Pharmaceutical composition for keratosis and use thereof

Abstract

Provided is a new pharmaceutical composition for keratosis such as Darier's disease and psoriasis.　A pharmaceutical composition for keratosis according to the present invention contains 5-methyl-2-(1-piperazinyl) benzenesulfonic acid. The compound is a hydrate thereof, a salt thereof, a hydrate or a solvate thereof, or a hydrate or a solvate of a salt thereof. The hydrate is, e.g., 5-methyl-2-(1-piperazinyl) benzenesulfonic acid monohydrate. The keratosis is, e.g., Darier's disease or psoriasis.

Description

Pharmaceutical composition for keratosis and use thereof

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

[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 in areas of the flexion where sweating is common and secondary infections are likely to occur. Furthermore, Darier's disease can become chronic and recur, leading to a decline in the patient's quality of life (QOL) and potential social handicaps.

[0003] In particular, for Darier's disease, a type of keratosis, there is currently no fundamental treatment 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 found to be safe and consistently effective. Therefore, there remains a high unmet medical need for safe and effective treatments.

[0004] Susan M. Cooper and Susan M. Burge, Darier's Disease Epidemiology, Pathophysiology, and Management, Am J Clin Dermatol 2003; 4 (2): 97-105

[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.

[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.

[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.

[0011] Figure 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 is a graph showing the relationship between administration of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate and biotin diffusion, an indicator of acantholysis, in Example 2. Figure 3 is a graph showing the relationship between administration of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate and pyknotic nucleus frequency, an indicator of dyskeratinization, in Example 2. Figure 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 is a graph showing the relationship between administration of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate and expression of keratinocyte differentiation marker molecules associated with dyskeratinization, in Example 2. Figure 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 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 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 is a graph showing the relationship between administration of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate and the dorsal skin thickness of hyperkeratinized mice in Example 3. FIG. 8 is a graph showing the relationship between the 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.

[0012] The present invention can be exemplified by the following embodiments. [1] A pharmaceutical composition for 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, a salt, 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, a salt, a hydrate, 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] , wherein at least one selected from the group consisting of hyperkeratosis, dyskeratosis, and acantholysis in the keratosis is suppressed.

[12] The pharmaceutical composition according to

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

[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, calculated as the anhydride of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid.

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

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

[17] , wherein the number of doses per day is 2.

[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 a 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 an anhydride, a salt, 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, a salt, a hydrate, 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 anhydride of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid.

[27] The method according to any one of

[21] to

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

[28] The method according to any one of

[21] to

[27] , wherein the administration interval is every day.

[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, a salt, a hydrate, or a solvate of the 5-methyl-2-(1-piperazinyl)benzenesulfonic acid, or a hydrate or solvate of the 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, a salt, a hydrate, or a hydrate of the 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, a salt, 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, a salt, a hydrate, 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, a salt, 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, a salt, a hydrate, 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, for example, as suppression of a disease, and "treatment or prevention of symptoms caused by a disease" can also be referred to, for example, as suppression of 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 may be interpreted as "animal patient." Furthermore, if 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 an 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. It may contain only one of these, or two 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 include, for example, alkali metal salts, alkaline earth metal salts, amphoteric element salts, amine salts, inorganic acid salts, or organic acid salts. Examples of the alkali metal salts include sodium salts or potassium salts, examples of the alkaline earth metal salts include magnesium salts or calcium salts, and examples of the amphoteric element salts include aluminum salts. Examples of the amine salts include lower alkylamine salts such as triethylamine salts; hydroxy lower alkylamine salts such as 2-hydroxyethylamine salts, bis-(2-hydroxyethyl)amine salts, tris(hydroxymethyl)aminomethane salts, and N-methyl-D-glucamine salts; cycloalkylamine salts such as dicyclohexylamine salts; benzylamine salts such as N,N-dibenzylethylenediamine salts; and dibenzylamine salts. 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, 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, more preferably the hydrated MPBS, and specifically, 5-methyl-2-(1-piperazinyl)benzenesulfonic acid monohydrate (also referred to as MPBS monohydrate).

