasis vulgaris, one of the keratosis disorders in which IL-22 is known to be involved.

[0117] <Test cells and study design> As shown below, image acquisition and library preparation were performed from FFPE block samples with a diagnostic history (Auras Biosciences, Inc.). Patient-derived FFPE block samples were obtained from a chest skin biopsy specimen of a Black female in her 70s from a patient with psoriasis vulgaris, and from a back skin biopsy specimen of a White female in her 40s from a patient with Darier's disease. From the FFPE block samples, 5 μm thick sections were cut using a slicer, and a section adhesion test was performed using Visium Test Slides (10X Genomics). After confirming that there was little peeling or curling of the sections by the section adhesion test, slides were prepared using a commercially available kit (Visium Spatial for FFPE Gene Expression Kit, Human Transcriptome, 10X Genomics). After staining the slides with HE, digital slide data was acquired from the slides using a scanner (Nanozoomer, Hamamatsu Photonics K.K.). Furthermore, a library was prepared from the aforementioned slides using a commercially available kit (Visium Spatial for FFPE Gene Expression Starter Kit, Human Transcriptome, 10X Genomics). Next-generation sequencing analysis was then performed on the library.

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

[0119] These results are shown in Table 6. As shown in the IL22RA1 epidermal expression ratio in Table 6, patients with Darier's disease showed strong gene expression of IL22RA1 (IL-22 receptor) 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. From this, it was found that IL-22 is involved in Darier's disease, just as it is in psoriasis vulgaris.

[0120] [Table 6]

[0121] From the results above, the involvement of IL-22 was confirmed in both psoriasis vulgaris and Darier's disease, which are major keratosis disorders. In Example 1, it was demonstrated that IL-22 induced hyperkeratosis, and that compound A suppressed this hyperkeratosis. From this, it can be said that the inhibitory effect of compound A on hyperkeratosis in Example 1 is effective against both psoriasis vulgaris and Darier's disease.

[0122] [Example 5] Evaluation of pharmacokinetics The pharmacokinetics of compound A were evaluated using the aforementioned compound A. In this example, the amount of compound A used is shown in terms of the amount of MPBS anhydrous (hereinafter sometimes referred to as compound B in this example).

[0123] A Phase 1, double-blind, placebo-controlled, dose-escalation repeated oral administration study was conducted in healthy men and women. In this study, subjects were evaluated by orally administering compound A once or twice daily. The single dose of compound A was 50 mg or 100 mg, equivalent to the amount of compound B. Twenty-four subjects (16 men and 8 women) were enrolled in the study, and all 24 completed the study. Table 7 below shows the study groups and treatments. The study groups consisted of three groups, A to C, as shown in Table 7. Each study group consisted of eight subjects; six received compound A and two received placebo. For the administration of compound A, a tablet containing 5 mg of compound A (equivalent to the amount of compound B) was used (hereinafter referred to as compound A 5 mg tablet). Test groups A and B received 10 tablets of compound A 5 mg or placebo per dose, while test group C received 20 tablets of compound A 5 mg or placebo per dose.

[0124] [Table 7]

[0125] Each subject received a total of 16 doses of either compound A 5 mg tablet or placebo tablet during the study period. These 16 doses were administered once in the morning on Day 1, once in the morning and once in the evening from Day 3 to Day 9, and once in the morning on Day 10. From Day 3 to Day 10, compound A 5 mg tablet or placebo tablet was 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 provided approximately 2 hours, 4 hours, 7 hours, and 10 hours after the morning dose, respectively. Meals were not to exceed 2500 kilocalories for men and 2000 kilocalories for women. Water intake was prohibited for 2 hours after administration on Day 1 and Day 10, but it could be consumed freely at any other time. The investigational drug (compound A 5mg tablet or placebo tablet) was taken with 200mL of water while standing. On Day 1 and Day 10, lying on one's back was prohibited for 2 hours after administration, unless following the study procedure or instructed by a physician.

[0126] Blood samples were taken over time from test groups A and B (hereinafter referred to as the 50 mg group), in which compound A was administered once at a dose of 50 mg equivalent to compound B, and from test group C (hereinafter referred to as the 100 mg group), in which compound B was administered once at a dose of 100 mg equivalent to compound B, and the plasma concentration of compound B was measured. The average plasma concentrations from Day 9 to Day 10 are shown in Figure 8 and Table 8. Figure 8 is a graph showing the average plasma concentration of compound B, where the vertical axis represents the plasma concentration of compound B (ng / mL), and the horizontal axis represents the elapsed time since administration on Day 9, with the time immediately before administration on Day 9 set as 0 hours. The results for the 50 mg group are the average values ​​for 12 subjects from test groups A and B excluding the placebo group, and the results for the 100 mg group are the average values ​​for 6 subjects from test group C excluding the placebo group.