[0026] The MPBSs are known compounds that 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] In the formula, R １ is a hydrogen atom, C １ -C ６ alkyl group of C ３ -C ７ a cycloalkyl group of C １ -C ４ a halogenated alkyl group, a halogen atom, or C ６ -C １２ represents an aryl group represented by the formula: R ２ is a hydrogen atom, C １ -C ６ or an alkyl group of C ７ -C １２ The aralkyl group is an aralkyl group represented by the formula: １ -C ６ an alkoxy group, a halogen atom, C １ -C ６ and an amino group; n represents an integer of 1 to 4.

[0029] In the general formula (I), R １ C defined as １ -C ６ Examples of the alkyl group include a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, an isobutyl group, a sec-butyl group, a tert-butyl group, a pentyl group, an isopentyl group, a neopentyl group, a tert-pentyl group, a hexyl group, and an isohexyl group. ３ -C ７ Examples of the cycloalkyl group include a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, and a cycloheptyl group. １ -C ４Examples of the halogenated alkyl group include a trifluoromethyl group, a trifluoroethyl group, and a pentafluoroethyl group. Examples of the halogen atom include a fluorine atom, a chlorine atom, and a bromine atom. ６ -C １２ Examples of the aryl group include a phenyl group and a naphthyl group.

[0030] R １ Preferred examples of the group include a hydrogen atom, C １ -C ６ alkyl group of C ５ -C ６ A cycloalkyl group represented by the formula (I), a trifluoromethyl group, a halogen atom, or a phenyl group is preferred. １ -C ３ Examples of the alkyl group include the above-mentioned groups, a cyclohexyl group, a trifluoromethyl group, a chlorine atom, a bromine atom, and a phenyl group. More preferred examples include a methyl group or a propyl group, and a methyl group is particularly preferred.

[0031] R ２ C defined as １ -C ６ Examples of the alkyl group in R １ Examples of the alkyl group include those defined in ７ -C １２ Examples of the aralkyl group include a benzyl group, a phenethyl group, and a naphthylmethyl group. Examples of the aralkyl group include a C aryl 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, and a hexyloxy group. １ -C ６ an alkoxy group of the above R １ a halogen atom as defined above; １ and an amino group.

[0032] R ２ Preferred examples of the group include a hydrogen atom, C １-C ３ and alkyl groups of C ７ -C １２ The aralkyl group is an aralkyl group represented by the formula: １ -C ３ alkyl group of C １ -C ３ and a halogen atom. More preferred examples include a hydrogen atom, ７ -C １２ The aralkyl group is an aralkyl group represented by the formula: １ -C ３ In particular, a hydrogen atom is preferable. 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]

[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] Keratosis, which is the subject of the present invention, is a disease that causes hyperkeratosis. Hyperkeratosis means 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 fissures or blister formation. Hyperkeratosis, dyskeratinization, and acantholysis can be observed, for example, by taking a sample of lesional skin from a patient and observing it according to 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, with higher scores indicating 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 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] 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 improve the score. (2) The PGIC is a single questionnaire that evaluates the subject's overall improvement in health using a score, and the pharmaceutical composition for keratosis of the present invention can improve the PGIC and improve the score. (3) The CGIS is a single questionnaire that evaluates the subject's overall impression of severity using a score by a physician, and the pharmaceutical composition for keratosis of the present invention can improve the CGIS and improve the score. (4) The CGIC is a single questionnaire that evaluates the subject's overall improvement in health using a score by a physician, and the pharmaceutical composition for keratosis of the present invention can improve the CGIC and improve the score. (5) The DLQI is a questionnaire that evaluates each question regarding QOL using a score by a subject. 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 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 MPBS. The pharmaceutical composition for keratosis of the present invention may contain, for example, only the MPBS as an active ingredient, or may further contain, in addition to the MPBS, another active ingredient 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 dose of the MPBSs can be expressed, for example, as a dose converted into the anhydrous MPBS. The doses exemplified below are doses of the MPBSs converted into the anhydrous MPBS, and specifically, for the MPBS monohydrate, are doses converted into the anhydrous MPBS.