[0127] [Table 8]

[0128] In the results of the thapsigargin-induced human keratinocyte epidermal acantholysis model in Example 2, the effective drug concentration of compound A was approximately 1 μmol / L (approximately 256 ng / mL) to approximately 10 μmol / L (approximately 2560 ng / mL). Therefore, based on the concentration profile of compound B in human plasma after administration of compound A shown in Figure 8 (drug concentration profile) and the effective drug concentration in Example 2, the clinically effective dose of compound A is, for example, approximately 50 mg or more twice a day orally, equivalent to the amount of compound B, and furthermore, 50 to 200 mg twice a day orally.

[0129] [Example 6] In vivo clinical trial The compound A is administered to patients with Darier's disease, and a Phase 2 clinical trial is conducted. For the administration of compound A, a 50 mg tablet of compound A, which contains 50 mg of compound A equivalent to the amount of compound B, is used.

[0130] <Subjects> Darier disease patients aged 18 to 75 <Exam Overview> Randomized double-blind clinical trial • Placebo administration: 24 cases • 100mg dose group: 12 patients One 50 mg tablet of compound A is administered orally twice a day (100 mg / day of compound B). • 200mg dose group: 24 patients Two 50 mg tablets of compound A are administered orally twice a day (200 mg / day of compound B). • 400mg group: 24 patients Four 50 mg tablets of compound A are administered orally 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 will be conducted on the following: Investigator's Global Assessment (IGA) score, Body Surface Area, pruritus, pain and odor scores, 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 the placebo group, administration of compound A is expected to result in improvement in at least one of the aforementioned test items.

[0132] In the aforementioned Examples 1-4, the inhibitory effect on epidermal thickening in epidermal hyperkeratosis and the inhibitory effect on acantholysis and abnormal keratinization in epidermal acantholysis were confirmed. Therefore, it can be said that various scores related to symptoms in patients with Darier's disease are improved by the administration of compound A.

[0133] [Example 7] Evaluation of the effect of diet on pharmacokinetics The effect of food on the pharmacokinetics of compound A was evaluated by administering it under fasting or postprandial conditions.

[0134] A Phase 1, double-blind, placebo-controlled, four-phase crossover study was conducted in healthy men. In this study, subjects received a single dose of compound A in each administration period (1-4) and were evaluated. The single dose of compound A was equivalent to 25 mg of compound B. Nine subjects were enrolled in the study. Table 9 below shows the test groups and the details of the treatment. The test groups consisted of three groups, Test Groups 1-3, 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 tablet of compound A described in Example 5.

[0135] [Table 9]

[0136] Each subject received either a 5mg tablet of compound A or a placebo tablet orally once on the morning of Day 1 of the study, for each administration period. Participants were prohibited from eating until 4 hours after administration and from drinking water until 2 hours after administration. The investigational drug (5mg tablet of compound A or placebo tablet) was taken while standing with 240mL of water. In each administration period, dinner was provided on Day 0, the day before the study, and lunch, afternoon snack, and dinner were provided on Day 1 at 4.5, 7.5, and 11 hours after administration, respectively. However, only in the third administration period, a high-fat meal was provided on the morning of Day 1. The high-fat meal was provided 20 minutes before administration, consumed over 15 minutes, and finished 5 minutes before administration.

[0137] Table 10 shows the pharmacokinetic parameters when compound A was administered at a dose of 25 mg after fasting (first, second, and fourth administration periods) or after consuming a high-fat meal (third administration period). In Table 10, column A represents the type of parameter, column B (25 mg (post-meal)) shows the mean result for the third administration period, and column C (25 mg (fasting)) shows the mean result for the second administration period. In columns B and C, Tmax represents the median, minimum, and maximum values ​​(in parentheses), while other values ​​represent the arithmetic mean and standard deviation (in parentheses). In column D, Tmax represents the least squares mean difference and 95% confidence interval (in parentheses), while other values ​​represent the least squares mean ratio and 95% confidence interval (in parentheses).

[0138] [Table 10]

[0139] The area under the drug concentration curve (AUC) from 0 hours after administration to the final quantifiable time point is reduced by the intake of the aforementioned high-fat diet. 0-t ), and the area under the drug concentration curve (AUC) from 0 hours after administration to indefinite time. 0-∞ ), and peak blood concentration (C max The times decreased by 23%, 27%, and 21%, respectively. Time to reach peak concentration ( Tmax ) was slightly faster with the intake of the aforementioned high-fat diet compared to administration on a fasting diet.

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

[0141] This application claims priority based on Japanese Patent Application No. 2024-104616, filed on 28 June 2024, and incorporates all of its disclosures herein. [Industrial applicability]

[0142] According to the pharmaceutical composition of the present invention, keratosis can be treated or prevented by including 5-methyl-2-(1-piperazinyl)benzenesulfonic acid.

Claims

[Claim 1] The inventions described herein.

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