[0057] Daily dose of the MPBS (converted to 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, 300 mg, 400 mg, 500 mg, 600 mg, 800 mg, 1000 mg, or 1200 mg Range: For example, 1 to 1000 mg, 10 to 1000 mg, 20 to 1000 mg, 25 to 1000 mg, 30 to 1000 mg, 50 to 1000 mg, 60 to 1000 mg, 100 to 1000 mg, 1 to 600 mg, 10 to 600 mg, 25 to 600 mg, 30 to 600 mg, 50 to 600 mg, 100 to 600 mg, 10 to 500 mg, 20 to 500 mg, 25 to 500 mg , 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 00 mg, 300-600 mg, 300-800 mg, 300-1000 mg, 300-1200 mg, 400-600 mg, 400-800 mg, 400-1000 mg, 400-1200 mg, 500-600 mg, 500-800 mg, 500-1000 mg, 500-1200 mg, 600-800 mg, 600-1000 mg, or 600-1200 mg 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 administrations 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 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, and / or, for example, administration at least 1 hour, 2 hours, 3 hours, 4 hours, or 4.5 hours, preferably 1 hour, after ingestion, after 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, and keratosis 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 / preventing keratosis of the present invention. The method for treating / 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 methods 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 descriptions of the pharmaceutical composition for keratosis of the present invention can be used for the MPBSs, their compositions, and administration methods.

[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 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.

[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 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 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. Unless otherwise specified, 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. 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, 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, compositions thereof, 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."

[0071] The present invention also relates to the use of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid in the manufacture 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, compositions thereof, 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."

[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 acanthosis 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 thickening of the spinous layer, resulting in hyperkeratosis. 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 by the following method. Human primary keratinocytes (foreskin, Lonza, 00192906) pooled from three lots of Caucasian newborn 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 (Day 0) using an air-liquid interface culture insert and a 24-well plate, and in vitro epidermal tissue was prepared by culturing for 14 days (Day 0 to Day 14). The culture environment conditions were 37°C, 5% CO ２ The culture medium was maintained at 200°C. 14 days (Day 14) after the initiation of the three-dimensional culture (Day 0), the Compound A aqueous solution (solvent: water) and the 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 3 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. In the three-dimensional culture, the culture medium was basically changed every day from Day 0 to Day 13 (except on weekends), and from Day 14 to Day 17, the culture medium was changed every day, and the Compound A aqueous solution and the IL-22 solution were added at the same time. Seventeen days after the start of three-dimensional culture (Day 17), the in vitro epidermal tissue was sampled and used for evaluation. Note that, 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]

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

[0079] The epidermal tissue slides were stained with hematoxylin-eosin (HE staining) to prepare samples for pathological evaluation. 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). The epidermal thickness was measured for each of the three images per sample using image analysis with NIS-Elements AR software (Nikon), and the average value was used as the representative value for that sample. Note that IL-22-induced hyperkeratosis is a cause of acanthosis, and although hyperkeratosis of the stratum corneum occurs in 3D culture, it peels off each time, making quantitative evaluation difficult. Therefore, epidermal thickness measurements were performed excluding the stratum corneum. The number of samples per experiment (group) was 3 to 9. The results of the four example groups 1-1, 1-2, 1-3, and 1-4 and the control group 1-C in Table 1 were integrated and analyzed to evaluate the effect of Compound A. The mean value of the normal group 1-N to which IL-22 was not added was set as 100%, and the relative value (%), 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 action of Compound A was 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), and the Aspin-Welch t-test was used as the test method. The p-value was 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, and is expressed as a relative value (%), with normal group 1-N to which IL-22 was not added being set at 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 example 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 hyperkeratinization in a concentration-dependent manner at concentrations of at least 1 µmol / L or more. Furthermore, the hyperkeratinization-inhibiting effect of compound A was statistically significant at concentrations of 10 µmol / L or more.

[0082] Example 2: Inhibition of acantholysis and dyskeratosis in an in vitro epidermal acantholysis model The inhibitory effect of Compound A on acantholysis and dyskeratosis in an epidermal acantholysis model was 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 prepared using the following method. Human primary keratinocytes (Foreskin, Lonza, 00192906) pooled from three lots of Caucasian newborn donors were used as cells. EpiLife™ culture medium (Thermo Fisher Scientific, M-EPI-500-A) was used to culture the human primary keratinocytes. 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 conditions were 37°C, 5% CO ２ The culture medium was maintained at 20°C. 14 days (Day 14) after the initiation of the three-dimensional culture (Day 0), the Compound A aqueous solution (solvent: water) and the thapsigargin solution (solvent: DMSO) were added to the in vitro epidermal tissue culture medium, and the three-dimensional culture was continued for another 3 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 thapsigargin was 0 or 30 nmol / L. In the three-dimensional culture, the culture medium was basically changed every day from Day 0 to Day 17 (except on weekends), and from Day 14 to Day 17, the culture medium was changed every day, and the Compound A aqueous solution and the thapsigargin solution were added at the same time. Seventeen days after the initiation 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]

[0087] <Evaluation Method> As shown below, acantholysis was evaluated by cell adhesion force based on biotin diffusion, acantholysis score evaluation based on pathological observation, and immunofluorescence staining of cell adhesion marker proteins, and dyskeratinization was evaluated by the frequency of pyknotic nuclei based on pathological observation and immunofluorescence 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 diluted with PBS / CaCl ２After rinsing twice with a (1 mmol / L) solution (hereinafter the same), 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, in order to confirm cell adhesion in the spinous layer of the epidermal tissue in particular, 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 resuspended in PBS / CaCl ２ The tissue was then rinsed twice with a 1 mmol / L (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 these were fixed on glass slides to prepare epidermal tissue slides.

[0089] When acantholysis occurs, the amount of biotin in the epidermal layer increases relative to the cell adhesion due to a decrease in cell adhesion. Therefore, the epidermal tissue slide prepared was used to detect the biotin marker to evaluate the intercellular adhesion of the epidermis in the in vitro epidermal tissue. That is, the epidermal tissue slide was first deparaffinized and 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 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 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 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 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 ​​of each group were calculated.

[0091] (2) Pathological Evaluation The sampled in vitro epidermal tissue was treated with PBS / CaCl ２ After rinsing twice with the solution, the 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.

[0092] The epidermal tissue slides were stained with HE staining 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). Dyskeratosis and acantholysis were visually evaluated for each sample by image analysis of the captured images.

[0093] The occurrence of abnormal keratinization relatively increases the frequency of pyknotic nuclei. Therefore, for the pathological evaluation of abnormal keratinization, the pathological evaluation sample was used, and the number of pyknotic nuclei in the 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. The number of samples was nine epidermal tissue slides per experiment (one group). Furthermore, for the pathological evaluation of acantholysis, the pathological evaluation sample was used, and observations were made, and the presence or absence of findings of acantholysis was qualitatively recorded. The qualitative evaluation of findings of acantholysis was performed by scoring the number of fissures that occurred at the boundary between the basal layer and the spinous layer. The scoring criteria were as follows, and the average value and standard deviation of each group were calculated for the score values. 0: No fissures 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 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 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 PBS / 0.1% Tween 20 solution containing 5% normal goat serum (NGS) in a thermo-humidifier for 1 hour to avoid nonspecific labeling. The epidermal tissue slides were then left overnight in a thermo-humidifier at 4°C together with primary antibodies (anti-filaggrin (FLG) antibody and anti-cytokeratin 10 (K-10) antibody). The epidermal tissue slide was washed successively with PBS / 0.1% Tween 20 solution and PBS, and then placed in a thermo-humidistat chamber at room temperature for 1 hour with Alexa Fluor dye-conjugated anti-mouse and anti-rabbit secondary antibodies. 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]

[0097] (ii) The epidermal tissue slide was deparaffinized and rehydrated in the same manner as in (i), and then heated with pepsin solution (Sigma, R2283) at 56°C for 15 minutes to activate the antigen. The antigen-activated epidermal tissue slide was rinsed with PBS and then saturated with PBS / 0.1% Tween 20 solution containing 2% bovine serum albumin (BSA) in a thermo-humidifier for 1 hour to avoid nonspecific labeling. The epidermal tissue slide was then left overnight in a thermo-humidifier at 4°C together with primary antibodies (anti-desmoglein 1 (DSG1) antibody and anti-claudin 1 (CLDN1) antibody). The epidermal tissue slide was washed successively with PBS / 0.1% Tween 20 solution and PBS, and then placed in a thermo-humidistat chamber at room temperature for 1 hour with Alexa Fluor dye-conjugated anti-goat and anti-rabbit secondary antibodies. Finally, the epidermal tissue slide was rinsed with PBS. The primary and secondary antibodies used for co-immunostaining of the cell adhesion-related proteins DSG1 / CLDN1 were as shown in the table below.

[0098]

[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 washing, 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 an optical microscope (Nikon Eclipse (Ni-E)) connected to a digital camera (DS-Ri2) in a dark place at 4°C. Image analysis was performed using NIS-Elements Advanced Research Imaging software (Nikon), and the fluorescent staining 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 stained area (%), calculated by dividing the stained area B by the ROI surface area A (B / A), and the mean and standard deviation for each group were calculated. For each sample, the mean value of the biotin fluorescence intensity for each of the three images was calculated, and this mean value was used as the representative value for the three images. Nine epidermal tissue slides were used 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 an epidermal acantholysis model by the addition of 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 in which 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 / no 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 showing the relative value (%) of biotin amount. Figure 3 is a graph showing the results of pyknotic nucleus frequency, an indicator of dyskeratinization, with the vertical axis showing 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 showing 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 of FLG and Figure 5B showing the results of K-10, each with the vertical axis showing 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 of DSG1 and Figure 6B showing the results of CLDN1, each with the vertical axis showing 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, which is an indicator of acantholysis, was significantly increased in the control group 2-C, which received thapsigargin, compared to the normal group 2-N, which did not receive thapsigargin. 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 example group, in which thapsigargin and 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 example group, the addition of 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 cell adhesion-related proteins DSG1 and CLDN1, and 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 example 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 abnormality induced by thapsigargin.

[0104] From these results, it is found that the addition of compound A can improve the attenuation of intercellular adhesive force induced by thapsigargin in vitro epidermal tissue in a concentration-dependent manner, that is, inhibit acantholysis.In addition, as a specific example, it is found that compound A is statistically significant at 10 μmol / L for inhibiting the decrease of intercellular adhesive force, and is statistically significant from 1 μmol / L for inhibiting the increase of pathological evaluation score of acantholysis.

[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 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 abnormal keratinization and increased the number of pyknotic nuclei. In contrast, in the example 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 1 μmol / L or more of Compound A, compared to the control group 2-C. Furthermore, as shown in Figures 5A and 5B, in immunofluorescent staining of the differentiation marker FLG expressed in keratinocytes in the granular layer and stratum corneum, and the differentiation marker K-10 expressed in keratinocytes in the spinous layer and granular layer, the immunofluorescent stained area of ​​the control group 2-C was statistically significantly increased and decreased, respectively, by the addition of thapsigargin, compared to the normal group 2-N. 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 areas immunofluorescently stained for FLG and 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 abnormalities 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 abnormal keratinization. Specifically, Compound A was found to be statistically significant at concentrations of 3 μmol / L and above in inhibiting the increase in pyknotic nuclei, an index of abnormal keratinization.

[0107] In addition, from these results, it can be seen that compound A can suppress both abnormal keratinization and acantholysis.Therefore, among keratosis, for example, not only the disease that only causes abnormal keratinization or acantholysis, but also the disease that causes both (for example, Darier's disease), it can be seen that by simply administering compound A, both symptoms can be simultaneously suppressed.In addition, in Example 1, the same keratinocyte is used to prepare a hyperkeratinization model, and the suppression of hyperkeratinization by compound A is also confirmed.Therefore, it can be said that compound A shows the suppression effect on all three main pathological symptoms of Darier's disease, that is, hyperkeratinization, acantholysis and abnormal keratinization.

[0108] Example 3 Inhibition of Hyperplasia 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 the compound A on hyperkeratosis was confirmed.

[0110] <Test Cells and Experimental Design> Seven-week-old male BALB / c mice (Charles River Japan) were used. The general condition of the mice was observed for six days during the 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 one of the experimental groups was shaved. Three days later, imiquimod 5% cream (Mochida Pharmaceutical Co., Ltd.) was applied to the dorsal skin of the mice under 2% inhalation anesthesia using isoflurane (Mylan Pharmaceutical Co., Ltd.) as an anesthetic once daily for a total of four days to induce psoriasis-like dermatitis. Psoriasis-like dermatitis manifests as hyperkeratosis, a pathological symptom. The application amount 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 lightly 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 10 hours or more 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. In normal group 3-N, the back hair was shaved, and then the animals were not challenged with imiquimod, nor were they administered with Compound A. In control group 3-C, the animals were challenged with imiquimod in the same manner as in the example group, but were not administered with Compound A. Instead, 10 mL / kg of a 1% Tween 80 solution not containing Compound A was orally administered.

[0112]

[0113] <Evaluation Method> The first application of imiquimod 5% cream was designated as 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 with a digital caliper (Mitutoyo Corporation). The measured 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 example group, and 4 for the normal group, and the effect of Compound A was evaluated. The mean value 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 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 Gene expression analysis was performed on formalin-fixed, paraffin-embedded (FFPE) human skin samples from patients with Darier's disease and psoriasis vulgaris. Specifically, the involvement of IL-22 in Darier's disease, which is also a type of keratosis, was evaluated by comparing it with psoriasis vulgaris, a type of keratosis in which IL-22 is known to be involved.

[0117] <Test Cells and Test Design> As shown below, images were acquired and libraries were prepared from FFPE block samples (Aurus Biosciences, Inc.) with a diagnosis history. Patient-derived FFPE block samples were obtained from chest skin biopsies of a black female in her 70s with psoriasis vulgaris, and from back skin biopsies 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 and peeling of the sections using the section adhesion test, 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 Co., Ltd.). A library was then prepared from the slide 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 intensities were calculated by log2-transforming UMI counts normalized by the total UMI counts of the same barcode sequence. To estimate the contribution of IL-22 to the pathology, 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 epidermal in Visium increases, forming layers. Therefore, the epidermis was classified into upper and basal layers, 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, in which IL22RA1 expression is low, was then calculated.

[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]

[0121] The above results confirmed the involvement of IL-22 in both psoriasis vulgaris and Darier's disease, which are major keratosis. 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 exhibits an inhibitory effect on both psoriasis vulgaris and Darier's disease.

[0122] [Example 5] Evaluation of Pharmacokinetics The pharmacokinetics was evaluated using Compound A. In this example, the amount of Compound A used is expressed in terms of the amount of MPBS anhydrate (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, subjects were orally administered 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 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 consisted of three groups, A, B, and 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]

[0125] Each subject received a total of 16 doses of the Compound A 5 mg tablet or placebo tablet during the study period. The 16 doses were divided into one in the morning on Day 1 (Day 1), one in the morning and one in the evening from Day 3 (Day 3) to Day 9 (Day 9), and one in the morning on Day 10 (Day 10). From Day 3 to Day 10 (Day 3-Day 10), the 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, an afternoon snack, and dinner were provided approximately 2 hours before, approximately 4 hours after, approximately 7 hours after, and approximately 10 hours after the morning dose, respectively. Meals were limited to no more than 2,500 kcal for men and 2,000 kcal for women. Water was prohibited for 2 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 2 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, and the plasma concentration of compound B was measured. 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]

[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 Figure 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, or further 50 to 200 mg, orally administered twice a day.

[0129] [Example 6] In vivo clinical trial Compound A is 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 to the amount of compound B are used for administration of compound A.

[0130] <Subjects> Darier's disease patients aged 18 to 75 years <Study overview> Randomized double-blind clinical trial Placebo tablet administration: 24 cases 100 mg administration 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) 200 mg administration group: 24 cases Two 50 mg tablets of compound A were orally administered twice a day (200 mg / day as the amount of compound B) 400 mg group: 24 cases Four 50 mg tablets of compound A were orally administered twice a day (400 mg / day as the amount of compound B)

[0131] <Test Items> In clinical trials of Compound A in patients with Darier's disease, evaluations will be made of 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 tablet group, administration of Compound A is expected to result in improvement in 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 dyskeratinization in epidermal acantholysis have been confirmed. Therefore, it can be said that various symptom scores in Darier's disease patients are improved by administration of Compound A.

[0133] [Example 7] Evaluation of the effect of food on pharmacokinetics Compound A was administered in a fasted state or after a meal to evaluate the effect of food on 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 study groups and treatment details. The study groups were composed of three groups, Study Groups 1 to 3, as shown in Table 9. Each study 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 from Example 5.

[0135]

[0136] Each subject was orally administered the Compound A 5 mg tablet or placebo tablet once in the morning of the first day of the study (Day 1) for each administration period. Food intake was prohibited until 4 hours after administration, and water intake until 2 hours after administration. The study drug (the Compound A 5 mg tablet or placebo tablet) was taken with 240 mL of water while standing. In all administration periods, dinner was provided on 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 administration, respectively. However, only in the third administration period, a high-fat meal was provided in the morning of Day 1. The high-fat meal was provided 20 minutes before administration, ingested over 15 minutes, and finished 5 minutes before administration.

[0137] Table 10 shows the pharmacokinetic parameters when 25 mg of Compound A was administered after fasting (1st administration period, 2nd administration period, 4th administration period) or after ingestion of a high-fat meal (3rd administration period). In Table 10, column A indicates the type of parameter, column B (25 mg after meals) indicates the average result for the 3rd administration period, and column C (25 mg fasting) indicates the average result for the 2nd administration period. In columns B and C, Tmax is the median, minimum, and maximum values ​​(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]

[0139] The area under the drug concentration curve (AUC) from time 0 to the last quantifiable time point was measured by the ingestion of 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 ) were reduced 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 on June 28, 2024, the disclosure of which is incorporated herein in its entirety by reference.

[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. The pharmaceutical composition according to 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 according to claim 1, wherein the keratosis is Darier's disease.

4. A pharmaceutical composition according to any one of claims 1 to 3, which inhibits at least one selected from the group consisting of hyperkeratosis, dyskeratosis, and acantholysis in said keratosis.

5. A pharmaceutical composition according to any one of claims 1 to 4, wherein the dosage of 5-methyl-2-(1-piperazinyl)benzenesulfonic acid is 1 to 1000 mg / day.

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

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